Thèses : « Immunohistochemistry and histology »

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Russell, Katherine Margaret. « Intestinal responses to Clostridium perfringens in broilers ». Thesis, University of Edinburgh, 2016. http://hdl.handle.net/1842/25514.

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Clostridium perfringens is the aetiological agent of Necrotic enteritis (NE); a disease that impacts on the health and welfare of broilers. This disease is a large cost to the industry and presents as lesions in the small intestine hindering productivity. Antibiotics are commonly used to treat NE but as pressure increases to limit their use further information about disease onset and broiler responses to the bacteria and it’s virulence factors during infection is required to implement new preventative measures and treatments. NetB is a secreted toxin from C. perfringens which has an important role in NE onset. Using an in situ intestinal loop model we have been able to characterise: I) temporal broiler responses to NetB positive bacterial culture supernatant (Chapter 2), ii) early host responses to different isolates possessing NetB (virulent) or not (avirulent) in the presence or absence of bacterial cells (Chapter 3) and iii) the responses of two commercial broiler breeds (Chapter 4) four hours post exposure. Samples collected from these experiments have been used for histology, mRNA expression and immunohistology. We have shown differences in mRNA expression in the duodenum of broilers after exposure to C. perfringens cells as well as the culture supernatant from the isolates used after four hours. The presence of bacteria cells resulted in up-regulation of pro-inflammatory cytokine, IFN-γ, mRNA, whereas it resulted in down-regulation of B-LA, mRNA a gene involved in presentation of pathogens to immune cells. IL-6 mRNA expression was also reduced in the presence of virulent isolates. This could indicate a possible evasion strategy for C. perfringens in broilers. Immunohistochemical analysis indicated that slower growing broilers have increased numbers of immune cells (macrophages and γδ T cells) in their duodenum compared with faster growing broilers, although this did not appear to have an effect on mRNA expression levels of pro-inflammatory cytokines, 4h post antigen infusion. Overall we detect greater changes when bacteria are included with culture supernatant and have highlighted possible mechanisms for C. perfringens to avoid the broiler immune system. Induction of NE in the literature requires pre-disposing factors, including co-infection with other intestinal pathogens and dietary manipulation of the host. The final experiment trialled protocols administering a virulent isolate of C. perfringens in-feed and via gavage along with an increased protein source to induce NE (Chapter 5). These models were not considered to be consistent for further investigation of NE in the future.

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Campbell, Thomas Mark. « Demographics and Posterior Knee Capsule Histologic and Genetic Characterization in Patients with Severe Knee Osteoarthritis : Comparing Those with Contracture to Those Without Contracture ». Thesis, Université d'Ottawa / University of Ottawa, 2012. http://hdl.handle.net/10393/23176.

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Introduction: Knee flexion contractures have a negative impact on function for patients with osteoarthritis (OA). Those with contracture treated with total knee arthroplasty (TKA) have more post-operative pain and worse outcome. Little knowledge is available about patient demographic factors or gene expression in the knee joint capsule in the setting of contracture and severe OA. // Methods: Subjects with primary severe knee OA awaiting a TKA were recruited. We collected subject demographic factors that may be associated with preoperative knee contracture. Subjects’ posterior knee capsule was harvested intraoperatively. Capsule histological analysis was performed using light microscopy. Gene expression analysis was performed using whole genome microarray and immunohistochemistry was used for protein production analysis comparing those with contracture to those without. // Results: Twenty subjects were recruited for the demographics portion of the study (13 contractures and 7 controls), and capsules from 12 subjects (6 contractures, 6 controls) were used for histology, microarray, and IHC analyses. Contracture subjects had longer duration of OA, reduced extension in the contralateral knee, and showed a trend toward elevated body mass index. Tissue cross-sectional areas of adipose, non-adipose and synovial tissues were not statistically different histologically between the two groups. There was increased expression in the contracture group for the genes CHAD, Cyr61, and Sox9. There was a corresponding increase in protein production for CHAD and Sox9. // Conclusions: Screening for OA duration and bilateral knee range of motion (ROM) could be functionally beneficial. When a knee joint contracture is present, correcting for the resulting leg length discrepancy pre- and post-operatively could improve patient outcome. Gene protein products linking capsular cells to the ECM can influence capsular fibrosis and potentially impact ROM.

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Forlani, A. « COMBINED DIAGNOSTIC APPROACHES FOR THE DIAGNOSIS OF CANINE SPLENIC NEOPLASM : CONTRAST-ENHANCED ULTRASONOGRAPHY, ULTRASOUND GUIDED CYTOLOGY, HISTOLOGY AND IMMUNOHISTOCHEMISTRY IN SELECTED LESIONS ». Doctoral thesis, Università degli Studi di Milano, 2014. http://hdl.handle.net/2434/246578.

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Focal splenic lesions are frequently identified during routine ultrasonographic examination. Differentiation between malignant and benign changes is crucial since it influence treatment and prognosis. Although B-mode ultrasound is very sensitive for the detection of splenic lesions, its specificity is low, whereas contrast enhanced ultrasound is used successfully to differentiate benign from malignant liver lesions in humans and dogs. Cytology represent another useful technique in the diagnosis of splenic disorders. However, few studies have addressed the accuracy of cytology in the evaluation of splenic lesions and specifically neoplasms. The aim of this study was to evaluate the diagnostic usefulness of contrast enhanced ultrasound and cytology in the diagnosis of splenic neoplasia, this study has been divided into a prospective and in retrospective part. The specific aim of the prospective part was to evaluate whether contrast-enhanced ultrasound can be used to more accurately characterize the perfusion of splenic focal abnormalities in small animals. Moreover, we attempted to establish diagnostic criteria useful to differentiate benign from malignant splenic lesions and to diagnose the various malignant tumors. Contrast enhanced ultrasound was performed to study focal to multifocal lesions of spleen using a second-generation microbubble contrast medium (Sonovue®) injected into the cephalic vein and enhancement patterns were described. Final diagnosis was obtained by fine needle cytology and/or histopathology after splenectomy. The aim of the retrospective study was to evaluate the role of cytology in primary and metastatic splenic tumour diagnosis using histology as the gold standard. Splenic cytological and corresponding histopathological samples obtained between 1998 and 2012 from the electronic archives of the Anatomical Pathology service of the School of Veterinary Medicine of the University of Milano, Italy. Concordance between cytology and histology was determined. Accuracy, sensibility, specificity, positive and negative predictive value of cytology for the diagnosis of splenic neoplasia was determined. Primary splenic lymphomas were collected and graded according to the WHO classification (cell size, diagnosis, mitotic index, phenotypes). A total of 26 cases of canine focal splenic masses analysed with contrast-enhanced ultrasound affected by 11 benign and 15 malignant and splenic lesions were included. In this series, most benign lesions (7/11) had a perfusion pattern similar to the adjacent parenchyma, so that the lesions were isoechoic or mildly hypoechoic. On the contrary all malignant lesions became completely or extensively hypoperfused during the wash-out phase. All hemangiosarcomas were characterized by hypoechogenicity trough all perfusion phases. Feeding vessels were present in 11/15 cases of malignancy and in 3/11 cases of benign lesions. In the retrospective study a total of 66 cases were collected. Thirty cytological samples were classified as non-neoplastic (12 true negatives, 18 false negatives compared with histopathology). Cytological diagnosis of neoplasia was obtained in 36 cases (35 true positives and 1 false positive). Cytological diagnosis was in agreement with histopathology in 71,2% (47/66) of cases. Cytology had a sensitivity of 66%, a specificity of 92%, a positive predictive value of 97%, and a negative predictive value of 40%. A comprehensive grading of 29 canine primary splenic lymphomas was performe. All cases of Marginal Zone Lymphoma (MZL), Mantle Cell Lymphoma (MCL) and follicular lymphoma with a nodular pattern of growth were of low grade. On the contrary, diffuse lymphomas were intermediate to high grade. Contrast-enhanced ultrasound can improve the characterization of focal to multifocal splenic lesions and cytology demonstrated to be a useful tool for splenic neoplasia diagnosis due to high specificity and a high positive predictive value highlighted in our study. Thus, they should be considered complementary techniques in the diagnosis of splenic lesions.

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Gondim, Roberta Marinho Falcão. « Avaliação da cicatrização cutânea : fluorescência e estereologia ». Universidade de São Paulo, 2012. http://www.teses.usp.br/teses/disponiveis/5/5160/tde-29102012-161842/.

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Objetivos: Recentemente, a Espectroscopia de Fluorescência (EF) tem sido estudada como método de análise de propriedades da pele de forma nãoinvasiva e em tempo real, utilizada em uma variedade de aplicações, incluindo avaliação e diagnóstico do tecido in vivo. Contudo, na cicatrização da pele, essa técnica não tem sido completamente explorada. Visto que a determinação da idade de uma lesão é um aspecto importante na medicina forense, esse trabalho tem por objetivo testar a aplicabilidade da medida da Intensidade de Fluorescência (IF) após o uso de metil-aminolevulinato (MAL) na estimativa da idade de lesão incisa, através da EF ao longo do tempo e fazer a correlação desta com os achados histológicos. Materiais e Métodos: Foram utilizados camundongos hairless como modelo experimental. Os animais foram divididos em dois grupos: com (+) e sem (-) o uso de MAL antes da EF. Incisões cirúrgicas lineares foram realizadas no dorso de cada animal. Espectros na faixa de 480 e 800 nm foram coletados da lesão e da pele normal adjacente, usando o sistema Ocean Optics, correspondendo a quatro condições: a) IF da lesão após MAL (+/+); b) IF da pele normal após MAL (-/+); c) IF da lesão sem MAL (+/-) e d) IF da pele normal sem MAL (-/-). Após a cirurgia, os animais foram monitorados periodicamente até 3 meses de pós-operatório e eutanaziados em grupos. Fragmentos de pele, contendo todo o ferimento, foram removidos e processados para análise histológica por métodos estereológicos. Vários cortes histológicos foram analisados para avaliar a organização da derme e da epiderme, deposição de colágeno e proliferação celular (imunoistoquímica por PCNA). Resultados: Nas fases iniciais da cicatrização, a EF in vivo mostrou acúmulo preferencial de protoporfirinas na lesão com uso de MAL (+/+), quando comparado à pele normal adjacente (-/+). Contudo, nas fases avançadas, ocorreu o inverso. Houve uma diferença estatisticamente significante neste grupo (+/+) ao longo do tempo (p < 0,0001), o que não aconteceu com os demais ((-/+); (+/-) e (-/-)). O modelo apresentado permitiu a estimativa da idade de lesão incisa no intervalo de dois meses, sendo possível estimar a fase de cicatrização em que esta se encontra. Achados histológicos confirmaram as fases da cicatrização, mostrando correlação com os achados de fluorescência. Conclusão: Os resultados mostraram que a técnica de EF usando MAL é um método promissor na análise dos estágios da cicatrização da pele
Background and Objective: In recent years, Fluorescence Spectroscopy (FS) has been explored as a novel noninvasive and real-time technique for analysis of skin properties, useful in a wide variety of applications, including tissue evaluation and diagnosis. However, the use of FS in skin wound healing has not been fully explored. Since aging of injuries on a victims body is an important aspect of forensic medicine, this paper intended to test the usefulness of collecting Fluorescence Intensity (FI) after topical MAL on age estimate of the incised lesion, through the study by FS over time and correlation with histological findings. Materials and Methods: As experimental model, was used hairless mice. The mice were divided into two groups: with (+) and without (-) use MAL before FS. Standardized linear wounds were made on the dorsum of each mice. Spectra in the 480-800 nm wavelength range were collected from normal and wound skin using Ocean Optics system, corresponding to four conditions: a) FI of skin wound after MAL (+/+); b) FI of normal skin after MAL (- /+); c) FI of skin wound without MAL (+/-) and d) FI of normal skin without MAL (- /-). After wounding, the animals were monitored periodically until 3 months and killed in groups. Tissue specimens, containing the whole wound, were removed and processed for histological analysis using stereological techniques. Several cross-sections were analyzed to evaluate the organization of the dermis and epidermis, collagen deposition and cellular proliferation (PCNA - imunohistochemistry antigen). Results: In vivo FS of skin wound healing with MAL (+/+) showed that there was a protoporphyrin preferential accumulation in healing site as compared to adjacent normal skin (+/-) in the early stage of healing. However, in the later stages, the reverse happened. There was statistically significant into this group (+/+) along the time (p < 0,0001); what not happened with another groups ((-/+); (+/-) and (-/-)). The model allows to estimate the age of an incised injury into interval of two months. Its possible to estimating his healing phase. Histological findings confirm the stages of healing and agree well with the fluorescence findings. Conclusion: The results showed that FS using MAL appears to be a promising approach for the analysis of stages of skin wound healing

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Rojo, Xicart Ernest. « Soft tissue volume gain around dental implants after abutment connection surgery using autogenous subepithelial connective tissue grafts harvested from the palate or tuberosity. A randomized prospective clinical study ». Doctoral thesis, Universitat Internacional de Catalunya, 2017. http://hdl.handle.net/10803/586354.

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The aim of the present study is to compare the volume gain around dental implants when a subepithelial connective tissue graft (SCTG) from palate or tuberosity is used randomly. The most studied donor area for soft tissue augmentation has been the autogenous connective tissue from the palate. However recent studies has affirmed that tuberosity tissue may possess better tissue qualities for soft tissue volume augmentation. It has been shown that tuberosity connective tissue is more dense with less fat and glandular tissue. Therefore, it could be speculated that this firmer tissue will have less shrinkage and achieve more soft tissue gain. In the present study 32 patients with 36 implants with localized volume deficiency has been included and received randomly a SCTG from palate or tuberosity. Measurements using an intraoral optical scan has been done at baseline and 3 months. Also 20 samples were obtained at baseline for immunohistochemistry and descriptive histological analysis. In conclusion both groups obtained volume gain at 3 months. No statistical significant differences were found. Even though a tendency of better results was observed for patients who received SCTG from tuberosity.
L’objectiu del present estudi es comparar el guany de volum al voltant d’implants dentals després d’haver utilitzat aleatoriament injert de teixit conectiu subepitelial de paladar o de tuberositat. L’àrea donant més utilitzada per realitzat procediments d’augment gingival ha estat sempre la zona del paladar. Tot i que estudis recents han demostrat que la zona de la tuberositat pot ser una bona alternativa degut a que pot tenir millors propietats per l’augment gingival. S’ha demostrat darrerament que el teixit conectiu de la tuberositat és més dens i conté menys teixit gras i glandular. Això pot comportar que aquest teixit no es contraigui tant i que per tant pugui aconseguir millors resultats en quant a guany de volum. En aquesta investigació 32 pacients portadors de 35 implants amb defecte de volum vestibular han rebut cirugía d’augment de teixit tou utilitzant injert de teixit conectiu de paladar o tuberositat. S’han realitzat mesures utilitzant un escáner intraoral a l’inici de l’estudi i 3 mesos després. També s’ha realitzat estudi histològic i d’immunohistoquímica de 20 mostres. Com a conclusió, els dos grups de l’estudi han aconseguit guanyar volum de teixit tou als 3 mesos. No s’han detectat diferencies estadísticament significatives entre els grups. Tot i així s’ha observat una tendencia a millors resultats en el grup de pacients que han rebut injert de teixit tou de la tuberositat.

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Gorricho, Camila Mario. « Vitrificação de tecido ovariano de gatas domésticas : o tamanho do fragmento influencia a viabilidade pós descongelação ? / ». Universidade Estadual Paulista (UNESP), 2018. http://hdl.handle.net/11449/154237.

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A criopreservação de ovário ou tecido ovariano permite a preservação do material genético de qualquer espécie animal que seja submetido à gonadectomia por indicação preventiva, terapêutica ou, até mesmo, por morte inesperada. O objetivo do presente trabalho foi avaliar se o tamanho do fragmento ovariano influencia ou não a resistência aos crioprotetores. Para tanto, os ovários foram colhidos de 34 gatas domésticas (várias raças, 1-5 anos de idade) por ovariectomia de rotina, transportados ao laboratório e depois seccionados em fragmentos de diferentes tamanhos (3 x 3 x 3mm, 5 x 3 x 3mm e 7 x 3 x 3mm) e destinados aleatoriamente aos grupos de controle (GC3, GC5 e GC7, respectivamente) ou vitrificados (GV3, GV5 e GV7, respectivamente). Os fragmentos vitrificados-aquecidos foram avaliados por histomorfologia e imunohistoquímica (para taxas de apoptose utilizando a caspase-3 clivada). A avaliação histológica demonstrou que 72,97% dos folículos presentes em GV3 e 72,58% nos fragmentos do grupo GV5 eram normais, enquanto que nos fragmentos do GV7 essa taxa foi de apenas 42,86%. A principal alteração morfológica foi o desprendimento das células epiteliais da membrana basal presentes em todos os grupos. Da mesma forma, a avaliação imunohistoquímica, utilizando a caspase 3, revelou uma pequena proporção de células apoptóticas nos fragmentos do grupo GV3 (53%), enquanto que no grupo GV7, 43,58% das células expressaram a caspase 3 clivada. Esses achados indicam que fragmentos seccionados em 3 x 3 x 3mm (27mm³) são mais adequados para a perfusão do crioprotetor, sem causar danos celular após o descongelamento.
Cryopreservation of ovary or ovarian tissue allows preservation of genetic material of any animal species that is submitted to gonadectomy for preventive, therapeutic or even by unexpected death. The aim of the present study was to investigate whether or not the size of the ovarian fragment influence its resistance to cryostorage. For that purpose, ovaries were collected from 34 queens (various breeds, age 1-5 y) by routine ovariectomy, transported to the laboratory and then sectioned in different sizes (3 x 3 x 3 mm, 5 x 3 x 3 mm and 7 x 3 x 3 mm) and randomly assigned to a control (GC3, GC5 and GC7, respectively) or vitrified (GV3, GV5 and GV7, respectively) groups. Vitrified-warmed fragments were evaluated by histomorphology and immunohistochemistry (for apoptotic rates by using cleaved caspase-3). Histological examination reveal that 72.97% of the follicles in GV3 and 72.58% in GV5 were normal while only 42.86% of the follicles in GV7. The main morphological alteration presented in all groups was a detachment of the epithelial cells by basement membrane. Similarly, immunohistochemistry evaluation using caspase 3 revealed a small proportion of apoptotic cells in GV3 (53%) while in GV7 43.58% of the cells expressed cleaved caspase-3. These findings indicate that fragments sectioned in 3 x 3 x 3 mm (27mm3) seems more adequate for perfusion of the cryoprotectant, causing less damage to the cell after vitrification-warming.

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Bassuino, Daniele Mariath. « Caracterização histológica, imuno-histoquímica e mapeamento de lesões da raiva em medula espinhal de bovinos e equinos ». reponame:Biblioteca Digital de Teses e Dissertações da UFRGS, 2015. http://hdl.handle.net/10183/115191.

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A raiva é uma doença zoonótica causada por um vírus RNA, envelopado, altamente neurotrópico, da família Rhabdoviridae, gênero Lyssavírus. Esta dissertação identificou e mapeou as lesões ocasionadas pelo vírus da raiva na medula espinhal de bovinos e equinos. Os casos foram obtidos no período entre janeiro de 2013 a novembro de 2014, através de necropsias realizadas pelo Setor de Patologia Veterinária da Universidade Federal do Rio Grande do Sul (SPV-UFRGS) de animais que apresentaram síndrome neurológica. Destes, 33 casos (26 bovinos e sete equinos) foram selecionados com diagnóstico final de raiva, confirmados através do exame histopatológico e imuno-histoquímico (IHQ). Todos os casos foram submetidos ao exame de imunofluorescência direta (IFD), e os negativos a este teste, submetidos à inoculação intracerebral em camundongos (ICC). O material foi processado rotineiramente para histologia e corados com hematoxilina e eosina (HE). O exame de IHQ foi realizado na medula espinhal de todos os animais com anticorpo primário policlonal anti-vírus rábico. A medula espinhal foi subdividida nas porções cervical (C), intumescência cervical (IC), torácica (T), lombar (L), intumescência lombar (IL) e sacral (S) para avaliação. As estatísticas descritivas foram feitas no software IBM SPSS Statistics 18® e um índice de severidade das lesões observadas foi criado, baseado na soma dos graus das lesões e posteriormente dividido pelo número total de animais de cada espécie. O índice agrupou manguitos perivasculares + microgliose + neuroniofagia e analisou separadamente a presença de células gitter e a intensidade de marcação IHQ. Utilizou-se, ainda, o teste exato de Fisher com nível de significância de 5% através do software Epi Info™ 7.1.4 para avaliação do risco relativo entre a presença ou ausência de lesão entre encéfalo e medula espinhal. Todos os casos obtiveram imunomarcação positiva no exame de IHQ (100%). Na IFD dois equinos foram positivos (28,5%). Todos os resultados negativos na IFD foram confirmados na ICC, como negativos. Nos bovinos, a IFD foi positiva em 20 casos (76,9%). Os resultados negativos foram submetidos à ICC, onde cinco casos foram negativos e um positivo. À avaliação da medula espinhal, todos os bovinos e equinos apresentaram lesões histológicas. Nos equinos, as lesões inflamatórias se revelaram discretas nas regiões C, IC; moderada em T, L e S e acentuada na secção de IL. Células gitter estiveram discretamente presentes em C, IC e T; em moderada quantidade nas regiões L e S e acentuada em IL. A marcação IHQ nos fragmentos medulares variou de moderada a acentuada. Nos bovinos as lesões inflamatórias foram moderadas em todos os segmentos medulares. Diferente do observado nos equinos, células gitter foram predominantemente discretas. A marcação IHQ foi acentuada e homogênea em todos os segmentos medulares. Nos equinos observou-se uma maior frequência de lesões a partir do segmento T, no entanto, a marcação IHQ revelou discreta variação entre os segmentos. Na distribuição lesional dos bovinos evidenciou-se variação apenas no quesito malacia, que se mostrou mais frequente nos segmentos L, IL e S. Identificou-se, através do método de Fisher, que a coleta da medula espinhal em equinos possibilita uma chance 3,5 vezes maior na detecção de lesões de raiva quando comparado à análise individual do encéfalo.
Rabies is a zoonotic disease caused by an enveloped ribonucleic acid (RNA) virus, highly neurotropic, from the Rhabdoviridae family, Lyssavirus genus. This thesis identified and mapped the lesions caused by the rabies virus in the spinal cord of cattle and horses. The samples were obtained within the period from January 2013 to November 2014 from necropsies carried out by the Setor de Patologia Veterinária of Universidade Federal do Rio Grande do Sul (SPV-UFRGS). We selected the thirty three animals (26 cattle and seven horses) that presented neurological syndrome and diagnosis of rabies. Samples from the brain and spinal cord were collected and routinely processed for hematoxylin and eosin (HE) and immunohistochemistry (IHC). The IHC was carried out at the spinal cord of all the animals, with a polyclonal anti-rabid virus as primary antibody. The spinal cord was sectioned at the cervical (C), cervical intumescence (CI), thoracic (T), lumbar (L), lumbar intumescence (LI) and sacral (S) portions to be evaluated. The descriptive statistics of this work was performed using the IBM SPSS Statistic 18® software and an index of the severity observed in the lesions was created based on the grades of the lesions and later divided by the total number of animals of each species. The index grouped cuffs of perivascular inflammatory cells, microgliosis and neuronophagia. Then, separately analyzed the presence of gitter cells and their IHC intensity. To evaluate the relative risk between the presence or absence of lesions between the encephalon and the spinal cord the exact Fisher test was carried out with 5% of significance level runned with the Epi Info™ 7.1.4 software. All the cases were positive for the IHC. At the DIF two horses (28.5%) and 20 cattle (76.9%) were positive. To all the samples negative to DIF, the test of inoculation was applied resulting in all horses and five cattle negative, and one cattle positive. Under the HE evaluating of the spinal cord, all the animals presented histologic lesions. In the horses, the inflammatory lesions were discrete in the C and IC sections; moderate in T, L and S; and accentuated in LI. Gitter cells were discretely present in the C, CI and T sections; moderate in L and S; and accentuated in LI. IHC of the spinal cord fragments varied from moderated to accentuate. The inflammatory lesions from cattle samples were moderate in all of the spinal cord sections. Differently from the observed in horses, gitter cells were predominantly discrete. The IHC staining was accentuated and homogenous in all spinal cord sections. In the horses lesions in the T section were frequently observed; however, IHC staining revealed discrete variation between the sections. The lesions distribution from cattle samples highlighted variation only in the issue concerning malacia, which was frequently demonstrated in the L, LI and S sections. The Fisher test identified that the gathering of horses spinal cords allows a 3.5 times higher chance of detection of rabies lesions than when the individual analysis of the brain is made.

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Poole, Kenneth. « The effects of stroke on the skeleton ». Thesis, University of Cambridge, 2006. https://www.repository.cam.ac.uk/handle/1810/244611.

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Stroke is now a well-recognised risk factor for hip fracture. The aim of this study was to elucidate the pathophysiological mechanisms by which hip bone loss occurs in hemiplegia and to test the efficacy of a novel pharmaceutical strategy for preserving bone in stroke patients. Patients who were admitted acutely with a first-ever stroke and who remained unable to walk one week later were studied prospectively for 12 months, with a series of bone mineral density measurements of the hips (dual energy X-ray absorptiometry) in the context of a randomised controlled trial. Untreated patients (n=13) experienced a decline in bone mineral density at the hemiplegic hip that was rapid, with the greatest losses in the trochanteric region of the affected side. This bone loss was prevented by the administration of a single 4 mg dose of the intravenous bisphosphonate, zoledronate (n=14) within 35 days of stroke onset. Computed tomography of the hips in 8 untreated patients more than a year after stroke confirmed that the greatest difference between sides was in the trochanteric region. Serum vitamin D measurements in 44 patients with acute stroke were substantially lower than healthy elderly controls, with 77% of patients in the insufficient range, suggesting that vitamin D insufficiency preceded stroke. Histomorphometric analysis of iliac bone biopsies from hemiplegic patients 10 weeks following stroke showed normal erosion parameters, but a striking decrease in the surface extent of osteoid when compared with healthy reference values. Unexpectedly, treatment with zoledronate was associated with a significantly higher osteoid surface compared with placebo treated subjects in cancellous, endocortical and cortical bone. Sclerostin, a newly discovered osteocyte-derived protein was studied using immunohistochemical staining of the bone biopsies. Sclerostin is known to be an inhibitor of active osteoblasts, which led to the hypothesis that in stroke, the proportion of osteocytes expressing sclerostin would be inversely associated with the surface extent of bone formation. Histological analysis revealed widespread expression of sclerostin in osteocytes and their canaliculi in all subjects. However, examining individual osteocytes in relation to bone forming surfaces revealed that newly embedded osteocytes did not express sclerostin until after primary mineralisation. It is proposed that this precise pattern and timing of sclerostin expression by osteocytes allows bone formation to continue locally (during remodelling), but prevents excessive new bone formation elsewhere, as seen in the single gene disorder sclerosteosis.

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Cavaglieri, Rita de Cássia. « Terapia com células-tronco na nefropatia crônica experimental : é possível bloquear a progressão da doença renal ? » Universidade de São Paulo, 2010. http://www.teses.usp.br/teses/disponiveis/5/5160/tde-19032010-125745/.

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Células-tronco (CT) apresentam potencial terapêutico para a doença renal pela possibilidade de regeneração tecidual e recuperação funcional, possivelmente por efeitos parácrinos. Diversos trabalhos mostraram seu efeito renoprotetor em modelo de insuficiência renal aguda. No entanto, existem poucos trabalhos que avaliaram o efeito da CT em doença renal crônica. Neste contexto, a via de inoculação e o número das CT na região da lesão podem desempenhar um papel crucial. Assim, o transplante de CT pela via EV não parece ser o mais apropriado para prover CT em número expressivo no órgão alvo. Uma técnica alternativa consiste em inocular as CT localmente, na região subcapsular renal. O objetivo do presente estudo foi analisar, em modelo experimental de doença renal crônica por nefrectomia 5/6 (Nx), a migração, a distribuição e o possível efeito renoprotetor da inoculação via subcapsular renal de 2 tipos de CT: derivadas da medula óssea (CTdmo) e mesenquimais (CTm). As CT foram coletadas de fêmur e tíbia de ratos doadores através da técnica de flushing. As CTdmo foram isoladas por gradiente de concentração e as CTm pela sua capacidade de aderência ao plástico e ambas marcadas com DAPI para a visualização no tecido. A caracterização das CT foi feita por citometria de fluxo e pela diferenciação celular in vitro. Foram realizados 2 protocolos experimentais. No protocolo I, CTdmo (1x106) foram inoculadas em ratos fêmeas e, no protocolo II, CTm (2x105) foram inoculadas em ratos machos. A região inoculada foi a subcapsula renal e os animais foram acompanhados por 15 e 30 dias. Os animais foram subdivididos nos grupos: Sham, ratos submetidos à cirurgia fictícia; Sham+CT, ratos submetidos à cirurgia fictícia que receberam CT (CTdmo ou CTm); Nx, ratos submetidos a nefrectomia 5/6; Nx+CT, Nx ratos que receberam CT (CTdmo ou CTm). Para avaliar a localização das CTdmo no tecido renal, utilizou-se a coloração de tricrômio de Masson e foi realizada uma análise semiquantitativa para avaliar o grau de infiltração. Foram analisadas a pressão arterial (PA), a albuminúria e a creatinina sérica. Para os animais que receberam CTm foi realizada a análise de parâmetros histológicos e a análise de marcadores inflamatórios, de células em atividade proliferativa, de miofibroblastos e de podócitos. Os resultados do Protocolo I avaliando a análise da infiltração no tecido renal das CTdmo marcadas com DAPI mostrou, em 5 dias, evidente infiltração das células da região subcapsular em sentido ao córtex e medula, inclusive presente em glomérulos. Ratos fêmeas Nx que receberam a inoculação das CTdmo na região da subcapsular renal não apresentaram melhora nos parâmetros que avaliaram a função renal. Protocolo II: as CTm cultivadas mostraram grande capacidade de aderência, crescimento em colônia e de diferenciação em células osteogênicas. A análise por citometria mostrou-se positiva para CD44 e CD90, com uma pequena população de células de CD34, CD45 e CD31, confirmando a presença preponderante de CTm. A inoculação de CTm em ratos Nx proporcionou um bloqueio da progressão da doença renal. Enquanto ratos Nx machos apresentaram elevada PA com 15 e 30 dias (149,6±9,1 e 191,7±2,8 mmHg) a inoculação de CTm promoveu significante redução após 30 dias (145,2±6,8 mmHg; p<0,05 vs Nx). Em ratos Nx foi observado um aumento na creatinina aos 15 e 30 dias (1,13±0,08 e 1,16±0,26 mg/dL) e a inoculação de CTm promoveu uma marcante redução aos 15 dias (0,58±0,03 mg/dL; p<0,05 vs Nx). A albuminúria foi elevada nos ratos Nx aos 15 e 30 dias (41,7±10,8 mg/24h e 138,7±33,6 mg/24h) enquanto os animais do grupo Nx+CTm aos 15 e 30 dias apresentaram diminuição significativa (4,6±1,5 mg/24h e 23,4±7,7 mg/24h; p<0,0001 vs Nx). A glomeruloesclerose do grupo Nx+CTm apresentou aos 30 dias uma redução significativa em relação ao grupo Nx (5,4±2,5% vs 22,0±6,1%, respectivamente; p<0,0001). A análise da fibrose intersticial não revelou diferença após 15 dias e 30 dias no grupo Nx+CTm em relação ao grupo Nx. Com relação ao número de macrófagos, linfócitos e de células em atividade proliferativa, os animais que receberam CTm apresentaram uma discreta diminuição de sua expressão no tecido renal. A expressão de -actina se reduziu significativamente no grupo Nx+CTm. Quanto à expressão de WT-1, específico para podócitos, os animais Nx+CTm tiveram aumento significativo da marcação em relação ao grupo Nx. Em resumo, após a inoculação de CT na região da subcapsula renal, houve marcante migração e distribuição das mesmas em direção à cortical e à medular. A inoculação de CTm proporcionou um efeito renoprotetor no modelo de nefrectomia 5/6. Sendo assim, a inoculação subcapsular renal pode representar uma importante via de inoculação, permitindo assim que um número maior de células atue na proteção da progressão da doença.
Stem cells (SCs) offer therapeutic potential for the treatment of renal diseases, due to the possibility of tissue regeneration and functional recovery. Various studies have shown renoprotection by SCs in experimental models of acute kidney disease. However, only a few studies have studied their effect in chronic kidney disease. The beneficial effect of SCs seems related to their capacity to differentiate or to secrete paracrine/endocrine factors. In this context, the inoculation route or the number of SCs homing in the injured region can play a crucial role. Therefore, transplantation of MSC through the intravenous route does not seem to be best suited for delivery of an important number of cells to the target organ. An alternative technique consists in local delivery of SCs in the subcapsular region of the kidney. The objective of the present study is to analyze the migration, distribution and potential renoprotective effect of the subcapsular inoculation of two types of SC - BSMC and mesenchymal stem cell (MSC) - in an experimental model of chronic kidney disease, the 5/6 nephrectomy (Nx). SCs were collected from the femur and tibia of donor rats by flushing. BSMC were isolated by centrifugation on a concentration gradient and MSCs were isolated by their capacity to adhere to plastic. Both types of SC were stained with DAPI to allow visualization in tissues. SC characterization was carried out by flow cytometry and differentiation in culture. Two experimental procedures were performed. In protocol I, BSMC (106 cells) were injected in female rats and in protocol II, MSCs (2x105 cells) were injected in male rats. Animals were divided into 4 groups: SHAM, sham-operated rats; SHAM+SC, sham-operated rats receiving BSMC or MSCs; Nx, rats undergoing 5/6 nephrectomy; Nx+SC, 5/6 Nx rats receiving BSMC or MSCs. We used Massons Trichrome staining and a semiquantitative analysis according to the degree of infiltration to follow the localization of BSMC in the renal tissue and to quantify their infiltration, respectively. The following parameters were studied: arterial blood pressure (AP), proteinuria (Uprot), albuminuria (Ualb) and serum creatinine (Screat). For the animals receiving SCs, analysis of histology, of inflammatory markers, of proliferating cells and of podocytes was performed. Results from Protocol I assessing DAPI-stained BSMC showed marked infiltration in 5 days from the subcapsular region to the cortex and the medulla, including presence in the glomeruli, over a period of 15 days. Female rats that received subcapsular injection of BSMC did not show improvement of the parameters used to assess kidney function. Protocol II: cultured MSCs demonstrated an ability to adhere to plastic, to grow in colonies and to differentiate in osteogenic cells. Quantitative analysis of cell markers by flow cytometry showed that isolated cells were positive for CD44 and CD90, with a small population of cells positive for CD31, CD34 and CD45, confirming a preponderant presence of MSCs. Inoculation of MSCs in Nx rats blocked the progression of the renal disease. Elevated AP in Nx rats at 15 and 30 days (149.6 ± 9.1 and 191.7 ± 2.8 mm Hg, respectively) was significantly reduced by inoculation of MSCs at 30 days (145.2 ± 6.8 mm Hg, p<0.05 vs Nx). Nx rats showed increased creatinine at 15 and 30 days (1.13 ± 0.08 and 1.16 ± 0.26 mg/dL, respectively) that was significantly reduced by injection of MSCs at 15 days (0.58 ± 0.03 mg/dL, p<0.05 vs Nx). Albuminuria was increased in Nx rats at 15 and 30 days (41.7 ± 10.8 and 138.7 ± 33.6 mg/24h, respectively) and was reduced in the Nx+MSC group at both time points (4.6 ± 1.5, and 23.4 ± 7.7 mg/24h, respectively; p<0.0001 vs Nx). Histologic analysis showed that glomerulosclerosis at 30 days in the Nx+MSC group was significantly reduced as compared to the Nx group (5.4 ± 2.5 % vs 22.0 ± 6.1 %, p<0.0001). Analysis of interstitial fibrosis did not show difference after 15 and 30 days in the Nx+MSC group compared to Nx group. Nx rats receiving MSCs showed slightly decreased inflammation markers, macrophages and lymphocytes, and proliferating cells in the renal tissue when compared to Nx rats. Analysis of myofibroblasts showed a significant decrease in expression of -smooth muscle actin in Nx+MSC rats compared to Nx rats. Podocyte number was analyzed by detection of WT-1, a specific marker. Nx rats receiving MSC had a significantly higher number of podocytes than Nx rats. In conclusion, our results show that after inoculation in the subcapsular region, SCs migrate throughout the cortex in direction of the medulla. Subcapsular inoculation of MSC provides a renoprotective effect in the model of 5/6 nephrectomy. Therefore, subcapsular inoculation could represent an important route of delivery of SCs to the kidney that allows a higher number of cells to act in the protection from progression of the disease.

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Karlsson, Christina. « Biomarkers in non-small cell lung carcinoma : methodological aspects and influence of gender, histology and smoking habits on estrogen receptor and epidermal growth factor family receptor signalling ». Doctoral thesis, Örebro universitet, Hälsoakademin, 2011. http://urn.kb.se/resolve?urn=urn:nbn:se:oru:diva-19725.

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Non-small cell lung carcinoma is a leading cause of cancer mortality worldwide. There are gender and smoking associated differences both in tumour types and clinical outcome. Squamous cell carcinomas (SCC) are more frequent among smoking men while females develop adenocarcinomas (ADCA). NSCLC among never smokers are mainly ADCA, and occurs mostly in females. The present thesis elucidates the role of estrogen receptor (ER) and epidermal growth factor receptor family (EGFR/HER2-4) in NSCLC in the perspective of gender and histology as well as the influence of smoking on those biomarkers. A recently developed technique, tissue micro array (TMA), was employed.The question of how much of a tumour tissue that needed to be included in a TMA for biomarker analysis was analyzed by a statistical approach. Data indicates a sample size of three cylinders of tumour tissue with a diameter of 0.6 mm each as being appropriate and cost-effective. In order to optimally use the up to thousands of different tumour samples within a TMA, it would be optimal to serially cut and store slides before performing in situ detection of proteins and nucleic acids. Applying up to date methodology, and by evaluation with image analysis, data are presented that shows that such handling of TMA slides would be possible without any loss of biomarker information. ERα is more frequently observed in ADCA and in females and a local estradiol synthesis is supported by the presence of aromatase. ERβ is identified as a positive prognostic marker in ADCA. Smoking is associated to increased levels of ERβ mRNA. EGFR over expression is associated with a ligand. Independent phosporylation of ERα. HER-4 intracellular domain may also act as a co-activator to ERα in ADCA, especially among neversmokers. The question of ER and EGFR family signalling crosstalk as a potential target for combined targeted therapy is raised.

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Kresak, Adam M. « The Technological History of Immunohistochemical Methods and Applications in Clinical Cancer Diagnosis and Research ». Case Western Reserve University School of Graduate Studies / OhioLINK, 2018. http://rave.ohiolink.edu/etdc/view?acc_num=case1531755545522738.

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Otali, Dennis. « The combined effect of formalin fixation and individual steps in tissue processing on immunorecognition ». Birmingham, Ala. : University of Alabama at Birmingham, 2007. https://www.mhsl.uab.edu/dt/2008r/otali.pdf.

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MILESI, GLORIA. « Temporal lobe epilepsy : a combined study with high field (7T) Magnetic Resonance Imaging and optical and ultrastructural histopathology ». Doctoral thesis, Università degli Studi di Milano-Bicocca, 2014. http://hdl.handle.net/10281/80943.

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Temporal lobe epilepsy (TLE) is the most common form of focal epilepsy with 60-75% of cases reported in surgical series. The hippocampus is involved in the genesis of the seizures and hippocampal sclerosis (HS) is the most frequent neuropathological finding observed in surgical samples. However extra-hippocampal neocortical structures may also be involved and the focal cortical dysplasia (FCD) is the most common second alteration frequently associated with HS. Aim of this project was to clarify, both at radiological and neuropathological level, the abnormalities frequently observed in the temporal pole of many TLE patients as the focal cortical dysplasia, the hippocampal sclerosis and the radiological alteration of grey/white matter boundary called “blurring”. The study was performed on surgical specimens of human temporal cortices and hippocampi from TLE patients and autoptic hippocampi at different stage of development submitted first to high resolution (7T) MRI and then processed for histological analysis. Comparison between 7T MRI data with corresponding histological and immunohistochemical features was done. The first part of this study demonstrates that thanks to differences in MRI signal intensities, depending on different density and distribution of cells and myelin fibres, high-resolution ex-vivo MRI enables to visualize the intracortical organization in normal and pathological areas and can detect the subtle architectural alterations in samples presenting dysplastic cortex. The temporo-polar blurring has demonstrated to be due to a great degeneration of fibre bundles in the white matter and, since the age at epilepsy onset is earlier in patients with blurring in comparison to those without blurring, these data suggest that it consists in a slowly evolving chronic degeneration process with the redistribution of the remaining fibres. The comparative imaging-histology investigation on healthy hippocampal specimens shows that 7T MRI can identify the main anatomical structures and the sublayers of the normal hippocampus along its anterior-posterior axis. In sclerotic surgical hippocampi high resolution MRI reveals areas of anomalous hyperintensity of signal associated to marked loss of neurons and intense gliosis as visualized by histological analysis for neuronal and glial cells. Moreover the investigation of intrahippocampal projections performed using fractional anisotropy and fiber tracking, demonstrates a disorganization of fibers in sclerotic samples in comparison to normal hippocampi. The same experimental protocol applied to healthy autoptic hippocampi at different stage of development reveals that during fetal period the MRI signal is related to the cellular density, in fact the regions characterized by densely packed neurons are easily recognizable on ex vivo high field MRI as hypointense areas, while in post-natal specimens the MRI signal is still linked to the cell density but, with the appearance and the progressive increase of the myelin fibers content, this correlation becomes less obvious and the myelin represent the histological component which mainly contributes on MRI signal intensity. This study shows the potential utility of ex-vivo high-field MRI in the understanding of temporal cortical and hippocampal abnormalities, included developmental hippocampal changes, and suggests its possible application in vivo in the near future for the study of several disorders.

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Jelena, Amidžić. « Histološke karakteristike i regeneratorni kapacitet endocervikalnih žlezda ». Phd thesis, Univerzitet u Novom Sadu, Medicinski fakultet u Novom Sadu, 2017. https://www.cris.uns.ac.rs/record.jsf?recordId=104710&source=NDLTD&language=en.

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U grliću materice se histološki razlikuju dva regiona: ektocerviks (vaginalna porcija) i endocervikalni deo. Osnovna histološka razlika između ovih delova se ogleda u epitelu koji oblaže sluznicu. Histološke osobenosti endocervikalnog dela grlića materice u dostupnoj literaturi nisu detaljno proučene i postoje mnoga neodgovorena pitanja u pogledu opštih kvantitativnih morfoloških karakteristika, kao i u pogledu dobno zavisnih promena ovih parametara. Takođe, podaci u vezi odeljka i tipa ćelijske populacije odgovorne za regeneracija epitela u endocerviksu su u dostupnoj literaturi heterogeni i nisu jasno precizirani. U ovoj studiji su morfometrijskim metodama (linearnim i stereološkim merenjem) analizirani histološki preparati grlića materice obojeni standarnim hematoksilin /eozin bojenjem i imunohistohemijskim metodama (Ki-67, p63 i CK17) s ciljem da se odrede prosečne i referentne vrednosti opštih histoloških parametara i ćelijske populacije odgovorne za regeneraciju epitela endocervikalnog dela grlića materice. Analiziran je broj endocervikalnih žlezda po santimetru dužine endocerviksa, dubina endocervikalnih žlezda, visina endocervikalnog epitela, volumenske gustine strome, žlezda, lumena žlezda i epitela, kao i volumenske gustine proliferativnih ćelija (p63+/CK17+ u odnosu na p63-/CK17-) u endocervikalnom epitelu. Obrađeno je ukupno 140 isečaka grlića materice, koji su podeljeni u dve grupe: grupa bez patoloških promena i grupa sa odabranim benignim procesima u endocerviksu. Isečci bez patoloških promena su podeljeni na osnovu godina starosti pacijentkinja u 5 podgrupa kako bi se utvrdilo da li postoje dobno zavisne razlike u vrednostima navedenih parametara. Na osnovu rezultata ovog istraživanja referentna vrednost broja endocervikalnih žlezda po santimetru dužine je 13 - 29 žl/cm, dubine endocervikalnih žlezda je 1,3 - 4 mm a visine epitelnih ćelija endocerviksa je 24 - 46 μm. Kod žena starosti preko 60 godina broj bazalnih ćelija endocervikalnog epitela opada u odnosu na mlađe pacijentkinje. Kod perimenopauzalnih žena, starosti od 40 do 49 godina, je nađena najveća prosečna vrednost dubine endocervikalnih žlezda, prosečne visine epitela endocerviksa, kao i najveći ukupni proliferativni indeks u endocervikalnom epitelu. U svim ispitivanim grupama, analizom volumenskih gustina regeneratornih ćelija endocervikalne sluznice (Ki-67 pozitivnih ćelija) u odnosu na druge imunohistomijske karakteristike (p63 i CK17 pozitivnost), rezultati dobijeni u ovoj disertaciji ukazuju na činjenicu da u endocervikalnom delu grlića materice postoje dva izvora regeneratornih ćelija.
In histologic terms, there are two different regions in the cervix: ectocervix (vaginal portion) and endocervical part. The main histologic difference between these two parts is reflected in the epithelium that coats the mucous. Histologic irregularities of the endocervical part of the cervix have not been studied in details in the available literature and there are many unanswered questions relating to general quantitative morphological features, as well as regarding age-dependent changes of such parameters. In addition, data from the available literature relating to the section and type of cell population responsible for epithelium regeneration in endocervix is heterogenous and not sufficiently precise. In this study, morphometric methods were used (by linear and stereologic measuring) to conduct histologic analysis of cervical preparations stained by standard hematoxylin/eosin stains, and immunohistochemical methods (Ki-67, p63 and CK17) with a goal to determine the average and reference values of general histologic parameters and cell populations responsible for regeneration of the epithelium of the endocervical part of cervix. The number of endocervical glands per centimetre of endocervix lenght, the depth of endocervical glands, the hight of endocervical epithelium, volumetric densities of stroma, glands, glands' lumen and epithelium, as well the volumetric densities of proliferative cells (p63+/CK17+ in relation to p63-/CK17-) in endocervical epithelium were analysed. A total of 140 cervical cuttings was processed, and they were divided into two groups: a group without pathologic changes and a group with chosen benign processes in the endocervix. Cuttings with no pathologic changes were divided into 5 subgroups on the basis of patients' age in order to determine if there are any age-dependent differences in the values of the mentioned parameters. On the basis of the results of this research, the reference value for the number of endocervical glands per centimetre of lenght is 13 - 29 gl/cm, for the depth of endocervical glands is 1.3 - 4 mm and for the hight of endocervical epithelium cells is 24 - 46 μm. With women older than 60, the number of basal cells of endocervical epithelium drops compared to younger patients. The biggest average value of the depth of endocervical glands, average hight of endocervical epithelium, as well as the biggest total proliferative index in the endocervical epithelium were found with women in perimenopause, age between 40 and 49. In all examined groups, by analysing the volumetric densities of regenerative cells of the endocervical mucus (Ki-67 positive cells) in relation to other immunohistochemical features (p63 and CK17 positivity), the results obtained in this dissertation indicate that there are two sources of regerative cells in the endocervical part of the cervix.

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Medeiros, Savi Flavia. « Bone responses to tissue engineered constructs (TECs) in critical large bone defects : Towards improved histological and immunohistochemical assessment ». Thesis, Queensland University of Technology, 2020. https://eprints.qut.edu.au/203906/1/Flavia_Medeiros%20Savi_Thesis.pdf.

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This research was focused on the development of advanced quantitative methods to evaluate the use of a 3D printed membrane scaffold, for the guidance and spatiotemporal delivery of recombinant human bone growth factor for the regeneration of a large bone defect, which still represents a major challenge in orthopaedic and reconstructive surgery. Two histomorphometric approaches were investigated and successfully applied. The combined histological, immunohistochemical and histomorphometric analyses outlined in this thesis further elucidated the mechanisms and patterns that govern the bone regeneration of a critical sized bone defect in a large experimental ovine model.

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Medrado, Andreone Teles. « An Atlas of catfish brain - Steindachneridion parahybae (Teleostei : Siluriformes) : a detailed cytoarchitectonic study of the different brain areas and nuclei as a basis for further morphological and functional studies ». Universidade de São Paulo, 2015. http://www.teses.usp.br/teses/disponiveis/41/41135/tde-04112015-150105/.

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In the present Master\'s Dissertation, a detailed cytoarchtectonic study of the brain of the juvenile catfish - Steindachneridion parahybae, has been performed. The animals used for this Atlas were juvenile specimens of one hundred days post-fertilization. The coronal (transverse) sections (5µm-thick) were obtained by using a rotary microtome, stained with cresyl-violet and examined under a photomicroscopy with the help of a digital system of analysis. Some criteria have been used to classify the different cell masses of the catfish brain: (i) characteristic size, shape and intensity of the staining from the perykarya; (ii) packing density and distribution pattern of the cell bodies; (iii) neuropil surrounding the cell groups and (iv) consistency of cell groups in both hemispheres and different brains of catfish. Thus, around one hundred and thirty nuclei have been described in the catfish brain, which are distributed in four main region that are from rostral to caudal: telencephalon, diencephalon, mesencephalon and rhombencephalon. Although we have observed important similarities between the brain of catfish and other teleosts, we have also noticed some differences in the characteristics and placement of several nuclei in relation to other teleosts, or even when compared to the brain of species of the same Order, the Siluriformes. Some of these differences could be related with the age of the animals studied here, but probably represent species-specific differences because the brain of adult catfish specimens has a great similarity in cytoarchitecture and overall organization compared to younger animals. The main outcome of this study has been the availability of a complete Atlas of the brain of catfish, which has been used to localize precisely the distribution of cells and fibers of the Gonadotropin-releasing hormone in the brain. This Atlas will also represent a valuable tool for future endocrine analyses, allowing the precise mapping of the different neurohormones in the brain of catfish, as well as for the study of neural connections among different brain areas
Esta Dissertação de Mestrado, apresenta-se estruturalmente como um Atlas, em que é apresentado um detalhado estudo citoarquitetônico do encéfalo de catfish- Steindachneridion parahybae. Para a realização deste, foram utilizados 7 juvenis de 100 dias após a eclosão, analisados por técnicas rotineiras de histologia, cujas secções coronais(transversais) - 5&um;m de espessura-, foram obtidas utilizando-se de um micrótomo rotativo, coradas com violeta de cresil e examinadas a partir de sistema digital de análise. Alguns critérios foram utilizados para classificar as diferentes massas de células do cérebro catfish, tais como: (i) o tamanho característico, forma e intensidade da coloração do pericário; (ii) padrão de densidade de agrupamento e distribuição dos corpos celulares; (iii) a presença de neurópilos ao redor dos desses agrupamentos celulares e (iv) a consistência/coerência destes agrupamentos em ambos os hemisférios dos diferentes encéfalos, então analisados. Dessa forma, são descritos aproximadamente130 massas celulares para o encéfalo de S. parahybae, as quais estão distribuídas em quatro principais regiões que, da parte rostral para caudal, são: telencéfalo, diencéfalo, mesencéfalo e rombencéfalo. Embora são observadas semelhanças entre o cérebro de S. parahybae e de outros teleósteos, nota-se, também, certas diferenças quanto às características e/ou localização das massas celulares em relação ao encéfalo de outros teleósteos, ou mesmo quando comparado com o cérebro de espécies da mesma ordem, Siluriformes. Algumas destas diferenças pode estar relacionada com a idade dos animais estudados, no entanto,também podem representar diferenças espécie-específicas, uma vez que o encéfalo adultos de S. parahybae apresentam grande similaridade citoarquitetônica, além da organização geral do encéfalo, previamente observadas em animais acima dos 100 dias após a eclosão. Portanto, como resultado deste estudo tem-se a disponibilidade de um Atlas completo do encéfalo de S. parahybae, o qual representa uma ferramenta valiosa para o estudo das conexões neurais entre diferentes áreas do encéfalo, bem como para futuras análises endócrinas, permitindo o mapeamento preciso de neuro-hormônios nesta espécie, como demonstrado ao longo deste estudo, para o hormônio liberador de gonadotropinas

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Tozzi, Brunela Bastos. « Análise histopatológica e imuno-histoquímica da erupção papular prurítica associada ao vírus da imunodeficiência humana ». Universidade Federal do Espírito Santo, 2012. http://repositorio.ufes.br/handle/10/5959.

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Made available in DSpace on 2016-12-23T13:56:13Z (GMT). No. of bitstreams: 1 Brunella Bastos Tozzi.pdf: 5530561 bytes, checksum: 4cb5717338550ea3614682134ea4a15f (MD5) Previous issue date: 2012-06-22
Introduction: Papular pruritic eruption (PPE) is characterized by the presence of pruritic erythematous papules as chronic disease that affects oftenly seropositive patients. The lesions are seen in different stages, preferably in the trunk and extremities, and end with post inflammatory hyperpigmentation and / or scars resulting from scratching. Objectives: To evaluate the hitopathological and immunohistochemical features of the PPE. Methods: The study enrolled HIV-positive patients with clinical diagnosis of PPE carried out at University Hospital Cassiano Antonio Moraes of the Federal University of Espirito Santo in Vitória, ES. Patients were dermatological examinated and skin lesions photographed. Skin biopsies were obtained to evaluate the histopathological and immunohistochemical features. Results: Thirty-nine patients with EPP were biopsied, 17 were women and 22 men, mean age was 40.2 years ranging from 25 to 63 years. Among the histopathological biopsies, 6 patterns were found and these often: 1) dermal perivascular inflammatory infiltrate, 2) inflammatory infiltrate affecting the follicular unit; 3) interstitial inflammatory infiltrate, 4) inflammatory infiltrate in the subcutaneous tissue, 5) inflammatory infiltrate affecting at least one nerve, 6) signal of the scratching in the epidermis. About immunohistochemistry features, Langerhans cells are found in normal amounts in the epidermis and relatively increased in dermal inflammatory infiltrate. There are predominantly CD8 + T lymphocyte and macrophages in the inflammatory infiltrate compared with CD4 + T lymphocytes. No microorganisms were found using the Ziehl-Neelsen staining and Grocott. Conclusions: It was identified six patterns present in biopsy, in order to obtain a set of histopathologic and immunohistochemical to describe and characterize the PPE histopathological diagnosis. The dermal perivascular inflammatory infiltrate is essential for diagnosis, and other tissue alterations are not mandatory that may be present together or separately, at different stages of the skin lesions.
Introdução: A Erupção Papular Prurítica (EPP) é caracterizada pela presença de pápulas eritematosas pruriginosas de evolução crônica que afetam, com frequência, pacientes HIV positivos. As lesões são observadas em diferentes estágios evolutivos, preferencialmente no tronco e extremidades, e evoluem com hiperpigmentação pós inflamatória e/ou cicatrizes decorrentes da coçadura. Objetivos: Descrever as características histopatológicas e imuno-histoquímicas de biópsias de lesões de EPP em pacientes HIV positivos. Metodologia: Foram selecionados pacientes HIV positivos com diagnóstico clínico de EPP atendidos no Hospital Universitário Cassiano Antônio Moraes (HUCAM) da Universidade Federal do Espírito Santo (UFES), em Vitória-ES. Os pacientes foram submetidos a um exame dermatológico criterioso, as lesões fotografadas e biopsiadas para a avaliação histopatológica e imuno-histoquímica. Resultados: Trinta e nove pacientes com EPP foram biopsiados; 17 eram mulheres e 22 homens; a média de idade foi 40,2 anos variando de 25 a 63 anos. Dentre os aspectos histopatológicos nas biópsias, 6 padrões foram encontrados com frequência sendo esses: 1) infiltrado inflamatório perivascular dérmico; 2) infiltrado inflamatório comprometendo a unidade folicular; 3) infiltrado inflamatório intersticial; 4)infiltrado inflamatório no subcutâneo; 5) infiltrado inflamatório comprometendo pelo menos um nervo; 6) sinal da coçadura na epiderme. Na imuno-histoquímica, as células de Langerhans encontram-se em quantidades normais na epiderme e relativamente aumentadas nos infiltrados inflamatórios dérmicos. Há predomínio de linfócitos T CD8+ e macrófagos nesses infiltrados, comparando com os linfócitos T CD4+. Não foram encontrados microorganismos nos exames, tanto utilizando-se a coloração Ziehl-Neelsen, quanto a coloração Grocott. Conclusões: Foram identificadas seis alterações mais frequentes presentes nas biópsias, afim de se obter um conjunto de alterações histopatológicas e imunohistoquímicas que descrevessem e caracterizassem o diagnóstico histopatológico da EPP. O infiltrado inflamatório perivascular dérmico é imprescindível para o diagnóstico, e as demais são alterações teciduais variáveis, que podem estar presentes juntas ou separadas, em estágios diferentes da doença

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Deshmukh, Abhay S. « Histological Characterization of Inter Ictal Epileptiform Discharges Generating Brain Regions using a Preclinical Model of Focal Cortical Dysplasia ». FIU Digital Commons, 2015. http://digitalcommons.fiu.edu/etd/2316.

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Current clinical practice of resective surgery in focal epilepsy involves electroencephalogram (EEG) brain source imaging to localize irritative brain areas from where Inter-ictal epileptiform discharges (IEDs) emerge, useful to localize the seizures-onset zones. Unfortunately, there are no previous systematic studies to characterize the pathophysiological mechanisms and abnormal cellular substrates in these irritative areas since histological data are available only from the final resective zones. To address this issue we applied a combination of EEG brain imaging described by Bae et al. (2015) using cutting-edge technology for high-density scalp EEG in rodents and histological analysis on a chronic rat model of focal cortical dysplasia. Post-mortem brain sections were stained for anatomical, functional and inflammatory biomarkers. Abnormal anatomical structures and increased expression of inflammatory biomarkers were found in the irritative regions. We conclude that IED-based brain source imaging can help to localize abnormal tissues highly prospective for epileptogenesis.

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Линдін, М. С. « Удосконалення роботи Наукового центру патоморфологічних досліджень Сумського державного університету в умовах реформування системи охорони здоров’я ». Master's thesis, Сумський державний університет, 2022. https://essuir.sumdu.edu.ua/handle/123456789/87718.

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У роботі проведено дослідження структури та функцій патологоанатомічної служби в Україні, її актуальні проблеми на сучасному етапі та органи при проведенні реформ. Проведено аналіз умов надання діагностичних послуг у Науковому центрі патоморфологічних досліджень СумДУ, як однієї зі складових патологоанатомічної служби, та проаналізовано основні показники його діяльності. На основі цього було виявлено основні проблеми та потенційні кризи у організації та управлінні роботи Наукового центру патоморфологічних досліджень. На основі виявлених проблем розроблені програми розвитку для подолання наявних та можливих перешкод в умовах реформування медицини.
В работе проведено исследование структуры и функций патологоанатомической службы в Украине, ее фактические проблемы на настоящем этапе и органы во время реформ. Был проведен анализ условий предоставления диагностических услуг в научном центре патоморфологических исследований СумГУ, как один из компонентов патлогоанатомической службы, и проанализированы основные показатели его деятельности. Исходя из этого, были выявлены основные проблемы и потенциальные кризисы в организации и управлении работой Научного центра патоморфологических исследований. На основании выявленных проблем были разработаны программы развития для преодоления доступных и возможных препятствий в условиях реформы медицины.
A study of the structure and functions of the pathoanatomical service in Ukraine as well as its actual problems at the present stage and organs during reforms have been conducted. The analysis of the conditions for the provision of diagnostic services at the Scientific Center of Pathomorphological Studies of SSU, as one of the components of the pathoanatomical service has been conducted, and the main indicators of its activity have been analyzed. On this basis, the main problems and potential crises in the organization and management of the scientific center of pathomorphological research have been identified. On the basis of identified problems, development programs have been developed to overcome available and possible obstacles in the conditions of medicine reform.

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Pinna, Fabio de Rezende. « Distribuição do neuroepitélio olfatório em concha média e superior em cadáveres humanos ». Universidade de São Paulo, 2008. http://www.teses.usp.br/teses/disponiveis/5/5143/tde-19112008-162629/.

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INTRODUÇÃO: A biópsia do neuroepitélio olfatório (NeuO) oferece perspectivas para aplicações terapêuticas tanto em doenças do olfato como doenças neurodegenerativas. Uma coleta bem sucedida desse tecido in vivo ainda não é rotina, devido á carência de estudos sobre a distribuição do NeuO em conchas superior (CS) e média (CM). Neste trabalho, descrevemos a distribuição do NeuO na CS e CM em cadáveres a partir da retirada integral dessas estruturas e posterior análise histológica por coloração de hematoxilina e eosina (HE) e imunoistoquímica. Além disso, também analisamos a influência do sexo, idade e lateralidade no grau de presença do NeuO nas CS e CM. CASUÍSTICA E MÉTODOS: Estudo anatômico prospectivo realizado de março de 2006 a janeiro de 2008. A CS e a CM foram endoscopicamente retiradas de um total de 25 cadáveres frescos com menos de 12 horas de óbito. Cada concha foi seccionada na metade de seu comprimento ântero-posterior. Assim, cada um dos 25 cadáveres deu origem a oito fragmentos de mucosa de regiões anatômicas distintas, totalizando 200 lâminas para análise tanto por coloração de HE como por reação de imunoistoquímica. Nas lâminas coradas por HE, classificamos a distribuição do NeuO em graus 0, 1, 2, 3, 4, sendo que a análise foi realizada por 3 patologistas de forma cega. Para imunoistoquímica, só obtivemos positividade com a proteína S-100. A concordância entre os três patologistas foi avaliada aos pares utilizando-se o coeficiente de Kappa. A distribuição do NeuO foi analisada de acordo com a idade, sexo, tempo de óbito, simetria entre as fossas nasais e acurácia da imunoistoquímica. RESULTADOS: Pela HE na CS, o NeuO esteve presente em 82,9% das vezes e, na CM, em 17,1%. Na CS, o NeuO foi detectado em 82,9 % das lâminas, 4,9 vezes a prevalência na CM, que foi de 17,1 % das lâminas (p < 0,001). Pela imunoistoquímica, foi possível encontrar NeuO em um total de 15 fragmentos. Desses, 10 (20%) eram da metade posterior da CS e cinco (7,6%) da metade anterior da CS. Pelo cálculo da razão de prevalência, temos que a chance de encontrar NeuO é 4,9 vezes maior na CS do que na CM (IC95%: 3,3 7,4). Dos 15 fragmentos com marcação positiva para proteína S-100, sete corresponderam aos que tinham uma distribuição grau 3 (>50% e 75%) pela HE e outros sete aos que tinham uma distribuição grau 4 (acima de 75%). Somente um fragmento teve marcação positiva para imunoistoquímica no grupo 2 (entre 26 e 50%) na HE. A proteína S-100 apresentou uma sensibilidade de 13,5% e especificidade de 100% para detecção de NeuO. Não houve diferença estatisticamente significante na prevalência de NeuO quando os fragmentos foram divididos de acordo com o sexo, idade de óbito e lado da fossa nasal. No entanto, ao analisarmos a presença de NeuO de acordo com o grau de distribuição entre cada lado, não se percebe uma concordância. CONCLUSÕES: A quantidade total de NeuO foi simetricamente distribuída entre as fossas nasais, mas não houve uma concordância entre os lados quanto à maneira como o NeuO está distribuído. O NeuO apresenta maior probabilidade de ser encontrado na metade posterior de CS. A HE é um método eficaz para distinção entre NeuO e epitélio respiratório, devido a grande concordância entre três patologistas distintos.
INTRODUCTION: Olfactory neuroepithelium (ON) biopsy provides perspectives for several therapeutic applications, both in disorders of olfaction and in neurodegenerative diseases. Successful in vivo collection of ON is still not routine, due to a dearth of studies on ON distribution in the superior and middle turbinate (ST and MT respectively). This study describes the distribution of ON in cadaver ST and MT as determined by complete endoscopic removal of turbinates and histological analysis with hematoxylin and eosin (H&E) and immunohistochemical staining. We also analyzed the influence of gender, age, and naris side on the extent to which ON is present in the superior and middle turbinate. CASE SELECTION AND METHODS: We conducted a prospective anatomical study from March 2006 to January 2008. The superior and middle turbinates of 25 fresh cadavers (less than 12 hours post-mortem) were removed endoscopically. Each turbinate was halved into anterior and posterior fragments. Eight anatomically distinct fragments were therefore obtained from each of the 25 cadavers for a total of 200 specimens, which were analyzed through H&E staining and immunohistochemistry. Hematoxylin and eosin-stained slides were subjected to blind examination by three independent pathologists; ON distribution was graded on a fivepoint numeric scale (grade 0, 1, 2, 3, or 4). Immunohistochemistry was only positive through S-100 staining. Pairwise agreement between pathologists was assessed by means of the Kappa coefficient. The distribution of ON was analyzed regarding age, gender, time elapsed between death and specimen harvesting, symmetry between nares, and accuracy of immunohistochemistry results. RESULTS: In H&E-stained slides, olfactory neuroepithelium was present in 82.9% of ST and 17.1% of MT specimens; prevalence in the superior turbinate was therefore 4.9-fold greater (p < 0.001). Immunohistochemical analysis was able to identify ON in 15 fragments, 10 of which (20%) were from the posterior half of the superior turbinate; the remaining five specimens (7.6%) were from the anterior ST. According to prevalence ratio, the odds of finding ON are 4.9 times greater in superior turbinate than in the middle turbinate (CI, 95%; 3.37.4). Of the 15 immunohistochemistry-positive fragments, seven were assigned distribution grade 3 (>50% and 75% presence of ON) on H&E staining seven others were graded 4 (>75% presence of ON). A single immunohistochemistrypositive fragment was found to have grade 2 ON distribution (i.e., it contained 26% to 50% olfactory neuroepithelium) on H&E staining. S-100 staining showed a sensitivity of 13.5% and specificity of 100% for ON detection. There was no statistically significant difference in ON prevalence when fragments were compared according to gender, age at time of death, and naris side. However, when we analyzed ON presence according to the degree of ON distribution in each side, we found no concordance. CONCLUSIONS: Total ON was distributed symmetrically between nares, but we found no concordance between sides in the manner in which ON is distributed. ON is most likely to be found in the posterior half of the superior turbinate. Hematoxylin and eosin (H&E) staining is an effective method for distinguishing ON from respiratory epithelium, as shown by high inter-rater agreement among three independent pathologists

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Tchokonte-Nana, Venant. « Cellular mechanisms involved in the recapitulation of endocrine development in the duct ligated pancreas ». Thesis, Stellenbosch : University of Stellenbosch, 2011. http://hdl.handle.net/10019.1/6828.

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Thesis (PhD)--University of Stellenbosch, 2011.
ENGLISH ABSTRACT: Diabetes mellitus is amongst the leading causes of morbidity and mortality in the world, affecting young, adult and old people. Beta cell replacement therapy for insulin delivery remains the ultimate remedy for diabetes. However, insufficient donor pancreas and the use of immunosuppressive drugs prevent the wide-spread of this therapy. Other avenues of self generated beta cells within the organ itself need to be explored. Therefore, understanding the chronobiology of cellular mechanisms in the lineage of beta cell induced neogenesis is a valuable tool in improving beta cell replacement in patients with diabetes. The aim of this study was to induce recapitulation of the morpho-genetic sequence of endocrine cells development in the pancreas of rats after the pancreatic duct ligation (PDL) procedure. Serial sections of PDL tissues of the pancreas were obtained from 78 Sprague- Dawley rats and were assessed morphologically. The immunofluorescent tissues were statistically analysed using a computerized morphometry technique. The protein expression indices of Caspase3, Insulin, Pdx1, Ngn3, NeuroD and Pax6 were quantified. The efficiency levels of coexpression of these homeodomain proteins separately with insulin were defined by the ratio of the mean value of insulin expression to the mean value of their respective protein expression. The morphological changes were characterized by the appearance of granulated acinar cells at 6 hours post-PDL and the proliferation of endocrine tissues from 84 hours through to 120 hours. The morpho-immunofluorescent evaluation showed the highest immunoreactivity of Caspase3 and Pdx1 at 6 hours, Ngn3 at 36 hours, Pax6 and insulin at 84 hours while NeuroD expression was at 120 hours. The immunohistofluorescent analysis showed that caspase3 and Pdx1 were the first to be expressed at 6 hours while the insulin and NeuroD expression appeared later at 84 hours and 120 hours, respectively. However, Pax6 expression was continuous across time periods post-PDL, while Ngn3 expression showed a peak at 36 hours. The efficiency (highest and earliest expression) of co-expression of all these homeodomain proteins with insulin was restricted between 12 hours and 24 hours. The optimal efficiency was at 12 hours by Ngn3 with insulin. A good efficiency was shown for Pdx1 with insulin, NeuroD with insulin and Pax6 with insulin at 12 hours and 24 hours, respectively. A low efficiency was observed for insulin and caspase3 co-expression at 24 hours. This study suggests that for transplantation, PDL tissues harvested at an early time post-PDL (between 12 and 24 hours) could yield a higher success rate; the study also provides evidence for a connection between morphological changes in the PDL pancreas and the protein synthesis necessary for the lineage of endocrine cell development.
AFRIKAANSE OPSOMMING: Diabetes Mellitus resorteer onder die vernaamste oorsake van morbiditeit en mortaliteit wêreldwyd, en tuister jongmense, volwassenes en bejaardes. Daar bestaan egter ‘n wêreldwye tekort aan skenkerorgane met immuun-onderdrukingsterapie as ondersteuningsbehandeling. Beta-sel vervangingsterapie, vir die voorsiening van insulien, bly daarom die voorkeur behandeling vir die siekte wat noodsaak dat die wetenskap kyk na alternatiewe behandelingsregimens wat meganismes rondom orgaanregenerasie insluit. Begrip van die chronobiologie van die sellulêre meganismes betrokke rondom beta-sel ontwikkeling mag waardevolle lig werp op die neogenese van beta-selle wat gevolglik daartoe mag lei dat beta-sel vervanging as ‘n moontlike behandelingsterapie oorweeg mag word vir pasiënte met suikersiekte. Die oogmerk van hierdie studie is om die rekapitulasie van die morfo-genetiese volgorde van die endokriene pankreas na afbinding van die pankreasbuis te bepaal. Pankreasbuis afbinding is op 78 Sprague-Dawley laboratorium rotte onder algemene narkose uitgevoer, die pankreas is na voorafbepaalde tydsvakke verwyder en in histologiese seriesnitte gesny. Snitte is immunositochemiese gekleur en morfometries assesseer. Die afskeidingsindeks vir selboodskappers vir Caspase3, Insulien, Pdx1, Ngn3, NeuroD en Pax6 is kwantifiseer. Die gelyktydige afskeiding van elk van bogenoemde boodskappers tesame met insulien is omskryf as ‘n verhouding tot mekaar en in terme van dié van insulien. Die morfologiese verandering in die weefsel bespeur is gekenmerk deur die verskyn van gegranuleerde asinêre selle ses (6) ure na buisafbinding en die proliferasie van endokriene weefsel vanaf vier-en-tagtig (84) ure deurlopend tot een-honderd-en-twintig (120) ure. Die morfo-immunofluoresserende evaluering toon dat Caspase3 en Pdx1 by 6 uur die hoogste is, die van Ngn3 by 36 ure, Pax6 en insulien by 84 ure en NeuroD by 120 ure. Verder toon die analise dat Caspase3 en Pdx1 rondom 6 ure hul verskyning gemaak het terwyl dié van insulien en NeuroD eers rondom 84 tot 120 uur verskyn het. Die verskyning van Pax6 het deurlopend regoor al die tydsduurtes verskyn en Ngn3 het rondom 36 uur sy hoogste vlak bereik. Die gelyktydige uitdrukking van homeodomein proteïene tesame met insulien het slegs tussen die tydperke van 12 en 24 ure plaasgevind. Die uitdrukking van Pdx1 met insulien, NeuroD met insulien en Pax6 met insulien het almal tussen 12 en 24 ure plaasgevind. Caspase3 tesame met insulien is slegs by die 24 uur tydsperiode bespeur. Vir die oorplant van pankreas weefsel wat aan buisafbinding onderwerp is suggereer hierdie studie dat die geskikste tyd vir die oes van endokriene weefsel liewer vroeër (12 to 24 ure) as later uitgevoer behoort te word. Verder wil dit voorkom of hierdie tydsperiode ook die hoogste seltelling lewer. Die studie lewer waardevolle inligting oor die verwantskap tussen die morfologiese veranderings wat na buisafbinding plaasvind en die proteïen sintese wat sel-opvolgontwikkeling bevorder.

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Dias, Fernando José. « Efeito do laser de baixa intensidade e da proteína purificada do látex natural (Hevea brasiliensis) sobre a lesão por esmagamento do nervo isquiático de ratos wistar : estudo morfológico, imunohistoquímico e ultraestrutural ». Universidade de São Paulo, 2014. http://www.teses.usp.br/teses/disponiveis/42/42131/tde-03062014-155359/.

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Esse estudo avaliou o laser de baixa intensidade (LBI-15J/cm2, 780nm) e a proteína do látex (P1; 0,1%) no esmagamento (15Kgf, axonotmese) do nervo isquiático de ratos Wistar. Grupos: GI-controle; GII-exposto; GIII-lesado; GIV-LBI; GV-P1; GVI-LBI e P1. Após 4 e 8 semanas realizou-se as análises morfológica, morfométrica, imunohistoquímica (VEGF e NGF) e ultraestrutural. Houve melhora com os tratamentos propostos. Após 4 semanas não foi possível eleger o melhor tratamento; após 8 semanas os animais que receberam a P1 associada ou não ao LBI apresentaram mais capilares e fibras nervosas mais organizadas. Após 4 semanas as expressões do NGF e VEGF estavam elevadas nos GIII, GIV e GVI e após 8 semanas elas estavam diminuídas. As características dos grupos lesados eram mais próximos dos controles após 8 semanas, revelando tempo-dependência; ainda nesse período a P1 apresentou melhor recuperação, pois o LBI não alcançou os mesmos resultados, e a associação dos tratamentos não potencializado a recuperação, sugerindo que a melhora observada nesse caso se refere à P1.
This study evaluated the low intensity laser (LBI-15J/cm2, 780nm) and latex protein (P1, 0.1%) in the crush (15 kgf, axonotmesis) of the sciatic nerve in rats. Groups: GIcontrol; GIIexposed; GIII-injured; GIV-LBI; GV-P1; GVI-LBI and P1. After 4 and 8 weeks were performed morphological, morphometric, immunohistochemical (VEGF and NGF) and ultrastructural analysis. There was improvement with the proposed treatments. After 4 weeks has not been possible to choose the best treatment, after 8 weeks, the animals that received the P1 associated or not to LBI had more capillaries and more organized nerve fibers. After 4 weeks the expressions of NGF and VEGF were higher in GIII, GIV and GVI and after 8 weeks they were decreased. The characteristics of the injured groups were closer to control after 8 weeks, revealing time-dependence, even in this period P1 showed better recovery because the LBI did not achieve the same results, and the association of treatments did not enhanced this recovery, suggesting that the improvement observed in this case refers to P1.

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Pellegrini, G. « ALVEOLAR SOCKET PRESERVATION TECHNIQUE : HISTOLOGICAL HEALING OF HARD AND SOFT TISSUES ». Doctoral thesis, Università degli Studi di Milano, 2012. http://hdl.handle.net/2434/173987.

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INTRODUCTION After tooth extraction, the use of grafting materials and absorbable membrane, alone or in association, were proposed for maintenance procedures of alveolar ridge volume. The preservation of bone volume in posterior maxillary area might be necessary to optimize the delayed implant placement without sinus augmentation procedure. In addition, importance of peri-implant soft tissue has been related to final aesthetic outcomes. The alveolar socket preservation technique may affect the healing of the overhanging mucosa. This research is composed by two studies (A and B). Aim of the study A was to evaluate histologically the integration and remodelling of porous bovine bone particles (PBBM) grafted in the extraction socket, and to compare the dimensional alterations of the alveolar ridge in the molar maxillary area after extraction with or without the graft of PBBM particles, at medium and advanced endpoints. Aim of the study B was to histologically compare soft tissue features of augmented alveolar sockets covered or not covered with collagen membrane. Methods Study A: This is a randomized double-blind controlled clinical trial. Sixteen patients who needed the extraction of one maxillary molar tooth were included in the study. After extraction seven sites were treated with PBBM particles (Bio-Oss Collagen) covered with collagen membrane (test), and nine sites were left untreated as control. At baseline, 3 and 6 months the alveolar ridge dimension changes were evaluated using an acrylic resin stent. At 3, 6 and 9 months one tissue sample was harvested in three selected patients and processed for histological analysis. At 9 months the authors also decided for implant placement with or without previous sinus floor augmentation. Study B. This is a randomized double-blind controlled split-mouth clinical trial. Twelve patients requiring extraction of 2 controlateral maxillary premolar teeth were included. Following extractions in each patient, one randomly selected site was augmented with graft material (Bio-Oss Collagen, Geistlich) (control group), the other site was augmented and covered with collagen membrane (Bio-Gide, Geistlich) (test group). Following 5 weeks (early endopint, n=7 patients) or 12 weeks (late endpoint, n=5 patients) samples of newly-formed soft tissue were harvested and processed for evaluation of micro vascular density (MVD) (immunohistochemistry with CD31), collagen content (AA%) and amount of inflammatory infiltrate (immunohistochemistry with CD3 for T-lymphocytes and CD20 for B-lymphocytes). Non-parametric statistical analyses were performed to compare data from test and control groups at early and late endpoint. Data from the same group were also compared between early and late endpoints. Pearson’s correlation was used to compare intra-patient data. RESULTS: Study A. Data on alveolar ridge volume changes did not show significant differences between test and control group. However in the control group sites that maintained the buccal bone showed a substantial loss of buccal bone compared to the test group. At 3 months histological evaluation grafting material was detected immersed in highly cellular connective tissue. At 6 and 9 months grafting material was mainly remodeled and remaining particles were in contact with newly formed lamellar bone. The odds ratio for receiving the sinus augmentation before implant placement resulted 2.25 times greater in control group than in test group (p=0.39). Study B. At 5 weeks after extraction, in both groups, epithelial and connective tissues appeared normally organized, without increased inflammatory infiltrate or cellular morphological alterations. Connective tissue of both groups presented remaining graft particles surrounded by mature collagen fibres. At 12 weeks a more mature structure of the epithelium and connective tissue was detected. No graft particles were observed. At 5 weeks the vascularization of tissue in samples sites treated with the graft+ resorbable membrane was significantly lower (MVD= 6.18) than in samples harvested from sites treated only with the filling material (MVD= 9.44) (Wilcoxon Matched Pair-Signed Rank, p<0.05). At this endpoint, data on Lymphocytes T and B, and collagen content showed no significant differences between test and control group. At 12 weeks no significant differences between test and control group were found for any of the considered parameter. Data on MVD and collagen content resulted increased at 12 weeks after surgery in both groups, however only data on collagen content in test group increased significantly. CONCLUSIONS: The alveolar ridge augmentation procedure increases the possibility to insert implants without sinus augmentation procedure. PBBM undergoes remodeling process, however particles included in newly formed bone still remain at 9 month. Soft tissue samples of the sites treated with PBBM particle and covered with collagen membrane showed an initial delayed healing process with a lower microvascular density and collagen content that sites treated with filling material alone. Subsequently sites treated with the membrane seem to gain the same maturation level than control sites, and were characterized by a strong augmentation of collagen fibers and microvascular density.

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Canciani, E. « ANALISI IMMUNOISTOCHIMICA E MOLECOLARE DEL PATTERN DI RIMODELLAMENTO OSSEO NELL¿ALVEOLO POST-ESTRATTIVO ». Doctoral thesis, Università degli Studi di Milano, 2014. http://hdl.handle.net/2434/229909.

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Analisi immunoistochimica e molecolare del pattern di rimodellamento osseo nell’alveolo post-estrattivo SCOPO Il sito dell’alveolo post-estrattivo è un modello ampiamente utilizzato nello studio del rimodellamento osseo fisiologico o patologico e del comportamento di biomateriali per la rigenerazione ossea. Lo scopo di questo progetto è stato quello di caratterizzare il pattern morfologico e molecolare dei marker coinvolti nel rimodellamento osseo alveolare, prima e dopo l’estrazione dentale, ed eventualmente in combinazione con un biomateriale utilizzato per preservare l’anatomia del sito ai fini riabilitativi. MATERIALI E METODI Per questo studio sono stati arruolati 36 pazienti, suddivisi in 3 gruppi indipendenti: osso alveolare (OA, n=14), guarigione spontanea (GS, n=10) e biomateriale (B, n=12). Durante la seduta di estrazione dentaria sono state raccolte 2 piccole biopsie per paziente per effettuare l’analisi morfologica (tessuto decalcificato) e molecolare (mRNA retro-trascritto in cDNA e Real Time-PCR) (gruppo OA). Dopo 4-6 mesi dall’estrazione dentale, sono state prelevate 3 biopsie per paziente, in concomitanza dell’inserimento dell’impianto dentale, nel sito guarito spontaneamente (GS) o mediante biomateriale (B) (idrossiapatite arricchita con magnesio). Per ogni gruppo sono state effettuate le stesse analisi del gruppo OA e l’analisi istomorfometrica (tessuto calcificato). L’analisi morfologica ha previsto: colorazione Ematossilina/Eosina per osservare un eventuale infiltrato infiammatorio, colorazione immunoistochimica dei fattori caratterizzanti il pattern di rimodellamento osseo (TNF-α, IL-6, RANK, RANKL e OPG), espressa come marcatura percentuale. L’analisi molecolare è stata effettuata sugli stessi marcatori normalizzati sul gene housekeeping 18S. Sui tessuti ottenuti per usura, è stata valutata l’osteointegrazione del biomateriale e la componente minerale mediante la valutazione delle frazioni di volume (stereologia). Per ciascun parametro sono state calcolate la media e la deviazione standard; è stato utilizzato il test di Kruskal Wallis (p<0,05) per valutare le eventuali differenze tra i 3 gruppi per ogni parametro e successivo test post hoc di Wilcoxon (p< 0,05/3). RISULTATI All’osservazione i campioni raccolti non presentavano né anomalie strutturali né infiltrato infiammatorio. Sia l’analisi immunoistochimica (AI) che molecolare (AM) ha mostrato un incremento per tutti i biomarker. AI: I tre gruppi erano significativamente differenti per il marcatore TNF-α (p<0.05), RANK (p<0.01), IL-6 e RANKL (p<0.001). Il test post hoc ha mostrato una differenza tra OA e B per RANK (p≤ 0.01), IL-6 e RANKL (p ≤0.001); tra GS e B per IL-6 (p≤0.01). AM: Il test di Kruskal Wallis ha mostrato differenze tra i gruppi per IL-6 (p≤0.05), RANKL e OPG (p≤0.001); il test post hoc ha riscontrato una significativa differenza tra OA e B per IL-6 (p≤0.01); OA e GS per RANKL e OPG (p≤0,001); tra GS e B per IL-6 e RANKL (p≤0,01), OPG (p≤0,001). Il rapporto RANKL/OPG ha mostrato una tendenza alla riduzione dell’osteoclastogenesi nel gruppo B. L’analisi istomorfometrica ha mostrato un’alta percentuale di componente mineralizzata nel gruppo con il biomateriale; tutte le particelle innestate erano circondate da osso rigenerato che creava ponti tra di esse. CONCLUSIONI Rispetto al rimodellamento fisiologico tutti i marker indagati hanno mostrato un incremento in entrambi i gruppi in fase di guarigione dell’alveolo post-estrattivo. Nei siti trattati con il biomateriale si è osservato un aumento della citochina IL-6 ed una diminuzione di RANK, RANKL e OPG rispetto alla guarigione spontanea. Questo può significare un rallentamento della ricostituzione del sito anatomico, dovuto ad una possibile down-regulation dell’osteoclastogenesi. Tuttavia, l’analisi della struttura dell’alveolo ha mostrato una buona rigenerazione ossea, in entrambi i gruppi; pertanto il biomateriale analizzato può essere considerato un valido ausilio per la preservazione dell’alveolo post-estrattivo.
Immunohistochemical and molecular analysis of bone remodelling pattern in alveolar socket. AIM Following tooth extraction, the alveolar bone remodelling process starts. The aim of this research project was to characterize the molecular and morphological aspects of the alveolar bone remodelling pattern before and after dental extraction and in conjunction with the use of hydroxyapatite enriched with magnesium (Mg-e HA) to preserve the post-extractive alveolar socket. MATERIALS AND METHODS Thirty-six patients, divided in three groups, were enrolled in this study. The studied groups were: alveolar bone (OA, n=14), spontaneous healing (GS, n=10) and biomaterial (B, n=12). The biopsies for morphological and molecular analyses were harvested during the extraction tooth session (group OA) or 4-6 months later (GS, B). In GS group, patients recovered spontaneously, while in B group Mg-e HA granules were grafted to avoid the collapse of post-extraction socket. For each group, one specimen was processed for Haematoxylin and Eosin staining and immunohistochemistry for TNF-α, IL-6, RANK, RANKL and OPG. Another specimen was processed to evaluate the gene expression of the same biomarkers by Real-Time PCR (mRNA retro-transcript into cDNA). Data were normalized on housekeeping gene 18s. In GS and B groups, ground sections were obtained to evaluate the mineral component by mean of stereological analyses. For each parameter, statistical analyses were performed to evaluate the differences among the three groups (Kruskal Wallis, significance for p<0.05); post hoc tests were made by Wilcoxon tests (p<0.05/3). RESULTS Samples harvested from each group presented a normal structure, without evident inflammatory infiltrate. Immunohistochemical (IA) and molecular analysis (MA) showed an increment for all biomarkers. IA: The three groups were significantly different for markers TNF-α (p<0.05), RANK (p<0.01), IL-6 e RANKL (p<0.001). Post hoc tests showed a difference between OA and B for RANK (p≤ 0.01), IL-6 and RANKL (p ≤0.001); between GS and B for IL-6 (p≤0.01). MA: Kruskal Wallis test showed a difference among groups for IL-6 (p≤0.05), RANKL and OPG (p≤0.001); post hoc tests revealed a significant difference between: OA and B for IL-6 (p≤0.01); OA and GS for RANKL and OPG (p≤0.001); GS and B for IL-6 and RANKL (p≤0.01), OPG (p≤0.001). RANKL/OPG ratio showed a tendency towards a reduced osteoclastogenesis in group B. Histomorphometry revealed a high percentage of mineralized tissue in the grafted sites; all the residual granules were surrounded by newly formed bone, consisting in regenerated bone bridges. CONCLUSION Both post-extractive groups had an increase of all analysed biomarkers in respect of the physiological remodelling pattern. In the grafted sites, IL-6 values were higher than in the spontaneous healing group; in contrast RANK, RANKL and OPG were slightly lower. The present data may suggest a slowing down in the restoration of the anatomical site, possibly due to a down regulation in the osteoclastogenesis. However, the tissue composition of the grafted alveolar socket showed a good bone regeneration in both groups. Therefore, the current biomaterial could be considered a useful tool for alveolar socket preservation.

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Gibelli, Nelson Elias Mendes. « "Fibrose portal e periportal na obstrução extra-hepática experimental em ratos jovens e adultos : contribuição para o estudo da atresia das vias biliares" ». Universidade de São Paulo, 2003. http://www.teses.usp.br/teses/disponiveis/5/5132/tde-11112005-112052/.

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A atresia das vias biliares é afecção hepática da infância. A etiologia é desconhecida, e o diagnóstico baseia-se na biópsia hepática, cujo achado é a proliferação ductular. A ligadura do ducto biliar comum em ratos é modelo utilizado para estudo das doenças colestáticas. A proposta do trabalho foi estudar, em modelo experimental de obstrução biliar, as alterações histológicas hepáticas em ratos jovens e compará-las com o animal adulto. Avaliou-se a semiquantificação da proliferação ductular e inflamação pelo HE; quantificação da fibrose portal e periportal pelo picrosírius; semiquantificação da expressão de desmina e a-actina de músculo liso pelas células estreladas e miofibroblastos. Apesar das respostas de proliferação ductular e inflamação mais lentas no rato jovem, a fibrose e a expressão de desmina foram mais intensas neste grupo
Biliary atresia is an hepatic disease of infancy. Etiology is unknown, and diagnosis is made by liver biopsy, with ductular proliferation being the main histological feature. Bile duct ligation in rats is an useful experimental model of biliary obstruction. The aim of this study of extra-hepatic cholestasis was analyse hepatic histological alterations in young rats compared to adult animals. The responses were studied by semiquantification of ductular proliferation and inflammatory infiltrated by HE stain; quantification of portal and periportal fibrosis with the sirius-red stain; semiquantification of the expression of desmin and a-smooth muscle actin by the hepatic stellated cells and myofibroblasts. In young animals, despite the very slow response of ductular proliferation and inflammation observed with HE, there were significantly more fibrosis and expression of desmin than in adult group

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Monteiro, Edison Daniel Schneider. « Quantificação digital da imunoexpressão de receptores adrenérgicos e terminações nervosas no detrusor de portadores da síndrome de prune belly ». Universidade de São Paulo, 2008. http://www.teses.usp.br/teses/disponiveis/5/5153/tde-28052008-143257/.

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INTRODUÇÃO: A síndrome de prune belly (PBS) é caracterizada por uma tríade com flacidez da parede abdominal, criptorquidia bilateral e malformações do trato urinário que compreende bexiga de capacidade aumentada, com complacência elevada, hipossensibilidade, hipocontratilidade, com divertículo ou fístula uracal e resíduo pós-miccional elevado. Alguns autores recomendam tratamento clínico, porém outros propõe correção cirúrgica, com reconstrução da via urinária incluindo ureteroplastia e cistoplastia redutoras, orquidopexia e abdominoplastia. Mesmo após a cirurgia, alguns doentes necessitam de cateterismo limpo intermitente. A inervação vesical determina seu funcionamento, mediado por neuroreceptores na junção neuromuscular. Os adrenoreceptores a1 estão relacionados à contratilidade detrusora e o b3 ao seu relaxamento, e certas condições como obstrução infravesical levam à hiperexpressão de receptores a1. O objetivo da presente pesquisa é verificar se no detrusor de doentes com PBS há alteração na densidade de terminações nervosas, hiper ou hipoexpressão de receptores adrenérgicos a1a, a1b, a1d e b3 e proporção anormal dos tecidos muscular e conectivo. MÉTODO Trata-se de estudo retrospectivo de caso-controle que envolveu 14 espécimes de detrusor de doentes com PBS operados entre 1985 a 2005 no Hospital das Clínicas da FMUSP. Dois grupos foram constituídos como controle: 13 fragmentos de bexiga de doentes submetidos à prostatectomia radical no Departamento de Urologia da Universidade de Mainz, com urodinâmica pré-operatória normal (GC1), e cinco fragmentos de bexiga de crianças submetidas à necrópsia no SVOC-USP, sem anomalias neurológicas e de trato urinário. A coloração de van Gieson foi realizada para análise da proporção músculo/tecido conectivo, e a reação imunohistoquímica para os anticorpos policlonais anti-proteína S100 e antiadrenoreceptores a1a, a1b, a1d e b3. A coloração castanho foi considerada evidência da expressão do adrenoreceptor na célula. Cinco a dez imagens digitais foram tomadas por meio de câmara digital e microscopia óptica. Estas foram analisadas pelo programa Adobe Photoshop CS2Ò. A quantidade relativa de receptores foi calculada e a análise estatística realizada pelos testes Kruskal-Wallis e Mann-Whitney. RESULTADOS A média de idade foi de 1,28 ± 1,14 ano no grupo caso (PBS), e de 64 ± 5,22 anos e 1,41 ± 1,11 ano, nos grupos GC1 e GC2, respectivamente. A mediana da relação músculo/tecido conectivo foi de 1,08 para o grupo PBS, 1,59 para o GC1 e para o GC2 de 1,28 (p=0,173). A mediana da proporção S100/tecido muscular foi de 0,21 para o grupo caso (PBS), de 0,20 para o GC1 e para o grupo GC2 de 0,01 (p=0,003). A mediana da relação a1a/tecido muscular foi de 0,06 para o grupo PBS, de 0,16 para o GC1 e para o grupo GC2 de 0,14 (p=0,026). Para a1b, as medianas foram 0,06 no grupo PBS, 0,006 no GC1 e 0,007 no GC2 (p=0,781). No a1d, as medianas foram 0,04 (PBS), 0,04 (GC1) e 0,05 (GC2) (p=0,618). Com relação ao b3, as medianas foram 0,07 no grupo PBS, 0,14 no GC1 e 0,10 no GC2 (p=0,378). CONCLUSÕES Comparando-se fragmentos de detrusor de doentes com PBS e bexigas normais não se observou alteração na densidade de terminações nervosas. Observou-se hipoexpressão do adrenoreceptor a1a, e não houve alteração dos adrenoreceptores a1b, a1d e b3. Também não se observou alteração entre a proporção de tecido muscular e conectivo no detrusor destes doentes. Investigações adicionais, com diferentes métodos e incluindo outros receptores, são necessárias antes de se aplicar esses conhecimentos na prática clínica.
INTRODUCTION: Prune belly syndrome (PBS) is charactherized by a triad of abdominal wall flaccidity, bilateral criptorchidism and urinary tract malformation, that includes a large-capacity bladder, with high detrusor compliance, low sensibility and contractility, associated to urachal diverticulum or fistula and elevated post void residual volumes. Some autors recommend clinical treatment, but others propose surgery correction, with urinary tract reconstruction, including reductive ureteroplasty and cystoplasty, orchidopexy and abdominoplasty. Even after surgery, some patients need intermittent catheterism. The detrusor innervation determines its function, mediated by neuroceptors at the neuromuscular junction. The a1 adrenoceptors are related to detrusor contractility and b3 to relaxation, and some conditions, like infravesical obstruction, lead to a1 adrenoceptor up-regulation. The objective of this work is to verify whether, in the detrusor from patients with PBS, there is altered nerve density, up or down-regulation of a1a, a1b, a1d and b3 adrenergic receptors and if there is an abnormal proportion between muscle and connective tissue. MATERIALS AND METHODS A retrospective case-control study was performed involving 14 detrusor specimens from patients with PBS, who underwent surgical treatment between 1985 an 2005 at University of São Paulo, Medical School Hospital. Two groups were taken as control: 13 bladder fragments from patients who underwent radical prostatectomy at Department of Urology of Mainz University, with normal urodynamic study prior to the surgery (GC1) and 5 bladder fragments from children submitted to autopsy at SVOC-USP, with no neurological or urinary tract malformation (GC2). Staining was performed using the van Gieson dye to analyse the proportion between muscle and connective tissue, and immunohistochemical reaction was employed, with polyclonal antibodies against S100 protein, as well as a1a, a1b, a1d and b3 adrenoceptors. Brown colour was considered as evidence of adrenoceptor cell expression. Five to ten digital images were captured on an optic microscope with a digital camera. These images were analysed with Adobe Photoshop CS2Ò software. The relative quantity of receptors was calculated and the statistic analysis was done with the Kruskal-Wallis and Mann-Whitney tests. RESULTS Mean age was 1.28 ± 1.14 year in PBS patients, and 64 ± 5.22 yrs. and 1.41 ± 1.11 yrs. in GC1 and GC2, respectively. The median proportion between muscle and connective tissue was 1.08 in PBS, 1.59 in GC1 and in GC2 of 1.28 (p=0.173). The median proportion of S100/muscle area was 0.21 in PBS, 0.20 in GC1 and in GC2 of 0.01 (p=0.003). The median relative quantity of receptors of a1a was 0.06 in PBS, 0.16 in GC1 and 0.14 in GC2 (p=0.026). In a1b, the median values were 0.06 in PBS group, 0.006 in GC1 and 0.007 in GC2 (p=0.781). In a1d, the median values were 0.04 (PBS), 0.04 (GC1) and 0.05 (GC2) (p=0.618). Regarding b3, the median values were 0.07 in PBS, 0.14 in GC1 and 0.10 in GC2 (p=0.378). CONCLUSION Comparing detrusor fragments from patients with PBS and normal bladders, there was no alteration in the density of nerve endings. We observed downregulation of a1a adrenoceptors, but no alteration in the a1b, a1d and b3 receptors. Furthermore, there was no alteration of the proportion between muscle and connective tissue areas. Further investigations, with different methods and including other receptors, are necessary to transfer this knowledge to clinical use.

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Minteer, Tanya E. « Detection of Collagen in Rat Abdominal Wound Healing : Contributions of Mesenchymal Stromal Cells and Platelet-Rich Plasma ». Youngstown State University / OhioLINK, 2012. http://rave.ohiolink.edu/etdc/view?acc_num=ysu1348848443.

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Shimomura, Juliana Zanini. « Comparação dos métodos de palpação retal, citologia, histologia e imunoistoquímica para o diagnóstico da hiperplasia prostática benigna no cão / ». Araçatuba : [s.n.], 2007. http://hdl.handle.net/11449/92185.

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Orientador: Flávia de Rezende Eugênio
Banca: Renée Laufer Amorin
Banca: Maria Cecília Rui Luvizotto
Resumo: Ahiperplasiaprostática benigna(HPB) é a afecção mais comum da próstata canina, porém, a comparação dos diferentes métodos diagnósticos como o exame de palpação retal, citologia, histologia e imunoistoquímica é pouco estudada nesta espécie, diferentemente do que ocorre no homem. Este trabalho teve por objetivo estudar, em vinte cães idosos, as alterações cito e histológicas com emprego de imunomarcadores, na glândula prostática. Em todas as glândulas observou-se HPB cística, associada ou não à hiperplasia glandular ou estromal. A imunomarcação com citoqueratina (CK) AE1/AE3 foi relevante em ácinos com epitélio achatado ou com proliferação acentuada. A Vimentina (VIM) V9 teve expressão moderada em áreas com hiperplasia estromal acentuada. O toque retal e o lavado prostático mostraram ser métodos de auxílio no diagnóstico das prostatopatias, sendo indispensável o uso da histopatologia para um diagnóstico definitivo. O emprego de imunomarcadores teciduais prostáticos infere que próstatas com HPB demonstram alterações metabólicas que respondem à imunomarcação em células hiperplásicas.
Abstract: Benign Prostatic Hyperplasia (BPH) is the most common affection of the canine prostate, however, the comparison of different diagnostic methods for BPH through, digital rectal examination, cytology, histology and immunohistochemistry has a few studies in this specie, unlike man. The purpose of this study was understood, in twenty old dogs, cytology, histology alterations using immunomarkers in the prostate gland. All the glands showed the presence of cystic BPH, associated or not with glandular or stromal hyperplasia. The immunomarking with AE1/AE3 cytokeratine was relevant in alveoli with flat epytelium or with a strong proliferation. V9 Vimentine showed to be moderate in areas with a deep stromal hyperplasia. Rectal palpation and the prostatic washing product compose auxiliary diagnostic methods for prostatic diseases, being undismissable the use of histopathology for a conclusive diagnosis. Prostatic tissue immunomarkers employment to conclude that the prostate with HPB demonstrate metabolic changes which react with immunomarking in hyperplasic cells.
Mestre

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Cymbalista, Natalia Cymrot. « Hipercromia cutânea idiopática da região orbital : avaliação clínica, histopatológica e imunohistoquímica antes e após tratamento com luz pulsada de alta energia ». Universidade de São Paulo, 2004. http://www.teses.usp.br/teses/disponiveis/5/5133/tde-04042007-130543/.

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Introdução: A hipercromia cutânea idiopática (HCIRO) não tem sua etiopatogenia bem esclarecida. Parecem estar envolvidos fatores genéticos (herança familiar autossômica dominante), aumento de melanina na derme, vasculatura proeminente e frouxidão da pele palpebral. Encontraram-se, na revisão da literatura, alguns artigos que contribuíram para o esclarecimento da etiopatogenia. Objetivos: Avaliar clínica e histologicamente indivíduos portadores de HCIRO, antes e após o tratamento com luz pulsada de alta energia (LPAE), considerando-se a quantidade de melanina na epiderme e na derme antes e após o tratamento e, assim, avaliar a eficácia da LPAE no clareamento da HCIRO. Avaliar possível diferença na quantidade de melanina da pálpebra inferior (afetada) em relação à pele pré-auricular (controle). Avaliar a qualidade das células dérmicas na pálpebra inferior, contendo melanina antes do tratamento (observação através de imuno-histoquímica para determinação do tipo de célula dérmica, se macrófagos ou melanócitos). Avaliar a presença ou não de hemossiderina na derme. Avaliar e seguir os indivíduos, clinicamente, após um ano do término do tratamento, para verificar a manutenção da melhora ou a recidiva da HCIRO. Casuística e Método: Selecionaram-se 12 indivíduos portadores de HCIRO, os quais foram submetidos à avaliação clínica através de fotografias e à avaliação histológica antes e após a aplicação de LPAE para o tratamento de HCIRO. Realizaram-se biopsias na pálpebra inferior, antes e após o tratamento com LPAE, e biopsia na pele pré-auricular esquerda para comparar a quantidade de melanina existente nela e na pálpebra inferior, antes do tratamento. Os indivíduos foram submetidos a uma série de uma a quatro sessões de aplicação de LPAE nas pálpebras inferiores, com intervalos de aproximadamente trinta dias entre as sessões. Sete observadores independentes, dermatologistas, analisaram fotografias da região palpebral inferior dos participantes antes e após o tratamento e classificaram os resultados como melhor, pior, ou inalterado. A reprodutibilidade dessa avaliação foi testada através do coeficiente Kappa. A quantificação de melanina, por área, antes e após o tratamento, foi realizada através de morfometria com análise de imagens por computador (método digital). A verificação de presença ou não de hemossiderina dérmica foi realizada pela coloração de Perls e a identificação da célula dérmica que contém melanina foi feita pela imunohistoquímica, com anticorpos monoclonais antimacrófagos humanos tipo CD68 e anticorpos monoclonais antimelanócitos humanos tipo Melan A. Resultados: Segundo os sete observadores, houve melhora clínica (clareamento da pele palpebral inferior) estastisticamente significativa (p=0,024) e o coeficiente Kappa mostrou boa concordância entre os observadores. Todos os indivíduos (100%) apresentaram hipercromia pós-inflamatória transitória, com tempo médio de desaparecimento de seis meses, enquanto 58,33% apresentaram hipocromia transitória, no local das biopsias palpebrais, com duração média de sete meses. A análise histológica mostrou ausência de hemossiderina dérmica na pele palpebral, antes e após o tratamento. A quantificação de melanina, por morfometria, mostrou diminuição da mesma após o tratamento tanto na epiderme como na derme, com significância estatística. A morfometria também mostrou, maior quantidade de melanina na epiderme e na derme da pele palpebral inferior, antes do tratamento, em relação à pele pré-auricular, com diferença estatisticamente significativa. A imunohistoquímica com anticorpos monoclonais antimacrófagos humanos tipo CD68 e anticorpos monoclonais antimelanócitos humanos tipo Melan A caracterizaram o macrófago como sendo a célula dérmica que contém melanina na HCIRO, não havendo melanócitos dérmicos. Conclusões: A quantidade de melanina, tanto da epiderme quanto da derme, diminuiu após o tratamento com LPAE na pálpebra inferior. Há maior quantidade de melanina, tanto na epiderme quanto na derme da pálpebra inferior em comparação com a pele pré-auricular, com diferença estatisticamente significativa. As células que contêm melanina na derme palpebral inferior dos indivíduos portadores de HCIRO são macrófagos (e não melanócitos). Nos espécimes examinados neste estudo, não houve presença de hemossiderina na derme da pálpebra inferior. Houve clareamento da pele palpebral inferior após o tratamento com LPAE, e o seguimento clínico após um ano do tratamento mostrou que esse clareamento se manteve, não havendo recidiva da HCIRO. No entanto, um seguimento mais longo é necessário para a avaliação de possível recidiva tardia da pigmentação palpebral.
Introduction: The cutaneous idiopathic hyperchromia (HCIRO) does not have a clear etiopathogeny. Genetic factors seem to be involved (familiar autossomic dominant heredity), increase of melanin in the dermis, proeminent vascularity and eyelid skin slackness. In the literature revision, a few articles were encountered, which contributed in clarifying the etiopathogeny. Objectives: Evaluate clinically and histologically individuals bearer of HCIRO, before and after the treatment with intense pulsed light (LPAE), considering the melanin quantity in the epidermis and dermis, before and after the treatment, in order to evaluate the efficacy of LPAE in the clearing up of HCIRO. Evaluate possible difference in the melanin quantity of the lower eyelid in relation to the pre-auricular skin. Evaluate the quality of the dermic cells in the lower eyelid, containing melanin before the treatment (observation through immunohistochemistry to determine the dermic cell type, whether macrophages or melanocytes). Evaluate the presence or not of hemossiderine in dermis. Evaluate and follow up clinically of the individuals after a year treatment to check the improvement maintenance or HCIRO recurrence. Casuistry and Method: There were selected 12 individuals, bearer of HCIRO, and these were submitted to a clinical evaluation through photographies and to histological evaluation before and after LPAE application for HCIRO treatment. Biopsies were made in the lower eyelid, before and after treatment with LPAE, and in the left pre-auricular skin to compare the melanin quantity with the lower eyelid, before the treatment. The individuals were submitted to one to four sessions of LPAE application in the lower eyelid with approximately thirty day intervals between sessions. Seven independent observers (dermatologists) analysed photos of the participants lower eyelids, before and after the treatment and classified the results as better, inaltered or worse. The reproducibility of this affraisal was tested through the Kappa coefficient. The quantification of melanin, per area, before and after the treatment, was made through morphometry, with image analysis by computer (digital method). The verification of the presence or not of dermic hemossiderin was made through the Perls colouring and the cell identification, which contains melanin, was made through immunohistochemistry with monoclonal antibodies anti human macrophages type CD68 and monoclonal antibodies anti human melanocytes type Melan-A. Results: According to seven observers, there was a clinical improvement (clearing up of the eyelid skin) statistically significant (p= 0.24) and the Kappa coefficient showed a good concordance between the observers. All of the of the individuals (100%) showed a temporary post-inflammatory hyperchromia with an average duration of six months, while 58.33% presents a transitory hypochromia, in the place of palpebral biopsies, of an average duration of seven months. The histological analysis showed absence of dermic hemossiderin in the palpebral skin, before and after the treatment. The melanin quantification, per morphometry, showed decrease of same after the treatment both in the epidermis and dermis, of statistical significance. Melanin quantity was higher in the lower eyelid (before treatment), comparing with pre-auricular skin, with significant difference statistically. Immunohistochemistry with monoclonal antibodies anti human macrophages type CD68 and monoclonal antibodies anti human melanocytes type Melan-A, characterized the macrophages as being the dermic cell which contains melanin in HCIRO, showing no dermic melanocytes. Conclusions: The melanin quantity, both in the epidermis and dermis, decreased after the treatment with LPAE in the lower eyelid. There was more melanin in the epidermis and dermis of the eyelid skin, comparing with the pre-auricular skin, with a statistically significant difference. The cells that contain melanin in the lower eyelid dermis of the individuals bearer of HCIRO, are macrophages (and not melanocytes). In the specimens examined in this study, there was absence of hemossiderin in the dermis of the lower eyelid. There was a clearing up of the lower eyelid skin after the treatment with LPAE, and the clinical follow-up after one year of treatment showed that this clearing up was maintained and no HCIRO reincidence occurred. In the meantime, a longer follow-up is necessary to evaluate a later possible reincidence of the palpebral pigmentation.

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Júnior, Renato Ambrósio. « "Estudo laboratorial da cicatrização de córneas humanas após debridamento epitelial" ». Universidade de São Paulo, 2004. http://www.teses.usp.br/teses/disponiveis/5/5149/tde-21102005-085247/.

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Objetivo: Verificar resposta após debridamento epitelial de córneas humanas. Métodos: Córneas normais foram submetidas a debridamento antes da cirurgia de enucleação. Realizou-se histologia, TUNEL, Ki67, SMA e microscopia eletrônica. Resultados: Seis córneas foram debridadas e preservadas entre ½ e 65 horas, apresentando apoptose nos ceratócitos do estroma anterior. Células estromais em proliferação foram observadas apenas no tempo de 65 horas. Miofibroblastos não foram encontrados. Uma córnea serviu de controle. Conclusões: Os eventos observados em córneas humanas após debridamento epitelial, apoptose e proliferação dos ceratócitos, foram semelhantes aos descritos em animais de experimentação
Purpose: To examine the early wound healing response to epithelial scrape in human corneas. Methods: Normal corneas had epithelial scrape prior to enucleation. Histology, TUNEL assay, Ki67, SMA and transmission electron microscopy were performed. Results: Epithelial scrape was performed in six corneas from ½ to 65 hours prior to preservation. Keratocyte apoptosis was detected in the anterior stroma in all scraped corneas. Keratocyte proliferation was detected exclusively 65 hours after scrape. No myofibroblast was detected. One cornea was not scraped (control). Conclusion: Results obtained in human corneas (keratocyte apoptosis and proliferation) were similar to animal models

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Shimomura, Juliana Zanini [UNESP]. « Comparação dos métodos de palpação retal, citologia, histologia e imunoistoquímica para o diagnóstico da hiperplasia prostática benigna no cão ». Universidade Estadual Paulista (UNESP), 2007. http://hdl.handle.net/11449/92185.

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Ahiperplasiaprostática benigna(HPB) é a afecção mais comum da próstata canina, porém, a comparação dos diferentes métodos diagnósticos como o exame de palpação retal, citologia, histologia e imunoistoquímica é pouco estudada nesta espécie, diferentemente do que ocorre no homem. Este trabalho teve por objetivo estudar, em vinte cães idosos, as alterações cito e histológicas com emprego de imunomarcadores, na glândula prostática. Em todas as glândulas observou-se HPB cística, associada ou não à hiperplasia glandular ou estromal. A imunomarcação com citoqueratina (CK) AE1/AE3 foi relevante em ácinos com epitélio achatado ou com proliferação acentuada. A Vimentina (VIM) V9 teve expressão moderada em áreas com hiperplasia estromal acentuada. O toque retal e o lavado prostático mostraram ser métodos de auxílio no diagnóstico das prostatopatias, sendo indispensável o uso da histopatologia para um diagnóstico definitivo. O emprego de imunomarcadores teciduais prostáticos infere que próstatas com HPB demonstram alterações metabólicas que respondem à imunomarcação em células hiperplásicas.
Benign Prostatic Hyperplasia (BPH) is the most common affection of the canine prostate, however, the comparison of different diagnostic methods for BPH through, digital rectal examination, cytology, histology and immunohistochemistry has a few studies in this specie, unlike man. The purpose of this study was understood, in twenty old dogs, cytology, histology alterations using immunomarkers in the prostate gland. All the glands showed the presence of cystic BPH, associated or not with glandular or stromal hyperplasia. The immunomarking with AE1/AE3 cytokeratine was relevant in alveoli with flat epytelium or with a strong proliferation. V9 Vimentine showed to be moderate in areas with a deep stromal hyperplasia. Rectal palpation and the prostatic washing product compose auxiliary diagnostic methods for prostatic diseases, being undismissable the use of histopathology for a conclusive diagnosis. Prostatic tissue immunomarkers employment to conclude that the prostate with HPB demonstrate metabolic changes which react with immunomarking in hyperplasic cells.

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Unhale, Sanket Anil. « Cryobiology of Cell and Tissue Cryopreservation : Experimental and Theoretical Analysis ». Diss., The University of Arizona, 2011. http://hdl.handle.net/10150/202974.

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Preservation of tissue structure, morphology and biomarkers is of utmost importance for pathological examination of biopsy specimens for diagnostic and therapeutic purposes. However current methods employed to evade tissue degradation and preserve biomarkers have several shortcomings that include irreproducibility, morphological artifacts and altered biomarker antigenicity. These artifacts may affect the analysis and subsequent diagnosis of the tissue pathology. This creates need for developing improved preservation methods that reproducibly maintain tissue morphology and biomarker antigenicity and are simple, rapid and inexpensive. Experiments conducted for testing the hypothesis that cryopreservation procedures yield high quality morphology and antigenicity showed that cryopreservation maintains tissue structure, morphology and antigenicity at equivalent or better levels compared to standard freezing techniques. In order to understand the mechanisms of osmotic transport in cellular systems upon exposure to multi-component solutions that are prevalent in virtification protocols, experimental studies were undertaken using microfluidics for single cell manipulation. The experimental data yielded permeability parameters in binary and ternary solutions for MC3T3-E1 murine osteoblasts for the first time. The hydraulic conductivity (L(p)) decreased with increasing concentrations but the solute permeability either increased or decreased with increasing solution concentration. The changes in hydraulic conductivity were consistent with previously published trends and conform to a functional relationship in the form of Arrhenius type relationship between L(p) and solution concentration. Further a theoretical model was developed from principles of linear irreversible thermodynamics to simulate multi--‐‑component mass transport across membrane. The model was successfully validated by comparison with experimental data for murine osteoblasts and showed good agreement between the numerical predictions and experimental observations. The modeling approach can be used to investigate the transport mechanisms, which show that in multicomponent osmotic transport response, the dynamics is dictated by slower moving solute.

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Milani, S. « DEPROTEINIZED BOVINE BONE GRAFT REMODELING PATTERN IN ALVEOLAR SOCKET. HISTOLOGIC AND IMMUNOHISTOLOGICAL EXPRESSION EVALUATION ». Doctoral thesis, Università degli Studi di Milano, 2014. http://hdl.handle.net/2434/229907.

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Deproteinized bovine bone (DBB) is a bone graft highly used in dentistry in bone regeneration and alveolar socket preservation techniques. Although there is a scientific consensus on the clinical benefit of this biomaterial, and several in vitro studies described its biologic effect on osteoblasts, in vivo analyses investigating its effect on bone dynamics on human are lacking. For this purpose 20 patients needing tooth extraction and implant placement were selected and randomized in test (alveolar socket preservation with DBB) and control (spontaneous healing). Bone specimens were collected during tooth extraction (T0) and, at 5 months, during implant placement (T1). The collected samples were processed for histologic and immunohistological analyses to reveal the presence of positive (BMP-2, BMP-7, ALP) and negative (Il-6, TNF-α) markers of bone remodeling. The sections were then micro-photographed, quantification was done and statistical analyses were performed to compare T0 and T1 in both groups and T1 test group versus T1 control group. The obtained results showed higher expression of BMP-2, BMP-7 and IL-6 at T1 in both groups (p<0.05), lower expression of ALP in both (p<0.05) at T1 and higher expression of TNF-α only in test group (p<0.05) while in control group it remained stable during time. When comparing T1 markers expression in control and test groups, a higher expression of BMP-2 (p<0.05) and lower expression of TNF-α (p<0.05) were found in the first one. These results are in concordance with the previous in vitro studies and show that DBB is able to maintain bone remodeling in active phases. As a matter of fact at 5 months a higher expression of the positive markers (BMP-2, BMP-7) was noticed compared to T0, but the presence of DBB resulted in lower expression of BMP-2 and higher expression of inflammatory factor (TNF-α) when comparing to spontaneous healing at the time of the implant placement.  For these reasons the use of DBB is suggested when clinical needs lead to a precise indication of alveolar socket preservation with biomaterial, while, if not clinically necessary, spontaneous healing is indicated because it shows more biological positive effects.

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França, Andréa Fernandes Eloy da Costa 1977. « Análise histopatológica e imunohistoquímica das lesões vitiligóides no lúpus eritematoso cutâneo ». [s.n.], 2011. http://repositorio.unicamp.br/jspui/handle/REPOSIP/309011.

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Orientador: Elemir Macedo de Souza
Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas
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Resumo: O lúpus eritematoso (LE) é uma doença auto-imune com espectro clínico variado. O lúpus eritematoso cutâneo (LEC) inclui o lúpus eritematoso agudo (LECA), o subagudo (LECSA) e o crônico (LECC). Lesões acrômicas podem ocorrer durante a evolução do LE, embora nunca tenham sido estudadas histologicamente. O objetivo deste trabalho é descrever os achados clínicos, laboratoriais e histológicos das lesões acrômicas no LE. Foram selecionados 12 pacientes com LE de um grupo de 220 atendidos no período de 2005 a 2008, sendo sete com LECC e cinco com LECSA. Doze pacientes com vitiligo e 10 controles de pele sã foram usados para comparação. As alterações histológicas encontradas foram: infiltrado inflamatório (75%); hiperceratose e espessamento da zona da membrana basal (ZMB) (66,7%); retificação da epiderme (58,3%); ceratinócitos apoptóticos epidérmicos, elastose e telangectasias (50%); fibrose (41,7%); degeneração vacuolar da ZMB (33,3%); rolhas córneas (16,7%). O diagnóstico histológico de LE foi possível em quatro casos. Melanina pela coloração de Fontana Masson (FM) foi vista em cinco casos e incontinência pigmentar em quatro. Melanócitos foram evidenciados em amostras de cinco doentes através da reação imunohistoquímica pelo HMB45 e Melan-A, com diferença estatística em relação aos controles. Quando comparado ao vitiligo, a diferença foi estatisticamente significante para os achados histológicos: ceratinócitos apoptóticos epidérmicos (p=0,014), espessamento da ZMB (p=0,009) e fibrose (p=0,037). Em relação à quantificação dos melanócitos, não houve diferença estatística entre o grupo LE e vitiligo usando os anticorpos Melan-A e HMB45. Concluímos que as lesões acrômicas no LE correspondem a lesões residuais, decorrentes de processo inflamatório liquenóide prévio que destrói os melanócitos. Não é possível diferenciar as lesões vitiligóides das duas dermatoses pela presença ou ausência de melanócitos, embora a repigmentação seja possível em ambas as doenças devido a presença de melanogênese ativa comprovada pela positividade pelo HMB45
Abstract: Lupus Erythematosus (LE) is an autoimmune disorder with multiple clinical manifestations. Skin damage is a hallmark of the disease. Cutaneous LE (CLE) includes acute LE (ACLE), subacute LE (SCLE) and chronic LE (CCLE). Although achromic lesions are often found in patients with LEC, there are no detailed data about the histological features of such lesions. Therefore, we designed this study to determine clinical, laboratorial and histological profile of patients with LEC presenting achromic lesions. Between 2005 and 2008, we identified 12 individuals with LEC and acromic lesions from a larger group of 220 patients with LEC that were followed at the Dermatology outpatient clinic. There were seven patients with LECC and five with LECSA. Twelve patients with chronic stable vitiligo and 10 controls of unaffected skin were used for comparison. The most frequent histological abnormalities found in LEC-related achromic lesions were inflammatory infiltrates (75%); hyperkeratosis and thickening of the basement membrane (BM) (66.7%); epidermal flattening (58.3%); apoptotic epidermal keratinocytes, elastosis and vasodilation (50%); fibrosis (41.7%); hydropic degeneration of the basal cells (33.3%); follicular plugging (16.7%). These achromic lesions retained histological features that enabled the diagnosis of CLE to be established in four patients. Fontana Masson (FM) staining was positive for melanin in five cases and revealed pigmentary incontinence in four. Immunohistochemistry for HMB45 and Melan-A identified melanocytes in five CLE-related achromic lesions. Melanocyte counts were significantly smaller in achromic lesions when compared to unaffected skin samples. When compared to vitiligo, CLE-related achromic lesions showed more frequently apoptotic epidermal keratinocytes (p=0,014), thickening of the BM (p=0,009) and fibrosis (p=0,037). Melanocyte counts according to immunohistochemistry were similar in CLE and vitiligo groups. Our results indicate that CLE-related achromic lesions represent residual scars due to chronic lichenoid inflammation that leads to melanocyte destruction. Melanocyte count does not help to distinguish CLE-related achromic lesions and true vitiligo lesions. Despite this, HMB45 staining was sometimes positive in both conditions, which indicates active melanogenesis and suggests that repigmentation may be possible at least for some individuals
Doutorado
Clinica Medica
Doutor em Clínica Médica

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FIRMIANO, Enely Maris da Silveira. « Descri??o histol?gica e detec??o imuno-histoqu?mica de c?lulas do sistema neuroend?crino difuso no oviduto de Phrynops geoffroanus (Testudines, Chelidae) ». Universidade Federal Rural do Rio de Janeiro, 2013. https://tede.ufrrj.br/jspui/handle/jspui/2501.

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Phrynops geoffroanus is a freshwater turtle popularly known as Geoffroy's toadhead turtle or Geoffroy's side-necked turtle. Like other species of the Testudines order, it is oviparous, meaning the females lay their eggs in the environment. The objectives of this article are to describe the morphology of the oviduct of P. geoffroanus as observed through light microscopy after performing standard histochemical techniques (AB pH 2.5, PAS, Mallory?s trichrome, xylidine Ponceau), besides identifying the endocrine cells in the various regions of the oviduct that produce motilin, serotonin and somatostatin through immunohistochemistry. The oviduct of this turtle is composed of five regions: the infundibulum, which receives the oocyte released at the moment of oocytation; the tube uterine (magnum), the spiraled region that produces the albumen; the isthmus, a transition region; the uterus, responsible for producing the egg shell; and the vagina, the final portion of the oviduct, which leads to the cloaca. The structure of the oviduct of P. geoffroanus is similar to that of other oviparous reptile species. Its description can be used for phylogenetic morphological comparisons. The immunohistochemistry study revealed the absence of neuroendocrine cells that produce motilin, serotonin and somatostatin in all regions of the oviduct. However, the existence of these cells in the gut of this turtle was verified, enabling the suggestion that the regulatory peptides produced by these cells are carried by the bloodstream and reach specific receptors on target cells located along the oviduct, to regulate the peristaltic movements of this organ.
Phrynops geoffronus ? uma esp?cie de r?ptil representante da ordem dos Testudines, popularmente conhecida como c?gado-de-barbichas ou c?gado-de-barbelas. Como os demais representantes desta ordem, ? uma esp?cie ov?para, ou seja, as f?meas colocam ovos no ambiente durante a reprodu??o. Os objetivos deste trabalho foram a descri??o histol?gica do oviduto das f?meas de P. geoffroanus com aux?lio da microscopia de luz, ap?s ser submetido ?s t?cnicas histol?gicas e histoqu?micas (AB pH 2,5, PAS, Tricr?mico de Mallory, Xylidine Ponceau), al?m de procurar identificar c?lulas end?crinas produtoras de motilina, serotonina e somatostatina atrav?s da imuno-histoqu?mica, ao longo dos diversos segmentos do oviduto. O oviduto deste c?gado ? composto por cinco diferentes regi?es: infund?bulo, que recebe o ov?cito liberado no momento da ovocita??o; a tuba uterina (magno), regi?o espiralada produtora do alb?men; o istmo, uma regi?o de transi??o; o ?tero, respons?vel pela produ??o da casca do ovo, e a vagina, por??o final do oviduto que leva ? cloaca. A estrutura do oviduto de P. geoffroanus ? semelhante ? de outras esp?cies de r?pteis ov?paros e pode ser utilizada para compara??es morfol?gicas filogen?ticas. O estudo imuno-histoqu?mico revelou aus?ncia de c?lulas neuroend?crinas produtoras de motilina, serotonina e somatostatina em todas as regi?es do oviduto da esp?cie estudada. Entretanto, a exist?ncia dessas c?lulas foi verificada no intestino deste c?gado (teste-controle), tornando poss?vel sugerir que provavelmente os pept?deos regulat?rios produzidos por tais c?lulas, sejam transportados pela corrente sangu?nea e atinjam receptores espec?ficos em c?lulas-alvo localizadas ao longo do oviduto para atuarem na regula??o dos movimentos perist?lticos deste ?rg?o.

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Godínez, Martínez José Manuel. « Detección de papiloma virus humano y genes supresores tumorales P16 y P53 en carcinomas de región genital extena ». Doctoral thesis, Universidad de Murcia, 2008. http://hdl.handle.net/10803/10723.

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La implicación del papiloma virus humano (HPV) en carcinomas de cérvix está totalmente demostrada, pero esta infección se puede producir en otras regiones anatómicas con similares consecuencias patológicas. El estudio presenta la detección de HPV en carcinomas de pene y vulva mediante PCR, analizando la sensibilidad de diferentes sistemas, primers GP5+/6* y MY09/11. También se lleva a cabo la detección inmunohistoquímica de proteínas celulares como posibles marcadores diagnóstico de infección por HPV, las proteínas P53 y P16 su expresión supuestamente se altera por la infección de HPV en carcinomas de pene (77,5%), mientras que en los carcinomas de vulva la tasa de detección está dentro de los rangos esperados según trabajos previos publicados (29, 7). El estudio de la relación entre la presencia de HPV y la expresión de los marcadores celulares muestra su no asociación en estas patologías.
The infection of human papilloma virus (HPV) is clearly related with cervical carcinomas, but this infection can be found in other regions whit PCR, analyzing the sensitivity of HPV in penile and vulvar carcinomas whit PCR, analyzing the sensitivity of two differents systems, MY09/11 and GP 5+/6+. Also the immunohistochemical detection of cell proteines as diagnostic markers for the HPV in penile carcinomas in the study populations (77,5%), whereas in vulgar carcinomas . The detection rates is similar to the previously published data ( 29, 7%). The study of the relatioship between HPV and the expression of the cell markers shows no asocciation with the pahologies.

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Nina, Vinicius José da Silva. « "Endotelização in vivo das biopróteses cardíacas porcinas : comparação entre a preservação convencional e a não-aldeídica" ». Universidade de São Paulo, 2003. http://www.teses.usp.br/teses/disponiveis/5/5156/tde-15102004-091024/.

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O revestimento completo das superfícies das biopróteses cardíacas por uma camada protetora de células endoteliais do hospedeiro ainda não foi relatado. As próteses cardíacas de uso corrente são comumente preservadas em glutaraldeído, o qual é citotóxico para as células do hospedeiro evitando a endotelização espontânea. O objetivo deste estudo é demonstrar o potencial para a endotelização in vivo das próteses cardíacas tratadas por um processo alternativo de preservação tecidual (L-Hydro). O processo de preservação L-Hydro consiste na extração controlada de substâncias antigênicas do tecido valvar e na incorporação de um agente antiinflamatório e anti-trombótico. Sete biopróteses porcinas com suporte tratadas pelo processo L-Hydro (grupo teste) e três fixadas em glutaraldeído (grupo controle) foram implantadas na posição mitral de ovelhas jovens. As próteses foram avaliadas pela ecocardiografia e angiografia antes do sacrifício aos cinco meses. As próteses explantadas foram avaliadas quanto ao aspecto radiológico, histológico e histoquímico. O teste-t não-pareado de Student e o teste exato de Fisher foram utilizados para a análise estatística. Não houve diferença hemodinâmica entre os grupos, exceto pela pressão capilar mais elevada no grupo controle. Entretanto, a microscopia óptica e a eletrônica de varredura e transmissão mostraram o revestimento quase completo de células endoteliais na superfície de todos os folhetos das próteses tratadas pelo L-Hydro após cinco meses de implante. As células endoteliais estavam em contato direto com a camada do colágeno subjacente e expressaram antígenos ligados ao fator von Willebrand. As superfícies das próteses tratadas pelo glutaraldeído encontravam-se cobertas por depósitos de fibrina, macrófagos, cálcio e material trombótico. Apenas células endoteliais esparsas foram observadas neste grupo, e o contato destas células com o colágeno subjacente foi incompleto. Estes dados indicam que a preservação tecidual não-aldeídica (L-Hydro) utilizada neste estudo é capaz de induzir a endotelização espontânea evidenciada pela boa adesividade celular do novo endotélio à matriz do colágeno, e pela maior resistência à trombose e à calcificação.
A protective layer of endothelial cells of host origin on the entire surfaces of bioprosthetic heart valves has never been reported. Current commercial bioprosthetic heart valves are commonly preserved in glutaraldehyde which is cytotoxic to host cells preventing spontaneous endothelialization. The aim of this study is to demonstrate the potential for in vivo endothelialization of heart valves treated by an alternative tissue preservation process (L-Hydro). L-Hydro preservation process consists of controlled extraction of antigenic substances from the valvular tissue and incorporation of an anti-inflammatory and a anti-thrombotic agent. Seven stented porcine heart valves treated by the L-Hydro process (test group) and three glutaraldehyde-fixed porcine heart valves (control group) were implanted in the mitral position of juvenile sheep. The valves were evaluated echocardiographically and angiographically prior to sacrifice at five months. Recovered valves were also radiologically, histologically and histochemically evaluated. Unpaired t-test of Student and Fishers exact test were performed for statistical analysis. There were no hemodynamic differences between groups except for a higher pulmonary artery wedge pressure in the control group. However, optical microscopy, scanning and transmission electron microscopy showed a nearly complete coverage of endothelial cells on the surfaces of all leaflets in the L-Hydro treated valves after five months of implantation. The endothelial cells were in direct contact with the underlying collagen layer and expressed von Willebrand-related antigens. The surfaces of the glutaraldehyde-treated valves were covered by fibrin deposition, macrophages, calcium and thrombotic material. Only sparse endothelial cells were observed in this group, and contact of the endothelial cells with the underlying tissue was incomplete. These data indicate that the non-aldehyde tissue preservation (L-Hydro) utilised in this study is capable of inducing spontaneous endothelialization with evidence of strong cell attachment of the new endothelium to the collagen tissue matrix, and it also provides a greater resistance to thrombosis and calcification.

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FERREIRA, Hugo Henrique. « Graduação histológica e expressão de metaloproteinases (MMP-9 e MMP-2) e seus inibidores (TIMP-1 e TIMP-2) na glândula mamária canina neoplásica ». Universidade Federal de Goiás, 2012. http://repositorio.bc.ufg.br/tede/handle/tde/898.

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Made available in DSpace on 2014-07-29T15:07:44Z (GMT). No. of bitstreams: 1 Dissertacao HUGO CAB.pdf: 2501194 bytes, checksum: 09df95fb673afe9494354f6ab1e4d5bd (MD5) Previous issue date: 2012-08-03
In bitchs, mammary glands are the most common sites for the occurrence of neoplasms. To evaluate the biological behavior of these lesions is routinely employ descriptive histological methods that, combined with histological grading, help predict prognosis of these diseases. The imbalance between the amount of tissue matrix metalloproteinases (MMPs), proteolytic enzymes involved in physiological and pathological processes, and their inhibitors (TIMP), plays an important role in development and tumor invasion. To evaluate the biological behavior of some common types of breast tumor bitches this research was divided in two steps. The first one was correlating histological classification and degree of mammary neoplasms in bitches, and to correlating the degree of malignancy and tumor size. For this, 119 samples were evaluated in canine mammary tumors, all tumors were considered disorders of the growth of canine mammary gland, including neoplastic (benign and malignant) and non-neoplastic (hyperplastic changes). All tumors were classified and only the malignant were graded histologically and correlated with tumor size. It was concluded that the complex carcinoma is the most common cancer histological type and which has the lowest histological grade compared to other malignant tumor types. Further, neoplasms larger than five centimeters were predominantly malignant (92,3% or 12/13) and no correlation was observed between tumor size and histologic grade. The relation between histological classification and graduation helped in microscopic description of malignant mammary tumors in bitches. The second step was to evaluate the expression of MMP-9, MMP-2 TIMP-1 and TIMP-2 in normal and neoplastic canine mammary tissue. For this purpose it was selected 48 samples of canine mammary tissue, 14 complex carcinomas (CC), 13 tubulopapillary carcinoma (TC), six simple mammary adenoma (MA) and 15 normal breast (MN), and submitted to technical immunohistochemistry for the evaluation of the intensity staining and the number of epithelial and stromal cells selected. Differences were observed in the expression of MMP-9 and TIMP-1 and TIMP-2 according to the mammary histomorphology. The MMP-9 showed higher expression in epithelial and stromal cells of the CC and CT compared to MN. It was found a lower expression of TIMP-1 in stromal cells of the CC and overexpression of TIMP-2 in epithelial cells of the CT. Also a positive correlation between MMP-9 and TIMP-1 and the intensity staining and number of epithelial and stromal cells marked in MN and between MMP-9 and TIMP-2 on TC neoplastic epithelial cells. It was concluded that there is variation between expression of MMP-9, TIMP-1 and TIMP-2 among the tumor types, and a positive correlation between TIMP-1 and MMP-9 in normal mammary gland and between MMP-9 and TIMP-2 in tubulopapilar carcinoma. The balanced expression of MMP-9, MMP-2, TIMP-1 and TIMP-2 is essential for the homeostasis of canine normal mammary gland, while the overexpression of MMP-9 and TIMP-2, and the reduced expression of TIMP-1 in carcinomas represents an appropriate condition to the tumor development.
Nas fêmeas caninas, as glândulas mamárias são os locais mais comuns para a ocorrência de neoplasias. Para avaliar o comportamento biológico dessas lesões empregam-se rotineiramente métodos histológicos descritivos que, associados à graduação histológica, auxiliam na predição prognóstica dessas enfermidades. Ainda, o desequilíbrio entre a quantidade tissular de metaloproteinases de matriz (MMP), enzimas proteolíticas implicadas em processos fisiológicos e patológicos, e seus inibidores (TIMP), desempenha papel importante na evolução e invasão tumoral. Para avaliar o comportamento biológico de alguns tipos tumorais frequentes da mama das cadelas esta pesquisa foi dividida em duas etapas. A primeira teve como objetivo relacionar a classificação e a graduação histológicas das neoplasias mamárias de cadelas, bem como correlacionar a graduação dos tumores malignos com o tamanho tumoral. Para isso, avaliaram-se 119 amostras de tumores mamários caninos, sendo considerados tumores todos os distúrbios do crescimento da glândula mamária canina, incluindo os neoplásicos (benignos e malignos) e os não neoplásicos (alterações hiperplásicas). Todos os tumores foram classificados e apenas os malignos graduados histologicamente e correlacionados ao tamanho tumoral. Concluiu-se que o carcinoma complexo é o tipo histológico mais comum e que possui menor grau histológico em relação aos demais tipos tumorais de classificação maligna. Ainda, neoplasias maiores que cinco centímetros foram predominantemente malignas (92,3% ou 12/13) e não foi observada correlação entre tamanho tumoral e grau histológico. A relação entre a classificação e a graduação histológica compreendeu instrumento auxiliar na descrição microscópica dos tumores mamários malignos das cadelas. A segunda etapa teve como objetivo avaliar a expressão de MMP-9, MMP-2, TIMP-1 e TIMP-2 no tecido mamário canino normal e neoplásico. Para isso, foram selecionadas 48 amostras de glândula mamária canina, sendo 14 de carcinoma complexo (CC), 13 de carcinoma tubulopapilar (CT), seis de adenoma mamário simples (AM) e 15 de mama normal (MN), submetidas à técnica de imuno-histoquímica para a avaliação da intensidade de marcação e do número de células epiteliais e estromais marcadas. Houve diferença na expressão de MMP-9, TIMP-1 e TIMP-2 de acordo com a histomorfologia mamária. A MMP-9 apresentou maior expressão nas células epiteliais e estromais dos CC e CT em relação às MN. Constatou-se menor expressão de TIMP-1 nas células estromais do CC e superexpressão de TIMP-2 nas células epiteliais do CT. Ainda, houve correlação positiva entre MMP-9 e TIMP-1 quanto a intensidade de marcação e número de células epiteliais e estromais marcadas na MN, e entre MMP-9 e TIMP-2 nas células epiteliais neoplásicas do CT. Concluiu-se que há variação na expressão da MMP-9, TIMP-1 e TIMP-2 entre os tipos tumorais, além de correlação positiva entre TIMP-1 e MMP-9 nas mamas normais e entre MMP-9 e TIMP-2 no carcinoma tubulopapilar. A atividade equilibrada entre MMP-9, MMP-2, TIMP-1 e TIMP-2 é fundamental à homeostase da glândula mamária canina normal enquanto a superexpressão de MMP-9 e TIMP-2, e a expressão reduzida de TIMP-1 nos carcinomas pode representar condição adequada à evolução tumoral.

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Hawthorne, Ana Carolina. « Avaliação comparativa entre enxertos alógenos e autógenos \'onlay\'. Estudo histológico, imunohistoquímico e tomográfico em coelhos ». Universidade de São Paulo, 2010. http://www.teses.usp.br/teses/disponiveis/58/58136/tde-03022011-171303/.

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A reconstrução dos maxilares em implantodontia através de métodos de enxertia óssea constitui o procedimento cirúrgico mais utilizado frente à perda fisiológica ou traumática a que estes ossos estão sujeitos. Os enxertos autógenos mostram vantagens em relação às demais técnicas de reconstrução no que se refere ao potencial regenerador ósseo, entretanto, a sua remoção implica obrigatoriamente na necessidade de áreas doadoras. Nas últimas décadas tem ocorrido um grande interesse pelos enxertos alógenos de banco de tecidos músculo-esquelético (BTME) como alternativa às enxertias autógenas, como forma de evitar morbidade do sítio doador e redução de tempo e custos da cirurgia. O propósito do estudo foi comparar o comportamento dos enxertos alógenos com autógenos avaliados por métodos imunohistoquímicos, histológicos e tomográficos. Trinta e seis coelhos da linhagem New Zealand White foram submetidos a cirurgias para enxertia ″onlay″ de osso autógeno (grupo controle) e osso alógeno em lados diferentes da mandíbula de forma aleatória. Seis animais de cada grupo foram sacrificados aos 03, 05, 07, 10, 20 e 60 dias após as cirurgias. Cortes histológicos foram corados com Tricrômio de Mallory para as análises histológicas. As imuno marcações foram realizadas com osteoprotegerina (OPG); receptor activator of nuclear factor-kβ ligand (RANKL); fosfatase alcalina (ALP); osteopontina (OPN); vascular endothelial growth factor (VEGF); tartrate-resistant acid phosphatase (TRAP); colágeno tipo I (Col I) e osteocalcina (OC). A manutenção do volume e densidade dos enxertos foi avaliada por meio de tomografias obtidas após as cirurgias e após os sacrifícios. Os enxertos autógenos e alógenos exibiram padrões de preservação de volume e densidade similares; os dados histológicos mostram que a remodelação óssea no grupo alógeno ocorreu de modo mais intenso que no grupo autógeno; a avaliação por microscopia de luz mostra que a incorporação do osso autógeno ao leito receptor foi mais eficiente que no grupo alógeno; no grupo alógeno os resultados de imunohistoquímica demonstraram um quadro típico de intensa remodelação dos enxertos.
The reconstruction of jaws in implantology using methods of bone grafting constitutes is becoming the most popular surgical procedure due to the physiologic bone loss that follows teeth extraction or trauma. The autogenous grafts show advantages in relation to the other reconstruction techniques because its potential as bone regenerator. However, its removal implicates obligatorily in the areas donor areas. In the last decades aroused the interest for the bone bank (BTME) as alternative to the autogenous grafting, as a manner to avoid donor sites morbidity and reduction of time and surgery costs. The purpose of the study was to compare the behavior of allografts with autogenous using methods of immunochemistry, histology and tomography. Thirty six rabbits of the lineage New Zealand White were submitted to surgeries for onlay grafting of autogenous bone (group control) and allogenous bone randomly placed bilaterally in the mandible. Six animals of each group were sacrificed to the 03, 05, 07, 10, 20 and 60 days after the surgeries. Paraffin sections were stained with Mallorys Trichrome for histologics analyses. Immuno labeling accomplished with osteoprotegerin (OPG); receptor activator of nuclear factor-kβ ligand (RANKL); alkaline fosfatase (ALP); osteopontin (OPN); vascular endothelial growth factor (VEGF); tartrate-resistant acid phosphatase (TRAP); collagen type I (COL I) and osteocalcin (OC). The maintenance of the volume and density of the grafts was evaluated on tomographs obtained after surgeries and sacrifices. The autogenous grafts and allografts exhibited patterns of volume preservation and similar density; the histological data show that the remodelation bone in the group allograft happened in a more intense way than in the autogenous group; the evaluation for light microscopic shows that the incorporation of the autogenous bone on donors bed was more efficient than in the allogenous group; in the allogenous group for immunohistochemical results demonstrated a typical picture of intense remodelation of the grafts.

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Neto, Patricio José de Oliveira. « Estudo comparativo da fixação e integração de enxertos ósseos \'onlay\' com o uso de n-Butil-2-Cianocrilato ou parafuso de titânio. Estudo histológico, imunohistoquímico e tomográfico em coelhos ». Universidade de São Paulo, 2010. http://www.teses.usp.br/teses/disponiveis/58/58136/tde-12082010-141918/.

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Alguns trabalhos sobre a resposta do tecido ósseo ao Cianocrilato podem ser encontrados na literatura, embora nenhum deles avalie a resposta histológica e a fixação de enxertos ósseos onlay com o N-Butil-2-Cianocrilato (Indermil™). O objetivo do estudo proposto foi (1) analisar a manutenção do volume de enxertos ósseos onlay fixados à mandíbula de coelhos usando N-Butil-2-Cianocrilato (NB-Cn) ou parafuso de titânio, assim como avaliar (2) a remodelação e incorporação desses enxertos ao leito receptor e também (3) observar a diferença do nível de expressão da proteína tartarato-resistente ácido fosfatase (Trap) envolvida na absorção dos mesmos na presença do NB-Cn e do parafuso de titânio. Dezoito coelhos adultos foram envolvidos nesse estudo. Dois blocos ósseos provenientes da calvária dos coelhos foram transplantados para a mandíbula, em que de um lado do leito receptor o osso autógeno foi fixado com parafuso de osteossíntese (Grupo I - controle), e do outro lado com NB-Cn (Grupo II). Após o procedimento cirúrgico, os animais foram submetidos a exame tomográfico. O sacrifício dos animais ocorreu após 1 (n=9) e 6 (n=9) semanas do procedimento cirúrgico inicial, quando então os animais foram submetidos à nova tomografia. As imagens de tomografia foram usadas para estimativa da manutenção do volume dos enxertos. Cortes histológicos das áreas enxertadas foram preparados para se avaliar o reparo dos enxertos ósseos no sítio receptor e o nível de expressão da proteína Trap. Os resultados tomográficos mostraram melhor manutenção do volume dos enxertos fixados com NB-Cn (p≤0,05) em comparação àqueles fixados com parafuso, em ambos os tempos experimentais. Na avaliação imunohistoquímica, observou-se que a marcação da proteína Trap no período de 6 semanas foi significativamente maior em comparação ao tempo de 1 semana, sem apresentar diferença significante entre os grupos. A análise histológica revelou que embora o NB-Cn tenha provocado a destruição do periósteo, a estabilidade promovida pela cola permitiu que a revascularização e incorporação do enxerto ocorresse de forma semelhante ao grupo controle. Esses resultados indicam que o NB-Cn se comportou de forma superior ao parafuso como material de osteossínte. No entanto, estudos adicionais são necessários para se investigar a toxicidade deste composto quando utilizado como meio de fixação óssea.
Some experimental studies on the bone tissue responses to cyanoacrylate can be found in the literature, although neither evaluating the histological response and the fixation of onlay bone grafts with Indermil™. The aim of the proposed study is (1) to analyze the bone grafts volume maintenance fixed to the mandible of rabbits using Indermil™ or titanium screw, as well as (2) assess the healing of onlay grafts to the recipient bed and (3) the differences of level expression of Trap protein involved in reabsortion of these bone grafts using Indermil™ or an osteosynthesis screw. Eighteen adult New Zealand White rabbits were submitted to calvaria onlay bone grafts on both sides of the mandible. On one side the onlay bone graft were fixated with Indermil™, and on the other side bone graft with an osteosynthesis screw. The wounds were then closed, and a computed tomography was realized immediately after this procedure. The animals were killed after 1 (n=9) and 6 weeks (n=9), then the animals were submitted to another computed tomography (CT). The CT images were used to estimate the bone grafts volume maintenance. Histological sections of the grafted areas were prepared to evaluate the healing of bone grafts in the receptor site and the expression level of Trap protein. The CT scan showed better volume maintenance of the bone grafts fixed with Indermil™ (p≤0.05) compared to those fixed with screws, in both the experimental times. The immunohistochemical evaluation showed that the protein Trap expression in 6-week period was significantly higher compared to the 1-week period, without showing significant difference between the groups. Histological analysis revealed that although the Indermil™ has caused the destruction of the periosteum, the stability provided by the glue allowed that the bone graft revascularization and incorporation has occurred in a similar manner to the control group. These results indicate that the Indermi™ behaved than the screw as osteosynthesys material. However, further studies are needed to investigate the toxicity of this compound when used as a means of bone fixation.

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Mirjana, Miladinović. « Proteinska ekspresija i genska amplifikacija receptora humanog epidermalnog faktora rasta 2 ( HER2) kod adenokarcinoma pluća ». Phd thesis, Univerzitet u Novom Sadu, Medicinski fakultet u Novom Sadu, 2019. https://www.cris.uns.ac.rs/record.jsf?recordId=107833&source=NDLTD&language=en.

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Receptor humanog epidermalnog faktora rasta 2 (HER2) pripada porodici receptora protein-tirozin kinaze čija je aktivacija povezana sa proliferacijom malignih ćelija, inhibicijom apoptoze, tumorskom angiogenezom i sposobnosti invazije i metastaziranja. Povećana proteinska ekspresija HER2 receptora može nastati kao posledica amplifikacije gena i/ili transkripcijskih promena. Ekspresija HER2 receptora u humanim tumorima povezuje se sa agresivnijim ponašanjem i lošijom prognozom. Učestalost povećane proteinske ekspresije HER2 receptora u nesitnoćelijskim karcinomima pluća (NSCLC) je najviše zastupljena u adenokarcinomu u odnosu na druge histološke tipove. Identifikacija HER2 pozitivnih NSCLC omogućava određivanje grupe pacijenata koji bi bili kandidati za specifičnu terapiju. Problem predstavlja izbor metode detekcije HER2 receptora i nepostojanje utvrđenog protokola za očitavanje rezultata kao što postoji kod karcinoma dojke i želuca. Osnovni ciljevi ove doktorske disertacije su bili: da se odredi učestalost povećane proteinske ekspresije HER2 receptora u adenokarcinomu pluća; da se uporede rezultati povećane proteinske ekspresije HER2 receptora dobijene korišćenjem HER2 antitela „Hercep Test Dako“ i „Ventana anti-HER2/neu (4B5)“ antitela; da se uporedi prisustvo amplifikacije HER2 gena pomoću in situ hibridizacije (ISH) (Dual IHC HER2 kit;Ventana Medical Systems) retestiranjem uzoraka kod kojih je povećana proteinska ekspresija HER2 receptora ocenjena sa 2+ i 3+ dobijena „Hercep Test Dako“ sa prisustnom amplifikacijom HER2 gena na uzorcima koji su pomoću „Ventana anti-HER2/neu (4B5)“ ocenjeni sa 2+ i 3+; da se uporedi učestalost povećane proteinske ekspresije HER2 receptora i prisustva HER2 genske amplifikacije kod različitih histoloških podtipova adenokarcinoma pluća; da se utvrdi da li je povećana proteinska ekspresija HER2 receptora u adenokarcinomu pluća i/ili prisustvo genske amplifikacije povezano sa demografskim (starost i pol pacijenta) parametrima, pušačkim statusom, pojavom metastaza u regionalnim limfnim čvorovima i udaljenim organima, infiltracijom pleure i okolnih struktura, odnosno stadijumom bolesti. Povećana proteinska ekspresija HER2 receptora u adenokarcinomu pluća iznosi 7,4% za Hercep Test Dako i 2,7% za Ventana anti-HER2/neu (4B5) antitelo. Kod pozitivne ekspresije slažu se u 2%, dok se kod negativne ekspresije slažu u 91,9% slučajeva, što je ukupno 93,9%. Učestalost amplifikacije HER2 gena kod adenokarcinoma pluća je 17,6%, od toga je kod 2,7% slučajeva prisutna high grade amplifikacija. Postoji statistički značajna povezanost između povećane proteinske ekspresije HER2 receptora dobijene upotrebom HercepTest Dako i Ventana anti-HER2/neu (4B5) antitela i amplifikacije HER2 gena. Amplifikacija HER2 gena prisutna je kod 90,9% pacijenata sa povećanom proteinskom ekspresijom HER2 receptora koja se dobije upotrebom HercepTest Dako i kod 75% upotrebom Ventana anti-HER2/neu (4B5) antitela. Povećana proteinska ekspresija HER2 receptora dobijena pomoću HercepTest Dako i Ventana anti-HER2/neu (4B5) antitela je najčešća kod solidnog predominantnog tipa adenokarcinoma u patološkom T2a deskriptoru i IB stadijumu i acinarnog predominantnog tipa adenokarcinoma u patološkom T1b deskriptoru i IA stadijumu. Amplifikacija HER2 gena je najčešća kod solidnog a zatim kod acinarnog i papilarnog predominantnog tipa adenokarcinoma. Povećana proteinska ekspresija HER2 receptora dobijena pomoću HercepTest Dako i Ventana anti-HER2/neu (4B5) antitela i amplifikacija HER2 gena se najčešće javljaju kod muškaraca, pušača, u starosnoj dobi od 61-70 godina, tumora veličine 31-50 mm, N0 i M0 statusu bolesti, bez prisustva tumorske infiltracije pleure i okolnih struktura.
Human epidermal growth factor 2 (HER2) is a member of the epidermal growth factor family having tyrosine kinase activity, which is directly linked to malignant cells proliferation, apoptosis inhibition, tumor angiogenesis and ability for invasion and metastasis. Increased protein expression of HER2 receptors can be the consequence of gene amplification and/or transcription changes. Expression of HER2 receptors in human tumors is associated with more aggressive behavior and worse prognosis. Incidence of increased protein expression of HER2 receptors in non-small-cell lung carcinoma (NSCLS) is mainly represented in adenocarcinoma, in comparison with other histological types. Identification of HER2 positive NSCLC enables determination of a group of patients who would be candidates for specific therapy. The problem occurs in choosing the method of detection of HER2 receptors and non-existence of determined protocol for reading the results, as the one ones which exist for breast and gastric carcinoma. The main objectives of this PhD dissertation were: to determine the incidence of increased protein expression of HER2 receptors in lung adenocarcinoma; to compare the results of the increased protein expression of HER2 receptors obtained by using HER2 antibodies "HercepTest Dako" and "Ventana anti-HER2/neu (4B5)" antibodies; to compare the presence of HER2 gene amplification by in situ hybridization (ISH) (Dual IHC HER2 kit: Ventana Medical Systems) by retesting the samples in which the increased protein expression of HER2 receptors was graded with 2+ and 3+, obtained by "HercepTest Dako" with present gene HER2 amplification on samples obtained by "Ventana anti-HER2/neu (4B5) and graded with 2+ and 3+; to compare the incidence of increased protein expression of HER2 receptors and presence of HER2 gene amplification in different histological subtypes of lung adenocarcinoma; to determine if the increased protein expression of HER2 receptors in lung adenocarcinoma and/or presence of gene amplification is related to demographic (age and sex of the patient) parameters, smoking status, appearance of metastases in regional lymphatic nodes, distant organs, infiltration of pleura and surrounding structures, and stage of the disease. Increased protein expression of HER2 in lung adenocarcinoma is 7.4% for HercepTest Dako and 2.7% for Ventana anti-HER2/neu (4B5) antibody. In positive expression they are correlated in 2%, while in negative expression they are correlated in 91.9% cases, which is overall 93.9%. The incidence of HER2 gene amplification in lung adenocarcinoma is 17.6%, from that in 2.7% of the cases high grade amplification is present. There is a statistically significant correlation between increased protein expression of HER2 receptors obtained by use of HercepTest Dako and Ventana anti-HER2 /neu (4B5) antibody and amplification of HER2 genes. Amplification of HER2 genes is present in 90.9% of patients with increased protein expression of HER2 receptors, which is obtained by using HercepTest Dako and in 75% patients by using Ventana anti-HER2/neu (4B5) antibody. Increased protein expression of HER2 receptors obtained by HercepTest Dako and Ventana anti-Her2/neu (4B5) antibody is most common in solid predominant type of adenocarcinoma in pathological T2a descriptor and IB stadium and acinar predominant type of adenocarcinoma in pathological T1b descriptor and IA stadium. Amplification of HER2 genes is most common in solid, and then in acinar and papillary predominant type of adenocarcinoma. Increased protein expression of HER2 receptors obtained by HercepTest Dako and Ventana anti-HER2/neu (4B5) antibody and amplification of HER2 genes most commonly occurs in men, smokers, at the age of 61-70 years, tumor size 31-50 mm, NO and MO disease status, without presence of tumor infiltration of pleura and surrounding structures.

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Govender, Dhirendra. « An immunohistochemical and microsatellite analysis of nephroblastomas ». Thesis, 2008. http://hdl.handle.net/10413/9724.

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The aims of this study were: (i) to determine the association between p53, bcl-2, pRb, p21, cyclin A and p-glycoprotein immunoexpression and prognosis, and (ii) to determine the frequency of loss of heterozygosity and microsatellite instability at 11 p, 16q and mismatch repair gene loci and their association with prognosis, in nephroblastomas in South African children. There were 138 cases (111 of whom received preoperative chemotherapy) in the immunohistochemical study and, 70 cases (48 with preoperative chemotherapy) in the microsatellite study. The following monoclonal antibodies were used after heat induced epitope retrieval; p53, bcl-2, pRb, p21, cyclin A and p-glycoprotein. Six polymorphic microsatellite markers were selected from the 11p region, 5 from the 16q region and 6 from the loci of known mismatch repair genes. Automated fluorescent DNA technology was used in the analysis. The results of the immunohistochemical and microsatellite studies were correlated with patient age, gender, preoperative chemotherapy, SlOP histological classification, SlOP histological risk group, clinicopathological stage, patient outcome and survival using X2 , Fisher's exact test, Cox regression model and Kaplan-Meier estimates. The majority of patients presented with advanced disease. Anaplastic tumours and high-risk histology were associated with high disease stage. Mortality was directly related to increasing stage and histological risk group. Multivariate analysis showed that clinicopathological stage was the only factor significantly associated with survival (p<0.001) (hr=5.6, 95%CI: 2.1-14.9). High expression of p53 was more frequent in anaplastic tumours suggesting that p53 mutations are common events in this tumour type (p<0.001). Despite the strong association with tumour histology, there was no association with stage. Although p53 expression was found to be a predictor of survival in the univariate analysis this was not retained in the multivariate analysis. Tumours treated with preoperative chemotherapy showed higher bcl-2 immunoreactivity (p=0.027 but lower levels of pRb (p=0.040) and cyclin A expression (p<0.001). All anaplastic tumours showed high expression of pRb compared to the other histological types (p=0.003). Expression of xxii pRb was significantly associated with survival in the univariate analysis but not in the multivariate analysis. High cyclin A expression was associated with high risk histology (p<0.001). Cyclin A expression was found to be a significant predictor of survival in both the univariate (hr=1.7; 95%CI 1.2-2.4; p=0.002) and multivariate analyses (hr=1.7; 95%CI1.1-2.7; p=0.032). Although tumours with high risk histology were more likely to express high levels of p-glycoprotein, this did not reach significance. LOH at 11 p was seen in 64.7% of 68 informative cases. LOH at 11 p13 was more frequent than LOH at 11p15. LOH for both 11p13 and 11p15 was found in 39.7% of all tumours. MSI at 11 p was seen in 22.1 % of informative cases. The majority showed MSI for one marker only. LOH 16q was seen in 66.7% of 66 informative cases. MSI at 16q was seen in 16.7% of cases. LOH for 016S496 and 016S520 appear to be related to tumour histology and risk group. The most frequent locus for LOH was 16q21-22, which is known to harbour important genes, such as, E2F4 and E-cadherin. LOH for MMR markers was seen in 43.5% of 69 informative cases. MSI was seen in 11.6% of tumours. In the multivariate analysis there was no significant correlation between LOH at any of the loci studied and survival. There were no tumours with high frequency MSI. Low frequency MSI was of no clinicopathological significance. The following conclusions are made: (i) p53 mutations determined by high p53 expression is a frequent finding in anaplastic tumours, (ii) Bcl-2 may play a role in the chemoresistance of nephroblastomas, (iii) Rb gene alterations are not important in the development of nephroblastoma and anaplasia, (iv) Cyclin A expression is an independent predictor of survival, (v) p-glycoprotein may be responsible for the chemoresistance in a proportion of nephroblastomas, (vi) MSI is a rare occurrence in nephroblastoma and does not play a role in the development of nephroblastoma, (vii) LOH at 11 p and 16q are frequent findings in nephroblastomas, (viii) LOH for the specific 16q markers (016S496 and 016S520) may have an important prognostic role in nephroblastoma.
Thesis (Ph.D.)-University of KwaZulu-Natal, Durban, 2008.

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Hayes, DA. « Further immunohistochemical (IHC) characterisation of devil facial tumour disease 1 (DFT1) in the Tasmanian Devil (Sarcophilus harrisii) ». Thesis, 2019. https://eprints.utas.edu.au/31712/1/Hayes_whole_thesis_ex_pub_mat.pdf.

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As a veterinary histologist at Tasmania’s Animal Health Laboratory, Department of Primary Industries, Parks, Water and Environment (DPIPWE), I am positioned to respond to any emerging biosecurity risk. It was incomprehensible in 2002 to foresee how sectioning and staining the first of an extraordinary number of Tasmanian devil tumours would influence my professional life as a medical scientist. Tasmanian devil numbers plummeted and the likelihood of losing Tasmania’s iconic marsupial to ‘Devil Facial Tumour Disease’ (DFTD) became clearly evident. Research into the aetiology of the transmissible cancer became paramount. I was a co-author on two initial papers by Richmond Loh et al. describing the immunohistochemistry and pathology of DFTD followed by a broader paper by Stephen Pyecroft et al. integrating the histopathology, cytogenetics and epidemiology of this unusual transmissible tumour. Our immunohistochemical research continued to investigate tissue specific markers that define DFTD and its transmissible nature. I dovetailed my research at the Animal Health Laboratory through graduate research at the University of Tasmania where I secured two research grants offered by ‘Save the Tasmanian Devil Program’ (STDP), funding crucial to advancing our research findings. During my candidature, I have tested and validated over 150 commercially available anti-human antibodies on Tasmanian devil (marsupial) tissues, unfortunately sequence homology was not always one hundred percent with approximately half failing to recognise marsupial protein sequences. DFTD is a clonally evolved tumour of Schwann cell origin affecting Tasmanian devils (Sarcophilus harrisii) and is transferred by biting. This thesis presents my research as two major chapters. Firstly, a very comprehensive immunohistochemical approach investigating neural crest derived Schwann cell and melanocyte associated markers. These findings are novel and will contribute significantly to the understanding of the pathobiology of this unusual tumour. I have prepared the following manuscript pending submission, presented in chapter 1. Devil Facial Tumour 1 (DFT1) Immunophenotype Reveals a Progenitor-Like Cell with Schwann cell, Melanocyte and Self-Renewal Characteristics. In addition to gene and protein expression by DFTD, I wanted my immunohistochemical research findings to be applied in a practical way. I identified ERBB3 as a biomarker that can be utilised in both the early detection of DFTD and the identification of potential therapeutic regimes that could be applied therapeutically to wild Tasmanian devils to eradicate this disfiguring and terminal disease. This article was published in 2017 (chapter 2) ahead of the main body of my research (chapter 1) to expedite modes of DFT1 treatment, prevention and eradication which is a high priority of the STDP. ERBB3: A potential serum biomarker for early detection and therapeutic target for devil facial tumour 1 (DFT1). In summary, my research has comprehensively redefined the immunophenotype of DFTD, expressing novel proteins not available in the current literature. I show divergent origins of DFTD, a progenitor like cell with Schwann cell and melanocyte lineages, self-renewal characteristics that contributes to MHC down regulation and therefore enhanced transmissibility. I describe for the first time the histomorphology of the 5 strains of DFTD and extensively immunohistochemically stain these strains to determine any variation present. My findings are drafted for imminent manuscript submission. From my research, I published the first description of the expression of ERBB3 by DFTD and its possible early detection and possible therapeutic approach. I also first reported the expression of ERBB3 in cutaneous lymphoma in Tasmanian devils. It is reassuring that two subsequent publications, like ourselves, found ERBB3 to be important in DFTD tumourigenesis. As unbelievable as it seems researching a transmissible tumour in Tasmanian devils, it would be inconceivable to think I would be involved in a second transmissible tumour however, this is exactly what has happened. At DPIPWE I was involved in the Histology and immunohistochemistry of this newly described Tasmanian devil tumour. During my candidature I collaborated with fellow Tasmanian devil researchers Ruth Pye et al. and Max Stammnitz et al. through STDP by providing histological and immunohistochemical components of the research defining this second devil facial tumour. Our research findings have therefore renamed the original DFTD as DFT1, with the newly described tumour named DFT2, its intention to avoid confusion in the literature. My contribution to DFT2 is outlined in chapter 3 of my thesis. My future directions are to publish unfinished immunohistochemical work completed during my candidature and continue researching DFT1 and DFT2 by collaboration with Tasmanian devil researchers as well as continue histological and immunohistochemical support at the Animal Health Laboratory, DPIPWE.

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Vlčková, Linda. « Interakce vajíček a miracidií Trichobilharzia regenti s nosní sliznicí kachen ». Master's thesis, 2018. http://www.nusl.cz/ntk/nusl-388296.

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Trichobilharzia regenti is a nasal avian schistosome which has during the initial phase of infection an affinity to the nervous system. Larvae migrate through the central nervous system to the nasal mucosa of waterfowl, where they mature and reproduce. Until now this infection phase has been described only marginally. Adults are located in the nasal mucosa approximately from 13th to 24th day post infection. During this life phase, they migrate through the vascularized connective tissue and lay eggs, the presence of which has been detected in the tissue only. Maturation and hatching of miracidia occur in the tissue (unlike human schistosomes). The parasite causes inflammation, and the tissue is infiltrated with immune cells. Lymphocytes, granulocytes, macrophages, plasma cells and giant multi-nuclear cells were described by histological methods. The thesis is focused on a more detailed description of cellular immune response and histopathological changes of the tissue by means of histological stains, and antibody/lectin probes. The flukes were observed more frequently in the blood vessel lumen, together with a higher number of immune cells compared to the healthy duck. Infiltration by a high number of lymphocytes occurred in the tissue, the macrophages were frequently observed in clusters around the...

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Thèses sur le sujet « Immunohistochemistry and histology »

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