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McKenna, Mary K., Amanda Rosewell-Shaw und Masataka Suzuki. „Modeling the Efficacy of Oncolytic Adenoviruses In Vitro and In Vivo: Current and Future Perspectives“. Cancers 12, Nr. 3 (07.03.2020): 619. http://dx.doi.org/10.3390/cancers12030619.
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Oncolytic adenoviruses (OAd) selectively target and lyse tumor cells and enhance anti- tumor immune responses. OAds have been used as promising cancer gene therapies for many years and there are a multitude of encouraging pre-clinical studies. However, translating OAd therapies to the clinic has had limited success, in part due to the lack of realistic pre-clinical models to rigorously test the efficacy of OAds. Solid tumors have a heterogenous and hostile microenvironment that provides many barriers to OAd treatment, including structural and immunosuppressive components that cannot be modeled in two-dimensional tissue culture. To replicate these characteristics and bridge the gap between pre-clinical and clinical success, studies must test OAd therapy in three-dimensional culture and animal models. This review focuses on current methods to test OAd efficacy in vitro and in vivo and the development of new model systems to test both oncolysis and immune stimulatory components of oncolytic adenovirotherapy.
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Yaneva, N., M. Baycheva, P. Kostova, V. Papochieva, S. Mileva, D. Miteva, A. Savov und G. Petrova. „Preventable Hazards from in Vitro Fertilization – A Case Series of CF Patients from Bulgaria“. Balkan Journal of Medical Genetics 26, Nr. 1 (01.07.2023): 83–88. http://dx.doi.org/10.2478/bjmg-2023-0001.
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Abstract Pre-implantation genetic diagnosis (PGD) is not often performed when donor gametes are used, due to its high cost. This is with the presumption that the donors are healthy. We report on five cases of babies with confirmed cystic fibrosis (CF), being the result from in vitro fertilization (IVF) with donor (4 cases) or own gametes (one case). There has been no family history for CF in any of the families affected. The clinical presentation in the children ranged from meconium ileus to recurrent respiratory infections and severe nasal polyposis. The age of diagnosis also varied from birth until 9 years. Since one of the presented cases was discovered in a very renowned private IVF clinic, the clinic changed their own protocol, and currently they test every donor for CF carriership. The percentage of CF carriers in the donor population is roughly the same as the one predicted in the general population of Bulgaria – 1/33. Although PGD is costly, the costs for proper care for a CF patient are currently much higher. The more economical option would to screen every donor for CF carriership. IVF requires a lot of physical and psychological stamina. The couples that go through this procedure also require a great deal of hope. It is essential to be more preconscious for possible congenital diseases. We advocate every IVF center to test the donors for CF carriership or to provide PGD for their clients.
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Shafira, Nadia, Liva Wijaya, Ayang Halim, Boy Abidin, Tri Bowo Hasmoro und Budi Wiweko. „#40 : Correlation Between Pre-Trigger Estradiol Levels and the Number of Mature Oocytes in Patients Who Underwent In Vitro Fertilization“. Fertility & Reproduction 05, Nr. 04 (Dezember 2023): 336. http://dx.doi.org/10.1142/s2661318223741449.
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Background and Aims: Strong association between pre-trigger estradiol levels on the day of human Chorionic Gonadotropin (hCG) administration and the number of mature oocytes retrieved was widely reported. Consequently, higher estradiol levels measured might give a prediction of the higher number of mature oocytes retrieved on Ovum Pick Up (OPU) procedure. This study was done to investigate the correlation between pre-trigger estradiol levels and number of mature oocytes retrieved in patients who underwent In Vitro Fertilization (IVF) cycles in Indonesian population. Method: A cross sectional study was done at Mbrio IVF Clinic, Mitra Keluarga Hospital Kelapa Gading, Jakarta, Indonesia. Data was collected from the medical records of 77 patients who underwent IVF cycles between September 2021 and March 2023. The level of estradiol on the hCG administration day and the number of mature oocytes retrieved were analysed to find any correlation between both variables using the Spearman Test. Results: There were 77 women who underwent IVF procedures with the age range between 27 and 45-year-old (mean 35,5 ± 4,52). Most of them (83%) had primary infertility and had infertility between 1 to 14 years (mean 5,5 ± 3,48). These subjects came to the clinic mostly due to both male and female factors (49,4%), followed by male factor only (27,3%). E2 on hCG day ranged between 194 to 13.000 pg/ml with the median 2.709 pg/ml, and the mature oocytes retrieved ranged between 0 and 31 with a median of 8 oocytes. A statistically significant correlation (r = 0.827; p<0.01) was found between the estradiol level and mature oocytes retrieved. Conclusion: Pre-trigger Estradiol level can be used as a predictor for the number of mature oocytes retrieved in IVF cycle as these two variables are strongly correlated to each other. A multi-centre study with larger sample size is needed to validate current data.
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Irfan, Sarah, Seema Irfan, Mubassar Fida und Israr Ahmad. „Contamination assessment of orthodontic bands after different pre-cleaning methods at a tertiary care hospital“. Journal of Orthodontics 46, Nr. 3 (13.06.2019): 220–24. http://dx.doi.org/10.1177/1465312519855402.
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Introduction: Infection control in dentistry is a major concern due to risk of transmission of communicable diseases. The aim of this study is to evaluate and compare the efficacy of various pre-cleaning methods for the tried-in orthodontic bands. Material and methods: An in-vitro experimental study was conducted at the Central Sterilization Services Department (Dental Clinic) and the Microbiology lab at our university hospital. A total of 130 bands were included in our study which comprised 10 controls and the rest were equally divided into three groups according to the pre-cleaning methods, i.e. manual scrubbing, enzymatic solution and a combination of both. The orthodontic bands were incubated in the brain heart infusion broth at 37 °C for five days after pre-cleaning and sterilisation in a steam autoclave and were assessed for any bacterial growth. The chi-square test was applied to determine any significant association between the various pre-cleaning methods and the frequency of bands that showed growth. Effect size was calculated using the phi coefficient. Results: The enzyme method revealed 5% of the sample to exhibit bacterial growth, whereas manual scrubbing and the combination of both showed no growth. There was no statistically significant difference among the three methods ( P = 0.131). Further investigations showed the presence of Staphylococcus non-aureus bacterial species in contaminated bands from group II. Conclusions: All pre-cleaning methods were found to be equally effective in the decontamination of bands. Hence, the tried-in bands can be safely reused after pre-cleaning and sterilisation.
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Corey, Jacquelynne P., und Anil Gungor. „In vitro testing for immunoglobulin E-mediated food allergies“. Otolaryngology–Head and Neck Surgery 115, Nr. 4 (Oktober 1996): 312–18. http://dx.doi.org/10.1016/s0194-5998(96)70044-x.
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The role of immunoglobulin E-mediated food allergy in subjects with allergic disorders, especially in patients with rhinitis and sinusitis, is underestimated by clinicians because of the initial attribution of these disorders to immediate-type hypersensitivity reactions. The difficulties of diagnosing food-related reactions have caused further delay in their recognition and incorporation into the daily practice of diagnosing food allergy. Two of the diagnostic methods for food allergy are the in vitro assay of total immunoglobulin E and the measurement of food-specific immunoglobulin E levels in serum with the radioallergosorbent test. Measurement of specific immunoglobulin E level is the most commonly used but also one of the most controversial techniques. We examined 123 patients with rhinitis who were referred to our otolaryngology/allergy clinic between January and April 1995. All patients received an initial radioallergosorbent test screen, which included milk. We determined the positive predictive value of this positive screen and, in particular, of a positive test for milk in the diagnosis of immunoglobulin E-mediated food allergies in these patients. Conclusions were based on comparison with the result of an additional radioallergosorbent test food panel consisting of eight common and two investigational food allergens. (Otolaryngol Head Neck Surg 1996;115:312-8.)
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Greer, J. J., Z. al-Zubaidy und J. E. Carter. „Thyrotropin-releasing hormone stimulates perinatal rat respiration in vitro“. American Journal of Physiology-Regulatory, Integrative and Comparative Physiology 271, Nr. 5 (01.11.1996): R1160—R1164. http://dx.doi.org/10.1152/ajpregu.1996.271.5.r1160.
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In the present study, we test whether thyrotropin-releasing hormone (TRH) stimulates respiratory frequency in perinatal rats by acting at regions of the medulla responsible for respiratory rhythmogenesis, the pre-Botzinger complex. We also test whether TRH stimulates respiration in the fetal rat at a time shortly after the inception of respiratory rhythmogenesis [embryonic days (E) 17-18]. Two in vitro experimental models were utilized: the isolated brain stem-spinal cord preparation from fetal (E17-E18) and neonatal [postnatal days (P) 0-2] rats and the medullary slice preparation isolated from neonatal rats (P1-P2). Bath application of TRH caused a dose-dependent, reversible increase (maximum increase approximately 60%) in the frequency of respiratory rhythmic neural discharge generated by brain stem-spinal cord [half-maximal effective concentration (EC50) approximately 9 nM] and medullary slice (EC50 approximately 2.5 nM) neonatal rat preparations. Pressure injection of TRH unilaterally into the region of the pre-Botzinger complex of the neonatal medullary slice caused an approximately 28% increase in the frequency of respiratory discharge. Application of TRH to the medium bathing fetal rat brain stem-spinal cord preparations caused an approximately threefold increase in respiratory discharge frequency. We conclude that TRH stimulates respiratory discharge frequency from the time near inception of respiratory motor discharge and acts directly at the pre-Botzinger complex.
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Khan, Muhammad Ali, Andrew J. Kassianos, Wendy E. Hoy, AHM Khurshid Alam, Helen G. Healy und Glenda C. Gobe. „Promoting Plant-Based Therapies for Chronic Kidney Disease“. Journal of Evidence-Based Integrative Medicine 27 (Januar 2022): 2515690X2210796. http://dx.doi.org/10.1177/2515690x221079688.
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Chronic kidney disease (CKD) is debilitating, increasing in incidence worldwide, and a financial and social burden on health systems. Kidney failure, the final stage of CKD, is life-threatening if untreated with kidney replacement therapies. Current therapies using commercially-available drugs, such as angiotensin-converting enzyme inhibitors, angiotensin II receptor blockers and calcium channel blockers, generally only delay the progression of CKD. This review article focuses on effective alternative therapies to improve the prevention and treatment of CKD, using plants or plant extracts. Three mechanistic processes that are well-documented in CKD pathogenesis are inflammation, fibrosis, and oxidative stress. Many plants and their extracts are already known to ameliorate kidney dysfunction through antioxidant action, with subsequent benefits on inflammation and fibrosis. In vitro and in vivo experiments using plant-based therapies for pre-clinical research demonstrate some robust therapeutic benefits. In the CKD clinic, combination treatments of plant extracts with conventional therapies that are seen as relatively successful currently may confer additive or synergistic renoprotective effects. Therefore, the aim of recent research is to identify, rigorously test pre-clinically and clinically, and avoid any toxic outcomes to obtain optimal therapeutic benefit from medicinal plants. This review may prove to be a filtering tool to researchers into complementary and alternative medicines to find out the current trends of using plant-based therapies for the treatment of kidney diseases, including CKD.
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Sinjari, Bruna, Manlio Santilli, Gianmaria D’Addazio, Imena Rexhepi, Alessia Gigante, Sergio Caputi und Tonino Traini. „Influence of Dentine Pre-Treatment by Sandblasting with Aluminum Oxide in Adhesive Restorations. An In Vitro Study“. Materials 13, Nr. 13 (07.07.2020): 3026. http://dx.doi.org/10.3390/ma13133026.
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Dentine pretreatment through sandblasting procedures has been widely studied but no curve test results are currently available. Thus, the aim herein was to in vitro compare the adhesive strength in sandblasted or not samples using a universal testing machine. Thirty -two bovine teeth were divided into two groups, namely test (n = 16 bars), sandblasting with aluminum oxide particles (50 µm) was performed before the adhesion procedures), and control (n = 16 bars), where no sandblasting procedure was performed. A bi-material curve test was used to evaluate the characteristics of the dentine pretreatment in terms of tensile stress and fracture strength. A scanning electron microscope (SEM) was used to analyze the fracture topography in the composite, bonding, dentin, and at the relative interfaces. The results demonstrated a statistically significant difference between the two groups in terms of tensile stress at maximum load showing values of 84.300 ± 51.342 MPa and 35.071 ± 16.609 MPa, respectively for test and control groups (p = 0.033). Moreover, a fracture strength test showed values of 18.543 ± 8.145 MPa for test and 8.186 ± 2.833 MPa for control group (p = 0.008). In conclusion, the sandblasting treatment of the dentine significantly influenced the mechanical resistance of the adhesion in this in vitro study.
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Zhou, Feng, Liang Liang Wu, Yuan Yuan Cui, Ying Chen, Jie Yang und Nan Huang. „A Method to Measure the Station of Artificial Mechanical Heart Valves“. Advanced Materials Research 683 (April 2013): 712–15. http://dx.doi.org/10.4028/www.scientific.net/amr.683.712.
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The experiments of artificial heart valves were divided into in vivo and in vitro experiments; in vivo experiments provide accurate experimental parameters serving in vitro research. Simulation experiment used in vitro usually goes like this, firstly design a similar model or prototype phenomenon, then analysis the model working out the regular parameters related to the process, ruled out the possibility of impact on the study of individual exist in vivo experiment. In vitro experiments are likely designed; performance can be simplified and prominently concerned about contents, even designed some extreme conditions to test. A number of means related to fluid experimental measurement are included, such as the Particle Image Velocimetry（PIV）[1], Dual Catheter Method [2]，and ultrasonic method[3] and so on. However, these methods have different kinds of limitations, for example the Dual Catheter Method cannot be used as a routine determination for clinic due to its destructiveness, and PIV test requires expensive equipment. This study was designed by the image processing technology of high-speed photography aiming at the production of a reliable, simple, economic, injury-free and non-contact measurement method.
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Schuler, Kevin M., Brooke S. Rambally, Megan J. DiFurio, Paola A. Gehrig und Victoria Lin Bae-Jump. „A preoperative window study of metformin for the treatment of endometrial cancer.“ Journal of Clinical Oncology 31, Nr. 15_suppl (20.05.2013): 5519. http://dx.doi.org/10.1200/jco.2013.31.15_suppl.5519.
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5519 Background: Obesity and diabetes have been linked to poorer survival and increased recurrence rates in endometrial cancer. The anti-diabetic medication, metformin, has been shown to have anti-tumorigenic effects in vitro and in vivo, via AMPK activation and inhibition of the mTOR pathway. We conducted a pre-operative window clinical trial of metformin in obese endometrial cancer patients to evaluate short-term in vivo molecular changes. Methods: Women with endometrioid endometrial cancer who were obese (BMI>30) were recruited from a gynecologic oncology clinic. Once enrolled, patients had a repeat pre-treatment endometrial biopsy and then began metformin at a dose of 850 mg PO once daily for 1-4 weeks prior to hysterectomy/surgical staging. A tissue microarray, using triplicate cores from each specimen, was constructed from paired formalin-fixed, paraffin-embedded endometrial biopsy (pre-treatment) and hysterectomy (post-treatment) specimens. The expression of Ki-67, a marker of cell proliferation, was measured by immunohistochemistry. Individual slides were digitized using the Aperio ScanScope (Aperio Technologies, Vista, CA), and digital images were analyzed using Aperio ImageScope software. The Signed Rank Test was used for statistical analysis. Results: Sixteen patients have completed the protocol. The mean duration of treatment was 14.5 days. Percent Ki-67 staining decreased significantly with metformin treatment (mean of 19.5% decrease, p = 0.026). Two patients experienced grade 1 toxicities, including mild abdominal pain and loose stools. Ten of the 16 patients responded to metformin based on decreased proliferation from their pre- to post-treatment specimens. There were no differences in median age, BMI, HgbA1c, or number of doses taken between responders and non-responders to treatment. Pre-treatment Ki-67 levels were statistically higher in the women that responded to metformin treatment (52% versus 27.5%, p = 0.0067). Conclusions: Metformin significantly reduced proliferation in a pre-operative window study in obese endometrial cancer patients, providing further support for therapeutic clinical trials of metformin in this obesity-driven disease.
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Bassi, Julio, Tamara Tedesco, Daniela Raggio, Ana Maria Santos, Renata Bianchi und Giselle de Sant’Anna. „Is it necessary to pre-treat Dentine before GIC Restorations? Evidence from an in Vitro Study.“ Acta Odontológica Latinoamericana 33, Nr. 1 (Juni 2020): 27–32. http://dx.doi.org/10.54589/aol.33/1/027.
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The aim of this study was to assess the influence of different pre-treatment approaches on glass ionomer cement (GIC) bond strength (BS) to dentine. Sixty bovine incisors were allocated into six groups according to substrate - sound or caries-affected dentine; and substrate pre-treatment approach - no conditioning (control), polyacrylic acid for 10 s and phosphoric acid for 7 s. Teeth in the caries-affected dentine group were previously submitted to cariogenic pH-cycling challenge. After dentine pre-treatment, according to experimental groups, polyethylene tubes were placed on flat dentine surfaces and filled with GIC. Teeth were stored in distilled water for 24 h at 37 °C and then submitted to microshear test (0.5 mm/min). Failure pattern analysis was performed under stereomicroscope (400x). Data were analysed using two-way ANOVA and Tukey's test (α=5%). Statistically significant differences were found for the pre-treatment approach, regardless of substrate (p<0.001). The polyacrylic acid group and control group had similar BS values, and were both better than the phosphoric acid group. In general, GIC had better bonding performance in sound dentine than in caries-affected dentine. In conclusion, dentine pre-treatment with polyacrylic acid did not improve the performance of GIC restoration on clinically relevant substrates.
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Juntke, Jenny, Xabier Murgia, Nazende Günday Türeli, Akif Emre Türeli, Chelsea R. Thorn, Marc Schneider, Nicole Schneider-Daum, Cristiane de Souza Carvalho-Wodarz und Claus-Michael Lehr. „Testing of aerosolized ciprofloxacin nanocarriers on cystic fibrosis airway cells infected with P. aeruginosa biofilms“. Drug Delivery and Translational Research 11, Nr. 4 (28.05.2021): 1752–65. http://dx.doi.org/10.1007/s13346-021-01002-8.
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AbstractThe major pathogen found in the lungs of adult cystic fibrosis (CF) patients is Pseudomonas aeruginosa, which builds antibiotic-resistant biofilms. Pulmonary delivery of antibiotics by inhalation has already been proved advantageous in the clinic, but the development of novel anti-infective aerosol medicines is complex and could benefit from adequate in vitro test systems. This work describes the first in vitro model of human bronchial epithelial cells cultivated at the air–liquid interface (ALI) and infected with P. aeruginosa biofilm and its application to demonstrate the safety and efficacy of aerosolized anti-infective nanocarriers. Such a model may facilitate the translation of novel therapeutic modalities into the clinic, reducing animal experiments and the associated problems of species differences. A preformed biofilm of P. aeruginosa PAO1 was transferred to filter-grown monolayers of the human CF cell line (CFBE41o-) at ALI and additionally supplemented with human tracheobronchial mucus. This experimental protocol provides an appropriate time window to deposit aerosolized ciprofloxacin-loaded nanocarriers at the ALI. When applied 1 h post-infection, the nanocarriers eradicated all planktonic bacteria and reduced the biofilm fraction of the pathogen by log 6, while CFBE41o- viability and barrier properties were maintained. The here described complex in vitro model approach may open new avenues for preclinical safety and efficacy testing of aerosol medicines against P. aeruginosa lung infection. Graphical abstract
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Sinenko, Irina, Fabien Kuttler, Valentin Simeonov, Alexandre Moulin, Christina Stathopoulos, Gerardo Turcatti, Adeline Berger, Francis Munier und Paul Dyson. „Abstract 3541: Tumoroid-based screening platform to test focal, chemo- and combination therapy for retinoblastoma“. Cancer Research 83, Nr. 7_Supplement (04.04.2023): 3541. http://dx.doi.org/10.1158/1538-7445.am2023-3541.
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Abstract Current treatments for retinoblastoma, the most common intraocular malignancy, rely on a limited number of drugs repurposed from other pediatric cancer therapies. Drug toxicity and relapse of the disease necessitate new therapeutic strategies. The implementation of drug candidates in the clinic is usually followed by in vitro validation, which requires accurate and clinically relevant in vitro models. We report here a robust 3D tumoroid-based platform, for testing various clinical protocols, e.g. therapeutic agents following a single or repeated exposure, alone or in combination with focal therapy (thermotherapy). The platform, generated from the Y79 retinoblastoma cell line, consists of matrix-embedded tumoroids, which retain the key features of the disease and allow prolonged culture time. Following the optimization of growth conditions, the effect of single and repeated exposure to chemotherapeutic agents has been measured by high-throughput automated fluorescence imaging followed by quantification. The clinical conditions of chemothermotherapy have been recapitulated with a setup consisting of: an infrared diode laser (810 nm, 0.3 W) that heats the tumoroids in the presence of indocyanine green (50 µg/mL); a thermal camera to enable real-time temperature measurement; and an environment-controlled inverted microscope to ensure physiological conditions and precise focusing of the laser. Melphalan and carboplatin, two drugs currently used to treat retinoblastoma in clinical protocols of chemotherapy and chemothermotherapy respectively, have been applied to validate the system.Tumoroid viability and size progressively decreased over 4 weeks of repeated drug exposure with melphalan, reflecting the response of retinoblastoma in advanced clinical cases. Carboplatin treatment showed an activity enhancement in the tumoroid model similar to that observed in the clinic, when combined with focal therapy, demonstrating that the platform system closely recapitulates the clinical conditions. Inversely, the thermotherapy experimental protocol designed for 2D in vitro cell culture and commonly used to date, i.e. an incubation temperature increased to 42 °C for 1 h, did not show an improvement of the carboplatin cytotoxicity, confirming that our laser-based thermotherapy protocol provides a more accurate and clinically relevant drug response. Such an approach, combining physiologically relevant in vitro cancer models and investigation of drugs in well-established treatment modalities, can significantly reduce the number of new drug candidates that give false-positives or overestimated efficacies when evaluated in relevant in vivo models. It is expected that the platform will contribute to the implementation of new therapeutic strategies to treat retinoblastoma, and could also be adapted to other diseases for which repeated drug exposure and/or chemothermotherapy is relevant. Citation Format: Irina Sinenko, Fabien Kuttler, Valentin Simeonov, Alexandre Moulin, Christina Stathopoulos, Gerardo Turcatti, Adeline Berger, Francis Munier, Paul Dyson. Tumoroid-based screening platform to test focal, chemo- and combination therapy for retinoblastoma. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3541.
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Silva, D. S., P. Rodriguez, A. Galuppo, N. S. Arruda und J. L. Rodrigues. „Selection of bovine oocytes by brilliant cresyl blue staining: effect on meiosis progression, organelle distribution and embryo development“. Zygote 21, Nr. 3 (27.07.2011): 250–55. http://dx.doi.org/10.1017/s0967199411000487.
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SummaryThe selection of competent oocytes for in vitro maturation is still a major problem during bovine in vitro embryo production. Markers for in vitro cytoplasmic maturation, based on the organization of cortical granule and mitochondria, are lacking. We examined the pre-selection of immature bovine oocytes by brilliant cresyl blue stain (BCB test) based on glucose-6-phosphate dehydrogenase (G6PDH) activity during oocyte development. Oocytes were recovered from ovarian follicles exposed to 26 μM BCB stain and classified according to the aspect of their cytoplasm: BCB+ (oocytes with blue cytoplasm) and BCB− (unstained cytoplasm) and then in vitro matured into a conventional in vitro maturation (IVM) medium and standard procedure. In Experiment 1, nuclear maturation was determined by polar body identification, while cytoplasmic maturation was based on cortical granule (CG) migration (peripheral) and mitochondria distribution (central). Evidence of polar body, cortical granule migration and of centrally located mitochondria was significantly (p < 0.05) higher in BCB+ oocytes than in BCB− (polar body present: 65% vs 20%; peripheral CG: 72% vs. 14%; and central mitochondria: 85% vs. 19%, respectively). In Experiment 2, the efficiency pre-selection of bovine oocytes by BCB on embryo development in vitro was assessed. Cleavage rates were similar (75%) among control, BCB+ and BCB− groups, while blastocyst rates on D7 were (p < 0.05) higher (35%) in BCB+ vs BCB− (10%) or control (28%). We showed that the BCB test is efficient to identify competent immature bovine oocytes to undergo synchronous nuclear and cytoplasmic in vitro maturation thus yielding higher in vitro embryo development to blastocyst stage.
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Brogni, Cláudio Francisco, Lain Uriel Ohlweiler, Norton Klein, Joana Claudia Mezzalira, Jose Cristani und Alceu Mezzalira. „Pre-incubation of porcine semen reduces the incidence of polyspermy on embryos derived from low quality oocytes“. Ciência Rural 46, Nr. 6 (Juni 2016): 1113–18. http://dx.doi.org/10.1590/0103-8478cr20150700.
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ABSTRACT: The main cause of low efficiency of in vitro produced porcine embryos is the high polyspermic penetration rates at fertilization, which is aggravated in low quality oocytes. Experiment 1 evaluated the embryo development in high and low quality oocytes. Experiment 2 evaluated the embryo development and quality of low quality oocytes fertilized with sperm pre-incubated during 0h (control), 0.5h, 1h and 1.5h. Experiment 3 investigated fertilization and monospermic rates of the same groups of Experiment 2. Experiment 4 evaluated embryo development, cell density, fertilization and monospermic rates of high quality oocytes using semen pre incubated during the best time observed in the previous experiments. Cleavage and blastocyst rates were analyzed by chi-square test, and remaining data by ANOVA and Tukey test (P≤0.05). The cleavage (74.8 vs 51.7%) and blastocyst (33.7 vs 9.8%) rates were greater in oocytes of high versus low quality, with no differences in cell density. Fertilization rates (65.6 to 79.5%) were not influenced by pre-incubation time. However, semen pre-incubation during 1.5h increased monospermic penetration (53.3%) and cleavage rates (92.5%) in low quality oocytes. Blastocyst rate was improved with 1.5h of semen pre incubation; however they were still lower than that observed with high quality control oocytes. Ultimately, pre-incubation did not influence fertilization, monospermic penetration, embryo development rates, nor cell density in oocytes of high quality. Low-quality porcine oocytes resulted in better rates of embryo development if in vitro fertilized with sperm pre-incubated for 1.5 hour.
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Beata, Birta Orsolya, Cristina Bica, Farkas Hunor Pal, Bernadett Kerekes Mathe, Andreea Bors und Melinda Szekely. „The Impact of the Phosphoric Acid on Calcium Content of Dental Enamel - in vitro Examination“. Revista de Chimie 68, Nr. 9 (15.10.2017): 2066–69. http://dx.doi.org/10.37358/rc.17.9.5823.
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This study is intended for testing the way and extent how various acids influence dental erosion. Impacted third molar teeth removed under surgical exploration have been used in our experiments. The test samples have been prepared based on the Semmelweis University Dental Care Clinic protocol. Then the organic material has been removed using incubation solutions. Every 6 hours, the calcium concentration has been measured using a Radelkis OP- 274 Ph-ionometer and an OP-Ca-0711P-S type electrode sensitive to calcium ions, strictly following the manufacturer�s instructions. Data were statistically processed in Graphpad InStat 3 (Graphpad Software Inc, CA, USA) using paired T-test and ANOVA. The results contradict several articles published in the scientific literature dealing with this subject and describing the linear dependence between the dental erosion progress and time. Our study emphasized that the erosion process progressed in time, however it failed to show a linear dependence.
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Chang, Yuhan, Mel S. Lee, Jiann-Jong Liau, Yu-Liang Liu, Wen-Chuan Chen und Steve W. N. Ueng. „Polyethylene-Based Knee Spacer for Infection Control: Design Concept and Pre-Clinical In Vitro Validations“. Polymers 12, Nr. 10 (13.10.2020): 2334. http://dx.doi.org/10.3390/polym12102334.
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Antibiotic-loaded polymethyl methacrylate (PMMA) has been widely applied in the treatment of knee periprosthetic joint infections. However, problems with antibiotic-loaded PMMA-based spacers, such as structural fracture and implant dislocation, remain unresolved. A novel polyethylene-based spacer, designed with an ultra-congruent articulating surface and multiple fenestrations, was introduced in the current study. Validation tests for biomechanical safety, wear performance, and efficacy of antibiotic cement were reported. During cycle fatigue testing, no tibial spacer failures were observed, and less wear debris generation was reported compared to commercial PMMA-based spacers. The volumetric wear of the novel spacer was within the safety threshold for osteolysis-free volumetric wear. An effective infection control was demonstrated despite the application of lesser antibiotic cement in the 30-day antibiotic elution test. The tube dilution test confirmed adequate inhibitory capabilities against pathogens with the loaded antibiotic option utilized in the current study. The novel polyethylene-based knee spacer may offer sufficient biomechanical safety and serve as an adequate carrier of antibiotic-loaded cement for infection control. Further clinical trials shall be conducted for more comprehensive validation of the novel spacer for practical application.
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Sale, M., Kwaji M. und Wakawa, H. Y. „Incidence and Antibiogram of Uropathogens Isolated from Pregnant Women Attending Antenatal Clinic in a Public Hospitalin Yola, Adamawa State, Nigeria“. UMYU Journal of Microbiology Research (UJMR) 7, Nr. 2 (30.12.2022): 92–98. http://dx.doi.org/10.47430/ujmr.2272.014.
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Urinary tract infection during pregnancy including asymptomatic bacteriuria has been associated with some adverse outcomes for the mother and foetus. Studies have shown that pregnant women with urinary tract infections have a significantly higher rate of intra-uterine growth restriction, pre-eclampsia, caesarean and pre-term deliveries. This study was undertaken to determine the occurrence, distribution and antibacterial susceptibility pattern of uropathogens among pregnant women attending antenatal clinic in a public hospital in Yola. A total of 200 clean catch mid-stream urine samples were collected from pregnant women and inoculated on cysteine lactose electrolyte deficient agar for bacterial count and isolation of uropathogens. The isolates were also screened for ability to form biofilms using Congo Red Agar method and antibiotic susceptibility test was done on the isolates using agar disk diffusion method. The incidence of UTI among the pregnant women was found to be 90%. The predominant uropathogens were E. coli (46.0%), S. aureus (20.0%) and Klebsiella pneumoniae (17.0%) while the least occurring uropathogen was Proteus sp. (2.0%). Women in the third and first trimester of pregnancy had the highest occurrence of UTI. A total of 47.0% of the isolates produced biofilm in vitro with the highest biofilm production observed among Proteus sp. (75.0%) and Pseudomonas aeruginosa (60.0%) isolates. The occurrence of antibiotic resistance among the isolates was high, with greater susceptibility observed to quinolone and third generation cephalosporin. The E. coli isolates demonstrated varying levels of resistance to all the antibiotics tested. Because of the gravity of problems of UTI in pregnancy and that 9 in 10 pregnant women from this study have symptomatic or asymptomatic infection, it is concluded that pregnant women should be screened for urinary tract infection during antenatal visits especially during the first and third trimesters of pregnancy.
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Poempida, Fayka Putri, Jimmy Yanuar, Hamdani Lunardhi, Samsulhadi Samsulhadi und Relly Y. Primariawan. „Factors Affecting The Outcome of in Vitro Fertilization (IVF)“. Syntax Literate ; Jurnal Ilmiah Indonesia 7, Nr. 1 (10.01.2022): 444. http://dx.doi.org/10.36418/syntax-literate.v7i1.5780.
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The high prevalence of infertility motivated researchers to find a solution, henceforth In Vitro Fertilization was invented. Factors that affect the outcome of IVF may include sperm analysis, maternal Body Mass Index (BMI), maternal smoking habits, endometriosis, and maternal age. However, there are ongoing debates about the role of said factors regarding the outcome of IVF. The objective of this research is to analyze those factors. This research is a Case-Control study with an analytical observational design. Data were retrieved from patients’ medical records undergoing IVF in Graha Amerta Fertility Clinic from January 2019-October 2020. First, the Chi-Square Test revealed sperm abnormality (p=0.212), Maternal BMI (p=0.427), endometriosis (p=0.067), meaning there was no connection with the outcome of IVF. Simultaneously, maternal age (p=0.037) showed a connection with the outcome of IVF. From the Binary Logistic Regression Test, maternal age 36-40 years old (p=0.044) affects the outcome of IVF significantly. Concurrently maternal BMI, endometriosis, and sperm abnormality have p value>0.05 meaning it is insignificant to the outcome of IVF. This research concluded that sperm abnormality, maternal BMI, and endometriosis do not affect the outcome of IVF. There was no data about maternal smoking habits. Whilst maternal age affects the outcome of IVF. Conclusion: This research concluded that sperm abnormality, maternal BMI, and endometriosis do not affect the outcome of IVF. There was no data about maternal smoking habits. Whilst maternal age affects the outcome of IVF.
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Kleymann, Gerald, und Hans-Otto Werling. „A Generally Applicable, High-Throughput Screening–Compatible Assay to Identify, Evaluate, and Optimize Antimicrobial Agents for Drug Therapy“. Journal of Biomolecular Screening 9, Nr. 7 (Oktober 2004): 578–87. http://dx.doi.org/10.1177/1087057104265291.
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Efficacy and tolerability are the key criteria for a successful medication in the clinic. Therefore, a new test method to obtain selective and active lead molecules has been developed. Recently, this novel screening strategy enabled a breakthrough in drug discovery in the field of herpes viruses. Here the authors report that this assay is a generally applicable screening test, which allows not only for identifying tolerable and potent antimicrobial agents in compound libraries, but also covers all potential in vitro targets of both the pathogen and the host simultaneously. The test system mimics the smallest unit of a natural infection. Host cells are incubated in the presence of the test sample and are infected with microbes, such as viruses, bacteria, or fungi. Analogous to (lethal challenge) animal models, cell survival is determined. This assay maximizes the chances of success of anti-infective drug discovery, is sensitive, robust, time- and cost-efficient, and especially effective in optimizing screening hits to lead structures and development candidates. In addition to the minimal inhibitory concentration or dose, this test system simultaneously provides the selectivity index, a measure of tolerability in vitro. The authors propose the activity selectivity assay format as a new standard in anti-infective drug discovery and clinical development.
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Kim, Kwang Mahn, Sang Bae Lee, Se Ho Lee, Yong Keun Lee und Kyoung Nam Kim. „Comparison of Validity between WST-1 and MTT Test in Bioceramic Materials“. Key Engineering Materials 284-286 (April 2005): 585–88. http://dx.doi.org/10.4028/www.scientific.net/kem.284-286.585.
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Cytotoxicity test was essential for the pre-clinical evaluation of bioceramics. Proliferation assays such as MTT, XTT and WST-1 were commonly used for measuring biocompatibility. WST-1 was more convenient than MTT because of its water-solubility and storage condition. The calcium phosphate glass and β-TCP have been used for bone substitute, and some magnetic ferrites have been used for hyperthermic treatment. L929, mouse fibroblast cell, was the representative cell-line for in vitro biocompatibility test. The extracts of test samples were prepared by ISO10993-12:2002. The biocompatibilities of the extracts were measured by MTT and WST-1 assay and their pH were measured with pH meter. The cellular survival rate of CPG was the lowest and the results of the WST-1 test showed results similar to those of the MTT test. Thus, proliferation assays using WST-1 may be conveniently and routinely applicable to pre-clinical evaluation of bioceramics.
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Govindasamy, Niraimathi, Binyamin Duethorn, Hatice O. Oezgueldez, Yung S. Kim und Ivan Bedzhov. „Test-tube embryos - mouse and human development in vitro to blastocyst stage and beyond“. International Journal of Developmental Biology 63, Nr. 3-4-5 (2019): 203–15. http://dx.doi.org/10.1387/ijdb.180379ib.
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Mammalian embryogenesis is intrauterine and depends on support from the maternal environment. Therefore, in order to directly study and manipulate early mouse and human embryos, fine-tuned culture conditions have to be provided to maintain embryo growth in vitro. Over time, the establishment and implementation of embryo culture methods have come a long way, initially enabling the development of few pre-implantation stages, expanding later to support in vitro embryogenesis from fertilization until blastocyst and even ex utero development beyond the implantation stages. Designing culture conditions that enable near physiological development of early embryos without maternal input, especially during the peri- and post-implantation stages, requires overcoming numerous experimental challenges, and it is still far from optimal. Nevertheless, embryo culture methods are an essential cornerstone of both assisted reproductive technologies and basic research, and these methods provide a platform to understand life’s greatest miracle – the development of a new organism.
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Ni, Pei, Rong Rong Zhan, Yu Long Wei, Jing Hui und Feng Qing Hu. „Bioactivity Assay of Chitosan Microspheres Containing Pig Thymosin In Vitro“. Advanced Materials Research 830 (Oktober 2013): 455–58. http://dx.doi.org/10.4028/www.scientific.net/amr.830.455.
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Thymosin, composed of many peptides, is distributed in animal and has immunomodulating effects. However, many factors could affect its bioactivity. Chitosan microspheres containing pig thymosin (CMPT) were prepared through W/O emulsion cross-linking method. In this study, thymosin activity between pig-thymosin and CMPT was investigated through E-rosette test and mouse spleen lymphocyte proliferation (MPLP) experiment in vitro. Results showed that CMPT has the same immunomodulatory activity as pig thymosin. When pig-thymosin was 1.5 μg / ml, E-rosette binding rate was 63%, and E-rosette binding rate of CMPT reached 60%。MPLP rate was 73.3% and 68.5%, respectively. CMPT have the capacity to keep the E-rosette binding and MPLP activity of pig thymosin. This may help to improve traditional formulation and bioavailability of thymosin, and expand its scope of application in clinic.
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Sakha, Binesh M., Gyan P. Rai, Shambhu P. Dhital und Ram B. Nepal. „Disease-free Pre-Basic Seed Potato Production through Tissue Culture in Nepal“. Nepal Agriculture Research Journal 8 (18.11.2014): 7–13. http://dx.doi.org/10.3126/narj.v8i0.11564.
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Pre-basic seed potatoes are disease free potato minitubers produced by transplanting pathogen free in vitro potato plantlets under protected condition in aphid-proof glasshouse and/or screen house. Double antibody sand witched - enzyme linked immuno-sorbant assay is used to test six major potato viruses, namely PLRV, PVS, PVX, PVY, PVA and PVM. Thermotherapy cum meristem tip excision techniques are used to eliminate these viruses. Virus free in vitro potato plantlets are rapidly propagated by single nodal cuttings on modified MS media. For pre-basic seed production disease free in vitro potato plantlets are transplanted in the sterile sand soil substrate under glasshouse and screen house, once in autumn season and next in spring season. Since 1990, National Potato Research Program has been producing about 200,000 pre-basic seeds annually. So far, PBS of 19 different recommended and released potato cultivars has been produced. Till date 3,465,799 PBS had been produced and 3,217,666 pre-basic seeds distributed to the different seed potato growers groups, District Agriculture Development Offices, government farms/research stations, and NGOs/INGOs for subsequent basic seed potato production. After establishment of tissue culture facilities in National Potato Research Program, the productivity of potato has been increased by 71% due to utilization of pre-basic seed potatoes.Nepal Agric. Res. J. Vol. 8, 2007, pp. 7-13DOI: http://dx.doi.org/10.3126/narj.v8i0.11564
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Affatato, Saverio, Paolo Erani, Maurizio Fersini, Vincenzo Contaldi, Anna Rita Terrizzi und Antonio Licciulli. „Preliminary In Vitro Wear Assessment of Ceramic Cemented Femoral Components Coupled with Polyethylene Menisci“. Materials 14, Nr. 9 (22.04.2021): 2112. http://dx.doi.org/10.3390/ma14092112.
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Success of total knee replacement (TKR) depends on the prosthetic design and materials. The use of metal components is well established with the disadvantage of allergic reactions. Ceramics have been recently proposed because of high wear resistance, excellent biocompatibility, wettability, and suitable mechanical properties. This study was aimed at investigating in vitro wear resistance of Zirconia Toughened Alumina (ZTA)/Ultra-high-molecular-weight polyethylene (UHMWPE) of TKR femoral components. An in vitro protocol was designed with the application of relevant load profile, 6-degrees-of-freedom knee simulator, and 8 × 105 cycles on the ZTA/UHMWPE configuration under bovine calf serum. Before and after wear test, the femoral components were investigated by using the Scanning Electron Microscope (SEM) and the X-Ray Diffraction (XRD) analyses, and stylus surface roughness measurements. The proposed pre-clinical test yielded repeatable results. In particular, gravimetric results showed that, after 8 × 105 cycles, the mean weight loss of the polyethylene mobile components is 5.3 ± 1.1 mg. The surface roughness measurements (Ramax) performed after the wear test showed no significant variation on the UHMWPE menisci. A slight increase of roughness has been found on the ZTA (0.02 µm before wear test, 0.28 µm after the test). SEM observations did not show significant modification of the surface morphology. Tetragonal to monoclinic phase ratio was measured by XRD before and after wear test to evaluate stability of tetragonal ZrO2 phase. Minimal conversion of tetragonal to monoclinic phase was found from 5.4 to 8%. Although this study is a preliminary evaluation limited to in vitro tests, it provides novel pre-clinical indications about the potential of ceramic TKR femoral components.
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Zhu, Ruixin, Zhihong Fan, Yue Han, Shuang Li, Guojing Li, Linlin Wang, Ting Ye und Wenqi Zhao. „Acute Effects of Three Cooked Non-Cereal Starchy Foods on Postprandial Glycemic Responses and in Vitro Carbohydrate Digestion in Comparison with Whole Grains: A Randomized Trial“. Nutrients 11, Nr. 3 (15.03.2019): 634. http://dx.doi.org/10.3390/nu11030634.
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Plant origin, processing, and domestic preparation may affect the postprandial glycemic response (PGR) of starchy foods. The objective of this study was to examine the possibility of integrating domestically cooked non-cereal starchy foods commonly consumed in Northeast Asia into glycemic management diet, and compare their glycemic characteristics with those of waxy and non-waxy whole grains and starchy beans. In a randomized crossover trial, ten healthy subjects consumed dried lily bulb (LB), lotus seed (LS), adlay (AD), waxy black rice (BR), millet (MI), and adzuki bean (AB), pre-soaked and each cooked for two time durations. Acute PGR tests and in vitro carbohydrate digestion were carried out for each test food. Both the LS and AB meals achieved low glycemic index (GI 21–51), while the other starchy foods failed to show significant difference with rice (GI 83–109). The hydrolysis indexes of LS and AB were 37.7%–61.1%, significantly lower than other test foods. The in vitro tests indicated that pre-soaking resulted in high rapidly digestible starch (RDS) and low resistant starch (RS). Careful choice of whole grain materials, minimized pre-soaking, and moderate cooking may be critical factors for successful postprandial glycemic management for diabetic and pre-diabetic.
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Xiao, Yonghao, Hui Zhao, Xiaoyu Ma, Zongheng Gu, Xin Wu, Liang Zhao, Lin Ye und Zengguo Feng. „Hydrogel Dressing Containing Basic Fibroblast Growth Factor Accelerating Chronic Wound Healing in Aged Mouse Model“. Molecules 27, Nr. 19 (26.09.2022): 6361. http://dx.doi.org/10.3390/molecules27196361.
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Due to the decreasing self-repairing ability, elder people are easier to form chronic wounds and suffer from slow and difficult wound healing. It is desirable to develop a novel wound dressing that can accelerate chronic wound healing in elderly subjects to decrease the pain of patients and save medical resources. In this work, Heparin and basic fibroblast growth factor(bFGF) were dissolved in the mixing solution of 4-arm acrylated polyethylene glycol and dithiothreitol to form hydrogel dressing in vitro at room temperature without any catalysts, which is convenient and easy to handle in clinic application. In vitro re-lease test shows the bFGF could be continuously released for at least 7 days, whereas the dressing surface integrity maintained for 3 days degradation in PBS solution. Three groups of treatments including bFGF-Gel, bFGF-Sol and control without any treatment were applied on the full-thickness wound on the 22 months old mice back. The wound closure rate and histological and immunohistochemical staining all illustrated that bFGF-Gel displayed a better wound healing effect than the other two groups. Thus, as-prepared hydrogel dressing seems supe-rior to current clinical treatment and more effective in elderly subjects, which shows promising potential to be applied in the clinic.
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Thanusha, A. V., und Veena Koul. „Biocompatibility evaluation for the developed hydrogel wound dressing – ISO-10993-11 standards – in vitro and in vivo study“. Biomedical Physics & Engineering Express 8, Nr. 1 (30.11.2021): 015010. http://dx.doi.org/10.1088/2057-1976/ac3b2b.
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Abstract Assessment of biocompatibility for the developed wound dressing plays a significant role in translational studies. In the present research work, a wound dressing has been developed using gelatin, hyaluronic acid and chondroitin sulfate using EDC as crosslinker in a specific manner. The characterized hydrogel wound dressing was evaluated for its biocompatibility studies by means of ISO-10993-11 medical device rules and standards. Various parameters like skin sensitization test, acute systemic toxic test, implantation study, intracutaneous reactivity test, in vitro cytotoxicity test and bacterial reverse mutation test, were evaluated and the results demonstrated its safety for the pre-clinical investigation.
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Monteiro, C. A. S., G. R. Leal, H. F. R. A. Saraiva, A. J. R. Camargo, P. M. S. Rosa, A. L. R. Rodrigues, R. V. Serapião et al. „36 EFFECT OF cAMP MODULATORS DURING OOCYTE IN VITRO MATURATION ON GAP JUNCTIONAL ACTIVITY OF VITRIFIED BOVINE OOCYTES“. Reproduction, Fertility and Development 28, Nr. 2 (2016): 148. http://dx.doi.org/10.1071/rdv28n2ab36.
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Oocyte cryopreservation is a strategic tool for in vitro embryo production, but low rates of cryosurvival are reported for bovine oocytes. Simulated physiological oocyte maturation system (Albuz et al. 2010 Hum. Reprod. 25, 12) uses cAMP modulators to increase oocyte competence by the extension of meiosis block and gap junctional communications activity. The aim of this study was to investigate the effect of simulated physiological oocyte maturation system on gap junctional activity of vitrified bovine oocytes. Oocytes from slaughterhouse ovaries were divided into 4 groups: C (control: fresh immature oocytes); V (vitrified immature oocytes); PM-V (vitrified oocytes after a 2-h pre-in vitro maturation phase – in the presence of AMPc modulators, 100 μM Forskolin, and 500 μM IBMX); and PM (fresh immature oocytes subjected to pre-in vitro maturation). Viable oocytes (n = 404 obtained from 4 replicates) were stained with Calcein-AM using the protocol of Thomas et al. (2004 Biol. Reprod. 71(4), 1142–1149) in order to measure gap junctions activity. Images were captured in fluorescence microscope, and fluorescence intensity was analysed with ImageJ software. Mean fluorescence intensity of each group was normalized to control group to obtain relative intensity value. Means were compared by Kruskal-Wallis test and Dunn post-test. A second analysis was performed considering the percentage of each staining pattern (low, middle, and high intensity) for each group. Results were analysed using Fisher exact test. All statistical analysis were performed in GraphPad Instat program with 5% significance level. Results demonstrated that all treatments induced an increase (P < 0.05) in fluorescence intensity (V: 1.76 ± 1.13; PM-V: 1.58 ± 0.98; PM: 1.38 ± 0.94) compared with control (C: 1.00 ± 0.48). Regarding the staining patterns analyses, immature vitrified oocytes (V group) differed from control group in middle and low patterns (G1, calibrator – high: 11.2%ab, middle: 43.8%a, low: 44.9%a; G2 – high: 8.2%ab, middle: 63.9%b, low: 27.9%b; G3 – high: 16.3%a, middle: 42.3%a, low: 41.3%a; G4 – high: 6.7%b, middle: 53.9%ab, low: 39.3a). In conclusion, unexpectedly, vitrification also increased gap junctional activity, as was found for pre-in vitro maturation group. However, staining pattern analysis results showed only vitrified group was different from control, suggesting vitrified and pre-in vitro maturation groups could have gap activity affected by different ways. This research was supported by FAPERJ (E26/111.61/2013) and CAPES.
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Kim, Ji-Soo, Myung-Hwan Jeong, Heung-Sik Seo, Myeong-Kyu Park, Hee Ju Park und Seong-Soon Nah. „In vitro and in vivo evaluation of the genotoxicity of titanium dioxide, GST“. Environmental Analysis Health and Toxicology 38, Nr. 2 (03.05.2023): e2023008. http://dx.doi.org/10.5620/eaht.2023008.
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Titanium dioxide (TiO<sub>2</sub>) was used in various applications in a wide range of products including food, cosmetics and photocatalyst. General toxicity studies of titanium dioxide, GST (Green Sludge Titanium) have been investigated in several reports, whereas studies concerning mutagenicity and genotoxicity have not been elucidated. Herein, we investigated the potential mutagenicity and genotoxicity of GST by genetic toxicology testing. The bacterial reverse mutation test was conducted by the pre-incubation method in the presence and absence of metabolic activation system (S9 mixture). The chromosome aberration test was performed using cultured Chinese hamster lung cell line in the absence and presence of S9 mixture. The micronucleus test was performed by using specific pathogen-free male ICR mice. Genotoxicity tests were conducted following the test guidelines of the Organisation for Economic Cooperation and Development with application of Good Laboratory Practice. No statistically significant increases were found in the bacterial reverse mutation test, in vitro chromosome aberration test, and in vivo micronucleus test when tested for induction of genotoxicity in GST. These results suggest that GST did not induce mutagenicity and genotoxicity in both in vitro and in vivo system.
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Tenorio, Maria, Gemma Moreno Jiménez, Valentín García Gutiérrez, Ana Jiménez, Maria Jesús Blanchard, Ana Vallés, Sandra López, Pilar Herrera, Javier Lopez Jimenez und Anabelle Chinea. „Validation of Daraex to Resolve Daratumumab-Induced Interferences in Pre-Transfusion Screen Tests“. Blood 134, Supplement_1 (13.11.2019): 4983. http://dx.doi.org/10.1182/blood-2019-131345.
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Daratumumab is a CD38-directed antibody increasingly used for the treatment of adult patients with multiple mieloma. The membrane of red blood cells express CD38 and thus samples from patients treated with daratumumab show agglutination in red blood cell antibody screen tests performed prior to transfusion. This interference hinders the detection of red blood cell alloantibodies. Published literature has described a method to eliminate CD38 in red blood cells with DTT (Chapuy, 2016). This technique is cumbersome, requires positive and negative controls as DTT destroys Kell antigens and can produce in vitro hemolysis. The increasing number of multiple myeloma patients treated with daratumumab poses the need for a simple and straightforward technique with applicability in standard transfusion centers. DaraEx (Inno-Train) is a new anti-CD38 neutralizing agent that overcomes daratumumab-induced interferences detected in pre-transfusion tests without the major drawbacks associated with the DTT technique. Our aim was to validate and implement DaraEx as the method of choice to solve daratumumab interferences detected in pre-transfusion screen tests in a tertiary care center. A two-step approach using in vitro and in vivo samples was designed to validate the new method. First, we compared DaraEx efficacy in vitro to the reference DTT method in two samples spiked with daratumumab to achieve a concentration of 10mg/mL (Sample A: serum from a patient without known red blood cell alloantibodies; Sample B: serum from a patient with alloantibody anti-c). Red blood cells in the screen test (3 red blood cell screen; ID-DiaCell I-II-III) as well as positive (E+ red blood cells) and negative controls (K+ red blood cells) were treated with DTT 0.2M solution for 30 minutes at 37ºC and then washed four times with saline. In parallel, red blood cells in the screen test were incubated during 30 minutes at room temperature in a shaker (600rpm) with DaraEx. Red blood cells treated with each of these methods were used for indirect antiglobulin test with our gel card system (BioRad; IH-1000). Preference of method in terms of time needed and result interpretation was evaluated by three hematologists specialized in blood banking and four different technicians. Secondly, we tested pre-transfusion samples from patients treated with daratumumab with the DaraEx technique to check in vivo efficacy. There was a 100% concordance between both techniques (DDT reference method and DaraEx new method) in both in vitro samples. All hematologists and technicians found the DaraEx technique less cumbersome in terms of processing and time to result (2 hours with DTT versus 1 hour with DaraEx) and the interpretation straightforward. Twelve samples with daratumumab-induced interference in pre-transfusion screen tests belonging to 5 patients were tested between January and July 2019. All the interferences detected resolved with DaraEx regardless of time from last daratumumab administration (range: 7-145 days; mean: 57 days). Figure 1 shows screen test with and without treatment with DaraEx in a patient sample. In our experience, DaraEx technique is a simple, fast and efficacious method, regardless of time from last daratumumab administration, to resolve interferences secondary to daratumumab administration without the major disadvantages associated with DTT. Figure 1 Disclosures García Gutiérrez: Pfizer: Honoraria, Research Funding; Incyte: Honoraria, Research Funding; Novartis: Honoraria, Research Funding; BMS: Honoraria, Research Funding.
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Mussa, O. R. P. A., A. Kurnianto und I. P. Hermawan. „Detection of Toxocara cati from Fecal Samples of Domestic Pet Cats at Pet Clinic Surabaya and Durability of Toxocara cati Eggs with In Vitro Media“. Jurnal Sain Peternakan Indonesia 16, Nr. 4 (11.12.2021): 297–300. http://dx.doi.org/10.31186/jspi.id.16.4.297-300.
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The purpose of this study was to determine the presence of Toxocara cati eggs in the feces of Domestic pet cats at the Surabaya Animal Clinic and the durability of Toxocara cati eggs with in vitro media. Total fecal samples taken were 30 cat feces from 5 animal clinics in Surabaya, then detected Toxocara cati eggs using an enlargement microscope 100 times, followed by counting eggs per gram of feces. The eggs used were approximately 200 eggs in each petri dish, then positive feces of Toxocara cati with various in vitro media using 0.9% NaCl, 1% PBS and 1% CMC for 24 hours and 48 hours. The results showed that two fecal samples positive Toxocara cati from 30 samples fecal and durability Toxocara cati eggs on medium in vitro over 24 showed 28% NaCl; PBS 68%; CMC 5% and at 48 hours showed 23.7% NaCl; PBS 58%; CMC 17.3%. Based on these results, the conclusion is two fecal (6,67 %) positive T.cati from 30 fecal samples, and the best medium in the test of the durability of Toxocara cati egg is 1% PBS.
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Lima, Keila de Oliveira, Aline Valério de Lima, Darlan Augusto da Costa Rocha, Suely Carlos Ferreira Sampaio, Paola Cappellano und Jorge Luiz Mello Sampaio. „A simple disk pre-diffusion test to predict in vitro aztreonam/avibactam activity against NDM-producing Klebsiella pneumoniae complex“. Journal of Global Antimicrobial Resistance 28 (März 2022): 49–52. http://dx.doi.org/10.1016/j.jgar.2021.12.009.
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Mutiarawati, Diah Titik. „In Vitro Anthelmintic Activity of Acalypha Indica Leaves Extracts“. Health Notions 4, Nr. 3 (31.03.2020): 94–99. http://dx.doi.org/10.33846/hn40305.
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Anting-anting leaves (Acalypha indica L) is one of the wild plants (weeds) that are often used by the community to treat helminthic diseases such as ascariasis, containing several potentially anthelmintic compounds, namely saponins, tannins and flavonoids. The purpose of this study was to determine the effect of giving Anting-anting leaf extract (Acalypha indica L) as an anthelmintic to the time of death of pork roundworms (Ascaris suum Goeze). The method in this study was pre-experimental with a post test only group design. The subjects of the study were Ascaris suum. The study was conducted at the Parasitology Laboratory of the Surabaya Polytechnic Health Medical Laboratory Technologist Department in December-May 2018. This study used 5 treatment groups, 0.9% NaCl as negative control and 0.25% pirantel pamoate as positive control and 60%, 80% and 100%. Data were analyzed using the Kolmogrov-Smirnov test, then the Kruskal-Wallis test was then continued using a Post-Hoc test to determine the difference in anthelmintic power of ear leaf extract of each concentration with positive control over the time of worm death. The average time of Ascaris suum worm death at a concentration of a concentration of 60% for 228.5 minutes, a concentration of 80% for 174 minutes, and a concentration of 100% for 92 minutes. So it can be concluded that the leaf extract of Acalypha L has an anthelmintic effect on the Ascaris suum worm.
Keywords: anthelmintic; Ascaris suum; anting-anting leaf
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KU, In-Hoe. „Ethical Problems of Pre-implantation Diagnosis“. Korean Journal of Medical Ethics 6, Nr. 2 (Dezember 2003): 21–35. http://dx.doi.org/10.35301/ksme.2003.6.2.21.
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The ability to diagnose genetic diseases early in pregnancy has already demonstrated the power of remarkable forms of technology and stimulated innumerable ethical debates. Now the diagnosis of genetic disease even before implantation provides new possibilities and adds to the ethical problems we face. There is a considerable difference between the wish to have a child and the desire to have a perfect child. As long as a pre-implantation diagnosis continues to require testing after embryo-transfer by an additional prenatal diagnosis test in order to ensure its results, at this stage of technology, there is no ethically sufficient reason to utilize pre-implantation diagnosis. At a future date, this problem must be discussed in greater detail. In this discussion, there is one perspective that must not be lost from sight: after prenatal diagnosis, at least theoretically, there is the possibility that a couple will decide to carry a genetically abnormal fetus to full-term. In the context of pre-implantation diagnosis and in vitro fertilization the decision making process for couples is eliminated; the genetically abnormal embryo will not be implanted. The question of the moral status of human life in its early stages - of its biological, social and personal attributes continue to be philosophically controversial and in need of exploration.
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Elgendy, Ali Y., Aya A. Salama und Ahmed N. Elsherbini. „Effect of Obturation Technique on Stresses Induced in Telescopic Retained Partial Denture (In vitro Study)“. Open Access Macedonian Journal of Medical Sciences 11, Nr. D (07.04.2023): 73–77. http://dx.doi.org/10.3889/oamjms.2023.11603.
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AIM: The purpose of this study was to evaluate the effect of obturation techniques on the stresses induced on the abutment teeth and the supported prosthesis. MATERIALS AND METHODS: A unilateral bounded edentulous space case design was chosen for this study, with telescopic retained partial dentures (TRPDs) selected as the treatment of choice. Two TRPDs were fabricated, the first pre-molar and second molar were used as abutments for the partial dentures. In the first group, the pre-molar was treated with the lateral compaction obturation technique. In the second group, the first pre-molar was treated with a vertical compaction obturation technique. The second molar was a common abutment in both groups and was left untreated. 50 Newtons were applied on the prostheses, with a strain gauge attached, and data were collected. RESULTS: t-test showed the mean values of microstrain (με) induced in pre-molars treated with lateral compaction was 159.64 ± 10.46 με, whereas with vertical compaction was 12.14 ± 2.57 με, with statistical significance of p < 0.05. CONCLUSION: Vertical obturation of endodontically treated teeth propagates less stress to the abutment teeth and to the supporting prostheses.
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Ulloa, S. M. B., J. Heinzmann, D. Herrmann, U. Baulain, K. G. Hadeler, P. Aldag, A. Lucas-Hahn und H. Niemann. „197 EFFECTS OF PRE-IN VITRO MATURATION WITH CAFFEINE ON BOVINE OOCYTE DEVELOPMENTAL CAPACITY“. Reproduction, Fertility and Development 28, Nr. 2 (2016): 229. http://dx.doi.org/10.1071/rdv28n2ab197.
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High cyclic adenosine monophosphate (cAMP) concentrations are critical for maintaining oocyte meiotic arrest in vivo. For in vitro maturation (IVM), the oocyte is released mechanically from the follicle, which induces a significant drop in intra-oocyte cAMP levels, triggering non-physiological meiotic resumption. It has been proposed that modulation of cAMP before IVM can increase bovine blastocyst rates in vitro. Caffeine is a nonspecific competitive phosphodiesterases (PDE) inhibitor and can inhibit meiotic resumption of oocytes due to maintenance of cAMP levels. It has been reported that gamete treatment with caffeine can increase developmental potential. The current study evaluated the effects of pre-in vitro maturation culture with different concentrations of caffeine on meiotic progress, developmental rates and blastocyst cell numbers. Bovine ovaries were collected from a local abattoir. A total of 6648 cumulus-oocyte complexes were obtained by slicing. Caffeine was used in 5 different concentrations (1, 5, 10, 20, and 30 mM) during slicing, searching, and 2 h pre-IVM culture. A control group, with 2 h pre-IVM without caffeine (0 mM) and a standard control were also included. Oocytes were washed either after standard or pre-IVM treatments and cultured for 24 h in vitro without caffeine. After IVM, oocytes were fertilised in vitro for 19 h, and zygotes were cultured in vitro for 8 days until the blastocyst stage. Subsets of oocytes were fixed in 2% glutaraldehyde at 9, 20, and 24 h after IVM. Hoechst staining was performed to evaluate nuclear status of matured oocytes. Cleavage and blastocyst formation rates were evaluated at Days 3 and 8 after IVF. Expanded blastocysts from all treatments were submitted to differential staining. One-way ANOVA from R software was applied to evaluate differences in cleavage and blastocysts rates and blastocyst cell numbers. Fisher’s exact test complemented by Bonferroni correction was used to determine meiotic progress. Caffeine maintained oocytes in meiotic arrest after 9 h of IVM in a concentration-dependent manner (germinal vesicle: 79.0%, 92.2%, 66.7%, 55.1%, 56.9%, 43.9%, 30.2%, respectively, for 30, 20, 10, 5, 1, 0 mM and standard; P < 0.016). Cleavage rates were similar in all treatments; however 30 mM caffeine decreased blastocyst rates (Table 1; P < 0.05). The number of cells did not differ significantly among in vitro treatments (Table 1; P > 0.05). Developmental competence was not affected by 2 h pre-IVM culture. Caffeine supplementation before IVM delayed resumption of meiosis and affected embryo development. Table 1.In vitro developmental competence of oocytes treated with different caffeine concentrations before IVM
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Babják, Michal, Alžbeta Königová, Michaela Urda Dolinská, Tomas Kupčinskas, Jaroslav Vadlejch, Georg von Samson-Himmelstjerna, Saulius Petkevičius und Marián Várady. „Does the in vitro egg hatch test predict the failure of benzimidazole treatment in Haemonchus contortus?“ Parasite 28 (2021): 62. http://dx.doi.org/10.1051/parasite/2021059.
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Considerable research has been directed towards optimising in vitro tests that can diagnose resistance in pre-parasitic stages of parasites. The objective of this study was to compare the in vivo faecal egg count reduction test (FECRT), the in vitro egg hatch test (EHT), and the molecular determination of the frequency of a codon 200 allele of β-tubulin isotype 1 associated with benzimidazole resistance in larval stages of Haemonchus contortus obtained from infected goats. Animals were infected with composite infective doses representing 10, 20, 30, 40, 60, and 80% resistant alleles. Faecal samples for the EHT were collected on 28, 33, and 35 days post-infection. The results of the in vivo FECRT indicated that albendazole treatment reduced infections consisting of composite doses of 10, 20, 30, 40, 60, and 80% larvae of the resistant isolate by 91.3, 78.0, 63.3, 48.4, 36.5, and 41.4%, respectively. The drug concentration at which 50% of the eggs were prevented from developing hatching larvae (ED50) in the in vitro EHT varied from 0.09 ± 0.01 to 15.63 ± 12.10 μg/mL thiabendazole. The results of the in vitro EHT indicated that the test could estimate in vivo resistance well. The EHT could thus accurately estimate the in vivo efficacy of the drug and percentage of the resistance allele in the population using hatching parameters in delineation doses. This finding was also supported by comparing the FECRT data to the hatching percentages in the EHT on 30 goat farms in Slovakia with natural mixed infections of gastrointestinal parasites.
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Chen, Yi-Wen, Tao Chiang, I.-Hui Chen, Da-Yo Yuh, Hsiu-Yang Tseng, Chuang-Wei Wang und Hsin-Han Hou. „Titanium Surfaces with a Laser-Produced Microchannel Structure Enhance Pre-Osteoblast Proliferation, Maturation, and Extracellular Mineralization In Vitro“. International Journal of Molecular Sciences 25, Nr. 6 (16.03.2024): 3388. http://dx.doi.org/10.3390/ijms25063388.
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The clinical success of dental titanium implants is profoundly linked to implant stability and osseointegration, which comprises pre-osteoblast proliferation, osteogenic differentiation, and extracellular mineralization. Because of the bio-inert nature of titanium, surface processing using subtractive or additive methods enhances osseointegration ability but limits the benefit due to accompanying surface contamination. By contrast, laser processing methods increase the roughness of the implant surface without contamination. However, the effects of laser-mediated distinct surface structures on the osteointegration level of osteoblasts are controversial. The role of a titanium surface with a laser-mediated microchannel structure in pre-osteoblast maturation remains unclear. This study aimed to elucidate the effect of laser-produced microchannels on pre-osteoblast maturation. Pre-osteoblast human embryonic palatal mesenchymal cells were seeded on a titanium plate treated with grinding (G), sandblasting with large grit and acid etching (SLA), or laser irradiation (L) for 3–18 days. The proliferation and morphology of pre-osteoblasts were evaluated using a Trypan Blue dye exclusion test and fluorescence microscopy. The mRNA expression, protein expression, and protein secretion of osteogenic differentiation markers in pre-osteoblasts were evaluated using reverse transcriptase quantitative polymerase chain reaction, a Western blot assay, and a multiplex assay, respectively. The extracellular calcium precipitation of pre-osteoblast was measured using Alizarin red S staining. Compared to G- and SLA-treated titanium surfaces, the laser-produced microchannel surfaces enhanced pre-osteoblast proliferation, the expression/secretion of osteogenic differentiation markers, and extracellular calcium precipitation. Laser-treated titanium implants may enhance the pre-osteoblast maturation process and provide extra benefits in clinical application.
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Budai, Péter, József Lehel, Judit Tavaszi und Éva Kormos. „HET-CAM test for determining the possible eye irritancy of pesticides“. Acta Veterinaria Hungarica 58, Nr. 3 (01.09.2010): 369–77. http://dx.doi.org/10.1556/avet.58.2010.3.9.
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Agrochemicals and veterinary products must undergo numerous toxicological tests before registration. The use of animals in these studies is a controversial issue. The Draize eye irritation test is one of the most criticised methods because of the injuries inflicted on the test animals. Several in vitro methods have been used to investigate the toxicity of potential eye irritants with a view to replacing in vivo eye irritation testing. One of these alternative methods is the Hen’s Egg Test — Chorioallantoic Membrane (HET-CAM) test. In the present studies comparative screening was performed with a set of agrochemicals to establish parallel data on in vitro (HET-CAM) and in vivo (Draize) results. The examined materials were: Totril (ioxynil), Omite 57 E (propargit), Actellic 50 EC (pyrimiphos-methyl), Stomp 330 EC (pendimethalin), Mospilan 3 EC (acetamiprid), Alirox 80 EC (EPTC), Match 050 EC (lufenuron), Nonit (dioctyl sulphosuccinate sodium), Perenal (haloxyfop-R methyl ester), Pyrinex 48 EC (chlorpyrifos). These experiments showed good correlation between results obtained by the HET-CAM test and those of the Draize rabbit eye test in most cases. The present form of the HET-CAM test can be proposed as a pre-screening method for the determination of eye irritative potential, therefore the number of test animals can be reduced and/or experimental animals can be replaced.
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Shipunova, Victoria O., Elena N. Komedchikova, Polina A. Kotelnikova, Maxim P. Nikitin und Sergey M. Deyev. „Targeted Two-Step Delivery of Oncotheranostic Nano-PLGA for HER2-Positive Tumor Imaging and Therapy In Vivo: Improved Effectiveness Compared to One-Step Strategy“. Pharmaceutics 15, Nr. 3 (03.03.2023): 833. http://dx.doi.org/10.3390/pharmaceutics15030833.
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Therapy for aggressive metastatic breast cancer remains a great challenge for modern biomedicine. Biocompatible polymer nanoparticles have been successfully used in clinic and are seen as a potential solution. Specifically, researchers are exploring the development of chemotherapeutic nanoagents targeting the membrane-associated receptors of cancer cells, such as HER2. However, there are no targeting nanomedications that have been approved for human cancer therapy. Novel strategies are being developed to alter the architecture of agents and optimize their systemic administration. Here, we describe a combination of these approaches, namely, the design of a targeted polymer nanocarrier and a method for its systemic delivery to the tumor site. Namely, PLGA nanocapsules loaded with a diagnostic dye, Nile Blue, and a chemotherapeutic compound, doxorubicin, are used for two-step targeted delivery using the concept of tumor pre-targeting through the barnase/barstar protein “bacterial superglue”. The first pre-targeting component consists of an anti-HER2 scaffold protein, DARPin9_29 fused with barstar, Bs-DARPin9_29, and the second component comprises chemotherapeutic PLGA nanocapsules conjugated to barnase, PLGA-Bn. The efficacy of this system was evaluated in vivo. To this aim, we developed an immunocompetent BALB/c mouse tumor model with a stable expression of human HER2 oncomarkers to test the potential of two-step delivery of oncotheranostic nano-PLGA. In vitro and ex vivo studies confirmed HER2 receptor stable expression in the tumor, making it a feasible tool for HER2-targeted drug evaluation. We demonstrated that two-step delivery was more effective than one-step delivery for both imaging and tumor therapy: two-step delivery had higher imaging capabilities than one-step and a tumor growth inhibition of 94.9% in comparison to 68.4% for the one-step strategy. The barnase*barstar protein pair has been proven to possess excellent biocompatibility, as evidenced by the successful completion of biosafety tests assessing immunogenicity and hemotoxicity. This renders the protein pair a highly versatile tool for pre-targeting tumors with various molecular profiles, thereby enabling the development of personalized medicine.
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Ulva, Prima, Idral Purnakarya und Arymbi Pudjiastuty. „EFFECT OF ENERGY DRINK ON MICROHARDNESS OF DENTAL ENAMEL (IN VITRO)“. Andalas Dental Journal 6, Nr. 1 (10.07.2020): 32–41. http://dx.doi.org/10.25077/adj.v6i1.88.
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A prolonged acidic ambience of oral environment will lead to demineralization process. It caused erosion to enamel surface after long time of exposure. Acidic beverages become the external factor of dental erosion. Energy drink is one of popular beverages which has acid ingredients and pH value below the critical pH (5.5). The objective is to determine the effect of energy drink on microhardness of dental enamel (in vitro). Methods that were used in this study was experimental pre test - post test with control group design. A total of 36 premolar were used as samples which were divided into energy drink group and control group of artificial saliva. The treatment group were immersed for 8 cycle for 7 days. Dental enamel microhardness were measured by Vickers Hardness Tester before and after treatment. The result of this study was microhardness mean of enamel surface decreased from 306 VHN to 262 VHN after immersion for energy drink group and no significant difference in artificial saliva group. Independent T-test analysis showed p value <0,005, thus concluding there was significant difference between two groups. The conclusion of this study there was significant effect of energy drink immersion to dental enamel microhardness.
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Sensi, Francesca, Edoardo D’Angelo, Martina Piccoli, Piero Pavan, Francesca Mastrotto, Paolo Caliceti, Andrea Biccari et al. „Recellularized Colorectal Cancer Patient-Derived Scaffolds as In Vitro Pre-Clinical 3D Model for Drug Screening“. Cancers 12, Nr. 3 (13.03.2020): 681. http://dx.doi.org/10.3390/cancers12030681.
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Colorectal cancer (CRC) shows highly ineffective therapeutic management. An urgent unmet need is the random assignment to adjuvant chemotherapy of high-risk stage II and stage III CRC patients without any predictive factor of efficacy. In the field of drug discovery, a critical step is the preclinical evaluation of drug cytotoxicity, efficacy, and efficiency. We proposed a patient-derived 3D preclinical model for drug evaluation that could mimic in vitro the patient’s disease. Surgically resected CRC tissue and adjacent healthy colon mucosa were decellularized by a detergent-enzymatic treatment. Scaffolds were recellularized with HT29 and HCT116 cells. Qualitative and quantitative characterization of matched recellularized samples were evaluated through histology, immunofluorescences, scanning electron microscopy, and DNA amount quantification. A chemosensitivity test was performed using an increasing concentration of 5-fluorouracil (5FU). In vivo studies were carried out using zebrafish (Danio rerio) animal model. Permeability test and drug absorption were also determined. The decellularization protocol allowed the preservation of the original structure and ultrastructure. Five days after recellularization with HT29 and HCT116 cell lines, the 3D CRC model exhibited reduced sensitivity to 5FU treatments compared with conventional 2D cultures. Calculated the half maximal inhibitory concentration (IC50) for HT29 treated with 5FU resulted in 11.5 µM in 3D and 1.3 µM in 2D, and for HCT116, 9.87 µM in 3D and 1.7 µM in 2D. In xenograft experiments, HT29 extravasation was detected after 4 days post-injection, and we obtained a 5FU IC50 fully comparable to that observed in the 3D CRC model. Using confocal microscopy, we demonstrated that the drug diffused through the repopulated 3D CRC scaffolds and co-localized with the cell nuclei. The bioengineered CRC 3D model could be a reliable preclinical patient-specific platform to bridge the gap between in vitro and in vivo drug testing assays and provide effective cancer treatment.
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Devi Thamizhanban, Gampa Tulja Rani und Kathiresan krishnasamy. „Development of bio-relevant dissolution method for prognosis of In-vivo performance of Dipyridamole from modified release capsules“. International Journal of Research in Pharmaceutical Sciences 11, Nr. 2 (07.05.2020): 2340–49. http://dx.doi.org/10.26452/ijrps.v11i2.2211.
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In-vitro biorelevant dissolution test method was developed for Dipyridamole in modified release multi-particulate dosage form, to simulate the product drug release after oral administration to human. The Dipyridamole concentration in blood plasma achieved after oral administration under pre-prandial (fasting) condition were used for deriving the target dissolution profile deconvoluting the plasma concentration using numerical deconvolution technique. The fraction of drug absorbed was considered as the target dissolution profile. The drug product was tested by using the dissolution method recommended by office of generic drugs, and the dissolution results observed are not comparable with the target dissolution profile. Dissolution method was developed using reciprocating cylinder, Bio-Dis (apparatus -3 as per USP), by simulating the pre-prandial conditions. A full factorial design of experiment was carried out to achieve the target dissolution profile. Media volume and dips per minute (DPM) are considered as main factors, in design of experiment. The dissolution results achieved with media volume 250ml and 10DPM were found to be comparable with target dissolution profile and observed with the F2 value (similarity factor) of 92%. The developed dissolution method demonstrates a very good in-vitro/in-vivo correlation under pre-prandial condition, and shall be used as a predictive in-vitro tool for evaluation Dipyridamole extended release capsules.
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Jiang, Su-zhen, Jia-jia Zheng, Xiang-Mei Chen, Ting Zhang, Qiang Xu, Hui Zhuang und Fengmin Lu. „Original Article. Hepatitis B Virus S Promoter Deletion in Hepatocellular Carcinoma“. Infection International 1, Nr. 1 (01.03.2012): 14–24. http://dx.doi.org/10.1515/ii-2017-0003.
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Abstract Objective To identify the difference and significance of dominant types of hepatitis B virus (HBV) pre-S mutation between liver tumor tissues and paired adjacent non-tumor tissues and to test if the mutations were tumor tissue specific. Methods HBV DNA isolated from 34 paired intratumoral and peritumoral tissues of hepatocellular carcinoma (HCC) patients were screened by PCR and direct sequencing. All patients carried HBV with genotype C, except for one B/C heterozygote. The expression, localization and excretion of LHBs mutant carrying pre-S deletions were characterized in vitro. The expression of endoplasmic reticulum (ER) GRP78 mRNA was assayed. Results Four patterns of pre-S mutations were identified: pre-S1 in-frame deletion involving the first start codon; pre-S2 in-frame deletion; pre-S2 start codon mutation with or without in-frame deletion; and S promoter in-frame deletion (ΔSP). The first two types were evenly found in both tumor and non-tumor tissues. They were rarely present as dominant strains. The last two types were frequently found in the dominant strains in tumor tissues. The overall prevalence of HBV carrying ΔSP was 17.64% (6/34) in tumor tissues, but none were dominant in nontumor tissues. HBV carrying ΔSP was unable to produce S protein in vitro. Immunocytofluorescence assay showed that the ΔSP LHBs mutant aggregated in the cytoplasm, accumulating mainly in the ER. Transient transfection and expression of ΔSP mutant caused GRP78 up-regulation in vitro. Conclusions HBV S promoter deletion was found dominantly in HCC tumor tissue. The aggregation of mutant large surface proteins in the ER possibly involved in HBV-related HCC.
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Tait, Robert Campbell, Annielle Hung und Roy S. Gardner. „Performance of the LumiraDx Platform INR Test in an Anticoagulation Clinic Point-of-Care Setting Compared With an Established Laboratory Reference Method“. Clinical and Applied Thrombosis/Hemostasis 25 (01.01.2019): 107602961989042. http://dx.doi.org/10.1177/1076029619890423.
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Vitamin K antagonists, such as warfarin, have a narrow therapeutic window; patients on these therapies therefore require regular international normalized ratio (INR) monitoring to maintain optimal dosing. This involves periodic checks and laboratory testing using venepuncture, which are often perceived as a burden. This study aimed to determine the accuracy and precision of the LumiraDx INR Test, a new point-of-care in vitro diagnostic platform, in an anticoagulation clinic setting. In this observational, cross-sectional study, precision of the LumiraDx INR Test was assessed using paired replicate samples (n = 366) and 3 test strip lots. Accuracy was determined by comparing capillary blood INR, ascertained by the LumiraDx INR Test, with venous plasma INR, measured by the laboratory reference instrument, the IL ACL ELITE Pro. Furthermore, INR was assessed across a range of hematocrit (25%-55%). In addition, feedback was collected from health-care professionals via a self-completed questionnaire. This trial was registered at ClinicalTrials.gov (NCT03682419). The precision (% coefficient of variation) of the LumiraDx INR Test was <4 when samples were applied by direct application or via a capillary transfer pipette, as well as between test strip lots. Accuracy of the LumiraDx INR Test, across the INR range of 0.8 to 7.5, was confirmed by a strong correlation of 0.965 (95% confidence interval: 0.959-0.970) when compared with the IL ACL ELITE Pro, which was maintained across the hematocrit range. Feedback from health-care professionals indicated that the instructions given by the system were easy to follow. In conclusion, the strong agreement between the LumiraDx Platform INR point-of-care test and the IL ACL ELITE Pro laboratory reference system, as well as between the different application methods and test lots, indicates that it can provide a rapid, accurate, and reliable INR analysis.
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Corey, Jacquelynne R., Saba Kaiseruddin und Anil Gungor. „Prevalence of Mold-Specific Immunoglobulins in a Midwestern Allergy Practice“. Otolaryngology–Head and Neck Surgery 117, Nr. 5 (November 1997): 516–20. http://dx.doi.org/10.1016/s0194-59989770024-x.
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Mold allergy surveys are an important part of the correct identification and treatment of mold allergies. This study included 100 patients who were referred to a Midwestern allergy clinic for the evaluation of rhinitis, suspected to be of allergic origin. An in vitro screening test for allergen-specific IgE (ImmunoCAP) comprised of 10 allergens, including Candida, Aspergillus, Helminthosporium, and Alternaria, was used. To assess the seasonal distribution of mold allergies, we randomly selected 8 patients out of the 100 from each season during which the clinical contact occurred, and we tested them for 14 varieties of mold. The overall incidence of mold allergy in atopic subjects was 44%. The most common molds were (in descending order of frequency) Alternaria, Helminthosporium, Aspergillus, Candida, and Curvularia. Mold allergy was diagnosed most frequently in the winter; the second highest period was the fall. Population surveys of IgE antibody sensitization by in vitro techniques can provide useful information about fungal allergy.
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Kandarova, Helena, Jamin A. Willoughby, Wim H. De Jong, Silvia Letasiova, Tatiana Milasova, Michael A. Bachelor, Bridget Breyfogle, Yuki Handa, Liset De la Fonteyne und Kelly P. Coleman. „Pre-validation of an in vitro skin irritation test for medical devices using the reconstructed human tissue model EpiDerm™“. Toxicology in Vitro 50 (August 2018): 407–17. http://dx.doi.org/10.1016/j.tiv.2018.02.007.
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Ariantini, Dyah, Mohammad Lutfi und Diah Rumekti Hadiati. „Kadar Hormon LH Basal sebagai Prediktor Kebrhasilan Stimulasi Ovarium pada Program Bayi Tabung“. Jurnal Kesehatan Reproduksi 5, Nr. 1 (26.04.2018): 32. http://dx.doi.org/10.22146/jkr.37988.
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Background: Ovarian stimulation is part of assisted reproductive technology (ART) process, which aims to spur the growth of follicles to be developed, so that it will increase the chance of getting pregnant. LH surge cause the final follicular maturation, ovulation and becoming corpus luteum. So that an increase in LH may adversely affect the development of the follicle and eventually affect in-vitro fertilization.Objective To determine the effect of basal LH hormone to follicles on the stimulation of ovarian.Method: Cohort Retrospective. Research Location: Permata Hati Clinic of Sardjito Hospital, YogyakartaResult dan Discussion: The study included 70 cycles of 70 women who underwent ovarian stimulation for in-vitro fertilization and fulfill inclusion and exclusion criterias. The subjects were divided into 2 groups based on basal LH hormone levels i.e. LH ≤ 3 mIU/ml as the test group and LH >3 mIU/ml as the control group. From analysis, response to ovarian stimulation in group with LH ≤3 mIU/ml was significantly different than LH >3 mIU/ml (RR 1,875; 95% CI 1,275–2,757; p=0,00*).Conclusion: Low level of basal LH (≤ 3 mIU/ml) generating fewer number of mature follicles in patients who performed ovarian stimulation in in-vitro fertilization program.Keyword: basal LH, ovarian stimulation, in-vitro fertilization.
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Johnson, S. M., J. C. Smith und J. L. Feldman. „Modulation of respiratory rhythm in vitro: role of Gi/o protein-mediated mechanisms“. Journal of Applied Physiology 80, Nr. 6 (01.06.1996): 2120–33. http://dx.doi.org/10.1152/jappl.1996.80.6.2120.
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Slice preparations from neonatal rat medulla that generate respiratory rhythm in vitro were used to test for Gi/o protein-mediated mechanisms affecting breathing rhythm in mammals. The frequency of inspiratory motor discharge recorded from hypoglossal (XII) nerve roots decreased with bath application of gamma-aminobutyric acid (GABA) and norepinephrine, as well as agonists specific for GABAB, alpha 2-adrenergic, and mu-opioid receptors; 5-hydroxytryptamine had little effect on frequency. Microinjection of these specific agonists into the pre-Botzinger complex, the site of respiratory rhythm generation in vitro, also decreased frequency. In contrast, substance P (SP) increased frequency when it was bath applied or microinjected into the pre-Botzinger complex. To test for involvement of Gi/o proteins, pertussis toxin (PTX) was injected into the cerebrospinal fluid of newborn rats, and slices from these animals were tested 48 h later for block of drug effects on rhythm. In PTX-treated slices the frequency decrease due to GABAB, mu-opioid, and alpha 2-adrenergic receptor activation was attenuated (P < or = 0.05), whereas the SP receptor-mediated response was unaltered. To test for involvement of K+ conductances linked to Gi/o proteins Ba2+ (0.2 mM) was added to the bath before application of drugs. Ba2+ attenuated the decrease in frequency associated with GABAB (P < or = 0.05) and mu-opioid (0.10 < or = P < or = 0.05) receptor activation, whereas the alpha 2-adrenergic and SP responses were unaltered. We conclude that GABAB and mu-opioid, but not alpha 2-adrenergic and SP, receptor activation modulates respiratory frequency via a Gi/o protein-dependent Ba(2+)-sensitive ionic conductance mechanism on neurons within the medullary locus for rhythm generation. This mechanism may be a convergent pathway for control of respiratory frequency.