Journal articles on the topic 'Tunnel staining'
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Inagaki, Yusuke, Kota Uematsu, Manabu Akahane, Yusuke Morita, Munehiro Ogawa, Tomoyuki Ueha, Takamasa Shimizu, Tomohiko Kura, Kenji Kawate, and Yasuhito Tanaka. "Osteogenic Matrix Cell Sheet Transplantation Enhances Early Tendon Graft to Bone Tunnel Healing in Rabbits." BioMed Research International 2013 (2013): 1–8. http://dx.doi.org/10.1155/2013/842192.
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The objective of this study was to determine whether osteogenic matrix cell sheets (OMCS) could induce bone formation around grafted tendons, thereby enhancing early stage tendon to bone tunnel healing in skeletally mature male Japanese white rabbits. First, the osteogenic potential of rabbit OMCS was evaluated. Then, the OMCS were transplanted into the interface between the grafted tendon and the bone tunnel created at the tibia. Histological assessments and biomechanical tensile testing were performed after 3 weeks. The rabbit OMCS showed high alkaline phosphatase (ALP) activity, positive staining of ALP, and osteogenic potential when transplanted subcutaneously with beta tricalcium phosphate disks. Newly formed bony walls and positive collagen type I staining were seen around the grafted tendon with OMCS transplantation, whereas such bony walls were thinner or less frequent without OMCS transplantation. Micro-computed tomography images showed significantly higher bone volume in the OMCS transplantation group. The pullout strength was significantly higher with OMCS (0.74±0.23 N/mm2) than without OMCS (0.58±0.15 N/mm2). These results show that OMCS enhance early tendon to bone tunnel healing. This method can be applied to cases requiring early tendon to bone tunnel healing after ligament reconstruction surgery.
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Mbegbu, Edmund C., Ikechukwu R. Obidike, and Ali A. Fouladi-Nashta. "Immunohistochemical Detection of Vasa Antigen and Apoptosis-Related DNA Fragmentation in Ovaries of Sheep Fetuses Prenatally Exposed to Vitamin D Deficiency." Acta Veterinaria 69, no. 3 (September 1, 2019): 262–74. http://dx.doi.org/10.2478/acve-2019-0022.
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Abstract The primordial germ cells (PGCs) in female animals are comprised of diplotene oocytes arrested in the first meiotic prophase. Expression of Vasa is one of the key factors required for subsequent resumption of development and recruitment of PGCs into the growing follicle class. Since vitamin D regulates recruitment of PGCs and developmental competence of ovarian follicles, this study was designed to investigate the expression of Vasa and rate of apoptosis in foetal ovaries prenatally restricted from dietary vitamin D. Nineteen sexually mature Welsh mountain ewes were randomly assigned to vitamin D deficient (VDD) and vitamin D control (VDC) diets from 17d before mating, up to 125d of gestation, when fetal ovaries were collected and fixed in formalin for immunohistochemistry and TUNEL assay. VDD ovaries had fewer healthy oocytes that could stain positive for Vasa as well as a lower integrated density value for DAB staining intensity. Conversely, TUNNEL staining in VDD animals showed a higher integrated density value and percentage of affected area (P<0.05). The present findings indicate that Vasa expression is decreased, while the rate of apoptosis increased in VDD fetal ovaries, and this may adversely affect resumption of growth and development of PGCs reserve.
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Rao, RohitS, Charudutt Kalamkar, Amrita Mukherjee, and BhavinK Patel. "Novel tunnel staining technique to reduce premature entry in manual small-incision cataract surgery." Indian Journal of Ophthalmology 70, no. 11 (2022): 4041. http://dx.doi.org/10.4103/ijo.ijo_1562_22.
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Wang, Rui, Bin Xu, and Hong-Gang Xu. "Up-Regulation of TGF-β Promotes Tendon-to-Bone Healing after Anterior Cruciate Ligament Reconstruction using Bone Marrow-Derived Mesenchymal Stem Cells through the TGF-β/MAPK Signaling Pathway in a New Zealand White Rabbit Model." Cellular Physiology and Biochemistry 41, no. 1 (2017): 213–26. http://dx.doi.org/10.1159/000456046.
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Background/Aims: This study aimed to explore the role of TGF-β in tendon-to-bone healing after anterior cruciate ligament (ACL) reconstruction using bone marrow-derived mesenchymal stem cells (BMSCs) through the TGF-β/MAPK signaling pathway in a New Zealand white rabbit model. Methods: A total of 72 healthy male New Zealand white rabbits were selected for these experiments. Flow cytometry and immunofluorescence were used to detect the expression of BMSC surface markers, and qRT-PCR was performed to detect TGF-β mRNA expression. The ACL reconstruction model was established with autografts. The rabbits were randomly divided into the following groups: inhibition of TGF-β (inhibition), over-expression of TGF-β (over-expression), empty vector and untreated (n = 18 per group). Hematoxylineosin (HE) staining, toluidine blue staining and Masson trichrome staining were conducted to observe any chondrocyte-like cell growth, and biomechanical tests were used to calculate the maximum load and rigidity. Three-dimensional CT imaging and Western blotting were applied to detect changes in bone tunnel size and bone density and the expression levels of TGF-β/MAPK signaling pathway-related proteins, respectively. Results: CD90 and CD44 were positively expressed, while CD11b was not detected. Compared with the empty vector and untreated groups, TGF-β mRNA expression was significantly decreased in the inhibition group but increased in the over-expression group; the latter group had a larger number of fibroblasts, a tighter tendon-bone interface, an increased number of chondrocyte-like cells and fibrochondrocytes, and more collagen fibers than the inhibition, empty vector and untreated groups. Compared with the empty vector and untreated groups, the maximum load and rigidity; the CT values of bone tunnel and bone tunnel margin; and the protein expression levels of TGF-β, p-ERK1/2, p-p38, p-JNK, c-jun and c-myc were significantly down-regulated in the inhibition group but up-regulated in the over-expression group. Conclusion: Our study indicated that up-regulating TGF-β expression in BMSCs from New Zealand white rabbits could promote tendon-to-bone healing after ACL reconstruction by regulating the TGF-β/MAPK signaling pathway.
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Khanna, Rohit K., Mark L. Rosenblum, Jack P. Rock, and Ghaus M. Malik. "Prolonged external ventricular drainage with percutaneous long-tunnel ventriculostomies." Journal of Neurosurgery 83, no. 5 (November 1995): 791–94. http://dx.doi.org/10.3171/jns.1995.83.5.0791.
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✓ External ventricular drainage has been used extensively for management of several neurosurgical disorders. The main limitation of this procedure has been the high risk of infection, especially with prolonged drainage. In an effort to minimize the risk of infection, the authors have used a new ventriculostomy technique that involves tunneling the ventricular catheter subcutaneously to an exit site in the lower chest or upper abdomen. This report describes the results of this procedure on 100 consecutive cases. Patients requiring emergency ventriculostomies had short-tunnel ventriculostomies placed at the bedside that were converted to long-tunnel ventriculostomies in the operating room within 5 days. The average duration of drainage was 18.3 days (range 5–40 days). Cerebrospinal fluid was routinely sent for Gram staining and culture to monitor for infection. Prophylactic antibiotic medications were administered only perioperatively. No infection was observed during the first 16 days of drainage in any patient. The overall incidence of infection was 4% and blockage occurred in 6% of the cases. In this series the incidence of ventricular infection was 2.37 per 1000 ventricular drainage days, one of the lowest reported incidences of infection in the literature. This procedure provides a simple and effective method of maintaining long-term ventricular drainage with a very low risk of infection or blockage.
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Wang, Xinxin, Peng Shen, Dezhi Tang, Hao Xu, Hongfu Qiu, Tao Wu, and Xiang Gao. "Effects of Qi-Fang-Xi-Bi-Granules on Cartilage Morphology and C/ebpαPromoter Methylation in Rats with Knee Osteoarthritis." Evidence-Based Complementary and Alternative Medicine 2018 (2018): 1–12. http://dx.doi.org/10.1155/2018/2074976.
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Objective. To investigate the effects of Qi-Fang-Xi-Bi-Granules (QFXBGs) on cartilage morphology and methylation of C/ebpα(CCAAT/enhancer binding proteinα) at the promoter region.Methods. Knee osteoarthritis (KOA) modeling was performed in rats in accordance with Hulth’s method, and control group received sham operation. Eight weeks after KOA modeling, the rats in the KOA modeling group were further divided into 6 groups. Each group was given the appropriate drug. After 8 weeks, half of the rats were used for Micro-CT scan, HE staining, ABH/OG staining, immunohistochemistry, and TUNNEL staining of the knee joint tissue, and the other half were used to examine C/ebpαpromoter methylation.Results. The three dose groups of QFXBGs all showed lower degrees of surface fissures and flaking, thicker cartilage layer, and restored chondrocyte and subchondral bone morphology, compared with the KOA model group. C/ebpα-22 promoter methylation levels in the high- and low-dose groups were significantly higher than that in the KOA modeling group (p<0.05), while C/ebpα-2 promoter methylation level in the medium-dose group was significantly higher than that in the KOA modeling group (p<0.05).Conclusions. QFXBGs may alleviate articular cartilage degeneration through promoting C/ebpα-2 or C/ebpα-22 methylation at specific promoter sites.
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Joseph, Oliver C., Oleg Uryasev, John P. McNamara, and Apostolos P. Dallas. "TIBIAL NERVE PERINEURAL INJECTIONS AT THE POSTERIOR TARSAL TUNNEL." Journal of Musculoskeletal Research 16, no. 03 (September 2013): 1350014. http://dx.doi.org/10.1142/s0218957713500140.
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Introduction: Posterior tarsal tunnel syndrome (PostTTS) refers to compression of the tibial nerve (TN) within this tunnel. PostTTS is most commonly secondary to entrapment with subsequent inflammation. As it is true with other entrapment-type neuropathies, corticosteroids could provide therapeutic relief. To the authors' knowledge, the feasibility of such injections using ultrasound guidance has not been described in the literature. We hypothesize that one can inject the TN perineural space immediately proximal to the posterior tarsal tunnel utilizing ultrasonography US-guidance. Methods: This research was a pilot study using four cadaveric models. US was utilized to image the proximal posterior tarsal tunnel. Perineural injections of methylene blue were performed with subsequent dissection. Injections were designated as accurate (referring to nerve staining) and precise (referring to dye localization). Results: One cadaver was precluded due to pronounced musculoskeletal abnormality. 5-of-6 (83%) injections were accurate and 6-of-6 (100%) precise. Conclusion: Initial attempt was inaccurate and precise, while later injections were both accurate and precise. The most apparent source of error was from one cadaver's pronounced musculoskeletal deformity, which precluded successful injections bilaterally. Of the three cadavers unaffected by musculoskeletal deformity, accuracy was 5-of-6 (83%) and precision was 6-of-6 (100%). While surgery is the definitive treatment for refractory PostTTS, therapeutic effect of corticosteroid injections has not been evaluated in this patient population. Such injections could provide symptomatic relief and postpone surgical intervention. Small sample size not withstanding the results suggest that TN perineural injections are feasible under US-guidance. This study suggests that US-guidance can increase accuracy and precision and is a potential adjunct to the treatment. Future study will expand the initial data set and categorize consistent protocol. Subsequent translational research will then be sought to evaluate therapeutic efficacy in this patient population.
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Kramer, Adriel A., Arnold-Peter C. Weiss, Hans-J. Barrach, and Edward Akelman. "VARIATIONS IN THE QUANTITY OF TYPE II COLLAGEN IN CARPAL TUNNEL SYNDROME AND TRIGGER FINGER." Hand Surgery 01, no. 02 (July 1996): 95–101. http://dx.doi.org/10.1142/s0218810496000178.
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Carpal tunnel syndrome (CTS) and trigger finger (TF) are two related disorders which involve alterations in the normal mechanical loading of tendon sheath pulleys. This study examines the presence of Type II collagen in the carpal ligament and A-1 pulley by Western blotting to determine the extent of cartilage metaplasia that may occur in these tissues. Cyanogen bromide peptides generated from tissue supplied from 77 patients were separated by SDS-PAGE and transferred to nitrocellulose. The membranes were stained with the E1E5 monoclonal antibody and collagen levels were quantified. All specimens were found to contain at least small amounts of Type II collagen. Approximately 15 percent of the samples from patients with carpal tunnel syndrome and trigger finger were found to contain significantly elevated levels of Type II collagen. All specimens with high levels of Type II collagen were found in female patients. These increases may indicate the presence of cartilage metaplasia enabling the tissue to counteract compressive loads. With further study, tissue staining techniques using monoclonal antibodies may provide useful clinical information on severity or long-term outcome prognosis.
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Wang, Joon Ho, and Byung Hoon Lee. "Mediolateral Differences of Proteoglycans Distribution at the ACL Tibial Footprint: Experimental Study of 16 Cadaveric Knees." BioMed Research International 2018 (2018): 1–6. http://dx.doi.org/10.1155/2018/3762580.
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This study aimed to identify the staining pattern of ACL attachment blended with cartilage of the medial tibial plateau at the tibial insertion and histologically characterize the tibial footprint. Sixteen fresh frozen cadaveric knees (mean age: 52.0±6.2 years) were used for this study. The specimens were bisected in the coronal plane, in accordance with the fiber orientation of the ACL tibial attachment. Adjacent sections were then stained with hematoxylin and eosin (H&E) to observe the morphology of the ACL insertion and with fast green and Safranin-O protocols to evaluate for collagen and proteoglycans (PG). The insertion area on the tibial footprint was divided into five zones in the medial to lateral direction, which was determined by division of the section from most prominent medial tibial spine to most lateral margin of ACL attachment. Then rectangular area with a vertical length that is twice the width of respective five zones was set. Stained areas of all images were quantified positively by using ImageJ software, and the value for staining area measured was defined in percentage by multiplying whole image area by 100. The mean proportion of Safranin-O staining is significantly greater nearer to the medial tibial spine (59% in zone 1, 32% in zone 2, 13% in zone 3, 13% in zone 4, and 4% in zone 5, P<0.001). The medial section of the tibial insertion area grew in size and increased in PG staining with more densely organized collagen arrangement with more fibrocartilage cells. The ACL tibial insertion showed a medially eccentric staining pattern by histological evaluation of the ACL attachment to cartilage. Our histological results of the eccentric biomaterial property in the medial tibial spine of ACL insertion area can be considered in making a more functional anatomic tibial tunnel placement.
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Zhao, YuHan, and QingHong Cheng. "Exogenous H2S Protects against Septic Cardiomyopathy by Inhibiting Autophagy through the AMPK/mTOR Pathway." Contrast Media & Molecular Imaging 2022 (June 10, 2022): 1–8. http://dx.doi.org/10.1155/2022/8464082.
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Background. Given the cardioprotective role of autophagy, this study aimed to investigate the protective effect of exogenous H2S (NaHS) on infectious cardiomyopathy through the inhibition of the AMPK/mTOR pathway. Methods. In this study, sepsis models were established by cecal ligation and puncture (CLP) induction in vivo and intraperitoneal injection of NaHS was performed. Autophagy- and apoptosis-related proteins were observed by western blot, isolated myocardial tissue morphology was observed by hematoxylin-eosin (H&E) staining, and myocardial apoptosis was evaluated by the tunnel method. The ultrastructure of autophagy was observed by using an electron transmission electron microscope. Results. In an SD rat model of cecum ligation puncture-induced sepsis, the level of autophagy-related proteins was significantly increased, and hematoxylin and eosin staining showed irregular myocardial bands and swollen cardiomyocytes. Following NaHS treatment, the level of autophagy-related proteins decreased, and electron transmission microscopy revealed decreased autophagosomes. Echocardiography suggested an increase in ejection fraction and significant relief of myocardial inhibition. Conclusions. Our results suggest that NaHS treatment can attenuate the cellular damage caused by excessive autophagy through the AMPK/mTOR pathway.
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Srivastava, Monika. "Comparative Study of Coronary Artery Proximal to Myocardial Bridge Segment." Journal of Advanced Research in Medical Science & Technology 08, no. 03 (August 26, 2021): 5–9. http://dx.doi.org/10.24321/2394.6539.202112.
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Introduction: The muscular segments which overlie the epicardial arteries are termed as myocardial bridges and the artery which travels through them are termed as tunnel arteries. These tunnel arteries get compressed during the systolic compression of the heart, thus partially or completely blocking the blood supply to the corresponding areas. Aim & Objectives: To assess the impact of these myocardial bridges on the proximal segment of the myocardial arteries. Methodology: The present study was cadaveric-based cross-sectional study. A total of 22 hearts which showed the presence of myocardial bridges were collected from two sources namely: cadaver dissections, autopsy. The hearts were clean and numbered. This was followed by fixation, dehydration, clearing, embedding, block formation, section cutting and staining. Result: The present study showed that there is a significant thickening in the tunica intima of the proximal to bridge segment of the coronary artery. The present study also noted that there is a marked thinning of the tunica media of the same segment. Conclusion: The present study concludes that there is a marked hyperplasia in the proximal segment of the myocardial bridges under tunica intima.
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Tachi, Masahito, Hideshi Okada, Nobuhisa Matsuhashi, Genzou Takemura, Kodai Suzuki, Hirotsugu Fukuda, Ayumi Niwa, et al. "Human Colorectal Cancer Infrastructure Constructed by the Glycocalyx." Journal of Clinical Medicine 8, no. 9 (August 22, 2019): 1270. http://dx.doi.org/10.3390/jcm8091270.
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Cancer cells can survive and grow via angiogenesis. An alternative but controversial theory is cancer cells may grow via vasculogenic mimicry (VM), in which the cancer cells themselves construct vessel-like channels that are considered a leading cause of drug resistance. The dynamic functions of the glycocalyx (GCX), a meshwork composed of proteoglycans and glycoproteins that surrounds cell membranes, have been observed in endothelial cells within tumors. However, the actual structural shape formed by the GCX in human patients remains unclear. Here, we visualized the three-dimensional (3D) network structure constructed by bulky GCX in human colorectal cancer (CRC) patients using scanning electron microscopy with lanthanum nitrate staining. The network structure extended throughout the cancer cell nest, opening into capillaries, with a tunnel channel that exhibited a net- and spongy-like ultrastructure. The expression of endothelial and cancer-specific GCX-binding lectins was dramatically increased in the interstitial spaces between cancer cells. Even accounting for the presence of artifacts resulting from sample preparation methods, the intercellular tunnels appeared to be coated with the bulky GCX. Further, this 3D network structure was also observed in the tumors of ApcMin/+ mice. In conclusion, the bulky GCX modifies the network structure of CRCs in human and mice.
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Fu, Haiying, Junjie Zhang, and Mayu Huang. "Topiroxostat ameliorates oxidative stress and inflammation in sepsis-induced lung injury." Zeitschrift für Naturforschung C 75, no. 11-12 (November 26, 2020): 425–31. http://dx.doi.org/10.1515/znc-2020-0074.
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AbstractSepsis-induced lung injury was the most common cause of death in patients. Topiroxostat, a novel xanthine oxidoreductase inhibitors, possessed obvious organ protectives effects. Xanthine oxidase played a vital role in acute lung injury. The study aimed to investigate the roles of Topiroxostat in sepsis-induced lung injury. The sepsis rats were established using cecum ligation and perforation. The lung damage induced by sepsis was evaluated by Hematoxylin and Eosin staining and lung tissue wet to dry ratio. The oxidative stress was detected by measurement of reactive oxygen species, malondialdehyde, myeloperoxidase and superoxide dismutase (SOD). The pro-inflammatory mediators, tumor necrosis factor-α, interleukin (IL)-1β, IL-6 and monocyte chemotactic protein 1, were measured by Enzyme-Linked Immunosorbent Assay. The cell apoptosis in lung was detected by TUNNEL staining and western blot analysis of apoptosis-related proteins including pro-apoptosis proteins, Bax, cleaved caspase9, cleaved caspase3 and anti-apoptosis protein Bcl2. The results showed that Topiroxostat significantly reduced lung damage, along with decreased oxidative stress, inflammation response and apoptosis in sepsis rats. Topiroxostat exerted markedly protective effects in sepsis-induced lung injury and could be an antioxidant in treating sepsis-induced lung injury.
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Yun, Damin, Liwei Zhou, Jie Shi, Xinyao Li, Xiaolong Wu, and Fei Sun. "G3BP2, a stress granule assembly factor, is dispensable for spermatogenesis in mice." PeerJ 10 (June 28, 2022): e13532. http://dx.doi.org/10.7717/peerj.13532.
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Background Spermatogenesis is a complex process that includes mitosis, meiosis, and spermiogenesis. During spermatogenesis, genetic factors play a vital role inthe formation of properly functioning sperm. GTPase-activating protein (SH3 domain)-binding protein 2 (G3BP2) is known to take part in immune responses, mRNA transport, and stress-granule assembly. However, its role in male fertility is unclear. Here, we generated a G3bp2 conditional knockout (cKO) mouse model to explore the function of G3BP2 in male fertility. Methods Polymerase chain reaction (PCR) and western blotting (WB) were used to confirm testis-specific G3bp2 knockout. Hematoxylin-eosin (HE) staining to observe testicular morphology and epididymal structure. Computer-aided sperm analysis (CASA) to detect sperm concentration and motility. Terminal deoxynucleotidyl transferase-dUTP nick-end labeling (TUNEL) assay was used to detect apoptotic cells. Results We found that cKO male mice are fertile with the normal morphology of the testis and sperm. Additionally, CASA of the semen from cKO mice showed that they all had a similar sperm concentration and motility. In addition, sperm from these mice exhibited a similar morphology. But the tunnel assay revealed increased apoptosis in their testes relative to the level in the wild type (WT). Conclusion Together, our data demonstrate that G3BP2 is dispensable for spermatogenesis and male fertility in mice albeit with the increased germ-cell apoptosis.
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Gabryś, Tobiasz, Beata Fryczkowska, Joanna Grzybowska-Pietras, and Dorota Biniaś. "Modification and Properties of Cellulose Nonwoven Fabric—Multifunctional Mulching Material for Agricultural Applications." Materials 14, no. 15 (August 3, 2021): 4335. http://dx.doi.org/10.3390/ma14154335.
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The paper describes a method of modifying a commercial viscose nonwoven fabric and its use as a modern mulching material in agriculture. The conducted research confirmed that the proposed modification of the viscose nonwoven fabric could be successfully used as a multipurpose and, above all, completely biodegradable nonwoven crop cover, which will eliminate the problem of disposal after the harvest period. Modified cellulose nonwoven fabric was obtained by staining with NB—BT helion brown, then padding with potassium nitrate (KNO3) solution (used as a fertilizer) and finally coating with polylactide (PLA) solution. The characterisation of the nonwoven fabric included structural analysis, physicochemical properties and mechanical tests. The modified cellulose nonwovens were used in the tunnel cultivation of tomatoes as a heat-retardant, water-absorbing, antiweed mulching material that prevents soil infestation and slowly releases fertilizers.
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Hamidah, Masyitah, Aty Widyawaruyanti, Widjiati Widjiati, and Budi Prasetyo. "Sambiloto (AS201-01) is better than standard antimalarial drug (DHP) in reducing Toll-Like Receptor 2 (TLR2) on placenta malaria model." Majalah Obstetri & Ginekologi 26, no. 2 (August 23, 2018): 74. http://dx.doi.org/10.20473/mog.v26i22018.74-82.
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Objectives: To compare the TLR2 expression in the placenta between treated by sambiloto EA-96 fraction tablet (AS201-01) and dihydroartemisinin-piperaquine phosphate (DHP)Materials and Methods: Experimental study using 24 pregnant mice. All sample divided into 4 groups with randomization are uninfected group, Plasmodium infected group and given placebo, sambiloto (AS201-01) and DHP. Then performed surgery and placental sampling were staining with adopting in tunnel assay method to measure the TLR2 expression of placental.Results: The expression of TLR2 in uninfected group has the lowest rate compared to other groups. The infected and placebo treated group has the highest TLR2 expression campared with sambiloto and DHP. The sambiloto group has not differ signi-ficantly with the group uninfected and lower than DHP.Conclusion: Tablet of sambiloto EA-96 fraction (AS201-01) decreased TLR2 expession better than with DHP tablet.
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Hara, Yukinori, Yasuhito Tajiri, Kenichi Kawano, Shinya Hoshikawa, and Yusuke Kita. "The Tenosynovitis of Fingers Associated with Transthyretin Amyloidosis." Journal of Hand Surgery (Asian-Pacific Volume) 25, no. 03 (July 28, 2020): 340–44. http://dx.doi.org/10.1142/s2424835520500381.
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Background: Amyloidosis treatment has advanced rapidly along with the discovery of drugs to prevent amyloid deposition. Therefore, it is vital to detect amyloidosis at an early stage. Wild-type transthyretin, which can cause carpal tunnel syndrome, may also cause finger tenosynovitis. However, the correlation between wild-type transthyretin amyloid and finger tenosynovitis is unclear. Here, we investigated pathological and clinical findings for 20 patients with finger tenosynovitis who underwent operation at our hospital to determine the frequency of transthyretin amyloid deposition in idiopathic finger tenosynovitis. Methods: To check for the presence of amyloid deposition, all specimens (tendon synovium tissue or flexor tendon sheath) resected during the operation were stained by the direct fast scarlet method. Amyloid-positive specimens were evaluated by immunohistochemical staining using an anti-transthyretin antibody. Patient characteristics were evaluated with respect to amyloid presence. Results: Thirteen (65%) of 20 finger tenosynovitis cases had amyloid deposition. Nine (69.2%) of the 13 amyloid-positive cases exhibited extensive transthyretin staining and were considered to have transthyretin amyloid. Amyloid deposition was more frequent in men. The mean number of fingers with tenosynovitis was significantly higher in amyloid-positive cases (3.8 fingers) than in amyloid-negative cases (2.0 fingers). Conclusions: Men with multiple finger tenosynovitis tended to have transthyretin amyloid deposition. Our results support that multiple finger tenosynovitis may serve as an initial indication of evaluation for transthyretin amyloidosis.
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Tao, Yi, Bing Zhou, Jie Zou, Yixian Yu, Jing Zhao, Naijia Xu, and Qiong Wang. "Nicorandil alleviates inflammation and oxidation in diabetic cardiomyopathy." Tropical Journal of Pharmaceutical Research 22, no. 2 (April 27, 2022): 273–79. http://dx.doi.org/10.4314/tjpr.v22i2.9.
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Purpose: To examine the effect of nicorandil on high glucose-induced cardiomyocyte inflammation and oxidative stress.Methods: H9C2 cardiomyocytes were divided into control group, high glucose group and nicorandil group. The survival rate of cardiomyocytes was determined using the CCK-8 method. The contents of reactive oxygen species (ROS) of cardiomyocytes were determined by flow cytometry. The contents of MDA and LDH in cell supernatant were determined by kit. Western blot and real-time PCR were used to assess oxidative stress, inflammation and apoptosis related factors in each group of cardiomyocytes. The expression levels of IL-1β were determined by immunofluorescence. Tunnel staining was used to determine the apoptosis level of each group.Results: The expressions of SOD1 and SOD2 in the high glucose group were significantly decreased (p < 0.05). Also, the contents of MDA and LDH were significantly increased (p < 0.05). Furthermore, IL-1β, TNF-α, caspase 3 and Bax expressions were increased, while Bcl-2 expression was inhibited. IL-1β and Tunnel fluorescence also increased significantly. NF-κB and Ikkα were significantly increased, while IκB-α was inhibited. Furthermore, nicorandil inhibited oxidative stress and apoptosis, as well as NF-κB pathway and downstream factor Ikkα.Conclusion: Nicorandil ameliorates the inflammation and oxidative damage of cardiomyocytes induced by high glucose, by inhibiting NF-κB pathway, thereby lowering apoptosis. Thus, the findings provide new insight into the development of new agents for the treatment of diabetic cardiomyopathy.
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Zhang, Jun, Ziming Liu, Yuwan Li, Qi You, Jibin Yang, Ying Jin, Gang Zou, Jingfeng Tang, Zhen Ge, and Yi Liu. "FGF-2-Induced Human Amniotic Mesenchymal Stem Cells Seeded on a Human Acellular Amniotic Membrane Scaffold Accelerated Tendon-to-Bone Healing in a Rabbit Extra-Articular Model." Stem Cells International 2020 (January 6, 2020): 1–14. http://dx.doi.org/10.1155/2020/4701476.
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Background. FGF-2 (basic fibroblast growth factor) has a positive effect on the proliferation and differentiation of many kinds of MSCs. Therefore, it represents an ideal molecule to facilitate tendon-to-bone healing. Nonetheless, no studies have investigated the application of FGF-2-induced human amniotic mesenchymal stem cells (hAMSCs) to accelerate tendon-to-bone healing in vivo. Objective. The purpose of this study was to explore the effect of FGF-2 on chondrogenic differentiation of hAMSCs in vitro and the effect of FGF-2-induced hAMSCs combined with a human acellular amniotic membrane (HAAM) scaffold on tendon-to-bone healing in vivo. Methods. In vitro, hAMSCs were transfected with a lentivirus carrying the FGF-2 gene, and the potential for chondrogenic differentiation of hAMSCs induced by the FGF-2 gene was assessed using immunofluorescence and toluidine blue (TB) staining. HAAM scaffold was prepared, and hematoxylin and eosin (HE) staining and scanning electron microscopy (SEM) were used to observe the microstructure of the HAAM scaffold. hAMSCs transfected with and without FGF-2 were seeded on the HAAM scaffold at a density of 3×105 cells/well. Immunofluorescence staining of vimentin and phalloidin staining were used to confirm cell adherence and growth on the HAAM scaffold. In vivo, the rabbit extra-articular tendon-to-bone healing model was created using the right hind limb of 40 New Zealand White rabbits. Grafts mimicking tendon-to-bone interface (TBI) injury were created and subjected to treatment with the HAAM scaffold loaded with FGF-2-induced hAMSCs, HAAM scaffold loaded with hAMSCs only, HAAM scaffold, and no special treatment. Macroscopic observation, imageological analysis, histological assessment, and biomechanical analysis were conducted to evaluate tendon-to-bone healing after 3 months. Results. In vitro, cartilage-specific marker staining was positive for the FGF-2 overexpression group. The HAAM scaffold displayed a netted structure and mass extracellular matrix structure. hAMSCs or hAMSCs transfected with FGF-2 survived on the HAAM scaffold and grew well. In vivo, the group treated with HAAM scaffold loaded with FGF-2-induced hAMSCs had the narrowest bone tunnel after three months as compared with other groups. In addition, macroscopic and histological scores were higher for this group than for the other groups, along with the best mechanical strength. Conclusion. hAMSCs transfected with FGF-2 combined with the HAAM scaffold could accelerate tendon-to-bone healing in a rabbit extra-articular model.
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Tsai, Karen, Alice Chen Yu, Masha J. Livhits, Dipti Sajed, Angela M. Leung, and Dianne S. Cheung. "Systemic light-chain amyloidosis incidentally diagnosed after subtotal parathyroidectomy and thyroid lobectomy." BMJ Case Reports 14, no. 4 (April 2021): e241282. http://dx.doi.org/10.1136/bcr-2020-241282.
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A 74-year-old woman with a history of primary hyperparathyroidism, thyroid nodules, atrial fibrillation and pacemaker placement for sick sinus syndrome presented with fatigue, constipation and persistent lower extremity oedema. She underwent subtotal parathyroidectomy and left thyroid lobectomy. Histopathology revealed amyloidosis affecting the thyroidand parathyroids confirmed by Congo Red Staining with Mayo Clinic subtyping of light chain kappa-type amyloidosis. She was found to have combined systolic and diastolic cardiac dysfunction, carpal tunnel neuropathy and pre-diabetes suggestive of systemic amyloidosis with involvement of the heart, nerves and pancreas. Congo red stain was positive for amyloidosis on bone marrow biopsy suggestive of a diagnosis of systemic amyloidosis. She was treated with daratumumab with good clinical response. This case illustrates the necessity of considering systemic amyloidosis in patients with incidentally discovered diffuse amyloid deposits on biopsy of an endocrine organ, as endocrine effects are a rare but likely underdiagnosed consequence of systemic amyloidosis.
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JEMEC, B., A. O. GROBBELAAR, G. D. WILSON, P. J. SMITH, R. SANDERS, and D. A. McGROUTHER. "Is Dupuytren’s Disease Caused by an Imbalance between Proliferation and Cell Death?" Journal of Hand Surgery 24, no. 5 (October 1999): 511–14. http://dx.doi.org/10.1054/jhsb.1999.0251.
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Dupuytren’s contracture shares certain properties with malignant tumours, characterized by proliferation and lack of apoptosis, which may be induced by the c- myc oncogene. Because of these similarities, the relationship between the c- myc oncogene expression, bcl-2 oncogene (antiapoptotic gene) and proliferation was investigated in Dupuytren’s disease. Proliferation was assessed by immunohistochemical staining of the mib-1 antibody. Results were compared with those from fibrosarcoma specimens, representing a related malignant tumour. Non-diseased fascia from Dupuytren patients and flexor retinaculum from patients undergoing carpal tunnel release without Dupuytren’s disease were used as controls. Expression of c- myc was elevated in primary Dupuytren’s disease and fibrosarcoma specimens, whilst recurrent Dupuytren’s disease, non-diseased Dupuytren fascia and flexor retinaculum exhibited significantly lower levels. Neither bcl-2 nor mib-1 were detected in Dupuytren’s disease, non-diseased fascia or flexor retinaculum, in contrast to fibrosarcoma. The imbalance between proliferation and apoptosis, producing malignant growth was thus confirmed for fibrosarcoma, but not for Dupuytren’s disease.
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Niu, Manlan, and Jingming Yan. "Tissue Engineering Properties of Nanomaterials and Their Performance Evaluation for Repairing Athletic Ligament Injuries in Sports Dance." Journal of Nanomaterials 2022 (July 16, 2022): 1–8. http://dx.doi.org/10.1155/2022/9902466.
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With the fast growth of nanotechnology, the usage of nuclear materials is becoming increasingly widespread, and the exposure of people, plants, and fauna to nanomaterials has become unavoidable. As scaffolds of biomaterials, nanomaterials are widely used in tissue engineering because of their good biocompatibility, noncytotoxicity, and noninflammatory reaction. In this paper, the tissue engineering properties of nanomaterials and their performance evaluation for repairing sports ligament injuries in dance sports were investigated. Sports ligament injury is a common sports disease, and ligament injury has a very serious impact on sports performance and sports life of athletes. In this paper, we takes football as an example, establishes a human body model of tendon-bone repair after anterior cruciate ligament reconstruction, evaluated the effect of injectable rhBMP-2 nanocontrolled release capsule on ligament bone tunnel interface repair, and evaluated from the scientific and biomechanical point of view. In addition, human ligament research and rabbit ligament supplementary experiment were carried out. In the study of human ligament, we selected 30 patients with unilateral polyarticular ligament injury caused by sports dance as the research subjects and randomly divided them into control group, experimental group, and blank group. The experimental group underwent the repair and reconstruction of rhBMP-2 nanocontrolled release capsule ligament. The results of human ligament study showed that the injectable rhBMP-2 nanocontrolled release capsules showed positive staining and uniform staining for ligament repair. The stiffness of the tender bony temple junction was 11.73%, 15.65%, and 50.59% greater in the test group than in the control group at 2, 4, and 8 weeks postoperatively, respectively.
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Zhu, Jingxian, Xin Zhang, Zhenxing Shao, Linghui Dai, La Li, Xiaoqing Hu, Xiaokun Wang, Chunyan Zhou, and Yingfang Ao. "In VivoStudy of Ligament-Bone Healing after Anterior Cruciate Ligament Reconstruction Using Autologous Tendons with Mesenchymal Stem Cells Affinity Peptide Conjugated Electrospun Nanofibrous Scaffold." Journal of Nanomaterials 2013 (2013): 1–11. http://dx.doi.org/10.1155/2013/831873.
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Electrospinning nanofibrous scaffold was commonly used in tissue regeneration recently. Nanofibers with specific topological characteristics were reported to be able to induce osteogenic differentiation of MSCs. In thisin vivostudy, autologous tendon grafts with lattice-like nanofibrous scaffold wrapping at two ends of autologous tendon were used to promote early stage of ligament-bone healing after rabbit ACL reconstruction. To utilize native MSCs from bone marrow, an MSCs specific affinity peptide E7 was conjugated to nanofibrous meshes. After 3 months, H-E assessment and specific staining of collagen type I, II, and III showed direct ligament-bone insertion with typical four zones (bone, calcified fibrocartilage, fibrocartilage, and ligament) in bioactive scaffold reconstruction group. Diameters of bone tunnel were smaller in nanofibrous scaffold conjugated E7 peptide group than those in control group. The failure load of substitution complex also indicated a stronger ligament-bone insertion healing using bioactive scaffold. In conclusion, lattice-like nanofibrous scaffold with specific MSCs affinity peptide has great potential in promoting early stage of ligament-bone healing after ACL reconstruction.
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Zhang, Q. L., L. Jia, X. Jiao, W. L. Guo, J. W. Ji, H. L. Yang, and Q. Niu. "APP/PS1 Transgenic Mice Treated with Aluminum: An Update of Alzheimer's Disease Model." International Journal of Immunopathology and Pharmacology 25, no. 1 (January 2012): 49–58. http://dx.doi.org/10.1177/039463201202500107.
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There is still no animal model available that can mimic all the cognitive, behavioral, biochemical, and histopathological abnormalities observed in patients with Alzheimer's disease (AD). We undertook to consider the interaction between genetic factors, including amyloid precursor protein (APP) and presenilin-1 (PS1), and environmental factors, such as Aluminum (Al) in determining susceptibility outcomes when studying the pathogenesis of AD. In this article, we provide an AD model in APP/PS1 transgenic mice triggered by Al. The animal model was established via intracerebral ventricular microinjection of aluminum chloride once a day for 5 days in APP/PS1 transgenic mice. Twenty wild type (WT) mice and 20 APP/PS1 transgenic (TG) mice were separately divided into 2 groups (control and Al group), and a stainless steel injector with stopper was used for microinjection into the left-lateral cerebral ventricle of each mouse. The Morris water maze task was used to evaluate behavioral function of learning and memory ability on the 20th day after the last injection. This AD model's brain was analyzed by: (1) amyloid β immunohistochemical staining; (2) Tunnel staining; (3) apoptotic rates; (4) caspase-3 gene expression. Here, decrease of cognitive ability and neural cells loss were shown in APP/PS1 transgenic mice exposed to Al, which were more extensive than those in APP/PS1 TG alone and WT mice exposed to Al alone. These findings indicate that there is a close relationship between over-expression of APP and PS1 genes and Al overload. It is also suggested that APP/PS1 TG mice exposed to Al have potential value for improving AD models.
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Al-Harbi, Laila Naif, Pandurangan Subash-Babu, Manal Abdulaziz Binobead, Maha Hussain Alhussain, Sahar Abdulaziz AlSedairy, Amal A. Aloud, and Ali A. Alshatwi. "Potential Metabolite Nymphayol Isolated from Water Lily (Nymphaea stellata) Flower Inhibits MCF-7 Human Breast Cancer Cell Growth via Upregulation of Cdkn2a, pRb2, p53 and Downregulation of PCNA mRNA Expressions." Metabolites 10, no. 7 (July 8, 2020): 280. http://dx.doi.org/10.3390/metabo10070280.
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Controlled production of cyclin dependent kinases (CDK) and stabilization of tumor suppressor genes are the most important factors involved in preventing carcinogenesis. The present study aimed to explore the cyclin dependent apoptotic effect of nymphayol on breast cancer MCF-7 cells. In our previous study, we isolated the crystal from a chloroform extract of Nymphaea stellata flower petals and it was confirmed as nymphayol (17-(hexan-2-yl)-10,13-dimethylhexadecahydro-1H-cyclopenta[a]phenanthren-3-ol) using x-ray diffraction (XRD), Fourier transform infrared (FTIR), and mass spectroscopy (MS) methods. The cytotoxic effect of nymphayol on MCF-7 cells were analyzed using the 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. The cellular and nuclear damage was determined using propidium iodide (PI) and acridine orange/ethidium bromide (AO/ErBr) staining. Tumor suppressor and apoptosis related mRNA transcript levels were determined using real-time polymerase chain reaction (RT-PCR). Nymphayol potentially inhibits MCF-7 cell viability up to 78%, and the IC50 value was observed as 2.8 µM in 24 h and 1.4 µM in 48 h. Treatment with nymphayol significantly increased reactive oxygen species (ROS) level and the tunnel assay confirmed DNA damage. We found characteristically 76% apoptotic cells and 9% necrotic cells in PI and AO/ErBr staining after 48 h treatment with 2.8 µM of nymphayol. Gene expression analysis confirmed significantly (p ≤ 0.001) increased mRNA levels of cyclin dependent kinase inhibitor 2A (Cdkn2a), retinoblastoma protein 2 (pRb2), p53, nuclear factor erythroid 2-factor 2 (Nrf2), caspase-3, and decreased B-cell lymphoma 2 (Bcl-2), murine double minute 2 (mdm2), and proliferating cell nuclear antigen (PCNA) expression after 48 h. Nymphayol effectively inhibited breast cancer cell viability, and is associated with early expression of Cdkn2a, pRb2, and activation of p53 and caspases.
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Romanenko, Svetlana A., Vladimir G. Malikov, Ahmad Mahmoudi, Feodor N. Golenishchev, Natalya A. Lemskaya, Jorge C. Pereira, Vladimir A. Trifonov, et al. "New Data on Comparative Cytogenetics of the Mouse-Like Hamsters (Calomyscus Thomas, 1905) from Iran and Turkmenistan." Genes 12, no. 7 (June 24, 2021): 964. http://dx.doi.org/10.3390/genes12070964.
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The taxonomy of the genus Calomyscus remains controversial. According to the latest systematics the genus includes eight species with great karyotypic variation. Here, we studied karyotypes of 14 Calomyscus individuals from different regions of Iran and Turkmenistan using a new set of chromosome painting probes from a Calomyscus sp. male (2n = 46, XY; Shahr-e-Kord-Soreshjan-Cheshme Maiak Province). We showed the retention of large syntenic blocks in karyotypes of individuals with identical chromosome numbers. The only rearrangement (fusion 2/21) differentiated Calomyscus elburzensis, Calomyscus mystax mystax, and Calomyscus sp. from Isfahan Province with 2n = 44 from karyotypes of C. bailwardi, Calomyscus sp. from Shahr-e-Kord, Chahar Mahal and Bakhtiari-Aloni, and Khuzestan-Izeh Provinces with 2n = 46. The individuals from Shahdad tunnel, Kerman Province with 2n = 51–52 demonstrated non-centric fissions of chromosomes 4, 5, and 6 of the 46-chromosomal form with the formation of separate small acrocentrics. A heteromorphic pair of chromosomes in a specimen with 2n = 51 resulted from a fusion of two autosomes. C-banding and chromomycin A3-DAPI staining after G-banding showed extensive heterochromatin variation between individuals.
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Zishiri, Rutendo M., Charles S. Mutengwa, Liliane N. Tandzi, and Alen Manyevere. "Growth Response and Dry Matter Partitioning of Quality Protein Maize (Zea mays L.) Genotypes under Aluminum Toxicity." Agronomy 12, no. 6 (May 25, 2022): 1262. http://dx.doi.org/10.3390/agronomy12061262.
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In this work, 75 quality protein maize (QPM) inbred lines were evaluated for aluminum tolerance using a nutrient solution assay in a laboratory and a soil-based technique in a greenhouse tunnel. The experiment was set up in a completely randomized design with three replications in the laboratory, and a randomized complete block design was used in the greenhouse. Aluminum toxicity was generated by amending a nutrient solution with 600 µM of aluminum sulfate (Al2 [SO4]3) in the laboratory, and Al2 [SO4]3 was applied at a rate of 24 mg kg−1 of soil in the greenhouse experiment. Relative root length (RRL) and hematoxylin staining (HS) scores were used to identify tolerant genotypes in the laboratory. According to RRL, 94.7% of genotypes were tolerant and 5.3% were sensitive, while Hematoxylin (HS) classified 77.9% of the genotypes as tolerant, and 22.1% as sensitive. RRL and HS presented a very strong negative association (−0.788). In the soil-based method, the experiments were conducted twice in successive summer seasons of 2019 and 2020. Several growth traits were measured and most genotypes that exhibited tolerance in the nutrient solution also had similar tolerance in the soil-based screening technique. Genetic variability for tolerance was identified, revealing potentially useful donors of tolerance genes in breeding programs.
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Ju, XiaoChen, Hao Chai, Sasirekha Krishnan, Abinaya Jaisankar, Murugan Ramalingam, and Lei Zhang. "Effect of Remnant-Preserving Reconstruction of Acute Anterior Cruciate Ligament Injuries in a Rabbit Model: Histological and Biomechanical Analysis." Journal of Biomaterials and Tissue Engineering 12, no. 5 (May 1, 2022): 897–906. http://dx.doi.org/10.1166/jbt.2022.2992.
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Acute anterior cruciate ligament (ACL) is a key structure that stabilizes knee joints. The objective of this research is to investigate the influence of ligament remnants preserved on the tendon-bone healing following ACL reconstruction and to examine postoperative articular cartilage degeneration in rabbit as a model animal. Sixty New Zealand rabbits are randomly divided into an ACL reconstruction without remnant preservation group (Group A; n = 30) or ACL reconstruction with remnant preservation group (Group B; n = 30). The expression of HIF-1α, VEGF, and micro vessel density (MVD) in the transplanted tendon was detected by immunohistochemical staining at week 6 and 12 after the operation. The signal intensity of the transplanted tendon was observed by MRI scanning, and the width of the bone tunnel was measured by CT scanning at week 6 and 12 after the operation. The graft biomechanics was tested 12 weeks after the operation. The JNK and MMP-13 expression levels were compared to analyze the cartilage degeneration of the knee at week 12 after the operation. The experimental results were analyzed and showed that the remnant-preserving ACL reconstruction is beneficial for bone healing of the tendon in rabbits, but ACL reconstruction with or without ligament remnants preserved will not affect knee articular cartilage degeneration post-surgery.
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Haidinger, W., M. P. Szostak, W. Jechlinger, and W. Lubitz. "Online Monitoring of Escherichia coli Ghost Production." Applied and Environmental Microbiology 69, no. 1 (January 2003): 468–74. http://dx.doi.org/10.1128/aem.69.1.468-474.2003.
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ABSTRACT Controlled expression of cloned φX174 gene E in gram-negative bacteria results in lysis of the bacteria by the formation of a transmembrane tunnel structure built through the cell envelope complex. Production of bacterial ghosts is routinely monitored by classical microbiological procedures. These include determination of the turbidity of the culture and the total number of cells and the number of reproductive cells present during the time course of growth and lysis. Although conceptually simple, these methods are labor intensive and time consuming, providing a complete set of results after the determination of viable cell counts. To avoid culturing methods for bacterial growth, an alternative flow cytometric procedure is presented for the quantification of ghosts and polarized, as well as depolarized, nonlysed cells within a culture. For this method, which is based on the discriminatory power of the membrane potential-sensitive dye bis-(1,3-dibutylbarbituric acid) trimethine oxonol, a staining protocol was developed and optimized for the maximum discrepancy in fluorescence between bacterial ghosts and viable cells. The total quantitative analysis procedure takes less than 2 min. The results derived from classical or cytometric analyses correlate with respect to the total cell numbers and the viability of the culture.
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Choi, Hyuk Soon, Hoon Jai Chun, In Kyung Yoo, Jae Min Lee, Seung Han Kim, Eun Sun Kim, Bora Keum, Yoon Tae Jeen, Hong Sik Lee, and Chang Duck Kim. "The therapeutic effect of irreversible electroporation ablation in mouse model of gastrointestinal cancer." Journal of Clinical Oncology 34, no. 4_suppl (February 1, 2016): 62. http://dx.doi.org/10.1200/jco.2016.34.4_suppl.62.
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62 Background: Irreversible electroporation (IRE) is a promising novel technique for the ablation of tumors. IRE has an advantage over other ablation technique in its mechanism to remove undesired cells by affecting the cell membrane without thermally destructing blood vessels, nerves and the surrounding tissues. Studies regarding the clinical application of IRE have been performed in humans, as well as in animals, for organs such as the liver, kidney, prostate, etc. and IRE is now accepted as a novel anti-cancer ablation modality. The aim of this study was to evaluate the therapeutic effect of IRE in mouse model of gastrointestinal cancer for the first time. Methods: The Caco2 cells (ATCC) were cultured in petri-dishes. Male nude mice (Immunodeficient (CAnN.Cg-Foxn1 nu/CrljBgi) 6 weeks old, Orient Inc., Korea) were introduced. Caco2 cells were each visually injected at 1.0 x 107cells/ml into both flakes (one for control, the other for IRE). We performed in vivo IRE procedures in the tumors of nude mouse model. Electrical pulses were applied to the tumor of nude mouse using a DC generator at 1~2kV/cm amplitude, 20~50 pulses, 100 µA length, with 1mm separation between two needle type electrodes. We analyzed the tissues with H&E staining and TUNEL assay immediately afterwards, and then 10 hours, 24 hours. Results: All mice were preserved during the experiment without significant complications. There was complete cell death within the IRE lesions without intervening live cells in 2KV after 24 hours. H& E statin and Tunnel stain at 10hr after 2KV IRE ablation revealed more severe apoptotic cell death comparing with control group. Apoptotic index peaks at 10 hours after IRE ablation, and decreases in 24hours. The framework of extracellular matrix and blood vessels were not affected by IRE. Conclusions: The present study demonstrated that IRE ablated colon cancer tissue very effectively through the induction of cellular apoptosis. This study suggests that IRE has the potentiality in treatment of gastrointestinal cancer patients.
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Matsumoto, Tatsuaki, Yuiko Sato, Tami Kobayashi, Kunika Suzuki, Atsushi Kimura, Tomoya Soma, Eri Ito, et al. "Adipose-Derived Stem Cell Sheets Improve Early Biomechanical Graft Strength in Rabbits After Anterior Cruciate Ligament Reconstruction." American Journal of Sports Medicine 49, no. 13 (October 13, 2021): 3508–18. http://dx.doi.org/10.1177/03635465211041582.
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Background: Although various reconstruction techniques are available for anterior cruciate ligament (ACL) injuries, a long recovery time is required before patients return to sports activities, as the reconstructed ACL requires time to regain strength. To date, several studies have reported use of mesenchymal stem cells in orthopaedic surgery; however, no studies have used adipose-derived stem cell (ADSC) sheets in ACL reconstruction (ACLR). Hypothesis: ADSC sheet transplantation can improve biomechanical strength of the autograft used in ACLR. Study Design: Controlled laboratory study. Methods: A total of 68 healthy Japanese white rabbits underwent unilateral ACLR with a semitendinosus tendon autograft after random enrollment into a control group (no sheet; n = 34) and a sheet group (ADSC sheet; n = 34). At 2, 4, 8, 16, and 24 weeks after surgery, rabbits in each group were sacrificed to evaluate tendon-bone healing using histological staining, micro–computed tomography, and biomechanical testing. At 24 weeks, scanning transmission electron microscopy of the graft midsubstance was performed. Results: The ultimate failure load for the control and sheet groups, respectively, was as follows: 17.2 ± 5.5 versus 37.3 ± 10.3 ( P = .01) at 2 weeks, 28.6 ± 1.9 versus 47.4 ± 10.4 ( P = .003) at 4 weeks, 53.0 ± 14.3 versus 48.1 ± 9.3 ( P = .59) at 8 weeks, 66.2 ± 9.3 versus 95.2 ± 43.1 ( P = .24) at 16 weeks, and 66.7 ± 27.3 versus 85.3 ± 29.5 ( P = .39) at 24 weeks. The histological score was also significantly higher in the sheet group compared with the control group at early stages up to 8 weeks. On micro–computed tomography, relative to the control group, the bone tunnel area was significantly narrower in the sheet group at 4 weeks, and the bone volume/tissue volume of the tendon-bone interface was significantly greater at 24 weeks. Scanning transmission electron microscopy at 24 weeks indicated that the mean collagen fiber diameter in the midsubstance was significantly greater, as was the occupation ratio of collagen fibers per field of view, in the sheet group. Conclusion: ADSC sheets improved biomechanical strength, prevented bone tunnel enlargement, and promoted tendon-bone interface healing and graft midsubstance healing in an in vivo rabbit model. Clinical Relevance: ADSC sheets may be useful for early tendon-bone healing and graft maturation in ACLR.
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Matsumoto, Tatsuaki, Yuiko Sato, Tami Kobayashi, Kunika Suzuki, Atsushi Kimura, Tomoya Soma, Eri Ito, et al. "Adipose-Derived Stem Cell Sheets Improve Early Biomechanical Graft Strength in Rabbits After Anterior Cruciate Ligament Reconstruction." American Journal of Sports Medicine 49, no. 13 (October 13, 2021): 3508–18. http://dx.doi.org/10.1177/03635465211041582.
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Background: Although various reconstruction techniques are available for anterior cruciate ligament (ACL) injuries, a long recovery time is required before patients return to sports activities, as the reconstructed ACL requires time to regain strength. To date, several studies have reported use of mesenchymal stem cells in orthopaedic surgery; however, no studies have used adipose-derived stem cell (ADSC) sheets in ACL reconstruction (ACLR). Hypothesis: ADSC sheet transplantation can improve biomechanical strength of the autograft used in ACLR. Study Design: Controlled laboratory study. Methods: A total of 68 healthy Japanese white rabbits underwent unilateral ACLR with a semitendinosus tendon autograft after random enrollment into a control group (no sheet; n = 34) and a sheet group (ADSC sheet; n = 34). At 2, 4, 8, 16, and 24 weeks after surgery, rabbits in each group were sacrificed to evaluate tendon-bone healing using histological staining, micro–computed tomography, and biomechanical testing. At 24 weeks, scanning transmission electron microscopy of the graft midsubstance was performed. Results: The ultimate failure load for the control and sheet groups, respectively, was as follows: 17.2 ± 5.5 versus 37.3 ± 10.3 ( P = .01) at 2 weeks, 28.6 ± 1.9 versus 47.4 ± 10.4 ( P = .003) at 4 weeks, 53.0 ± 14.3 versus 48.1 ± 9.3 ( P = .59) at 8 weeks, 66.2 ± 9.3 versus 95.2 ± 43.1 ( P = .24) at 16 weeks, and 66.7 ± 27.3 versus 85.3 ± 29.5 ( P = .39) at 24 weeks. The histological score was also significantly higher in the sheet group compared with the control group at early stages up to 8 weeks. On micro–computed tomography, relative to the control group, the bone tunnel area was significantly narrower in the sheet group at 4 weeks, and the bone volume/tissue volume of the tendon-bone interface was significantly greater at 24 weeks. Scanning transmission electron microscopy at 24 weeks indicated that the mean collagen fiber diameter in the midsubstance was significantly greater, as was the occupation ratio of collagen fibers per field of view, in the sheet group. Conclusion: ADSC sheets improved biomechanical strength, prevented bone tunnel enlargement, and promoted tendon-bone interface healing and graft midsubstance healing in an in vivo rabbit model. Clinical Relevance: ADSC sheets may be useful for early tendon-bone healing and graft maturation in ACLR.
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Wang, Kai, Liangqun Rong, Xiu’e Wei, and Qingxiu Zhang. "Analysis of Antiapoptosis Effect of Netrin-1 on Ischemic Stroke and Its Molecular Mechanism under Deleted in Colon Cancer/Extracellular Signal-Regulated Kinase Signaling Pathway." BioMed Research International 2020 (November 14, 2020): 1–9. http://dx.doi.org/10.1155/2020/8855949.
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To analyze the regulatory effect of Netrin-1 in ischemic stroke and its influence on Deleted in Colon Cancer (DCC)/Extracellular Signal-regulated Kinase (ERK) signaling pathway, 20 male rats were selected to construct the rat model of middle cerebral artery occlusion (MCAO), 10 normal rats were selected as healthy controls (Normal Saline (NS)), and they were divided into the MCAO+Netrin-1 group, MCAO group, and NS group according to different treatment schemes. The positive expression of Netrin-1 was detected by immunostaining, magnetic resonance imaging (MRI) was adopted to detect the percentage of rat cerebral infarct volume in the cerebral hemispheres, and Modified Neurological Severity Score (mNSS) was adopted to evaluate postoperative neurological function in rats. Besides, a tunnel staining experiment was applied to detect the apoptosis rate of rat neurons, the sticker removal test was applied to evaluate the postoperative sensory function of rats, and fluorescence staining was adopted to detect the expression of DCC and ERK in rats. The results showed that the percentage of cerebral infarction volume in the cerebral hemispheres of the MCAO+Netrin-1 group was higher than that of the MCAO and NS groups ( P < 0.05 ); in the MCAO+Netrin-1 group, the MCAO mNSS scoring and the time spent in the sticker removal test were lower than the MCAO group ( P < 0.05 ); the apoptosis rate of rats in the MCAO+Netrin-1 group was lower than that in the MCAO group ( P < 0.05 ); the average fluorescence intensity of DCC and p-ERK in the MCAO+Netrin-1 group was higher than that in the MCAO group ( P < 0.05 ); the average fluorescence intensity of p-ERK in the MCAO+Netrin-1 group was higher than that in the MCAO group ( P < 0.05 ). In short, Netrin-1 can effectively reduce the brain tissue damage in rats with ischemic stroke, improve the nerve function and sensory function of rats, and inhibit neuronal cell apoptosis. Netrin-1 can promote DCC expression and ERK phosphorylation, and the EPK signaling pathway may be involved in the antiapoptotic effect of Netrin-1.
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Fujiwara, Toshihiro, Ken Matsuda, Tateki Kubo, Koichi Tomita, Ryo Hattori, Takeshi Masuoka, Kenji Yano, and Ko Hosokawa. "Axonal supercharging technique using reverse end-to-side neurorrhaphy in peripheral nerve repair: an experimental study in the rat model." Journal of Neurosurgery 107, no. 4 (October 2007): 821–29. http://dx.doi.org/10.3171/jns-07/10/0821.
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Object In an attempt to improve peripheral nerve repair, the influence of the addition of reverse end-to-side neu-rorrhaphy for an injured peripheral nerve was investigated in the rat sciatic nerve transection model. Methods Twelve Sprague–Dawley rats were divided into two groups (six rats in each group). In Group I, the right sciatic nerve was cut at a point distal to the gluteal notch and repaired using end-to-end neurorrhaphy with four 10-0 nylon epineurial sutures. In Group II, after performing the same procedure as in Group I, the left sciatic nerve was cut distally and passed through a subcutaneous tunnel to the right side. The proximal stump of the left sciatic nerve was coapted to the epineurial window of the right sciatic nerve distal to the injured point in an end-to-side fashion using 10-0 nylon epineurial sutures. The effects were evaluated using analgesimeter recordings for the hind paw, electrophysiological tests, measurement of the muscle contraction force, a double-labeling technique, weight measurement and histological examination of the gastrocnemius muscle, histological examination of the bilateral sciatic nerves, and immunofluorescent staining. Results Results from the many tests used to evaluate the reverse end-to-side neurorrhaphy technique indicated thatfunctional recovery of the denervated target organs was promoted by axonal augmentation. Conclusions The reverse end-to-side neurorrhaphy technique could be useful in peripheral nerve repair.
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Morgan, Hannah L., Isaac Ampong, Nader Eid, Charlène Rouillon, Helen R. Griffiths, and Adam J. Watkins. "Low protein diet and methyl-donor supplements modify testicular physiology in mice." Reproduction 159, no. 5 (May 2020): 627–41. http://dx.doi.org/10.1530/rep-19-0435.
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The link between male diet and sperm quality has received significant investigation. However, the impact diet and dietary supplements have on the testicular environment has been examined to a lesser extent. Here, we establish the impact of a sub-optimal low protein diet (LPD) on testicular morphology, apoptosis and serum fatty acid profiles. Furthermore, we define whether supplementing a LPD with specific methyl donors abrogates any detrimental effects of the LPD. Male C57BL6 mice were fed either a control normal protein diet (NPD; 18% protein; n = 8), an isocaloric LPD (LPD; 9% protein; n = 8) or an LPD supplemented with methyl donors (MD-LPD; choline chloride, betaine, methionine, folic acid, vitamin B12; n = 8) for a minimum of 7 weeks. Analysis of male serum fatty acid profiles by gas chromatography revealed elevated levels of saturated fatty acids and lower levels of mono- and polyunsaturated fatty acids in MD-LPD males when compared to NPD and/or LPD males. Testes of LPD males displayed larger seminiferous tubule cross section area when compared to NPD and MD-LPD males, while MD-LPD tubules displayed a larger luminal area. Furthermore, TUNNEL staining revealed LPD males possessed a reduced number of tubules positive for apoptosis, while gene expression analysis showed MD-LPD testes displayed decreased expression of the pro-apoptotic genes Bax, Csap1 and Fas when compared to NPD males. Finally, testes from MD-LPD males displayed a reduced telomere length but increased telomerase activity. These data reveal the significance of sub-optimal nutrition for paternal metabolic and reproductive physiology.
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Hart, Malcolm B., and Christopher W. Smart. "The 2007 recipient of the Brady Medal: Professor John W. Murray." Journal of Micropalaeontology 27, no. 1 (May 1, 2008): 95–96. http://dx.doi.org/10.1144/jm.27.1.95.
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Abstract. The first recipient of the Brady Medal, which was presented on 7 November 2007, is Professor John W. Murray of the National Oceanographic Centre, University of Southampton. The awardee is singularly appropriate as Henry Bowman Brady studied (mainly benthic) foraminifera. John was also one of the speakers at the first open meeting of the then British Micropalaeontological Group when it met at Sheffield University in March 1971 under the Chairmanship of Professor Leslie R. Moore (one of the founding fathers of the TMS). Other speakers on that day included Charles Downie, Alan Higgins, Keith Gueinn, Eric Robinson, Ron Austin, Geoff Warrington, Gerry Orbell, Malcolm Hart, Geoff Eaton and Mike Boulter.John was born in London in 1937 and was evacuated to the north of England during the Second World War. The family returned south to Worthing in 1953. As a teenager John asked his parents for a biological microscope with which he developed his interest in microscopy (both biological and geological). In 1956 he enrolled on the BSc (Hons) Geology programme at Imperial College (London University) in the first year of which he was introduced to foraminifera by David Carter, who also supervised a final year project on the planktonic foraminifera recovered from one of the early boreholes for the Channel Tunnel Site Investigation. For his PhD research (again supervised by David Carter) John elected to study the benthic foraminifera of Christchurch Harbour in Dorset. This involved regular sampling of the harbour sediments, staining the living foraminifera and taking measurements . . .
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Zamani, M., J. Hassanshahi, M. Soleimani, and F. Zamani. "Neuroprotective effect of olive oil in the hippocampus CA1 neurons following ischemia: Reperfusion in mice." Journal of Neurosciences in Rural Practice 04, no. 02 (April 2013): 164–70. http://dx.doi.org/10.4103/0976-3147.112753.
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ABSTRACT Introduction: Transient global ischemia induces selective, delayed neuronal death of pyramidal neurons in the hippocampal CA1. Oxidative Stress is considered to be involved in a number of human diseases including ischemia. Preliminary studies confirmed reduction of cell death in brain following treatment with antioxidants. Aim: According to this finding, we study the relationship between consumption of olive oil on cell death and memory disorder in brain ischemia. We studied the protective effect of olive oil against ischemia‑reperfusion. Material and Methods: Experimental design includes three groups: Intact (n = 8), ischemic control (n = 8) and treatment groups with olive oil (n = 8). The mice treated with olive oil as pre‑treatment for a week. Then, ischemia induced by common carotid artery ligation and following the reduction of inflammation [a week after ischemia], the mice post‑treated with olive oil. Nissl staining applied for counting necrotic cells in hippocampus CA1. Tunnel kit was used to quantify apoptotic cell death while to short term memory scale, we apply y‑maze and shuttle box tests and for detection the rate of apoptotic and treated cell, we used western blotting test for bax and bcl2 proteins. Results: High rate of apoptosis was seen in ischemic group that significantly associated with short‑term memory loss. Cell death was significantly lower when mice treated with olive oil. The memory test results were adjusted with cell death results and bax and bcl2 expression in all groups’ comparison. Ischemia for 15 min induced cell death in hippocampus with more potent effect on CA1. Conclusion: Olive oil intake significantly reduced cell death and decreased memory loss.
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Chalcev, B., A. Torgashina, E. Sokol, and J. Khvan. "AB1298 AL-AMYLOIDOSIS MIMICKING IgG4-RELATED DISEASE." Annals of the Rheumatic Diseases 81, Suppl 1 (May 23, 2022): 1755.2–1755. http://dx.doi.org/10.1136/annrheumdis-2022-eular.3264.
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BackgroundHematological diseases such as multiple myeloma (MM) and POEMS-syndrome can be accompanied by high serum IgG4 level and thus mimic an IgG4-related disease (IgG4-RD) [1, 2]. However, there are no descriptions of AL-amyloidosis with increased secretion of IgG4 in the literature.Objectivesto present a clinical case of AL-amyloidosis mimicking IgG4-RD.MethodsAt the age of 42, patient S. developed nasal congestion, and 2 years later bilateral symmetrical swelling of submandibular salivary glands developed. Chest CT scan revealed hydrothorax and pericarditis, interstitial changes in both lungs and intrathoracic lymphadenopathy. According to blood tests, ANF, RF and CRP were within normal values, an increase in the serum IgG4 (32.9 g/l) and IgG (34.1 g/l) levels was detected, and the patient was admitted to our clinic with suspicion of IgG4-RD.ResultsOn examination, we noticed a symmetrical enlargement of submandibular salivary glands and macroglossia. The patient also complained of tingling in the fingers, we performed electroneuromyography and revealed bilateral carpal tunnel syndrome. According to the immunofixation of blood and urine proteins, IgG-lambda paraproteinemia (18,3 g/l) and a trace amount of Bence-Jones-lambda protein in the urine were detected. AL-amyloidosis was suspected and a biopsy of the submandibular salivary gland was performed, followed by Congo red staining and darkfield microscopy that confirmed amyloidosis (Figure 1). There were no signs of IgG4-RD (storiform fibrosis or obliterative phlebitis) in the biopsy specimens. Echocardiography revealed thickening of the interventricular septum and hypertrophy of the ventricular myocardium. MM and other malignancies were excluded on the basis of bone marrow trephine biopsy and PET-CT (also revealed severe hepatomegaly). The diagnosis of IgG4-secreting AL-amyloidosis affecting salivary glands, tongue, heart, lungs, liver, nervous system was made and polychemotherapy was started, the patient’s further life remained unknown.Figure 1.Congo red staining and darkfield microscopy demonstrating amyloid deposits.ConclusionAL-amyloidosis can be accompanied by high serum IgG4 level and mimic IgG4-RD. The serum level of IgG4 should not be used as a criterion for IgG4-RD, and in all cases of suspected IgG4-RD the diagnosis should be confirmed morphologically.References[1]Geyer JT, Niesvizky R, Jayabalan DS, et al. IgG4 plasma cell myeloma: new insights into the pathogenesis of IgG4-related disease. Mod Pathol. 2014;27(3):375-381. doi:10.1038/modpathol.2013.159[2]Zheng M, Zhou P, Zheng K, et al. A special subtype of POEMS syndrome: IgG4 subtype. Am J Transl Res. 2016;8(2):588-596. Published 2016 Feb 15.Disclosure of InterestsNone declared
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Wang, Yao, Fan Yang, Fang-Zhou Jiao, Qian Chen, Wen-Bin Zhang, Lu-Wen Wang, and Zuo-Jiong Gong. "Modulations of Histone Deacetylase 2 Offer a Protective Effect through the Mitochondrial Apoptosis Pathway in Acute Liver Failure." Oxidative Medicine and Cellular Longevity 2019 (April 28, 2019): 1–17. http://dx.doi.org/10.1155/2019/8173016.
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The purpose of this study was to investigate the modulation of histone deacetylase 2 (HDAC2) on mitochondrial apoptosis in acute liver failure (ALF). The cellular model was established with LO2 cells stimulated by tumor necrosis factor alpha (TNF-α)/D-galactosamine (D-gal). Rats were administrated by lipopolysaccharide (LPS)/D-gal as animal model. The cell and animal models were then treated by HDAC2 inhibitor CAY10683. HDAC2 was regulated up or down by lentiviral vector transfection in LO2 cells. The mRNA levels of bcl2 and bax were detected by real-time PCR. The protein levels of HDAC2, bcl2, bax, cytochrome c (cyt c) in mitochondrion and cytosol, apoptosis protease activating factor 1 (apaf1), caspase 3, cleaved-caspase 3, caspase 9, cleaved-caspase 9, acetylated histone H3 (AH3), and histone H3 (H3) were assayed by western blot. Apoptosis was detected by flow cytometry. The serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), and total bilirubin (TBIL) levels were also assayed. The openness degree of the mitochondrial permeability transition pore (MPTP) was detected by ultraviolet spectrophotometry. The apoptosis of hepatocytes in liver tissues was determined by tunnel staining. The liver tissue pathology was detected by hematoxylin eosin (HE) staining. The ultrastructure of liver tissue was observed by electron microscopy. Compared with cell and rat model groups, the bax mRNA level was decreased, and bcl2 mRNA was increased in the CAY10683 treatment group. The protein levels of HDAC2, bax, cyt c in cytosol, apaf1, cleaved-caspase 3, and cleaved-caspase 9 were decreased, and the apoptosis rate was decreased (P<0.05), whereas the protein level of bcl2 and cyt c in the mitochondrion was elevated (P<0.05) in the CAY10683 treatment group. In the HDAC2 down- or upregulated LO2 cells, the mitochondrial apoptosis pathway was inhibited or activated, respectively. After being treated with TNF-α/D-gal in HDAC2 down- or upregulated LO2 cells, the mitochondrial apoptosis pathway was further suppressed or activated, respectively. The MPTP value was elevated in CAY10683-treated groups compared with the rat model group (P<0.05). Liver tissue pathological damage and apoptotic index in the CAY10683-treated group were significantly reduced. In addition, AH3 was elevated in both cell and animal model groups (P<0.05). Downregulated or overexpressed HDAC2 could accordingly increase or decrease the AH3 level, and TNF-α/D-gal could enhance the acetylation effect. These results suggested that modulations of histone deacetylase 2 offer a protective effect through the mitochondrial apoptosis pathway in acute liver failure.
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Almatroodi, Saleh A., Faris Alrumaihi, Mohammed A. Alsahli, Mazen Fahad Alhommrani, Arif Khan, and Arshad Husain Rahmani. "Curcumin, an Active Constituent of Turmeric Spice: Implication in the Prevention of Lung Injury Induced by Benzo(a) Pyrene (BaP) in Rats." Molecules 25, no. 3 (February 7, 2020): 724. http://dx.doi.org/10.3390/molecules25030724.
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Benzo(a)pyrene (BaP) is a well-known carcinogen and enhances oxidative stress and apoptosis and also alters several molecular pathways. Curcumin is an active ingredient of Curcuma longa, and it has potent anti-inflammatory, antioxidant activity that defends cells from oxidative stress and cell death. The objectives of the present study were to explore the protective effects of curcumin against long-term administration of BaP induced disturbances in lungs of rats. Male rats were randomly divided into four groups: saline control, BaP only, BaP + curcumin, and curcumin only. Lung histopathology, electron microscopy, inflammatory cytokine release, antioxidant levels, apoptosis, and cell cycle were examined. Instillation of BaP significantly increased infiltration of inflammatory cells in alveolar space and inflammatory cytokine in blood. BaP induced lung tissue alterations including mild bronchitis, scant chronic inflammatory cell infiltrate in the wall of the respiratory bronchiole, and mild intra-alveolar haemorrhage. However, these alterations were found to be significantly less as mild inflammatory cell infiltrate in curcumin plus BaP treated group. Furthermore, electron microscopy results also showed necrotic changes and broken cell membrane of Type-II epithelial cell of alveoli in BaP group, which was reduced after adding curcumin treatment. In addition, we found BaP plus curcumin treatment effectively reduced inflammatory cytokines Tumour Necrosis Factor alpha (TNF-α), Interleukin 6 (IL-6), and C-reactive protein (CRP) levels in blood serum. Moreover, the levels of tunnel staining and p53 expression were significantly increased by BaP, whereas these changes were noticeably modulated after curcumin treatment. BaP also interferes in normal cell cycle, which was significantly improved with curcumin treatment. Overall, our findings suggest that curcumin attenuates BaP -induced lung injury, probably through inhibiting inflammation, oxidative stress and apoptosis in lung epithelial cells, and improving cell proliferation and antioxidants level. Thus, curcumin may be an alternative therapy for improving the outcomes of Benzo(a)pyrene-induced lung injury.
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Ismail, Can, Aksak Karamese Selina, Findik Guvendi Gulname, Can Serpil, Kocaaslan Ramazan, and Unal Bunyami. "Effects of Estrogen and Exercise Stress on Adrenal Glands of Male Rats." International Journal of Medical Science and Clinical Invention 8, no. 06 (June 6, 2021): 5449–57. http://dx.doi.org/10.18535/ijmsci/v8i06.04.
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Consciously done exercises are beneficial for human health. However, unconscious and wrong practices (medication use, etc…) can cause various injuries and permanent damage to the human body. For reasons such as increasing muscle mass and/or to getting more performance, taking steroid hormones disrupts overall body hormone balance. Effects of exercise on oxygen and energy metabolism and estrogen as an exogenous steroid have significant effects on the adrenal gland. The aim of this study was to see the effects of 17b-estradiol on surrenal glands of rats that is put through regular physical exercises.
36 male Sprague-Dawley rats between the weights 220-250 gr, were put into thermal-controlled room with day-night cycles to stimulate an optimal day for the subjects. The experiment was modeled by dividing 36 animals into 6 groups in total according to the control, exercising and estrogen administration criteria. Experimental group animals received daily doses of 10 μg/kg/day 17-β-estradiol during 30 days. Also exercises group animals ran at 20 m/min on a 15% grade for 90 minutes and rest 34 minutes. Then, TUNNEL and Hematoxylen & Eosin staining were performed to measure the damage on the adrenal glands.
In group 2, dense presence of degenerative fibroblasts and inflammatuary cells infiltration in zona fasciculata were significantly different. In group 3, the degenerative areas were significant in all adrenal cortex zones. In group 4, necrotic areas were determined in zona reticularis. In group 5, zona fasciculata was severely degenerated. With group 6, Sinusoidal features were completely lost in zona reticularis. The results strongly show that exercise may affect the zona glomerulase in short time period. As a result, exposure to exercise and exercise stress with external administration of estradiol may cause cellular degeneration especially zona fasciculata and zona reticularis in the adrenal gland.
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Yang, Mei, Wen Jin, Wei Shi, Bo Wang, Qing Li, and Chunfeng Guan. "Effect of α-momordicine on proliferation and apoptosis of liver cancer, and its associated mechanisms of action." Tropical Journal of Pharmaceutical Research 18, no. 9 (June 29, 2021): 1935–41. http://dx.doi.org/10.4314/tjpr.v18i9.22.
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Purpose: To investigate the effect of α-momordicine (α-MMC) on liver cancer cell proliferation and apoptosis, and to elucidate the mechanisms of action involved.
Methods: In in vitro experiments, hepatoma cell lines were used, while nude mice with hepatocellular carcinoma were used for in vivo studies. Cancer cell proliferation was determined using MTT assay while apoptosis was assayed by flow cytometry and TUNNEL staining. Gene expression was determined with real-time polymerase chain reaction (RT-PCR), while protein expression levels were assayed by Western blot, immunohistochemistry and immunofluorescence.
Results: Alpha-MMC decreased HCC cell viability dose-dependently (p < 0.05). In HepG2 cells, G2/M cell cycle was halted after 48 h intervention with 1.24 mg/mL α-MMC. However, at G0/G1 phase, αMMC at doses of 1.06 and 0.92 mg/mL caused cell cycle arrest of HCC-LM3 and SMMC-7721 cells. In vivo studies showed that after establishment of the nude mice liver cancer model, exposure to α-MMC at a dose of 0.70 mg/kg or 2.08mg/kg for 4 weeks reduced the size of liver cancer in the treatment group, relative to control group; mean diameter of liver cancer decreased from 2.16 to 0.51 cm, while mean volume decreased from 1.185 to 0.085 cm3 . Moreover, α-MMC increased apoptosis level in liver cancer tissues in nude mice, and down-regulated the expressions of P-AKT, RAGE, MMP-9 and HMGB1, but upregulated Bax/Bcl2 ratio (p < 0.05).
Conclusion: α-MMC inhibits cancer cell growth and proliferation, and facilitates their apoptosis by positively regulating the ratio of Bax/Bcl2. The anti-liver cancer effect of α-MMC is mediated via HMGB1-RAGE and AKT signaling pathways
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Tegginmani, Veeresh S., Beenarani Goel, Virendra Uppin, LS Vijay Kumar, and Abhinav Nainani. "Comparison of Antibacterial Activity of Glass-ionomer Cement and Amalgam in Class Two Restorations by Streptococcus mutans Count Analysis at Fixed Intervals: An in vivo Study." Journal of Contemporary Dental Practice 14, no. 3 (2013): 381–86. http://dx.doi.org/10.5005/jp-journals-10024-1332.
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ABSTRACT Aim The purpose of the present study was to determine the influence of glass ionomer cement and amalgam restoration on the level of Streptococcus mutans in the interproximal plaque at periodic intervals and also to compare these values. Materials and methods Seventeen adult patients having two proximal carious lesions on any quadrant of the jaw (either opposing or contralateral) were selected for this study. Carious lesions were diagnosed clinically and from bitewing radiographs. Of the two carious lesions, one was restored with glass ionomer cermet cement and another with amalgam. Plaque samples were collected from interproximal areas before and at 1 month and 3 months post-treatment in a test tube containing 5 ml of modified Stuart's liquid transport fluid. Identification of organisms in the colony was done after Gram staining. Results Comparison of values before restoration and after restoration at 1 month interval showed a statistically significant decrease (p < 0.001). Similarly, comparison of values before and after restorations at 3 months also showed statistically significant decrease (p < 0.02). But comparison of restorations of 1 and 3 months intervals showed no statistical significant difference (p > 0.05). Conclusion Glass ionomer restorations have definite advantage over the amalgam, as the tunnel preparation is more conservative and fluoride release from the glass ionomer inhibits the growth of S. mutans in the plaque. Clinical significance Glass ionomer cement should be preferred over amalgam in conservatively prepared restorations as it reduces the microbial activities due to fluoride release. How to cite this article Tegginmani VS, Goel B, Uppin V, Horatti P, Kumar LSV, Nainani A. Comparison of Antibacterial Activity of Glass-ionomer Cement and Amalgam in Class Two Restorations by Streptococcus mutans Count Analysis at Fixed Intervals: An in vivo Study. J Contemp Dent Pract 2013;14(3):381-386.
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Kasibhatla, Shailaja, Gustavo P. Amarante-Mendes, Deborah Finucane, Thomas Brunner, Ella Bossy-Wetzel, and Douglas R. Green. "TUNEL Staining of Adherent Cells to Detect Apoptosis." Cold Spring Harbor Protocols 2006, no. 1 (January 1, 2006): pdb.prot4433. http://dx.doi.org/10.1101/pdb.prot4433.
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Kasibhatla, Shailaja, Gustavo P. Amarante-Mendes, Deborah Finucane, Thomas Brunner, Ella Bossy-Wetzel, and Douglas R. Green. "TUNEL Staining of Tissue Sections to Detect Apoptosis." Cold Spring Harbor Protocols 2006, no. 2 (July 2006): pdb.prot4496. http://dx.doi.org/10.1101/pdb.prot4496.
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Austgulen, Rigmor, Lisa Chedwick, Christina Vogt Isaksen, Lars Vatten, and Catherine Craven. "Trophoblast Apoptosis in Human Placenta at Term as Detected by Expression of a Cytokeratin 18 Degradation Product of Caspase." Archives of Pathology & Laboratory Medicine 126, no. 12 (December 1, 2002): 1480–86. http://dx.doi.org/10.5858/2002-126-1480-taihpa.
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Abstract Context.—Apoptosis occurs in the normal placenta. The monoclonal antibody M30 is directed against a novel epitope of cytokeratin 18 (CK18) that is formed by caspase cleavage early in the apoptotic cascade, and this antibody may therefore be useful for evaluating trophoblast apoptosis. Objective.—We undertook the present study to evaluate the use of monoclonal antibody M30 to assess trophoblast apoptosis in placenta at term. Methods.—We stained paraffin-embedded placental tissues from 15 deliveries at term with M30. We compared positive M30 staining and CK18 staining (as detected by a monoclonal antibody directed against CK18) of trophoblasts in serial slides. We also compared apoptotic rates as detected by M30 and TUNEL (terminal deoxynucleotidyl transferase–mediated deoxyuridine triphosphate nick end labeling) in 7 of the placentas. Results.—In fields of villous tissue, most M30-positive cells were CK18-positive syncytiotrophoblasts. Approximately half of M30-positive cells occurred as focal positive staining in the syncytial layer, and half occurred as abundant staining of syncytiotrophoblasts in areas with increased intervillous or perivillous fibrinoid. We found very few M30-positive cells in villous stroma. In decidual/basal plate tissues, most (two thirds) of the M30-positive cells were CK18-positive extravillous trophoblasts, whereas one third were syncytiotrophoblasts of anchoring villi. Since TUNEL detects apoptosis in both epithelial and nonepithelial cells, more cells were positively stained with TUNEL than with M30 in some tissue fields. However, our observations suggest that M30 was more sensitive than TUNEL in recognizing apoptotic trophoblasts and had less nonspecific staining than TUNEL. Conclusion.—We recommend the use of monoclonal antibody M30 for apoptosis studies in placental tissues. This antibody is easy to handle, the staining obtained seems specific, and the nonspecific staining seems negligible.
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Lu, Cheng-Chang, Cheng-Jung Ho, Hsuan-Ti Huang, Sung-Yen Lin, Shih-Hsiang Chou, Pei-Hsi Chou, Mei-Ling Ho, and Yin-Chun Tien. "Effect of Freshly Isolated Bone Marrow Mononuclear Cells and Cultured Bone Marrow Stromal Cells in Graft Cell Repopulation and Tendon-Bone Healing after Allograft Anterior Cruciate Ligament Reconstruction." International Journal of Molecular Sciences 22, no. 6 (March 10, 2021): 2791. http://dx.doi.org/10.3390/ijms22062791.
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Graft cell repopulation and tendon-bone tunnel healing are important after allograft anterior cruciate ligament reconstruction (ACLR). Freshly isolated bone marrow mononuclear cells (BMMNCs) have the advantage of short isolation time during surgery and may enhance tissue regeneration. Thus, we hypothesized that the effect of intra-articular BMMNCs in post-allograft ACLR treatment is comparable to that of cultured bone marrow stromal cells (BMSCs). A rabbit model of hamstring allograft ACLR was used in this study. Animals were randomly assigned to the BMMNC, BMSC, and control groups. Fresh BMMNCs isolated from the iliac crest during surgery and cultured BMSCs at passage four were used in this study. A total of 1 × 107 BMMNCs or BMSCs in 100 µL phosphate-buffered saline were injected into the knee joint immediately after ACLR. The control group was not injected with cells. At two and six weeks post operation, we assessed graft cell repopulation with histological and cell tracking staining (PKH26), and tendon-bone healing with histological micro-computed tomography and immunohistochemical analyses for collagen I and monocyte chemoattractant protein-1 (MCP1). At two weeks post operation, there was no significant difference in the total cell population within the allograft among the three groups. However, the control group showed significantly higher cell population within the allograft than that of BM cell groups at six weeks. Histological examination of proximal tibia revealed that the intra-articular delivered cells infiltrated into the tendon-bone interface. Compared to the control group, the BM cell groups showed broader gaps with interfacial fibrocartilage healing, similar collagen I level, and higher MCP1 expression in the early stage. Micro-CT did not reveal any significant difference among the three groups. BMMNCs and BMSCs had comparable effects on cell repopulation and interfacial allograft-bone healing. Intra-articular BM cells delivery had limited benefits on graft cell repopulation and caused higher inflammation than that in the control group in the early stage, with fibrocartilage formation in the tendon-bone interface after allograft ACLR.
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Zolotukhin, Irene, Brett Palaschak, David M. Markusic, and Roland Herzog. "Clotting Factor VIII Overexpression Shows Signs of ER Stress but Does Not Cause Toxicity upon Gene Transfer." Blood 126, no. 23 (December 3, 2015): 3238. http://dx.doi.org/10.1182/blood.v126.23.3238.3238.
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Abstract Hemophilia A is the X-linked bleeding disorder resulting from the loss of functional clotting factor VIII (FVIII). Hemophilia A patients with severe disease (<1% residual FVIII activity) experience spontaneous bleeds into the joints and closed spaces with severe morbidity. Restoration of hemostasis is managed by repeated infusions (2-3 times per week) of plasma derived or recombinant FVIII protein. While a standard treatment is available for patients, life-long infusions of FVIII protein is very expensive, has a negative impact on the patient's quality of life, and FVIII protein products are not available worldwide. Hence, there is a need to develop a more robust and cost effective treatment for hemophilia A patients. Liver-directed gene therapy using adeno-associated virus (AAV) represents a promising approach to treat hemophilia A. However, previous studies have shown that overexpression of human FVIII protein in the context of hydrodynamic delivery of plasmid vectors induces ER stress mediated through the unfolded protein response (UPR). Because human FVIII protein is inefficiently secreted into circulation, high AAV vector doses will be required to obtain therapeutic expression levels. Therefore, we sought to determine if AAV-FVIII gene delivery also triggers cellular UPR in hepatocytes in vivo. To this end, we selected to use a codon-optimized FVIII cDNA, which has been shown by our lab to significantly increase FVIII protein expression, and a high vector dose of AAV8-ApoE-hAAT-cohF8, containing a hepatocyte-specific enhancer/promoter combination. We evaluated this vector at doses of 1 x 1011 vg (4x1012 vg/kg) and 1 x 1012 vg (4x1013 vg/kg) in hemophilia A mice on a 129/BL6 background. Intravenous administration of the highest vector dose completely restored hemostasis, which was sustained and achieved super-physiological levels in some animals. Importantly, none of these mice developed inhibitors against FVIII. Next, we administered the vector at the same 2 doses to C67BL/6 mice, which show higher hepatic transduction efficiency than other strains. Experimental controls consisted of mice, with no vector or 1x1012 vg of AAV8-ApoE-hAAT-F9, expressing human factor IX (FIX) protein. Injection of tunicamycin, a potent inducer of the ER stress response, served as a positive control for all assays. Vector-treated mice were studied 2 and 4 weeks after gene transfer (n=3-4 per group). First, we evaluated the status of key molecular chaperones, known to be the mediators of the UPR: Bip, p-PERK, and p-eIF2a. Western blotting performed on the liver lysates indicated modest up-regulation of all three markers compared to normal control, but that effect was neither dose nor gene dependent. In addition, we tested the splicing of Xbp1 mRNA by PCR assay and observed low level of the 26 bp spliced fragment, indicative of the UPR, at the high vector dose. Immunohistochemistry on liver sections from each of our experimental groups including H&E staining, Tunnel staining for apoptotic cells, and reactive oxygen species staining. None of the stains yielded evidence for liver damage even in the 1x1012 AAV8-cohF8 treated mice compared to untreated controls. There was also no elevation of liver enzyme levels in plasma samples. Analysis of plasma from vector injected mice showed systemic levels of human FVIII and FIX proteins at ~30 ng/ml and ~6300 ng/ml, respectively (these ELISA-based measurements likely underestimate FVIII levels due to interference by von Willebrand factor). Overall our results suggest that over-expression of coagulation factors in hepatocytes from AAV vectors causes a mild cell stress response that is not strong enough to cause liver toxicity, is not specific for FVIII, and does impact expression or immunogenicity. Disclosures No relevant conflicts of interest to declare.
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McCall, Cordero L., James Stinson, Ryan W. Dobbs, Neil Mistry, Adrian Rosenberg, Oluwarotimi Nettey, Pooja Sharma, et al. "Abstract A079: Genetic ancestry and vitamin D may predict degree of prostatic apoptosis in prostate tumor and benign epithelium among men undergoing radical prostatectomy." Cancer Epidemiology, Biomarkers & Prevention 32, no. 1_Supplement (January 1, 2023): A079. http://dx.doi.org/10.1158/1538-7755.disp22-a079.
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Abstract There is evidence that vitamin D metabolites such as 25 hydroxyvitamin D [25(OH)D], may be protective against prostate cancer (PCa). Vitamin D and its analogs are hypothesized to have pro-apoptotic effects in multiple cell lines. Blacks have higher rates of vitamin D deficiency, higher tumor progression rates 1,2, and higher apoptosis rates3 compared to Whites. West African ancestry (WAA) may confound this relationship if it is associated with both vitamin D status and apoptosis rates. Our objective is to assess for independent associations between in vivo vitamin D status, genetic ancestry, and degree of apoptosis using prostatic epithelial Terminal deoxynucleotide transferase dUTP Nick End Labeling (TUNEL) staining. Radical prostatectomy specimens of men with clinically localized PCa were stained for TUNEL. Prostatic and serum levels of 25(OH)D and serum levels of 1,25 hydroxyvitamin D were assessed at the time of surgery. Ancestry informative markers were used to estimate the percentage of genetic West African, Native American, and European ancestry. Continuous variables were compared using medians tests across three groups of vitamin D status. Categorical variables were compared using the Chi-Squared test. Correlations between the degree of TUNEL staining and vitamin D metabolites were assessed using Spearman correlations. Linear regressions were used to access the bivariant associations between tumor and benign epithelial TUNEL staining with quartiles of ancestry, and serum and prostatic vitamin D metabolite levels. Multivariate linear regressions for the percentage of benign and tumor TUNEL staining were used to test the independent associations of vitamin D metabolites and genetic ancestry. Overall, 121 men, age 40-79, who underwent radical prostatectomy were enrolled between 2013-2018. Serum 25(OH)D correlated positively with both tumor (ρ = 0.17, p = 0.03), and benign (ρ = 0.16, p = 0.04) prostatic epithelial TUNEL staining. Similarly, prostatic 25(OH)D correlated positively with both tumor (ρ = 0.31, p < 0.001) and benign (ρ = 0.20, p = 0.03) prostatic tissue TUNEL staining. Relative to West African ancestry, Native American ancestry was more strongly positively correlated with tumor (ρ = 0.22, p = 0.05) and benign (ρ = 0.27, p= 0.02) TUNEL staining. In multivariate regression models, increasing quartiles of prostatic 25(OH)D (β = 0.25, p = 0.04) and Native American ancestry (β = 0.327, p = 0.004) were significant predictors of tumor TUNEL staining. Physiologic levels of serum and prostatic 25(OH)D and percentage of Native American ancestry are positively associated with the degree of apoptosis in tumor and benign prostatic epithelium from men with clinically localized PCa. Vitamin D may have secondary chemoprevention benefits in preventing PCa progression in localized disease. Citation Format: Cordero L. McCall, James Stinson, Ryan W. Dobbs, Neil Mistry, Adrian Rosenberg, Oluwarotimi Nettey, Pooja Sharma, Michael Dixon, Jamila Sweis, Virgilia Macias, Roohollah Sharifi, Rick A. Kittles, Andre Kajdacsy Balla, Adam B. Murphy. Genetic ancestry and vitamin D may predict degree of prostatic apoptosis in prostate tumor and benign epithelium among men undergoing radical prostatectomy [abstract]. In: Proceedings of the 15th AACR Conference on the Science of Cancer Health Disparities in Racial/Ethnic Minorities and the Medically Underserved; 2022 Sep 16-19; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2022;31(1 Suppl):Abstract nr A079.
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Sakic, Boris, Irina Maric, Paulo D. Koeberle, Jason M. Millward, Henry Szechtman, Dragan Maric, and Judah A. Denburg. "Increased TUNEL staining in brains of autoimmune Fas-deficient mice." Journal of Neuroimmunology 104, no. 2 (May 2000): 147–54. http://dx.doi.org/10.1016/s0165-5728(99)00277-5.
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