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        A novel low-cost method for Mycobacterium avium subsp. paratuberculosis DNA extraction from an automated broth culture system for real-time PCR analysis

        Miguel Salgado,Cristobal Verdugo,Cord Heuer,Pedro Castillo,Patricia Zamorano 대한수의학회 2014 Journal of Veterinary Science Vol.15 No.2

        PCR is a highly accurate technique for confirming thepresence of Mycobacterium avium subsp. paratuberculosis(Map) in broth culture. In this study, a simple, efficient, andlow-cost method of harvesting DNA from Map cultured inliquid medium was developed. The proposed protocol(Universidad Austral de Chile [UACH]) was evaluated bycomparing its performance to that of two traditionaltechniques (a QIAamp DNA Stool Mini Kit andcethyltrimethylammonium bromide [CTAB] method). Theresults were statistically assessed by agreement analysis forwhich differences in the number of cycles to positive (CP)were compared by Student’s t-test for paired samples andregression analysis. Twelve out of 104 fecal pools culturedwere positive. The final PCR results for 11 samples analyzedwith the QIAamp and UACH methods or ones examinedwith the QIAamp and CTAB methods were in agreement. Complete (100%) agreement was observed between datafrom the CTAB and UACH methods. CP values for theUACH and CTAB techniques were not significantlydifferent, while the UACH method yielded significantlylower CP values compared to the QIAamp kit. The proposedextraction method combines reliability and efficiency withsimplicity and lower cost.

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        Experimental Tracheal Replacement: Angiogenesis and Null Apoptosis Promote Stenosis

        J. Alfredo Santibáñez-Salgado,Avelina Sotres-Vega,Miguel O. Gaxiola-Gaxiola,,Jaime Villalba-Caloca,Karen Bobadilla Lozoya,Joaquín A. Zúñiga-Ramos 대한흉부외과학회 2021 Journal of Chest Surgery (J Chest Surg) Vol.54 No.3

        Background: Tracheal replacement is a challenge for thoracic surgeons due to stenosis in the trachea-prosthesis anastomosis. We propose that stenosis occurs due to fibrosis as a result of an abnormal healing process, characterized by an increased expression of wound healing growth factors (vascular endothelial growth factor [VEGF], survivin, and CD31), which promote angiogenesis and decrease apoptosis. We analyzed the immunoreactivity of VEGF, survivin, CD31, and caspase-3 in the development of fibrotic stenosis in prosthetic tracheal replacement. Methods: Fourteen dogs were operated on: group I (n=7) received a 6-ring cervical tra- cheal segment autograft, while in group II (n=7), a 6-ring segment of the cervical trachea was resected and tracheal continuity was restored with a Dacron prosthesis. The follow-up was 3 months. Immunoreactivity studies for VEGF, survivin, CD31, and caspase-3 were per- formed. A statistical analysis was done using the Wilcoxon signed rank test. Results: Four animals in group I were euthanized on the 10th postoperative day due to autograft necrosis. Three animals completed the study without anastomotic stenosis. Moderate expression of VEGF (p=0.038), survivin (p=0.038), and CD31 (p=0.038) was found. All group II animals developed stenosis in the trachea-prosthesis anastomotic sites. Micros- copy showed abundant collagen and neovascularization vessels. Statistically significant immunoreactive expression of VEGF (p=0.015), survivin (p=0.017), and CD31 (p=0.011) was observed. No expression of caspase-3 was found. Conclusion: We found a strong correlation between fibrosis in trachea-prosthesis anas- tomoses and excessive angiogenesis, moderate to intense VEGF, CD31, and survivin ex- pression, and null apoptotic activity. These factors led to uncontrolled collagen production.

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        Eryngium carlinae Ethanol Extract Corrects Lipid Abnormalities in Wistar Rats with Experimental Diabetes

        Ruth Noriega-Cisneros,Donovan J. Pena-Montes,Maribel Huerta-Cervantes,Rafael Torres-Martınez,Miguel Huerta,Salvador Manzo-Avalos,Rafael Salgado-Garciglia,Alfredo Saavedra-Molina 한국식품영양과학회 2020 Journal of medicinal food Vol.23 No.8

        Abnormalities in lipid metabolism, associated with increased risk of cardiovascular disease (CVD), frequently occur in people with diabetes. Eryngium carlinae is a plant used in traditional medicine to treat lipid abnormalities. The chemical composition and hypolipidemic activity of the ethanolic extract of E. carlinae were analyzed to broaden our knowledge of its mechanism of action. The ethanolic extract of E. carlinae was tested for hypolipidemic activity by oral administration for 40 days. Atorvastatin, a widely used statin, was also administered to compare its effect with that of the extract. Serum was used for analysis of the lipid profile and liver microsomes to assess 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase activity and low-density lipoprotein receptor (LDL-r) levels. The extract was able to reduce total cholesterol and non-high-density lipoprotein cholesterol (C-HDL) levels and increase the C-HDL levels reduced in diabetes, decreasing the atherogenic index and therefore the risk of suffering CVD at the same level as atorvastatin. The HMG-CoA reductase activity and LDL-r levels were not modified by the administration of E. carlinae. The results demonstrate the hypolipidemic potential of ethanol extract of E. carlinae and support its use in traditional medicine as a hypolipidemic agent.

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