Novel Apoplastic Antifreeze Proteins of Deschampsia antarctica as Enhancer of Common Cell Freezing Media for Cryobanking of Genetic Resources, a Preliminary Study.

Antifreeze proteins (AFPs) are natural biomolecules found in cold-adapted organisms that lower the freezing point of water, allowing survival in icy conditions. These proteins have the potential to improve cryopreservation techniques by enhancing the quality of genetic material postthaw. Deschampsia antarctica, a freezing-tolerant plant, possesses AFPs and is a promising candidate for cryopreservation applications. In this study, we investigated the cryoprotective properties of AFPs from D. antarctica extracts on Atlantic salmon spermatozoa. Apoplastic extracts were used to determine ice recrystallization inhibition (IRI), thermal hysteresis (TH) activities and ice crystal morphology. Spermatozoa were cryopreserved using a standard cryoprotectant medium (C+) and three alternative media supplemented with apoplastic extracts. Flow cytometry was employed to measure plasma membrane integrity (PMI) and mitochondrial membrane potential (MMP) postthaw. Results showed that a low concentration of AFPs (0.05 mg/mL) provided significant IRI activity. Apoplastic extracts from D. antarctica demonstrated a cryoprotective effect on salmon spermatozoa, with PMI comparable to the standard medium. Moreover, samples treated with apoplastic extracts exhibited a higher percentage of cells with high MMP. These findings represent the first and preliminary report that suggests that AFPs derived from apoplastic extracts of D. antarctica have the potential to serve as cryoprotectants and could allow the development of novel freezing media.

Preparation and extraction of chorion proteins from Salmo salar embryos at the pigmented eye stage for electrophoresis with SDS-polyacrylamide gel.

The chorion fulfills important functions in fish embryos, including protecting the embryo during development. The characterization of the protein profile of this envelope could be used as a bioindicator in the evaluation of the quality of embryonic development. The object of this work was to validate a standardized protocol for protein extraction from chorion of Salmo salar embryos at 280 accumulated thermal units (ATU) by comparing and combining existing methods. The protocol consists of consecutive washing of the chorion samples followed by protein extraction with the solution that was named SDS solution (Tris-HCl 100 mM (pH 8), Urea 8 M, 1% SDS, ß-mercaptoethanol 300 mM and EGTA 10 Mm, and 1% protease inhibitor cocktail) and mechanical methods. Protein extraction is enhanced by a working temperature of 75 °C and use of a disperser. The protein concentration was quantified by Bradford Assay. After extraction, the samples were diluted (dilution factor 10) before reading against the calibration curve. After gel electrophoresis with a load of 3 µg of protein, staining showed more than 4 bands, with molecular weights between 25 kDa and 180 kDa.•The protein profile of fish chorion was between 25 kDa and 180 kDa.•Solution containing 1% SDS allows a higher extraction of proteins from the chorion of Atlantic salmon embryos with 280 ATU.•Chorion protein identification is a valuable tool in determining gamete and embryo quality in fish.

Delayed hemorrhagic gastritis caused by immunotherapy in a patient With metastatic melanoma.

Pembrolizumab is a monoclonal antibody which targets the programmed cell death protein 1 (PD-1) receptor of lymphocytes. It is commonly used to treat many types of malignancies. Immunotherapy-related adverse events are relatively common and include pneumonitis, colitis and hepatitis. A rare side effect of immunotherapy is gastrointestinal (GI) bleeding secondary to hemorrhagic gastritis. Side effects from immunotherapy most commonly occur eight to twelve weeks after initiation of therapy but can vary from days after the first dose to even months later. We present a rare case of a patient with metastatic melanoma who had confirmed immune-mediated hemorrhagic gastritis which occurred after 23 cycles of Pembrolizumab. Biopsies for Heliobacter Pylori (H. pylori) and cytomegalovirus (CMV) were negative. The patient's immunotherapy was discontinued, and he was started on high dose steroids. The symptoms (nausea, vomiting, and abdominal pain) improved dramatically with a long steroid taper. An esophagogastroduodenoscopy (EGD) performed three months after hospital discharge showed improvement in gastric mucosa, but biopsies continued to show evidence of acute and chronic gastritis. As cancer patients continue to live longer with immunotherapy, it is important for all providers to be aware of the less common side effects of newer agents such as pembrolizumab.

Malnutrition and Alcohol-Associated Hepatitis.

Malnutrition is common in alcohol-associated hepatitis (AH); almost all patients with severe AH have some component of malnutrition. The classic phenotype of malnutrition in AH is sarcopenia, but this has become more difficult to discern clinically as patients have become more obese. Patients with AH are often drinking 10 to 15 standard drinks per day. This substantial alcohol consumption becomes a major source of calories, but these are considered "empty" calories that contain little nutritional value. Malnutrition is associated with liver complications, such as hepatic encephalopathy, and worse liver outcomes. Nutrition support can improve nutrition status and reduce complications.

Occlusal trauma is associated with periodontitis: A retrospective case-control study.

BACKGROUND: Excessive occlusal forces and occlusal trauma have been implicated as co-destructive factors for periodontitis in animal models. The objective of this study was to assess the association between occlusal trauma and periodontitis. METHODS: A total of 167 cases and 205 controls were included for analysis. Occlusal trauma was the exposures of interest and was registered for cases and controls. Additional clinical co-variables were registered as well for further analysis. The association of the exposure and periodontitis was determined by the odds ratio and logistic regression analysis adjusted for confounders. RESULTS: Amalgam restorations, pathogenic occlusion and occlusal trauma were more frequent in cases as compared to controls (P ≤ 0.05). The distribution of pathogenic occlusion and occlusal trauma was similar according to the stage and class of periodontitis. Logistic regression analysis showed a significant association for cigarette smoking, systemic conditions, amalgam restorations, pathogenic occlusion, and occlusal trauma with periodontitis (P ≤ 0.05). After adjusting for confounding variables in the model, amalgam restorations and occlusal trauma remained strongly associated with periodontitis (P ≤ 0.05). CONCLUSIONS: Occlusal trauma is strongly associated with periodontitis. Additional long-term prospective studies are necessary to further understand the impact of the occlusal condition and periodontitis.

Association between traumatic occlusal forces and periodontitis: A systematic review.

OBJECTIVE: Occlusal adjustment is commonly recommended for patients with periodontitis and traumatic occlusion. The objective of this systematic review was to analyze available evidence for the association between traumatic occlusal forces and periodontitis. METHODS: Two focused questions were proposed: What is the effect of traumatic occlusal forces on periodontal parameters in patients with and without periodontitis? And what is the effect of occlusal interventions on periodontal parameters in patients with periodontitis? A systematic review of clinical and observational studies was performed and presented in narrative form. RESULTS: After title and abstract review a total of 30 articles were retrieved and of these 14 full-text articles were retrieved for analysis. Two RCTs, 1 cohort, 4 retrospective and 7 cross-sectional studies were included. Cross-sectional studies reported a significant association between occlusal discrepancies and probing depth and clinical attachment level. However, the magnitude of the effect is negligible when groups with and without occlusal discrepancies are compared. Intervention studies reported a minimal effect on probing depth and clinical attachment level after occlusal adjustment in patients with periodontitis as compared to teeth without occlusal adjustment. CONCLUSIONS: Available human studies showed that there is limited evidence that traumatic occlusion is associated with periodontitis and to support the implementation of occlusal adjustment to significantly improve the periodontal condition in patients with periodontitis.

Reproductive failures associated with Trypanosoma (Duttonella) vivax.

Trypanosomosis is a disease of a high economic impact in different productive livestock systems, especially in Africa and South America; cattle, sheep, goats, horses and donkeys can be prone to infection. Trypanosoma vivax is a blood parasite normally associated with a case of anemia and persistent febrile status, which causes a reduction in production, weakness of affected animals and occasionally death. However, the reproductive disorders which include abortion, disruption of the estrous cycle, placental retention, perinatal mortality, among others, can be very severe, especially when they occur during the last third of gestation. In addition, the transplacental transmission has been proved in this agent with serious consequences in the fetus. Additionally, the infection reduces the fertilizing capacity of the spermatozoa by degenerative effects at a testicular and epididymal level. These reproductive alterations can lead to temporal or permanent infertility problems and in some cases to sterility. The capacity of T. vivax to evade the host's immune system, allows it to persist in animals for months or years. The development of diagnostic tools is very important in order to increase the sensibility and specificity in parasite detection, but it is of paramount importance to prepare field professionals to consider this agent in the differential diagnosis of reproductive failure. The rational and strategic use of the therapeutic drugs will enable to use them in the future, thus avoiding resistance problems.

GABA induction of the Saccharomyces cerevisiae UGA4 gene depends on the quality of the carbon source: role of the key transcription factors acting in this process.

Yeast cells are able to adapt their metabolism according to the quality of both carbon and nitrogen sources available in the environment. Saccharomyces cerevisiae UGA4 gene encodes a permease capable of transporting γ-aminobutyric acid (GABA) into the cells. Yeast uses this amino acid as a nitrogen source or as a carbon skeleton that enters the tricarboxylic acid cycle. The quality of the carbon source modulates UGA4 expression through two parallel pathways, each one acting on different regulatory elements, the UAS(GATA) and the UAS(GABA). In the presence of a fermentable carbon source, UGA4 expression is induced by GABA while in the presence of a non-fermentable carbon source this expression is GABA-independent. The aim of this work was to study the mechanisms responsible for the differences in the profiles of UGA4 expression in both growth conditions. We found that although the subcellular localization of Gln3 depends on the carbon source and UGA4 expression depends on Tor1 and Snf1, Gln3 localization does not depend on these kinases. We also found that the phosphorylation of Gln3 is mediated by two systems activated by a non-fermentable carbon source, involving the Snf1 kinase and an unidentified TORC1-regulated kinase. We also found that the activity of the main transcription factors responsible for UGA4 induction by GABA varies depending on the quality of the carbon source. In a fermentable carbon source such as glucose, the negative GATA factor Dal80 binds to UGA4 promoter; only after the addition of the inducer, the positive factors Uga3, Dal81 and Gln3 interact with the promoter removing Dal80 and leading to gene induction. In contrast, in the non-fermentable carbon source acetate the negative GATA factor remains bound to UGA4 promoter in the presence or absence of GABA, the positive factors are not detected bound in any of these conditions and in consequence, UGA4 is not induced.

Dosis util de la yodo-povidona in vitro; estudio experimental / Useful dose of povidone-iodine in vitro; experimental study

Se estudió la máxima dilución útil de yodo-povidona contra diversas especies bacterianas con diluciones en agua destilada de 1/2000 a solución pura (al 10%). Se efectuaron pruebas contra cepas hospitalarias de Escherichia coli, Pseudomona aeruginosa, Klebsiella ozanae, Citrobacter cloacae, Shigella sonnei, Clostridium y un bacilo anaerobio gram negativo no tipificado. Resultaron eficaces las diluciones al 2% ó mayores. No hubo desarrollo en las pruebas con suero humano al 10 y 20% y la dilución al 2,5% del antiséptico. Para comprobar la tolerancia se inyectaron 40 pollos y 7 ratas Wistar a una dosis de 10 cc/Kg de la dilución al 2,5% equivalente a 25 mg/Kg de yodo, con buena tolerancia y sin hallazgos patológicos en las necropsias efectuadas.

Dosis util de la yodo-povidona in vitro; estudio experimental / Useful dose of povidone-iodine in vitro; experimental study

Se estudió la máxima dilución útil de yodo-povidona contra diversas especies bacterianas con diluciones en agua destilada de 1/2000 a solución pura (al 10%). Se efectuaron pruebas contra cepas hospitalarias de Escherichia coli, Pseudomona aeruginosa, Klebsiella ozanae, Citrobacter cloacae, Shigella sonnei, Clostridium y un bacilo anaerobio gram negativo no tipificado. Resultaron eficaces las diluciones al 2% ó mayores. No hubo desarrollo en las pruebas con suero humano al 10 y 20% y la dilución al 2,5% del antiséptico. Para comprobar la tolerancia se inyectaron 40 pollos y 7 ratas Wistar a una dosis de 10 cc/Kg de la dilución al 2,5% equivalente a 25 mg/Kg de yodo, con buena tolerancia y sin hallazgos patológicos en las necropsias efectuadas. (AU)

Evaluación experimental de un método de mantenimiento de la asepsia en el colon durante el postoperatorio / Experimental evaluation of a method of maintainance of colonic asepsis during postoperative period

Se evaluaron los efectos bacteriológicos y sistémicos de la utilización de una solución de yodo-povidona al 5% durante 24 hs en goteo continuo intracolónico en 18 perros a los que se había efectuado una anastomosis. El método dio como resultado una disminución muy importante de los recuentos con cultivos estériles en un 71,64% para germenes aerobios y 89,32% para anaerobios, con diferencias significativas con respecto a sus controles con solución fisiológica y una serie tratada con cloranfenicol y gentamicina endovenosos. Se encontró una elevada excresión de yodo libre en orina. Los dosajes hormonales no demostraron variación de la tirotrofina, mientras que hubo descenso transitorio de T3 y T4. Se requerirá más investigación para precisar el tiempo que se deberá mantener el goteo

Evaluación experimental de un método de mantenimiento de la asepsia en el colon durante el postoperatorio / Experimental evaluation of a method of maintainance of colonic asepsis during postoperative period

Se evaluaron los efectos bacteriológicos y sistémicos de la utilización de una solución de yodo-povidona al 5% durante 24 hs en goteo continuo intracolónico en 18 perros a los que se había efectuado una anastomosis. El método dio como resultado una disminución muy importante de los recuentos con cultivos estériles en un 71,64% para germenes aerobios y 89,32% para anaerobios, con diferencias significativas con respecto a sus controles con solución fisiológica y una serie tratada con cloranfenicol y gentamicina endovenosos. Se encontró una elevada excresión de yodo libre en orina. Los dosajes hormonales no demostraron variación de la tirotrofina, mientras que hubo descenso transitorio de T3 y T4. Se requerirá más investigación para precisar el tiempo que se deberá mantener el goteo (AU)

Toxicidad de la yodo-povidona intraperitoneal: estudio experimental / Toxicity of the intraperitoneal iodo-povidone

Se instiló una solución al 10% de yodo-povidona en la cavidad abdominal de diversas especies de animales. En todas se demostró un efecto letal en breve plazo con dosis entre 10 y 40 cc/kg. Se obtuvo dosis letal-50 en ratones C57-CFW que correspondió a 5 cc/kg. Se concluye que la yodo-povidona por vía intraperitoneal es tóxica y que su uso debe ser limitado a cavidades bloqueadas. Se requerirán más estudios para determinar su utilidad e inocuidad en lavados peritoneales de toda la cavidad

Toxicidad de la yodo-povidona intraperitoneal: estudio experimental / Toxicity of the intraperitoneal iodo-povidone

Se instiló una solución al 10% de yodo-povidona en la cavidad abdominal de diversas especies de animales. En todas se demostró un efecto letal en breve plazo con dosis entre 10 y 40 cc/kg. Se obtuvo dosis letal-50 en ratones C57-CFW que correspondió a 5 cc/kg. Se concluye que la yodo-povidona por vía intraperitoneal es tóxica y que su uso debe ser limitado a cavidades bloqueadas. Se requerirán más estudios para determinar su utilidad e inocuidad en lavados peritoneales de toda la cavidad (AU)