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Cholestasis, which is impaired bile flow from the liver into the intestine, can be caused by cholangitis and/or bile duct obstruction. Cholangitis can arise from bacterial infections and cholelithiasis, however, immune-mediated cholangitis in primary biliary cholangitis (PBC) and primary sclerosing cholangitis (PSC) is characterized by a strong immune response targeting the biliary epithelial cells (BECs). Persistent biliary inflammation further represents a risk for biliary neoplasia, cholangiocarcinoma (CCA) by driving chronic cellular stress in the BECs. Currently, immune-mediated cholangitis is considered a Th1-Th17-dominant disease, however, the presence of Th2-related mast cells (MCs) in tissue samples from PBC, PSC and CCA patients has been described, showing that these MCs are active players in these diseases. Here, we reviewed and discussed experimental and clinical data supporting a pro-fibrotic role for MCs in immune-mediated cholangitis as well as their participation in supporting tumor growth acting as angiogenesis promoters. Thus, although MCs have classically been identified as downstream effectors of Th2 responses in allergies and parasitic infections, evidence suggests that these MCs are relevant players in biliary inflammation and neoplasia. The availability of strategies to prevent MCs' activation represents a therapeutic opportunity in biliary diseases.

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Cells from different origins behave differently regarding the incorporation of exogenous DNA and formation of transgenic cells. Milk production of recombinant antibody may benefit from efficient transfection protocols to produce transgenic animals. In this context, the objective of this study was to verify the transfection potential of bovine mesenchymal stem cells from Wharton's jelly (MSC-WJ) and adipose tissue (MSC-AT), comparing co-transfection protocols with vectors pBC1-anti-CD3 and pEF-NEO-GFP, using transfection reagents Lipofectamine LTX with Plus Reagent or Xfect. Skin fibroblasts (FIB) were used as the control group. Forty-eight hours after transfection, neomycin was added and cells cultured for 2 weeks. Treated cells were submitted to fluorescence microscopy, flow cytometry, and PCR evaluations. Wharton's jelly cells were sensitive to treatments and started necrosis. In the flow cytometry assay, the median fluorescence was higher in adipocytes than fibroblasts, for both the Xfect (20.057 ± 1.620,7 and 10.601 ± 702,86, respectively, p < 0.05) and LTX (19.590 ± 113,84 and 10.518 ± 442,65, respectively, p < 0.05). These results, associated with evaluation of epifluorescence, demonstrated that adipocytes presented a better response to transfection than other cells, independent of the kit used. Performing PCR on co-transfected cells demonstrated the presence of anti-CD3, making this approach feasible for future experiments.

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INTRODUCTION: Acoustic Radiation Force Impulse (ARFI) elastography evaluates hepatic fibrosis non-invasively and has been mainly validated in viral hepatitis. Data on rare liver diseases such as autoimmune hepatitis (AIH), overlap syndrome, primary biliary cholangitis (PBC) or primary sclerosing cholangitis (PSC) are sparse. MATERIAL AND METHODS: 85 patients (including 31 AIH, 26 PBC, 16 PSC and 3 PSC-and 9 PBC-AIH-overlap syndromes) were retrospectively analysed pointing at ARFI elastography of the liver and the correlation with histologic Ishak fibrosis score (F0-6). Results of shear wave velocities (m/s) were expressed as mean ± standard deviation. RESULTS: The mean shear wave velocity of all 85 patients showed 1.80 ± 0.84 m/s (0.74-3.98). The ARFI elastography values correlated with the degree of fibrosis in all patients overall and in patients with AIH, overlap syndrome and PSC, respectively. The subgroup of 26 patients with PBC (only with Ishak F > 3) revealed no correlation between ARFI and these early fibrosis stages (r = 0.019, p = 0.927). ARFI elastography correlated with bilirubin, AST, but not with patient age, body mass index or measurement depth. The cut-off of 2.04 m/s for detecting cirrhosis (Ishak F > 5) leads to a sensitivity of 90.0% and specificity of 74.7% (AUROC 87.2%). CONCLUSION: ARFI elastography can evaluate fibrosis in AIH, PSC and PSC-/PBC-AIH-overlap syndrome with good accuracy for the detection of hepatic cirrhosis. Shear wave velocities in PBC should be interpreted with caution in early stages of fibrosis.

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Infections by Candida albicans in immune compromised patients cause significant morbidity and mortality. In the search for potential molecular targets for drug development, the family of agglutinin-like proteins (Als) in C. albicans have been identified due to numerous attributes associated with high virulence, most prominently due to their role in adherence. Here, molecular models of individual members of the Als family illustrated common and unique structure features. Additionally, dynamic simulations were performed to display regions of high mobility. The results showed variations between Als members in the fluctuation of the A1B1 protein loop, which is located at the entrance to the peptide binding cavity, suggesting that this feature may be a factor contributing to observed differences in affinities to ligands and adhesion properties. Molecular docking results further suggested that ligand affinity could be influenced by movements in the A1B1 loop. In addition, a new site was identified in Als in an area adjacent to the peptide binding cavity that could serve as a new binding site for the design of future anti-adhesion ligands that provide increased specificity inhibiting Als proteins from C. albicans.

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Single-crystal X-ray diffraction and quantum mechanical theories were used to examine in detail the subtle nature of non-covalent interactions in the [2:1:1] multicomponent crystal of 1,1-aminocyclopentanecarboxylic acid:oxalic acid:water. The crystal, which is a hydrate salt of the amino acid with the hydrogen-oxalate ion, also contains the zwitterion of the amino acid in equal proportions. It was found that a dimeric cation [Acc5(Z)...Acc5(C)]+ bonded by an O-H...O hydrogen bond exists due to a charge transfer between acid and carboxylate groups. The three-dimensional crystal is built by blocks stacked along the [101] direction by dispersion interactions, with each block growing along two directions: a hydrogen oxalate HOX-...HOX- catameric supramolecular structure along the [010] direction; and double ...HOX--W-[Acc5(Z)... Acc5(C)]+... chains related by inversion centers along the [1 0 {\bar 1}] direction. A PBE-DFT optimization, under periodic boundary conditions, was carried out. The fully optimized structure obtained was used to extract the coordinates to calculate the stabilization energy between the dimers under the crystal field, employing the M062X/aug-cc-pVTZ level of theory. The non-covalent index isosurfaces employed here allow the visualization of where the hydrogen bond and dispersion interactions contribute within the crystal. The crystal atomic arrangements are analyzed by employing the Atoms in Molecules and electron localization function theories. Within this context, the presence of density bond critical points is employed as a criterion for proving the existence of the hydrogen bond and it was found that these results agree with those rendered by the crystallographic geometrical analysis, with only three exceptions, for which bond critical points were not found.

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OBJETIVOS: Comparar el deterioro psicosocial experimentado por los usuarios de pasta base de cocaína (PBC) y clorhidrato de cocaína en función de los datos recolectados en las fichas de admisión de un centro de tratamiento ambulatorio de la ciudad de Buenos Aires. METODO: Se analizaron estadísticamente las diferencias entre ambos grupos de usuarios respecto a la frecuencia de consumo, tenencia de causas penales, nivel educativo alcanzado, cantidad de tratamientos previos al momento de la entrevista de admisión, situación laboral, lugar de residencia y tenencia de obra social. Se calculó la intensidad de la asociación entre estas tres últimas y el resto de las variables, así como la intensidad de asociación entre la sustancia de preferencia y todas las demás variables. RESULTADOS: Se hallaron diferencias estadísticamente significativas entre los usuarios de PBC y cocaína respecto a algunas de las variables seleccionadas. Se observan diferencias semejantes entre los pacientes con empleo y los desempleados, independientemente del tipo de sustancia consumida. CONCLUSIONES: Los resultados permiten inferir un mayor nivel de deterioro psicosocial en los usuarios de PBC en comparación con los consumidores de cocaína. No obstante, esta diferencia es leve y podría atribuirse a ciertas condiciones sociales de exclusión. Además, los resultados permiten cuestionar ciertos prejuicios comunes asociados al consumo de PBC.(AU)

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OBJETIVOS: Comparar el deterioro psicosocial experimentado por los usuarios de pasta base de cocaína (PBC) y clorhidrato de cocaína en función de los datos recolectados en las fichas de admisión de un centro de tratamiento ambulatorio de la ciudad de Buenos Aires. MÉTODO: Se analizaron estadísticamente las diferencias entre ambos grupos de usuarios respecto a la frecuencia de consumo, tenencia de causas penales, nivel educativo alcanzado, cantidad de tratamientos previos al momento de la entrevista de admisión, situación laboral, lugar de residencia y tenencia de obra social. Se calculó la intensidad de la asociación entre estas tres últimas y el resto de las variables, así como la intensidad de asociación entre la sustancia de preferencia y todas las demás variables. RESULTADOS: Se hallaron diferencias estadísticamente significativas entre los usuarios de PBC y cocaína respecto a algunas de las variables seleccionadas. Se observan diferencias semejantes entre los pacientes con empleo y los desempleados, independientemente del tipo de sustancia consumida. CONCLUSIONES: Los resultados permiten inferir un mayor nivel de deterioro psicosocial en los usuarios de PBC en comparación con los consumidores de cocaína. No obstante, esta diferencia es leve y podría atribuirse a ciertas condiciones sociales de exclusión. Además, los resultados permiten cuestionar ciertos prejuicios comunes asociados al consumo de PBC.

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BACKGROUND: The peptide mastoparan 7 (MST7) stimulated ATP release in human erythrocytes. We explored intra- and extracellular processes governing the time-dependent accumulation of extracellular ATP (i.e., ATPe kinetics). METHODS: Human erythrocytes were treated with MST7 in the presence or absence of two blockers of pannexin 1. ATPe concentration was monitored by luciferin-luciferase based real-time luminometry. RESULTS: Exposure of human erythrocytes to MST7 led to an acute increase in [ATPe], followed by a slower increase phase. ATPe kinetics reflected a strong activation of ATP efflux and a low rate of ATPe hydrolysis by ectoATPase activity. Enhancement of [ATPe] by MST7 required adhesion of erythrocytes to poly-D-lysin-coated coverslips, and correlated with a 31% increase of cAMP and 10% cell swelling. However, when MST7 was dissolved in a hyperosmotic medium to block cell swelling, ATPe accumulation was inhibited by 49%. Erythrocytes pre-exposure to 10µM of either carbenoxolone or probenecid, two blockers of pannexin 1, exhibited a partial reduction of ATP efflux. Erythrocytes from pannexin 1 knockout mice exhibited similar ATPe kinetics as those of wild type mice erythrocytes exposed to pannexin 1 blockers. CONCLUSIONS: MST7 induced release of ATP required either cell adhesion or strong activation of cAMP synthesis. Part of this release required cell swelling. Kinetic analysis and a data driven model suggested that ATP efflux is mediated by two ATP conduits displaying different kinetics, with one conduit being fully blocked by pannexin 1 blockers. GENERAL SIGNIFICANCE: Kinetic analysis of extracellular ATP accumulation from human erythrocytes and potential effects on microcirculation.

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En la investigación "Alcances de la aplicación del modelo de reducción de daños al campo asistencial en las adicciones" (Alicia Donghi), ubicamos la siguiente particularidad: ciertos usuarios de sustancias "resistían" con sus cuerpos el Ideal de la ciencia del siglo XXI: la salud, el bienestar. Algunos pacientes adictos se presentaban con niveles de deterioro mental y corporal, debido a la ingesta o la abstinencia de sustancias duras. Nuestra tarea en la función pública y en la atención por obras sociales nos exige el diagnóstico según el DSM-IV. En muchas becas de derivación para internación de usuarios de PBC el rotulo de "Psicosis" abunda, a pesar de haber sido entrevistados bajo efectos"tóxicos" del consumo. Así comenzó nuestra investigación sobre diagnóstico diferencial de psicosis según el psicoanálisis y el DSM-IV, a in de garantizar al consultante la derivación a un dispositivo acorde a su patología de base, para un tratamiento posible.

In the research "Scope of the application of the model of harm reduction to the health care ield in addiction" (Alicia Donghi), are located the following peculiarity: certain users of substances "resisted" with their bodies the Ideal of the science of the 21st century: health, well-being. Some addicted patients presented with levels of mental and physical, deterioration due to the intake or the withdrawal of harsh substances. Our task in the public service and care by social work requires us according to the DSM-IV diagnosis. In many scholarships for placement of PBC users bypass the tagline of "Psycho" abounds, despite having been interviewed under "toxic" effects of consumption. Thus began our research on differential diagnosis of psychosis in psychoanalysis and the DSM-IV, in order to ensure the consultant referral to a device according to their pathology of basis, for a possible treatment.