Effect of Melatonin on Chemoresistance Exhibited by Spheres Derived from Canine Mammary Carcinoma Cells.

Mammary cancer is a frequent disease in female dogs, where a high proportion of cases correspond to malignant tumors that may exhibit drug resistance. Within the mammary tumor microenvironment, there is a cell subpopulation called cancer stem cells (CSCs), which are capable of forming spheres in vitro and resisting anti-tumor treatments, partly explaining the recurrence of some tumors. Previously, it has been described that spheres derived from canine mammary carcinoma cells CF41.Mg and REM 134 exhibit stemness characteristics. Melatonin has shown anti-tumor effects on mammary tumor cells; however, its effects have been poorly evaluated in canine mammary CSCs. This study aimed to analyze the effect of melatonin on the chemoresistance exhibited by stem-like neoplastic cells derived from canine mammary carcinoma to cytotoxic drugs such as doxorubicin and mitoxantrone. CF41.Mg and REM 134 cells were cultured in high-glucose DMEM supplemented with fetal bovine serum and L-glutamine. The spheres were cultured in ultra-low attachment plates in DMEM/F12 medium without fetal bovine serum and with different growth factors. The CD44+/CD24-/low phenotype was analyzed by flow cytometry. The viability of sphere-derived cells (MTS reduction) was studied in the presence of melatonin (0.1 or 1 mM), doxorubicin, mitoxantrone, and luzindole. In addition, the gene (RT-qPCR) of the multidrug resistance bombs MDR1 and ABCG2 were analyzed in the presence of melatonin. Both cell types expressed the MT1 gene, which encodes the melatonin receptor MT1. Melatonin 1 mM does not modify the CD44+/CD24-/low phenotype; however, the hormone reduced viability (p < 0.0001) only in CF41.Mg spheres, without inducing an additive effect when co-incubated with cytotoxic drugs. These effects were independent of the binding of the hormone to its receptor MT1, since, by pharmacologically inhibiting them, the effect of melatonin was not blocked. In CF41.Mg spheres, the relative gene expression of ABCG2 and MDR1 was decreased in response to the hormone (p < 0.001). These results indicate that melatonin negatively modulates the cell survival of spheres derived from CF41.Mg cells, in a way that is independent of its MT1 receptor. These effects did not counteract the resistance to doxorubicin and mitoxantrone, even though the hormone negatively regulates the gene expression of MDR1 and ABCG2.

Pathological Characterization and Risk Factors of Splenic Nodular Lesions in Dogs (Canis lupus familiaris).

In dogs, the spleen is a secondary lymphoid organ that can be affected by both neoplastic and non-neoplastic nodules. In general, few studies relate histopathological diagnosis to tumor size and the number of nodules in spleen biopsies. Some of these studies are inconclusive regarding the difference between neoplastic and non-neoplastic lesions and have small sample sizes or do not consider all splenic lesions. This study aimed to characterize splenic masses and determine risk factors for spleen tumors in dogs. A total of 507 histological reports corresponding to the diagnosis of splenic lesions in dogs from a private laboratory of animal pathology in the Metropolitan Region, Chile, were used. Data were analyzed by descriptive statistics and multiple logistic regression. The most frequent neoplastic and non-neoplastic diagnoses were hemangiosarcoma and hyperplasia, respectively. Most of the cases occurred in male (265 cases, 52.3%), senior (421 cases, 83%), and purebred individuals (342 cases, 67.5%). The most affected breeds were the Cocker Spaniel, German Shepherd, and Labrador Retriever. The most frequent lesion was a single nodule. The variables that exhibited a greater risk for the presentation of splenic neoplasia were male sex (odds ratio (OR) = 16.21; 95% confidence interval (CI) 1.741-150.879; p = 0.014), the presence of two or more splenic nodules (OR = 3.94; 95% CI 2.168-7.177; p < 0.001), an increase in nodule size greater than 2 cm (OR for quartiles 2, 3 and 4 of 2.2; 95% CI 1.036-4.941; p = 0.041, 2.9; 95% CI 1.331-6.576; p = 0.008, and 3.6; 95% CI 1.562-8.499; p = 0.003, respectively), and increasing age (OR = 1.23; 95% CI 1.048-1.436; p = 0.011). On the other hand, males exhibited a lower risk as age increases (OR = 0.76; 95% CI 0.615-0.928; p = 0.008). In conclusion, this study identified that males, multinodular presentation, nodule size, and age are risk factors for the occurrence of splenic neoplasia in dogs, knowledge that will contribute to the diagnostic management of dogs with spleen lesions.

Bovine adipose tissue-derived mesenchymal stem cells self-assemble with testicular cells and integrates and modifies the structure of a testicular organoids.

Mesenchymal stem cells (MSC) display self-renewal and mesodermal differentiation potentials. These characteristics make them potentially useful for in vitro derivation of gametes, which may constitute experimental therapies for human and animal reproduction. Organoids provide a spatial support and may simulate a cellular niche for in vitro studies. In this study, we aimed at evaluating the potential integration of fetal bovine MSCs derived from adipose tissue (AT-MSCs) in testicular organoids (TOs), their spatial distribution with testicular cells during TO formation and their potential for germ cell differentiation. TOs were developed using Leydig, Sertoli, and peritubular myoid cells that were previously isolated from bovine testes (n = 6). Thereafter, TOs were characterized using immunofluorescence and Q-PCR to detect testicular cell-specific markers. AT-MSCs were labeled with PKH26 and then cultured with testicular cells at a concentration of 1 × 106 cells per well in Ultra Low Attachment U-shape bottom (ULA) plates. TOs formed by testicular cells and AT-MSCs (TOs + AT-MSCs) maintained a rounded structure throughout the 28-day culture period and did not show significant differences in their diameters. Conversely, control TOs exhibited a compact structure until day 7 of culture, while on day 28 they displayed cellular extensions around their structure. Control TOs had greater (P < 0.05) diameters compared to TOs + AT-MSCs. AT-MSCs induced an increase in proportion of Leydig and peritubular myoid cells in TOs + AT-MSCs; however, did not induce changes in the overall gene expression of testicular cell-specific markers. STAR immunolabelling detected Leydig cells that migrated from the central area to the periphery and formed brunches in control TOs. However, in TOs + AT-MSCs, Leydig cells formed a compact peripheral layer. Sertoli cells immunodetected using WT1 marker were observed within the central area forming clusters of cells in TOs + AT-MSCs. The expression of COL1A associated to peritubular myoids cells was restricted to the central region in TOs + AT-MSCs. Thus, during a 28-day culture period, fetal bovine AT-MSCs integrated and modified the structure of the TOs, by restricting formation of branches, limiting the overall increase in diameters and increasing the proportions of Leydig and peritubular myoid cells. AT-MSCs also induced a reorganization of testicular cells, changing their distribution and particularly the location of Leydig cells.

Multivariate prediction of Saliva Precipitation Index for relating selected chemical parameters of red wines to the sensory perception of astringency.

Astringency is an essential sensory attribute of red wine closely related to the saliva precipitation upon contact with the wine. In this study a data matrix of 52 physico-chemical parameters was used to predict the Saliva Precipitation Index (SPI) in 110 Italian mono-varietal red wines using partial least squares regression (PLSr) with variable selection by Variable Importance for Projection (VIP) and the significance of regression coefficients. The final PLSr model, evaluated using a test data set, had 3 components and yielded an R2test of 0.630 and an RMSEtest of 0.994, with 19 independent variables whose regression coefficients were all significant at p < 0.05. Variables selected in the final model according to the decreasing magnitude of their absolute regression coefficient include the following: Procyanidin B1, Epicatechin terminal unit, Total aldehydes, Protein content, Vanillin assay, 520 nm, Polysaccharide content, Epigallocatechin PHL, Tartaric acid, Volatile acidity, Titratable acidity, Catechin terminal unit, Proanthocyanidin assay, pH, Tannin-Fe/Anthocyanin, Buffer capacity, Epigallocatechin PHL gallate, Catechin + epicatechin PHL, and Tannin-Fe. These results can be used to better understand the physico-chemical relationship underlying astringency in red wine.

Mind the Curve: Dose-Response Fitting Biases the Synergy Scores across Software Used for Chemotherapy Combination Studies.

Drug combinations are increasingly studied in the field of anticancer agents. Mathematical models, such as Loewe, Bliss, and HSA, are used to interpret drug combinations, while informatics tools help cancer researchers identify the most effective combinations. However, the different algorithms each software uses lead to results that do not always correlate. This study compared the performance of Combenefit (Ver. 2.021) and SynergyFinder (Ver. 3.6) in analyzing drug synergy by studying combinations involving non-steroidal analgesics (celecoxib and indomethacin) and antitumor drugs (carboplatin, gemcitabine, and vinorelbine) on two canine mammary tumor cell lines. The drugs were characterized, their optimal concentration-response ranges were determined, and nine concentrations of each drug were used to make combination matrices. Viability data were analyzed under the HSA, Loewe, and Bliss models. Celecoxib-based combinations showed the most consistent synergistic effect among software and reference models. Combination heatmaps revealed that Combenefit gave stronger synergy signals, while SynergyFinder produced better concentration-response fitting. When the average values of the combination matrices were compared, some combinations shifted from synergistic to antagonistic due to differences in the curve fitting. We also used a simulated dataset to normalize each software's synergy scores, finding that Combenefit tends to increase the distance between synergistic and antagonistic combinations. We conclude that concentration-response data fitting biases the direction of the combination (synergistic or antagonistic). In contrast, the scoring from each software increases the differences among synergistic or antagonistic combinations in Combenefit when compared to SynergyFinder. We strongly recommend using multiple reference models and reporting complete data analysis for synergy claiming in combination studies.

Conocimientos de enfermería sobre medidas de prevención en neumonía asociada a ventilación mecánica / Nursing knowledge about preventive measures in pneumonia associated with mechanical ventilation

Determinar el conocimiento del personal de enfermería respecto a medidas de prevención de neumonía asociada a ventilación mecánica y manejo de alimentación enteral en pacientes ingresados a la UCI de un hospital público. Metodología: estudio descriptivo, de corte transversal. La muestra fue representada por personal de enfermería de terapia intensiva de un hospital público de Resistencia- Chaco (n=75), seleccionados mediante muestreo no probabilístico por conveniencia durante el periodo de estudio. Se diseñó un instrumento de recolección de datos con las variables en estudio. Se respetaron los principios bioéticos de la Declaración de Helsinki. Resultados: Respecto a los conocimientos sobre las medidas de precauciones estándar, se observó que referente a la utilización de EPP en la asistencia a pacientes ventilados, un 48% de los mismos presento un conocimiento regular, en los momentos de aplicación del lavado de manos, el 44% mostró un conocimiento deficiente. Sobre el manejo de tubo endotraqueal y ventilador mecánico, se observó un nivel de regular en cuanto a conocimientos sobre la correcta fijación del TET (56%); sobre el manejo del respirador y sus conexiones (64%); manejo de humidificadores (60%)y recambios de los circuitos (64%), en contraposición con Dos Santos8 y Granizo-Taboada, Wagner Thomas6 quienes revelaron buenas prácticas de enfermería para el mantenimiento de la oxigenación en pacientes bajo ventilación mecánica. Conclusión: Se destaca en líneas generales, que el personal de enfermería presento conocimiento regular, enfatizando la necesidad de capacitación de los mismos sobre las medidas de prevención de neumonía asociada a ventilación mecánica[AU]

To determine the knowledge of nursing staff regarding prevention measures for pneumonia associated with mechanical ventilation and management of enteral feeding in patients admitted to the ICU of a public hospital. Methodology: descriptive, cross-sectional study. The sample was represented by intensive care nursing staff from a public hospital in Resistencia-Chaco (n=75), selected by non-probabilistic convenience sampling during the study period. A data collection instrument was designed with the variables under study. The bioethical principles of the Declaration of Helsinki were respected. Results: Regarding the knowledge about standard precaution measures, it was observed that regarding the use of PPE in the care of ventilated patients, 48% of them presented regular knowledge, at the times of application of hand washing, 44% showed poor knowledge. Regarding the management of the endotracheal tube and mechanical ventilator, a fair level was observed in terms of knowledge about the correct fixation of the ETT (56%); about the management of the respirator and its connections (64%); management of humidifiers (60%) and replacement of circuits (64%), in contrast to Dos Santos8 and Granizo-Taboada, Wagner Thomas6 who revealed good nursing practices for maintaining oxygenation in patients under mechanical ventilation. Conclusion: It stands out in general lines, that the nursing staff presented regular knowledge, emphasizing the need for their training on the prevention measures of pneumonia associated with mechanical ventilation[AU]

Verificar o conhecimento da equipe de enfermagem sobre as medidas de prevenção de pneumonia associada à ventilação mecânica e manejo da alimentação enteral em pacientes internados na UTI de um hospital público. Metodologia: estudo descritivo, transversal. A amostra foi representada por enfermeiros intensivistas de um hospital público de Resistencia-Chaco (n=75), selecionados por amostragem não probabilística por conveniência durante o período do estudo. Foi elaborado um instrumento de coleta de dados com as variáveis em estudo. Os princípios bioéticos da Declaração de Helsinque foram respeitados. Resultados: Em relação ao conhecimento sobre medidas de precaução padrão, observou-se que quanto ao uso de EPI no cuidado de pacientes ventilados, 48% deles apresentaram conhecimento regular, nos momentos de aplicação da lavagem das mãos, 44% apresentaram conhecimento ruim. Em relação ao manejo do tubo endotraqueal e do ventilador mecânico, observou-se nível razoável de conhecimento sobre a fixação correta do TET (56%); sobre o manejo do respirador e suas conexões (64%); manejo de umidificadores (60%) e troca de circuitos (64%), ao contrário de Dos Santos8 e Granizo-Taboada, Wagner Thomas6 que revelaram boas práticas de enfermagem para manutenção da oxigenação em pacientes sob ventilação mecânica. Conclusão: Destaca-se em linhas gerais, que a equipe de enfermagem apresentou conhecimento regular, ressaltando a necessidade de seu treinamento sobre as medidas de prevenção de pneumonia associada à ventilação mecánica[AU]

Evaluation of Plant-Based Byproducts as Green Fining Agents for Precision Winemaking.

Consumers are increasingly looking for foods, including wine, that are free of animal-derived proteins. This study seeks to evaluate patatin, a new, plant-based and allergen-free fining agent, by comparing it with the fining agents polyvinipolypyrrolidone, bovine serum albumin, and methylcellulose. Specifically, its effects on the phenolic profile of enological tannins were analyzed with four spectrophotometric assays: OD 280 nm, Folin−Ciocâlteu, Adams−Harbertson, and methylcellulose. In addition, changes in the polyphenol composition of Sangiovese red wine were determined by UV-Vis spectrophotometry and HPLC with adsorption trials, and the solid−liquid interaction in a wine solution was modeled by both Langmuir and Freundlich equations. Our findings highlight the occurrence of systematic proportional error between the selected spectrophotometric assays. As a result, direct comparisons of protein precipitation assays can be made only among results obtained with the same spectrophotometric method. However, it is clear that patatin has an impact on the phenolic profile of Sangiovese red wine: it removes simple phenolics (gallic acid, (+)-catechin, (−)-epicatechin, epicatechin gallate, syringic acid, fertaric acid, coutaric acid, and rutin) as well as both oligomeric and polymeric tannins to different extents. In concentrations of less than 1 g/L, the patatin isotherm showed a linear relation between the equilibrium concentration and the quantity absorbed, obeying the Freundlich model reasonably well (KF 1.46; 1/n 1.07; R2 0.996 with 1/n > 1). Thus, the adsorption process is strongly dependent on the fining dosage.

Hormonal Carcinogenesis in Canine Mammary Cancer: Molecular Mechanisms of Estradiol Involved in Malignant Progression.

Mammary cancer is a frequent neoplasia in female dogs, in which most important risk factors are hormonal. Sexual hormones as estradiol play an important role in mammary carcinogenesis, being able to induce carcinogenic initiation, promotion and progression. However, the molecular mechanisms involved are incompletely understood. Estradiol is synthesized mainly in the ovaries, nevertheless, high concentrations of estradiol and some of its hormonal precursors have also been described in malignant mammary tumor tissue. The mechanisms of action of estradiol include the classic genomic effects that modulate gene transcription, and non-genomic effects, which trigger quick effects after estradiol binds to its specific receptors. These responses modulate various intracellular signaling pathways, triggering post-translational modification of several proteins. This review will discuss the well-known underlying mechanisms associated with the action of estradiol in the malignant progression of canine mammary tumors.

Melatonin decreases in vitro viability and migration of spheres derived from CF41.Mg canine mammary carcinoma cells.

BACKGROUND: Mammary cancer is a common disease affecting female dogs, where approximately 50% of the cases are malignant. There is a subpopulation of cancer cells with stem cell-like features within the tumour microenvironment, which can form in vitro spheres, cell structures that grow in anchor-free conditions. This cell population shows resistance to conventional antitumor treatments explaining in part the recurrence of some type of cancers. It has been previously reported that spheres derived from CF41.Mg canine mammary carcinoma cells exhibit several stemness features. Melatonin has shown antitumor effects on cancer mammary cells; nevertheless, its effects have been poorly evaluated on canine mammary cancer stem-like cells. In this regard, it has described that melatonin decreases the expression of OCT-4 in CMT-U2229 mammary cancer cells, a transcription factor that participates in the modulation of self-renewal and drug resistance in cancer stem-like cells. The aim of this study was to compare the effects of melatonin on viability and migration of canine mammary carcinoma CF41.Mg-spheres, and CF41.Mg-parental cells. CF41.Mg cells were grown in DMEM high-glucose medium containing 10% bovine foetal serum. CF41.Mg-spheres were cultured in ultra-low attachment plates with serum-free DMEM/F12 containing several growth factors. Cell viability (MTS reduction) and migration (transwell) assays were conducted in presence of melatonin (0.01, 0.1 or 1 mM). RESULTS: Melatonin decreased cell viability at 1 mM (P < 0.05), with a significant reduction in spheres compared to parental cells at 24 and 48 h (P < 0.05). Cell migration was inhibited in response to non-cytotoxic concentration of melatonin (0.1 mM) (P < 0.05) in spheres and monolayer of cells, no significant differences were detected between both cell subtypes. CONCLUSIONS: These results indicate that melatonin reduces viability and migration of CF41.Mg cells, where spheres exhibit greater sensitivity to the hormone. Thus, melatonin represents a valuable potential agent against mammary cancer cells, especially cancer stem-like cells.

Oral masses in African pygmy hedgehogs.

African pygmy hedgehogs (Atelerix albiventris) frequently develop oral neoplasms, and most of these neoplasms are malignant. We characterized oral masses detected in hedgehogs at clinical examination. During a 1-y period, we diagnosed oral cavity masses in 27 privately owned hedgehogs; 16 were female and 11 were male, with ages of 2-7 y (mean: 4.3 y). Eight masses were non-neoplastic and were diagnosed as gingival hyperplasia (GH). Nineteen masses were neoplastic, of which 17 were squamous cell carcinomas (SCCs) and 2 were mesenchymal tumors (1 spindle cell tumor of probable neural origin, and 1 hemangiosarcoma). The GHs were noninvasive, exophytic, and did not recur after surgical excision. The SCCs were highly invasive tumors that induced facial deformation and were located in the caudal portion of the oral cavity, with 12 of them arising from the right-caudal maxilla. Thus, clinical signs, growth pattern, and anatomic location can be used to suspect a diagnosis of SCC among the other possible diagnoses, such as GH, in this location. However, histopathology is necessary for confirmation. Also, hemangiosarcoma should be considered among the differential diagnoses.

Caracterización de los modelos pedagógicos y su pertinencia en una educación contable crítica / Characterization of pedagogical models and their relevance in a critical accounting education / Caracterização de modelos pedagógicos e sua relevância em uma educação contábil crítica

Resumen En el presente documento se identifican los elementos, los actores y los intereses presentes en los modelos pedagógicos más relevantes en la literatura académica, en contraste con lo aplicado en la educación contable, con el fin de fundamentar un modelo pedagógico tendiente a una formación contable crítica, analítica, activa y propositiva. Para ello, se analizan las teorías y los modelos pedagógicos a través de los autores más citados por investigadores que describen las características de los procesos de aprendizaje desde distintas ciencias sociales, y en el ámbito de la educación contable se revisan artículos, libros y tesis que abordan las necesidades sociales del ejercicio profesional contable enfocando la revisión en las investigaciones dadas con posterioridad a la crisis económica del 2002 en Estados Unidos y aquellas publicaciones emitidas por organizaciones multilaterales que buscan guiar criterios de confiabilidad y comparabilidad de la profesión y la información contable a nivel internacional. Se concluye que los modelos pedagógicos que podrían tener un mayor impacto en la generación de contadores críticos, analíticos, proactivos y propositivos, son aquellos que otorgan un rol principal al estudiante y se enfocan en la formación de individuos que permitan el desarrollo social y económico de su entorno. Clasificación JEL: A22, I23.

Abstract This document identifies elements, actors and interests present in the most relevant pedagogical models in the academic literature, in contrast to what is applied in accounting education, in order to base a pedagogical model tending to a critical, analytical, active and proactive accounting education. For this, theories and pedagogical models are analyzed through of the authors most cited by researches books and thesis academics that investigate about the social necessities of the accounting professional activity focused in the researches that originate after the United States economic crisis the 2002 and the publications generated by multilateral organizations that guide the criteria's reliability and comparability of the accounting profession and information are reviewed, in order to support a pedagogical model tending to a critical, analytical, active and proactive accounting education. The conclusion of this study is that pedagogical models that could have a greater impact on the generation of critical, analytical, proactive and proactive accountants, are those that grant a main role to the student and focus on the training of individuals that allow the social and economic development of their environment. JEL Classification: A22, I23.

Resumo Este documento identifica os elementos, atores e interesses presentes nos modelos pedagógicos mais relevantes na literatura acadêmica, em contraste com o que é aplicado na educação contábil, a fim de fundamentar um modelo pedagógico tendendo a uma formação contábil crítica. , analítica, ativa e proativa. Para isso, as teorias e modelos pedagógicos são analisados através dos autores mais citados pelos pesquisadores que descrevem as características dos processos de aprendizagem das diferentes ciências sociais, e no campo da educação contábil, artigos, livros e teses são revisados. abordar as necessidades sociais da prática contábil focando revendo as investigações proferidas após a crise econômica de 2002 nos Estados Unidos e essas publicações emitidos por organismos multilaterais que buscam orientar critérios de fiabilidade e comparabilidade da profissão e as informações contábeis no nível internacional Conclui-se que os modelos pedagógicos que podem ter um grande impacto sobre a geração de contadores críticos, analíticos, pró-ativa e proposicional são aqueles que dão um papel de liderança para o aluno e se concentrar no treinamento de pessoas para permitir o desenvolvimento social e econômico seu ambiente. Classificações JEL: A22, I23.

Bovine fetal mesenchymal stem cells exert antiproliferative effect against mastitis causing pathogen Staphylococcus aureus.

Staphylococcus aureus is the most commonly isolated pathogen from clinical bovine mastitis samples and a difficult pathogen to combat. Mesenchymal stem cells (MSC) are multipotent progenitor cells equipped with a variety of factors that inhibit bacterial growth. The aim of the present study was to evaluate the in vitro antibacterial potential against S. aureus of conditioned medium (CM) from MSC derived from fetal bovine bone marrow (BM-MSC) and adipose tissue (AT-MSC). BM-MSC, AT-MSC and fetal fibroblasts (FB) cultures were activated by infection with S. aureus. Bacterial growth was evaluated in presence of CM, concentrated CM (CCM), activated CM (ACM) and concentrated ACM (CACM) from BM-MSC, AT-MSC and FB. Gene expression of ß-defensin 4A (bBD-4A), NK-lysine 1 (NK1), cathelicidin 2 (CATHL2), hepcidin (HEP) and indoleamine 2,3 dioxygenase (IDO) and protein expression of bBD-4A were determined in activated and non-activated cells. The majority of BM-MSC and AT-MSC expressed CD73, Oct4 and Nanog, and were negative for CD34. Growth of S. aureus decreased when it was exposed to CM from BM-MSC, AT-MSC and FB. Moreover, growth of S. aureus in CCM, ACM and CACM was lower compared to controls of CM from BM-MSC and AT-MSC. Activated AT-MSC increased mRNA levels of bBD4A and NK1, and protein levels of bBD4A in CM. Thus, CM from fetal bovine BM-MSC and AT-MSC has the capacity to reduce in average ~30% of S. aureus relative growth under in vitro conditions. The in vitro antibacterial effect of fetal bovine MSC may be mediated by bBD4A and NK1 activity.

Immunomodulatory and immunogenic properties of mesenchymal stem cells derived from bovine fetal bone marrow and adipose tissue.

Little information is currently available on therapeutic features of bovine mesenchymal stem cells (MSCs), despite the development of large animal experimental models including cattle may open alternative strategies for investigating MSC physiology and eventual applications for regenerative therapy. The aim of the present study was to compare in vitro immunomodulatory and immunogenic potentials of bovine fetal MSCs (bfMSCs) derived from bovine fetal bone marrow (BM-MSCs) and adipose tissue (AT-MSCs). Immunomodulatory analyses in bfMSCs were performed by determination of the effect of interferon-γ (IFNγ) on mRNA levels of indoleamine 2, 3-dioxygenase (IDO), transforming growth factor ß1 (TGFß1), prostaglandin E receptor 2 (PTGER2), interleukin-6 and -10 (IL-6 and IL-10), and IDO enzymatic activity. The effect of conditioned medium from IFNγ-stimulated bfMSCs on the proliferation of alloantigen-activated peripheral blood lymphocytes (PBLs) was assessed. Immunogenicity of bfMSCs was determined by quantification of mRNA levels of major histocompatibility complex I and II (MHC-I and -II), CD80 and CD86, and the proportion of cells positive for MHC-I and -II by flowcytometry (FACS) analyses. IFNγ treatment increased IL-6, PTGER2 and IDO gene expression and activity in bfMSCs but did not affect suppressive effect on proliferation of PBLs. Lower proportion of AT-MSCs expressed MHC-I and MHC-II in comparison to BM-MSCs. In conclusion, BM-MSCs and AT-MSCs upregulated expression of immunomodulatory genes in a similar way after IFNγ stimuli. BM-MSCs and AT-MSCs in basal condition and treated with IFNγ displayed similar in vitro immunomodulatory ability. Lower expression of MHC-I and MHC-II suggest that AT-MSCs might be less immunogenic compared to BM-MSCs.

Myogenic Differentiation Potential of Mesenchymal Stem Cells Derived from Fetal Bovine Bone Marrow.

The myogenic potential of bovine fetal MSC (bfMSC) derived from bone marrow (BM) remains unknown; despite its potential application for the study of myogenesis and its implications for livestock production. In the present study, three protocols for in vitro myogenic differentiation of bfMSC based on the use of DNA methyltransferase inhibitor 5-Aza-2'-deoxycytidine (5-Aza), myoblast-secreted factor Galectin-1 (Gal-1), and myoblast culture medium SkGM-2 BulletKit were used. Plastic-adherent bfMSC were isolated from fetal BM collected from abattoir-derived fetuses. Post-thaw viability analyses detected 85.6% bfMSC negative for propidium iodine (PI). Levels of muscle regulatory factors (MRF) MYF5, MYF6, MYOD, and DES mRNA were higher (P < 0.05) in bfMSC cultured under 100 µM of 5-Aza compared to 1 and 10 µM. Treatment of bfMSC with 10 µM of 5-Aza resulted in down-regulation of MYOD mRNA (Days 7 to 21) and up-regulation of MYF6 (Day 7), MYF5, and DES mRNA (Day 21). Gal-1 and SkGM-2 BulletKit induced sequential down-regulation of early MRF (MYF5) and up-regulation of intermediate (MYOD) and late MRF (DES) mRNA. Moreover, DES and MYF5 were immunodetected in differentiated bfMSC. In conclusion, protocols evaluated in bfMSC induced progress into myogenic differentiation until certain extent evidenced by changes in MRF gene expression.

[Segmental portal hypertension with splenic vein thrombosis caused by pancreatitis]. / Hipertensión portal segmentaria por trombosis de la vena esplénica causada por pancreatitis.

BACKGROUND: Splenic vein thrombosis is a complication of pancreatic carcinoma, pancreatitis or pancreatic pseudocyst. It may lead to segmental portal hypertension and bleeding from gastric varices. CLINICAL CASE: A 31 year-old man was diagnosed with pancreatitis of two weeks of evolution and was referred to our hospital in 2013. He had a history of alcohol consumption. Physical examination showed no stigmata of liver cirrhosis. Laboratory analyses revealed hemoglobin 9.5 g/dL, and leukocytes and platelets were normal. Liver function tests were normal as well. Abdominal CT showed a pseudocyst, which was drained by percutaneous puncture. By pseudocyst recurrence, drainage and necrosectomy by retroperitoneal laparascopy were performed. The patient presented hyperglycemia during his treatment in hospital. He was discharged, but he returned to emergency room because of gastrointestinal bleeding without hemodynamic instability. Gastroscopy showed bleeding gastric varices. The colonoscopy showed normal results. Liver biopsy was also normal. Abdominal CT angiography revealed blockage of the splenic vein. Patient underwent splenectomy and was discharged. CONCLUSION: This case is rare due to the high frequency of portal hypertension and cirrhosis. The isolated gastric varices with normal liver function are a sign of splenic thrombosis. The definitive treatment is splenectomy.

Introducción: la obstrucción aislada de la vena esplénica es una complicación de carcinoma pancreático, pancreatitis o pseudoquiste del páncreas. La trombosis de la vena esplénica puede conducir a hipertensión portal segmentaria y sangrado de várices gástricas. Caso clínico: un hombre de 31 años de edad fue referido a nuestro hospital en 2013 con el diagnóstico de pancreatitis de dos semanas de evolución. Tenía el antecedente de consumo de alcohol. El examen físico no mostró estigmas de cirrosis hepática. El laboratorio reveló hemoglobina de 9.5 g/dL con leucocitos y plaquetas normales. Las pruebas de función hepática fueron normales. La TAC abdominal mostró un pseudoquiste, el cual fue drenado por punción percutánea. Por recurrencia del pseudoquiste, se efectuó drenaje y necrosectomía por laparoscopia retroperitoneal. El paciente presentó hiperglucemia durante su estancia. Después de haber egresado, acudió a urgencias por sangrado gastrointestinal superior sin inestabilidad hemodinámica. La gastroscopia mostró várices gástricas sangrantes. La colonoscopia mostró resultados normales. La biopsia de hígado también resultó normal. La angio-TAC abdominal mostró obstrucción de la vena esplénica. Se sometió a esplenectomía y fue egresado. Conclusión: este caso es raro en nuestro medio debido a la alta frecuencia de hipertensión portal por cirrosis. Las várices gástricas aisladas con función hepática normal son un signo de trombosis de la vena esplénica. El tratamiento definitivo es la esplenectomía.

Meloxicam decreases the migration and invasion of CF41.Mg canine mammary carcinoma cells.

Cyclooxygenase (COX)-2 expression is positively correlated with malignant features in canine mammary carcinomas. Non-steroidal anti-inflammatory drugs (NSAIDs) inhibit COX activity and may therefore possess anticancer effects. Meloxicam is an NSAID that is widely used in human and veterinary medicine. High concentrations of meloxicam have been reported to be antitumorigenic in vitro; however, the effect of meloxicam at concentrations that are equivalent to those that can be obtained in vivo remains unknown. In the current study, the in vitro effects of low-dose meloxicam (0.25 µg/ml) on CF41.Mg canine mammary carcinoma cells were evaluated. The effects on cell proliferation, apoptosis, cell migration and invasion, in addition to the expression of different molecules associated with tumor invasiveness were analyzed. No effect on cell viability and apoptosis were observed. However, cell migration and invasion were significantly reduced following treatment with meloxicam. MMP-2 expression and activity were similarly reduced, explaining the impaired cell invasion. In addition, ß-catenin expression was downregulated, while its phosphorylation increased. These results indicate that 0.25 µg/ml meloxicam reduces cell migration and invasion, in part through modulating MMP-2 and ß-catenin expression. Additional studies are required to elucidate the mechanism associated with the anti-invasive effect of meloxicam on CF41.Mg cells. The results of the present study suggest that meloxicam has a potential adjunctive therapeutic application, which could be useful in controlling the invasion and metastasis of canine mammary carcinomas.

Pro-apoptotic and anti-angiogenic properties of the α /ß-thujone fraction from Thuja occidentalis on glioblastoma cells.

The most aggressive type of brain tumor is glioblastoma multiforme, which to date remains incurable. Thuja occidentalis is used in homeopathy for the treatment of cancer, however, its mechanism of action remains unknown. We set out to study the effects of thujone fractions of Thuja on glioblastoma using in vitro and in vivo models. We found that the α/ ß-thujone fraction decrease the cell viability and exhibit a potent anti-proliferative, pro-apoptotic and anti-angiogenic effects in vitro. In vivo assays showed that α /ß-thujone promotes the regression of neoplasia and inhibits the angiogenic markers VEGF, Ang-4 and CD31 into the tumor.

Implicancias de enfermería en accesos radiales y femorales / Nursing implications radial access an femoral preparation of drugs

El cateterismo cardiaco es una técnica de diagnóstico precoz y tratamiento rápido respectivamente, la enfermera debe ser capaz de transmitir al paciente la seguridad necesaria para enfrentar esta técnica para su diagnostico y tratamiento. Para ello debe conocer los cuidados que se derivan de este paciente sometido a cateterismo, la información y educacion sanitaria, imprescindibles para asegurar unos cuidados integrales y un proceso asistencial de calidad, tanto del punto de vista físico como emocional...

Endometrial expression of selected transcripts in postpartum of primiparous Holstein cows with clinical and subclinical endometritis.

Postpartum endometritis compromises milk production and fertility in high-producing dairy cows. Infection of the endometrium induces an inflammatory response with secretion of cytokines that lead to polymorphonuclear cells (PMN) influx and bacterial clearance. Considering that only a portion of cows with endometritis is eligible for clinical diagnosis, there is an increasing effort for developing reliable tools and protocols for diagnosis of subclinical endometritis. Recent reports have indicated that primiparous cows are at greater risk of uterine infection and primiparous cows with subclinical endometritis produce less milk compared to healthy cows. In the present study, gene expression profiles were compared for selected cytokine and hormone endometrial transcripts in the postpartum of primiparous Holstein cows with clinical and subclinical endometritis. Cows were classified as healthy (no signs of clinical endometritis), cows with subclinical endometritis (PMN<5% in the cytological sample) and cows with clinical endometritis (PMN>5%). Although, cows with clinical endometritis had greater (P<0.05) relative amounts of mRNA for the IL1A, IL6, IL17A, TNFα, PGES and PGHS2 genes compared to healthy cows; no significant differences were detected between clinical and subclinical endometritis groups. Spearman correlation coefficients were positive between relative amounts of gene expression as indicated by amount of these transcripts and PMN percentages and ranged from 0.74 to 0.93 (P<0.05). Relative amounts of cytokine mRNA suggest similar inflammatory response in the endometrium of cows with subclinical and clinical endometritis. Moreover, differential relative amounts of hormone transcripts suggest dysregulation of the luteolytic mechanism and PG synthases but not ERα in cows with endometritis.

Simvastatin exhibits antiproliferative effects on spheres derived from canine mammary carcinoma cells.

Mammary cancer is the most frequent type of tumor in the female canine. Treatments are mainly limited to surgery and chemotherapy; however, these tumors may develop clinical recurrence, metastasis and chemoresistance. The existence of a subpopulation of cancer cells with stemness features called cancer stem-like cells, may explain in part these characteristics of tumor progression. The statins, potent blockers of cholesterol synthesis, have also shown antitumor effects on cancer mammary cells, changes mediated by a decrease in the isoprenylation of specific proteins. Few studies have shown that simvastatin, a lipophilic statin, sensitizes cancer stem-like cells eliminating drug resistance. The aim of the present study was to evaluate the effects of simvastatin on spheres derived from CF41.Mg canine mammary tumor cells, which were characterized by phenotypic and functional analyses. Spheres exhibited characteristics of stemness, primarily expressing a CD44⁺/CD24⁻/low phenotype, displaying auto-renewal and relative chemoresistance. Exposure to simvastatin induced a decrease in the sphere-forming capacity and cell viability, accompanied by a concentration- and time-dependent increase in caspase-3/7 activity. In addition, modulation of ß-catenin and p53 expression was observed. Simvastatin triggered a synergistic effect with doxorubicin, sensitizing the spheres to the cytotoxic effect exerted by the drug. Invasiveness of spheres was decreased in response to simvastatin and this effect was counteracted by the presence of geranylgeranyl pyrophosphate. Our results suggest that simvastatin targets canine mammary cancer stem-like cells, supporting its therapeutical application as a novel agent to treat canine mammary cancer.