Co-infection by Toxoplasma gondii and Neospora caninum in Goats Reared in Extensive System of Mexico.

Background: The aim of the present study was to describe the presence of co-infection by Toxoplasma gondii and Neospora caninum in goats reared in extensive systems from Mexico. Materials and Methods: A cross-sectional study was conducted to determine the frequency of T. gondii and N. caninum, by detecting antibodies to each parasite by mean commercial ELISA kits. A total of 176 blood samples were randomly collected from mature females reared in extensive system herds from 20 municipalities of state of Guanajuato, Mexico. Results: The general seroprevalence was 23.9 and 21.0% for T. gondii and N. caninum, respectively, while co-infection rate was 3.6%. For geographic and environmental variables, no differences were observed among T. gondii and coinfection; however, it was observed that altitude, annual precipitation, annual average temperature, and rainy period showed significant differences with N. caninum seropositive goats. Conclusion: The seroprevalence of both parasites was appreciated in most of the studied herds. The present study is the first report of T. gondii and N. caninum co-infection in goats from extensive herds in Mexico.

Detection of Cyclomodulin CNF-1 Toxin-Producing Strains of Escherichia coli in Pig Kidneys at a Slaughterhouse.

Food is often contaminated with Escherichia coli (E. coli) bacteria strains, which have been associated with different diseases, including urinary tract infections. The consumption of meat by humans is a potential route of transmission of antimicrobial resistance, and food-producing animals have been associated as a major reservoir of resistant bacterial strains. The aim of this study was to determine the presence of the E. coli strains producing the CNF-1 toxin in pig kidneys. Pig kidneys were collected from a Mexican slaughterhouse and classified according to their coloration into reddish kidneys (RK) and yellowish kidneys (YK). A tissue sample from each kidney was processed for histological analysis, the presence of E. coli was determined by conventional PCR assay, and the CNF-1 toxin was detected by both conventional PCR and Western blotting. Herein, an inflammatory cell infiltrate was found in all collected kidneys, regardless of macroscopic differences. Surprisingly, E. coli and the CNF-1 toxin were detected in all kidney samples. We clearly demonstrate contamination by CNF-1 toxin-producing E. coli in pork kidneys from a slaughterhouse, even in those without apparent damage. This suggests that pork may serve as a reservoir for pathogens, representing an important risk to human health.

[Association of neutrophil, lymphocyte, platelet ratio with acute kidney injury in sepsis]. / Asociación de la relación neutrófilos, linfocitos y plaquetas con lesión renal aguda en sepsis.

Background: Acute kidney injury (AKI) is frequent in sepsis (25 to 51%), with high mortality (40 to 80%) and long-term complications. Despite its importance we do not have accessible markers in intensive care. In other entities (post-surgical and COVID-19) the neutrophil/lymphocyte and platelet (N/LP) ratio has been associated with acute kidney injury; however, this relationship has not been studied in a pathology with a severe inflammatory response such as sepsis. Objective: To demonstrate the association between N/LP with AKI secondary to sepsis in intensive care. Material and methods: Ambispective cohort study in patients over 18 years who were admitted to intensive care with a diagnosis of sepsis. The N/LP ratio was calculated from admission up to the seventh day and up to the diagnosis of AKI and outcome. Statistical analysis was performed with chi squared test, Cramer's V and multivariate logistic regression. Results: Out of the 239 patients studied, the incidence of AKI developed in 70%. 80.9% of patients with N/LP ratio > 3 had AKI (p < 0.0001, Cramer's V 0.458, OR 3.05, 95% CI 1.602-5.8) and increased renal replacement therapy (21.1 vs. 11.1%, p = 0.043). Conclusion: N/LP ratio > 3 has a moderate association with AKI secondary to sepsis in the intensive care unit.

Introducción: la lesión renal aguda (LRA) es frecuente en la sepsis (25 a 51%), tiene alta mortalidad (40 a 80%) y complicaciones a largo plazo. A pesar de su importancia, no contamos con marcadores accesibles en la terapia intensiva. En otras entidades (posquirúrgicos y COVID-19) la relación neutrófilos/linfocitos y plaquetas (N/LP) se ha asociado con lesión renal aguda; sin embargo, no se ha estudiado esta relación en una patología con una respuesta inflamatoria severa como la sepsis. Objetivo: demostrar la asociación entre la relación N/LP con la LRA secundaria a sepsis en la terapia intensiva. Material y métodos: estudio de cohorte ambispectiva en pacientes mayores de 18 años que ingresaron con diagnóstico de sepsis a la terapia intensiva. Se calculó la relación N/LP desde el ingreso hasta el séptimo día y hasta el diagnóstico de LRA y el desenlace. El análisis estadístico se realizó con chi cuadrada, V de Cramer y regresión logística multivariable. Resultados: de los 239 pacientes estudiados, la incidencia de LRA se desarrolló en el 70%. El 80.9% de los pacientes con relación N/LP > 3 tuvieron LRA (p = < 0.0001, V de Cramer 0.458, RM 3.05, IC al 95% 1.602-5.8) y mayor tratamiento sustitutivo renal (21.1 frente a 11.1%, p = 0.043). Conclusión: la relación N/LP > 3 tiene una asociación moderada con la LRA secundaria a sepsis en la unidad de terapia intensiva.

Relación neutrófilos, linfocitos y plaquetas como predictor de lesión renal aguda en sepsis por COVID-19 en la Unidad de Cuidados Intensivos / Neutrophiles, lymphocytes and platelets ratio as a predictor of acute kidney injury in sepsis by COVID-19 in the Intensive Care Unit / Relação de neutrófilos, linfócitos e plaquetas como preditor de lesão renal aguda na sepse por COVID-19 na Unidade de Terapia Intensiva

Resumen: Introducción: La lesión renal aguda se encuentra en 40% de los pacientes que presentan sepsis (S-LRA), ya que la inflamación es una de las causas fisiopatológicas de la lesión renal aguda. Durante la pandemia, la principal causa de sepsis en la unidad de cuidados intensivos (UCI) fue secundaria a enfermedad por coronavirus 2019 (COVID-19), en ésta se ha reportado incidencia de lesión renal de 36 a 75%. La fisiopatología de esta complicación aún no se conoce, pero se han demostrado mecanismos similares a la lesión renal séptica típica. La relación neutrófilos, linfocitos y plaquetas (RNLP) previamente se ha asociado con la presencia de lesión renal aguda en otros ámbitos (cirugía cardiaca y cirugía abdominal mayor), y en pacientes con sepsis secundaria a COVID-19 puede ser un marcador que identifique a los pacientes con riesgo de presentar esta complicación. Objetivo: Determinar si la relación neutrófilos, linfocitos y plaquetas es un predictor de lesión renal aguda en sepsis secundaria a COVID-19 en la UCI adultos. Material y métodos: Estudio de cohorte prospectiva, unicéntrico. En pacientes mayores de 18 años que ingresen a la UCI con diagnóstico de sepsis por COVID-19 se realizará el cálculo de la RNLP desde el día uno hasta el día siete. Se dividen en dos grupos: RNLP mayor de tres puntos y RNLP igual o menor de tres puntos, observando presencia o no de lesión renal aguda durante su estancia, y posterior al desenlace. Para el objetivo principal se hace prueba χ2, y se realiza prueba de regresión logística multivariable para valorar la asociación de las diferentes variables con el desenlace (OR IC95%). Resultados: Se estudió una población de 119 pacientes, se obtuvo una incidencia de lesión renal aguda inducida por sepsis (S-LRA) de 53.8% (IC95% 44-62%) en pacientes con sepsis secundaria a COVID-19, siendo la mayoría KDIGO I (53.2%). El grupo de RNLP mayor de tres tuvo una incidencia de 68.4% de S-LRA en comparación con el grupo de RNLP igual o menor de tres con 28% (p = 0.001, OR 4.255 IC95% 1.782-10.16), los pacientes con RNLP mayor de tres tuvieron estancia más prolongada en la UCI (12 versus 10 días, p = 0.018), y más tiempo de ventilación mecánica (11 versus ocho días, p = 0.003). Conclusión: El incremento de la relación neutrófilos, linfocitos y plaquetas es un factor de riesgo y puede ser pronóstico para la presencia de lesión renal aguda en sepsis por COVID-19 en la UCI.

Abstract: Introduction: Acute kidney injury is found in 40% of patients with sepsis (S-AKI), since inflammation is one of the pathophysiological causes of acute kidney injury. During the pandemic, the main cause of sepsis in the ICU was secondary to COVID-19, in which an incidence of kidney injury of 36 to 75% has been reported. The pathophysiology of this complication is not yet known, but mechanisms similar to typical septic kidney injury have been demonstrated. The neutrophil, lymphocyte and platelet ratio (RNLP) has previously been associated with the presence of acute kidney injury in other settings (cardiac surgery and major abdominal surgery) and in patients with sepsis secondary to COVID-19 it may be a marker that identifies the patients at risk of presenting this complication. Objective: To determine if the ratio of neutrophils, lymphocytes and platelets is a predictor of acute kidney injury in sepsis secondary to COVID-19 in the adult ICU. Material and methods: Prospective, single-center cohort study. In patients over 18 years of age who are admitted to the ICU with a diagnosis of sepsis due to COVID-19, the RNLP will be calculated from day 1 to day 7, it is divided into 2 groups: RNLP greater than 3 and RNLP equal or less than 3, observing the presence or not of acute kidney injury during their stay, and after the outcome, for the main objective a χ2 test is performed, and a multivariate logistic regression test is performed to assess the Association of the different variables with the outcome (OR with 95% CI). Results: A population of 119 patients was studied, there was an incidence of S-AKI of 53.8% (95% CI 44-62%) in patients with sepsis secondary to COIVD-19, the majority being KDIGO I (53.2%). In the RNLP group greater than 3 I had a 68.4% incidence of S-AKI compared to the RNLP group less than or equal to 3 with 28% (p = 0.001, OR 4.255 95% CI 1.72-10.16), the patients with RNLP greater than 3 had a longer stay in the ICU (12 vs 10 days, p = 0.018), and a longer time of mechanical ventilation (11 vs 8 days, p = 0.003). Conclusion: The increase in the neutrophil, lymphocyte and platelet ratio is a risk factor and can be a prognostic for the presence of acute kidney injury in sepsis due to COVID-19 in the ICU.

Resumo: Introdução: A lesão renal aguda é encontrada em 40% dos pacientes com sepse (S-LRA), uma vez que a inflamação é uma das causas fisiopatológicas da lesão renal aguda. Durante a pandemia, a principal causa de sepse na UTI foi secundária à COVID-19, na qual a incidência de lesão renal foi relatada de 36 a 75%. A fisiopatologia dessa complicação ainda não é conhecida, mas mecanismos semelhantes à lesão renal séptica típica foram demonstrados. A proporção de neutrófilos, linfócitos e plaquetas (RNLP) já foi associada à presença de lesão renal aguda em outros âmbitos (cirurgia cardíaca e cirurgia abdominal de grande porte) e em pacientes com sepse secundária à COVID-19 pode ser um marcador que identifica os pacientes em risco de apresentar esta complicação. Objetivo: Determinar se a proporção de neutrófilos, linfócitos e plaquetas é um preditor de lesão renal aguda na sepse secundária à COVID-19 na UTI adulto. Material e métodos: Estudo de coorte prospectivo, unicêntrico. Pacientes maiores de 18 anos admitidos na UTI com diagnóstico de sepse por COVID-19, o RNLP será calculado do dia 1 ao dia 7, dividido em 2 grupos: RNLP maior que 3 e RNLP igual ou inferior a 3, observando a presença ou não de lesão renal aguda durante sua internação, e posteriormente o desfecho, para o objetivo principal, é realizado um teste de χ2, e se realiza um teste de regressão logística multivariável para avaliar a associação das diferentes variáveis com o resultado (OR com 95% IC). Resultados: Estudou-se uma população de 119 pacientes, com incidência de S-LRA de 53.8% (IC 95% 44-62%) em pacientes com sepse secundária a COVID-19, sendo a maioria KDIGO I (53.2%). No grupo RNLP maior que 3, houve uma incidência de 68.4% de S-LRA comparado ao grupo RNLP menor ou igual a 3 com 28% (p = 0.001, OR 4.255, IC 95% 1.782-10.16), os pacientes com RNLP maior que 3 tiveram maior tempo de permanência na UTI (12 vs 10 dias, p = 0.018) e maior tempo em ventilação mecânica (11 vs 8 dias, p = 0.003). Conclusão: O aumento da proporção de neutrófilos, linfócitos e plaquetas é um fator de risco e pode ser prognóstico para a presença de lesão renal aguda na sepse por COVID-19 na UTI.

Antitarget Selectivity and Tolerability of Novel Pyrrolo[2,3-d]pyrimidine RET Inhibitors.

The selective inhibition of RET kinase as a treatment for relevant cancer types including lung adenocarcinoma has garnered considerable interest in recent years and prompted a variety of efforts toward the discovery of small-molecule therapeutics. Hits uncovered via the analysis of archival kinase data ultimately led to the identification of a promising pyrrolo[2,3-d]pyrimidine scaffold. The optimization of this pyrrolo[2,3-d]pyrimidine core resulted in compound 1, which demonstrated potent in vitro RET kinase inhibition and robust in vivo efficacy in RET-driven tumor xenografts upon multiday dosing in mice. The administration of 1 was well-tolerated at established efficacious doses (10 and 30 mg/kg, po, qd), and plasma exposure levels indicated a minimal risk of KDR or hERG inhibition in vivo, as evaluated by Miles assay and free plasma concentrations, respectively.

Both in vitro T cell proliferation and telomere length are decreased, but CD25 expression and IL-2 production are not affected in aged men.

INTRODUCTION: Aging is a natural process involving dysfunction of multiple organs and is characterized by increased susceptibility to infections, cancer and autoimmune diseases. The functionality of the immune system depends on the capacity of lymphocytes to proliferate in response to antigenic challenges, and telomere length has an important role regulating the number of cell divisions. The aim of this study was to determine the possible relationship between telomere length, interleukin 2 (IL-2) production, CD25 expression and proliferation of peripheral blood mononuclear cells (PBMCs) in aged men. MATERIAL AND METHODS: Telomere length was measured by RT-PCR in PBMCs from young and aged men. IL-2 production and CD25 expression were determined by ELISA and flow cytometry, respectively. Cell proliferation was measured by CFSE dilution assays upon in vitro stimulation with concanavalin A (Con A). RESULTS: PBMCs from aged men showed a shorter telomere length and a reduced capacity to proliferate in vitro, compared to young men. In contrast, no significant differences in the level of CD25 expression on T lymphocytes, and in vitro production of IL-2 were detected in both groups. In addition, no significant correlation was detected between levels of CD25 expression, IL-2 production, cell proliferation, and telomere length in aged men. CONCLUSIONS: In aged men the telomere length shortening and the reduced T cell proliferation are not related to the capacity of IL-2 production and CD25 expression on T lymphocytes.

Th17 and regulatory T cells in patients with different time of progression of type 2 diabetes mellitus.

INTRODUCTION: Type 2 diabetes mellitus (T2DM) is characterized by chronic inflammation, in which different types of immune cells participate, such as TH17 cells and Treg cells. The aim of this study was to determine the relationship between Treg and Th17 in patients with different times of T2DM progression. MATERIAL AND METHODS: Nineteen control subjects and 40 patients with T2DM were included. T2DM patients were classified into two groups: the first group consisted of twenty patients with less than10 years of disease progression (T2DM < 10), and the second group included 20 patients with a disease progression of 10 years or more (T2DM ≥ 10). Additionally, an analysis was performed according to the metabolic control, depending on HbA1c levels. The peripheral blood ratio of both Th17 and Treg cells was measured by standard flow cytometry protocols. RESULTS: No significant difference was found in Th17 cells of patients with T2DM < 10 or T2DM ≥ 10 and controls. With respect to CD4+CD25+FoxP3+ and CD4+CD25h Treg cells, a significant decrease was observed in patients with T2DM ≥ 10, mainly in patients with poor or moderate metabolic control. Statistical analysis performed in all patients with T2DM revealed a decrease in three cell subsets as well a negative correlation between Th17 cells and total cholesterol, CD4+CD25h cells with glucose and HbA1c levels, while a positive correlation was observed between CD4+CD25h cells and BMI. CONCLUSIONS: A decrease on both Treg and Th17 cell subsets in T2DM patients was observed suggesting that the metabolic decontrol and the progression time of T2DM could modify the proportions of Th17 and Treg cells.

Efficacy and Tolerability of Pyrazolo[1,5-a]pyrimidine RET Kinase Inhibitors for the Treatment of Lung Adenocarcinoma.

RET (REarranged during Transfection) kinase gain-of-function aberrancies have been identified as potential oncogenic drivers in lung adenocarcinoma, along with several other cancer types, prompting the discovery and assessment of selective inhibitors. Internal mining and analysis of relevant kinase data informed the decision to investigate a pyrazolo[1,5-a]pyrimidine scaffold, where subsequent optimization led to the identification of compound WF-47-JS03 (1), a potent RET kinase inhibitor with >500-fold selectivity against KDR (Kinase insert Domain Receptor) in cellular assays. In subsequent mouse in vivo studies, compound 1 demonstrated effective brain penetration and was found to induce strong regression of RET-driven tumor xenografts at a well-tolerated dose (10 mg/kg, po, qd). Higher doses of 1, however, were poorly tolerated in mice, similar to other pyrazolo[1,5-a]pyrimidine compounds at or near the efficacious dose, and indicative of the narrow therapeutic windows seen with this scaffold.

The Four Horsemen in Colon Cancer.

Worldwide, neoplasms of the gastrointestinal tract have a very high incidence and mortality. Among these, colorectal cancer, which includes colon and rectum malignancies, representing both highest incidence and mortality. While gallbladder cancer, another neoplasm associated to gastrointestinal tract occurs less frequently. Genetic factors, inflammation and nutrition are important risk factors associated with colorectal cancer development. Likewise, pathogenic microorganisms inducing intestinal dysbiosis have become an important scope to determine the role of bacterial infection on tumorigenesis. Interestingly, in human biopsies of different types of gastrointestinal tract cancer, the presence of different bacterial strains, such as Fusobacterium nucleatum, Escherichia coli, Bacteroides fragilis and Salmonella enterica have been detected, and it has been considered as a high-risk factor to cancer development. Therefore, pathogens infection could contribute to neoplastic development through different mechanisms; including intestinal dysbiosis, inflammation, evasion of tumoral immune response and activation of pro-tumoral signaling pathways, such as ß catenin. Here, we have reviewed the suggested bacterial molecular mechanisms and their possible role on development and progression of gastrointestinal neoplasms, focusing mainly on colon neoplasms, where the bacteria Fusobacterium nucleatum, Escherichia coli, Bacteroides fragilis and Salmonella enterica infect.

Diet-induced obese mice exhibit altered immune responses to early Salmonella Typhimurium oral infection.

Obesity is a chronic disease associated with different metabolic diseases as well as alterations in immune cell function. It is characterized by a chronic systemic low grade inflammation. There are several studies demonstrating the influence of obesity on the impaired immune response to infection. However, it is not completely clear whether the obese environment influences the development or maintenance of the immune response against infections. The aim of this study was to determine how obesity induced by a high-fat diet affects the immune response to an early oral Salmonella infection. Four groups of mice were kept in separate cages. Two of these designated as controls, fed with a normal diet; whereas other two groups were fed with a high fat diet for 10 weeks. Some mice were used for Salmonella oral infection. After 7 days of oral infection with S. Thypimurium the proportions of spleen cell subsets expressing activation markers in normal diet and HFD obese mice were stained with monoclonal antibodies and analyzed by flow cytometry. Also, mRNA levels of different cytokines were quantified by RT-PCR. It was found that obesity affects the function of the immune system against an early oral Salmonella infection, decreasing NK cells, altering the expression of activation molecules as well as cytokines mRNA levels. Interestingly, the expression some activation molecules on T lymphocytes was reestablished after Salmonella infection, but not the CD25 expression. Immune alterations could lead to immunosuppression or increased susceptibility to infections in HFD obese mice.

A PPARγ, NF-κB and AMPK-dependent mechanism may be involved in the beneficial effects of curcumin in the diabetic db/db mice liver.

Turmeric (Curcuma longa) is a rhizomatous herbaceous perennial plant of the ginger family which has been used to treat biliary disorders, anorexia, cough, rheumatism, cancer, sinusitis, hepatic disorders, hyperglycemia, obesity, and diabetes in both Ayurvedic and Traditional Chinese Medicine. Suggested mechanisms of action include the modulation of signal transduction cascades and effects on gene expression, however they remain to be elucidated. In this study, the expression of some proteins responsible for transcription factors, inflammation, and metabolic control were evaluated by western blot in 15-week-old db/db mice livers treated with curcumin 0.75% mixed in their diet for 8 weeks. In addition, nitrosative stress was evaluated. Curcumin increased the expression of AMPK and PPARγ, and diminished NF-κB protein in db/db mice. However, it did not modify the expression of PGC-1α or SIRT1. Nitrosative stress present in db/db mice livers was determined by a unique nitrotyrosylated protein band (75 kDa) and was not reverted with curcumin. In conclusion, curcumin regulates the expression of AMPK, PPARγ, and NF-κB; suggesting a beneficial effect for treatment of T2DM complications. In order to observe best beneficial effects it is desirable to administer curcumin in the earlier states of T2DM.

Effects of curcumin on brain-derived neurotrophic factor levels and oxidative damage in obesity and diabetes.

We evaluated the effects of curcumin treatment on protein oxidation (PO), lipid peroxidation (LP) and brain-derived neurotrophic factor (BDNF) levels in the hippocampus and frontal cortex (FC) of diabetic db/db mice (DM) and in sera of obese humans. Thus, DM were treated daily with 50 mg/kg of curcumin during an 8-week period. Obese human were treated daily with 500 and 750 mg of curcumin that was administered orally for 12 weeks; BDNF, PO and LP levels in sera were determined at in weeks 0, 2, 6 and 12 of treatment. BDNF levels decreased in hippocampus and FC of DM as compared with untreated wild-type mice. Curcumin improved or restored BDNF levels to normal levels in DM, but curcumin did not have any effect on BDNF levels in sera of obese humans. In hippocampus and FC of DM, hyperglycaemia and curcumin did not have effect on LP levels. Hyperglycaemia increased PO levels in hippocampus and FC, whereas curcumin decreased these levels in hippocampus but not in FC. In sera of obese humans, the 500-mg dose decreased LP levels in weeks 6 and 12 when compared with basal levels, but the 750-mg dose did not have any effect; both doses of curcumin decreased PO levels in weeks 2, 6 and 12 of treatment when compared with basal levels. Present results suggest a therapeutic potential of curcumin to decrease oxidation caused by obesity in humans and also show that curcumin restores BDNF levels in DM.

Curcumin restores mitochondrial functions and decreases lipid peroxidation in liver and kidneys of diabetic db/db mice.

BACKGROUND: Nitrosative and oxidative stress play a key role in obesity and diabetes-related mitochondrial dysfunction. The objective was to investigate the effect of curcumin treatment on state 3 and 4 oxygen consumption, nitric oxide (NO) synthesis, ATPase activity and lipid oxidation in mitochondria isolated from liver and kidneys of diabetic db/db mice. RESULTS: Hyperglycaemia increased oxygen consumption and decreased NO synthesis in liver mitochondria isolated from diabetic mice relative to the control mice. In kidney mitochondria, hyperglycaemia increased state 3 oxygen consumption and thiobarbituric acid-reactive substances (TBARS) levels in diabetic mice relative to control mice. Interestingly, treating db/db mice with curcumin improved or restored these parameters to normal levels; also curcumin increased liver mitochondrial ATPase activity in db/db mice relative to untreated db/db mice. CONCLUSIONS: These findings suggest that hyperglycaemia modifies oxygen consumption rate, NO synthesis and increases TBARS levels in mitochondria from the liver and kidneys of diabetic mice, whereas curcumin may have a protective role against these alterations.

Curcumin restores mitochondrial functions and decreases lipid peroxidation in liver and kidneys of diabetic db/db mice

BACKGROUND: Nitrosative and oxidative stress play a key role in obesity and diabetes-related mitochondrial dysfunction. The objective was to investigate the effect of curcumin treatment on state 3 and 4 oxygen consumption, nitric oxide (NO) synthesis, ATPase activity and lipid oxidation in mitochondria isolated from liver and kidneys of diabetic db/db mice. RESULTS: Hyperglycaemia increased oxygen consumption and decreased NO synthesis in liver mitochondria isolated from diabetic mice relative to the control mice. In kidney mitochondria, hyperglycaemia increased state 3 oxygen consumption and thiobarbituric acid-reactive substances (TBARS) levels in diabetic mice relative to control mice. Interestingly, treating db/db mice with curcumin improved or restored these parameters to normal levels; also curcumin increased liver mitochondrial ATPase activity in db/db mice relative to untreated db/db mice. CONCLUSIONS: These findings suggest that hyperglycaemia modifies oxygen consumption rate, NO synthesis and increases TBARS levels in mitochondria from the liver and kidneys of diabetic mice, whereas curcumin may have a protective role against these alterations.

[Neuroendocrine differentiation in prostate adenocarcinoma]. / Diferenciación neuroendocrina en adenocarcinoma de próstata.

The human prostate is a gland composed of many types of cells and extracellular components with specific functions. The stromal compartment includes nerve tissue, fibroblasts, lymphocytes, macrophages, endothelial cells, and smooth muscular cells. The epithelial compartment is composed of luminal epithelial cells, basal cells, and a lesser number of neuroendocrine cells, which are transcendental in growth regulation, differentiation, and secretory function. In prostate cancer, neuroendocrine cells replicate especially in high grade and advanced stage, and hormonally treated tumoral cells adopt characteristics that make them resistant to hormonal deprivation. Androgen receptors have a crucial role in tumorigenesis of prostate adenocarcinoma. Deprivation hormone therapy blocks the expression of androgen receptors in the prostatic epithelial cells. Neuroendocrine cells lack androgen receptors; their growth is hormonally independent and that is why deprivation hormonal therapy does not eliminate the neoplasic neuroendocrine cells. In contrast, these types of cells proliferate after therapy and make a paracrine network, stimulating the proliferation of androgen-independent neoplastic cells, which finally lead to tumoral recurrence. In this work we describe the neuroendocrine function in normal tissue and in prostatic adenocarcinoma, including neoplasic proliferation stimulation, invasion, apoptosis resistance, and angiogenesis, and describe some molecular pathways involved in this neuroendocrine differentiation.

Increased telomere length and proliferative potential in peripheral blood mononuclear cells of adults of different ages stimulated with concanavalin A.

BACKGROUND: Recently, a direct correlation with telomere length, proliferative potential and telomerase activity has been found in the process of aging in peripheral blood cells. The objective of the study was to evaluate telomere length and proliferative potential in peripheral blood mononuclear cells (PBMCs) after stimulation with Concanavalin A (ConA) of young adults compared with older adults. METHODS: Blood samples were obtained from 20 healthy young males (20-25 years old) (group Y) and 20 males (60-65 years old) (group O). We compared PBMC proliferation before and after stimulation with ConA. DNA was isolated from cells separated before and after culture with ConA for telomeric measurement by real-time polymerase chain reaction. RESULTS: In vitro stimulation of PBMCs from young subjects induced an increase of telomere length as well as a higher replicative capacity of cell proliferation. Samples from older adults showed higher loss of telomeric DNA (p = 0.03) and higher levels of senescent (≤6.2 kb) telomeric DNA (p = 0.02) and displayed a marked decrease of proliferation capacity. Viability cell counts and CFSE tracking in 72-h-old cell cultures indicated that group O PBMCs (CD8+ and CD4+ T cells) underwent fewer mitotic cycles and had shorter telomeres than group Y (p = 0.04). CONCLUSIONS: Our findings confirm that telomere length in older-age adults is shorter than in younger subjects. After stimulation with ConA, cells are not restored to the previous telomere length and undergo replicative senescence. This is in sharp contrast to the response observed in young adults after ConA stimulation where cells increase in telomere length and replicative capacity. The mechanisms involved in this phenomenon are not yet clear and merit further investigation.

[Cells of innate and adaptive immunity in type 2 diabetes and obesity]. / Células de la inmunidad innata y adaptativa en la diabetes mellitus tipo 2 y obesidad.

Both type 2 diabetes mellitus (T2DM) and obesity are a major public health problem in Mexico and around the world for increased incidence. In T2DM, insulin secretion, insulin action or both are altered. Also, in T2DM as well as in obesity a low grade chronic inflammation has been associated. In both conditions there is an important increase of visceral adipose tissue, which induces to an up-regulation of synthesis in proinflammatory molecules. This process involves different subsets of the immune system. The macrophages and monocytes are the best studied, but recently has been reported the involvement of other type of cells; such as neutrophils, mast cells, eosinophils, dendritic cells, NKs, NKT. Also, some T cells subsets, such as Th1, Th2, T regulatory, Th17 and B cells seems to be involved in the low grade chronic inflammation. This review focuses on recent evidences of the role of innate and adaptative immune system cells in the pathology of T2DM and obesity. We concluded with the general proposal of a theoretical model, how the immune cells may participate in inflammation of fat tissue, insulin resistance and T2DM.

Intercellular adhesion molecules and mortality for sepsis in infants younger than 1 year of life.

BACKGROUND: Adhesion molecules in sepsis syndrome are correlated with the severity of illness and may be considered as predictors of survival outcome in adults. However, only few studies have been performed in infants and none using international criteria for sepsis. OBJECTIVE: To determine whether adhesion molecules during the first 7 days of the disease could predict sepsis outcome and its severity. MATERIAL AND METHODS: We performed a prospective study in 88 infants with sepsis and 30 controls. Soluble intercellular adhesion molecule (ICAM)-1, vascular cell adhesion molecule (VCAM)-1, and E-selectin levels were determined at days 1, 3 and 7 of follow-up in those patients with sepsis and only one determination in the control group. The main outcome measure was mortality during 10 days of monitoring. RESULTS: Positive hemoculture was reported in 64(72.7%). ICAM-1, VCAM-1, and E-selectin levels were higher in the group of sepsis than in the control group. However, no association was found between ICAM-1, VCAM-1 or E-selectin levels with sepsis severity. Mortality linked to sepsis was observed in 9 patients (10.2%). In the logistic regression analysis, those variables positively associated with mortality were the increase in ICAM-1 levels > 250 ng/mL between day 1 to 3, number of amines and the baseline severity of sepsis. However, we did not identify in those patients who died a specific pattern in adhesion molecules levels during follow up. CONCLUSIONS: ICAM-1 levels, number of amines and severity of sepsis levels predict mortality during 10 days of monitoring in infants younger than 1 year of age with sepsis.

Normal mouse kidneys contain activated and CD3+CD4- CD8- double-negative T lymphocytes with a distinct TCR repertoire.

Healthy liver, intestine, lung, and skin harbor resident lymphocytes with conventional and unconventional phenotypes. Lymphocytes also have been detected in healthy mice kidneys; however, these cells have not been well studied and have been largely overlooked. To better characterize the intra-renal lymphocytes, we extensively perfused C57BL/6J mice with PBS and then isolated mononuclear cells for flow cytometry analysis. We observed T cells, B cells, and NK cells in normal mice kidneys after extensive perfusion. Approximately 50% of kidney T lymphocytes expressed intermediate levels of CD3 (CD3int T cells). Similar to liver and lung, a high percentage of unconventional CD3+CD4(-)CD8(-) double-negative T cells was observed in normal mice kidneys, from which 11% expressed B220 antigen. Unlike the spleen and blood, the classic CD4+ and CD8+ T lymphocytes in the kidney had a high proportion of activated CD69+ and effector/memory CD44- CD62L ligand phenotypes. Also, a small percentage of CD4+CD25+forkhead box p3+ and NKT cells was observed in perfused and exanguinated kidneys. In addition, a distinct TCR repertoire was found on intra-renal conventional and unconventional T cells compared with those from the spleen. Finally, after 24 h of renal ischemia reperfusion injury (IRI), increased production of cytokines IFN-gamma and TNF-alpha by CD4+ and CD8+ T cells, isolated from perfused kidneys, was observed. These data suggest that some of these cells harbored in the kidney could be implicated in the immune response of the IRI pathogenic process.

Pharmacological analysis of the interaction of antimuscarinic drugs at M(2) and M(3) muscarinic receptors in vivo using the pithed rat assay.

Muscarinic receptor antagonists form the mainstay of the therapeutic options for airway, bladder, and gastrointestinal smooth muscle disorders. Both M(2) and M(3) muscarinic receptors are involved in mediating smooth muscle contractility, although the relative functional contribution of each subtype, especially in the disease state, is unclear. Because the potency and selectivity of compounds for a given receptor in an in vivo setting can be dissimilar to that observed in an in vitro system, we developed an in vivo assay to simultaneously determine the absolute potency and selectivity of muscarinic receptor antagonists at M(2) and M(3) receptors using the pithed rat. Methacholine (MCh)-induced bradycardia and depressor responses were used as surrogate functional endpoints for M(2) and M(3) receptor activation, respectively. The influence of the muscarinic antagonists, tolterodine, oxybutynin, darifenacin, Ro 320-6206, solifenacin, or tiotropium on the MCh-induced responses were studied. The estimated DR(10) values (dose producing a tenfold shift in the MCh curve) of tolterodine, oxybutynin, darifenacin, Ro 320-6206, solifenacin, and tiotropium for the M(2) muscarinic receptor-mediated bradycardia were 0.22, 1.18, approximately 2.6, 0.025, 0.40, and 0.0026 mg/kg, respectively, and 0.14, 0.18, 0.11, 3.0, 0.18, and 0.0017 mg/kg, respectively, for the M(3) muscarinic receptor-mediated depressor response. In a separate set of experiments, a single intravenous dose of tiotropium was administered before a MCh curve at 1, 3, 6, or 9 h to determine if tiotropium exhibited time-dependent selectivity for the M(3) receptor as has been reported from in vitro studies. The results indicate a slight preference of tiotropium for the M(3) receptor at later time points. The pithed rat assay may serve useful for elucidating the functional contribution of M(2) and M(3) receptors to the in vivo pharmacological effects of antagonists in disease animal models.