Albumin, the responsible protein of the Cu2+-dependent hydrolysis of O-hexyl O-2,5-dichlorophenyl phosphoramidate (HDCP) by chicken serum "antagonistic stereoselectivity"

O-hexyl O-2,5-dichlorophenyl phosphoramidate (HDCP) is a chiral analogous compound of the methamidophos insecticide that induces delayed neuropathy, and the R-(+)-HDCP enantiomer is an inhibitor of neuropathy target esterase (NTE). This enantiomer is not hydrolized by Ca2+-dependent phosphotriesterases in mammal tissues. Our group had reported R-(+)-HDCP hydrolysis in chicken serum enhanced by 30–250 μM copper in ex vivo assays, which we call "antagonistic stereoselectivity". We checked the hypothesis of the role of cupper binding proteins. Two hundred micrograms of human serum ceruloplasmine or horse kidney methallotionein in 1 mL containing 400 μM HDCP for 60 min showed no significant Cu2+-dependent hydrolysis. However under the same conditions, 10 μL of chicken serum or 10 μL of buffer containing 216 μg of chicken serum albumin (CSA) (amount of albumin content in this serum volume) with 100 μM Cu2+ showed the same stereoselectivity and similar levels to the Cu2+-dependent R-(+)-HDCP hydrolysis. About 75% of R-(+)-HDCP were hydrolyzed after 120 min in the presence of 100 μM Cu2+ (inhibited by 5 mM EDTA). No effects was observed by divalent cations Cu2+, Zn2+, Fe2+, Ca2+, Mn2+ and Mg2+. These results confirm that albumin is the protein responsible for "antagonistic stereoselectivity" observed in chicken ser

DAEH N-terminal sequence of avian serum albumins as catalytic center of Cu (II)-dependent organophosphorus hydrolyzing A-esterase activity

O-hexyl O-2,5-dichlorophenyl phosphoramidate (HDCP) induces delayed neuropathy. The R (+)-HDCP inhibits and caused the so call “aging reaction” on inhibited-NTE. This enantiomer is not hydrolyzed by Ca(II)-dependent A-esterases in mammal tissues but is hydrolyzed by Cu(II)-dependent chicken serum albumin (CSA). With the aim of identifying HDCP hydrolysis by other vertebrate albumins, we incubated albumin with 400 μM racemic HDCP in the presence of 100 μM copper sulfate. HDCPase activity was assessed by measurement of HDCP with chiral chromatography. Human, sheep, dog, pig, lamprey or cobra serum albumin did not show a significant activity (~10%). Rabbit and bovine albumins hydrolyzed both enantiomers of HDCP (25% and 50% respectively). Turkey serum albumin had more HDCPase activity (~80 μM remaining) than the chicken albumin (~150 μM remaining). No animal albumins other than chicken showed stereoselective hydrolysis. Preincubation of chicken albumin with 1 mM the histidine modifying agents, 100 μM N-bromosuccinimide (NBS) and Zn(II), inhibited its Cu(II)-dependent R (+)-HDCPase activity, where as other mM amino acids modifiers had no inhibitory effects. . These results confirm that the stereoselective hydrolysis of (+)-HDCP is a specific A-esterase catalytic property of chicken albumin. The higher HDCPase activity by turkey albumin suggests the amino-terminal sequence of avian albumins (DAEHK) is the active center of this Cu(II)-dependent A-esterase activity

Intervalos de referencia del perfil de lípidos en trabajadores y estudiantes de la Universidad Autónoma del Estado de Morelos, México

Introduction. Interpreting medical data from clinical laboratory analyses is a key process in making clinical patient decisions. The decisions are based upon comparing patient results against the reference intervals that are calculated using results from human reference populations. Objective. Determine the reference intervals of the lipid profile of worker and student populations at the Autonomous University of Morelos State. Materials and Methods. The subjects were a volunteer group of 142 participants between 20 and 45 years old. The concentration of total cholesterol, triglycerides, HDL-cholesterol and LDLcholesterol was determined by endpoint methods of enzymic analyses. The reference intervals were established using non - parametric methods within the two tailed 2.5 and 97.5 percentiles. Results. The reference intervals were: total cholesterol 90.0-208.3 mg/dL (2.4-5.4 mmol/L); triglycerides 40.7-215.8 mg/dL (0.46-2.5 mmol/L); HDL-cholesterol 26.0-74.9 mg/dL (0.68-1.9 mmol/L) and LDL-cholesterol 31.4-126.7 mg/dL (0.82-3.3 mmol/L). Conclusions. Comparing our data with other populations found that our results are very similar to other reference populations, and we recommend the use of these values for the clinical interpretation of the university population of the Morelos State, where food, genetics and environmental factors are characteristic of this population.Introducción. La interpretación médica de los datos de laboratorio clínico resulta ser un proceso fundamental para las decisiones clínicas sobre el paciente y se basan en la comparación de los resultados obtenidos frente al intervalo de referencia calculado dentro de la misma población considerada de referencia. Objetivo. Determinar los intervalos de referencia del perfil de lípidos en una población de trabajadores y estudiantes de la Universidad Autónoma del Estado de Morelos (UAEM), México. Materiales y Métodos. Se incluyeron 142 participantes sanos de entre 20 y 45 años; se determinaron las concentraciones de colesterol total, triglicéridos, HDL-colesterol y LDLcolesterol, mediante técnicas enzimáticas de punto final. Los intervalos de referencia se establecieron con base en métodos no paramétricos, dentro del percentil 2.5% y 97.5%. Resultados. Se obtuvieron los siguientes intervalos de referencia: colesterol total 90.0-208.3 mg/ dL (2.4-5.4 mmol/L); triglicéridos 40.7-215.8 mg/ dL (0.46-2.5 mmol/L); HDL-colesterol 26.0-74.9 mg/dL (0.68-1.9 mmol/L) y LDL-colesterol 31.4- 126.7 mg/dL (0.82-3.3 mmol/L). Conclusiones. Al comparar nuestros datos con otras poblaciones, encontramos que los resultados son muy semejantes y recomendamos el uso de estos valores para la interpretación clínica de la población universitaria del Estado de Morelos, donde la alimentación, la genética y los factores ambientales son característicos de esta población

EVALUACIÓN DE ASPECTOS CLÍNICOS RELACIONADOS CON LA AMPUTACIÓN DE MIEMBROS INFERIORES EN PERSONAS QUE VIVEN CON DIABETES MELLITUS TIPO 2 EN MÉXICO: EVALUATION OF CLINICAL ASPECTS RELATED TO LOWER LIMB AMPUTATION AMONG INDIVIDUALS LIVING WITH TYPE 2 DIABETES MELLITUS IN MEXICO

Introduction: Diabetes continues to be one of the leading causes of disability and death in the global population. It is estimated that approximately 25% of people living with diabetes will develop an ulcer on one of their lower pelvic limbs. Objective: The present study evaluates the clinical aspects related to lower pelvic limb amputation in a cohort of patients with diabetes mellitus. Materials and Methods: A retrospective cross-sectional study was conducted consisting in a review of the total census of patients with type 2 diabetes. They received medical attention in the setting of the study in the period January-May 2022. According to the ICD-10 a total of 2021 patients with multiple complications of diabetes were identified through their clinical records. One hundred patients were diagnosed with diabetic foot. Results: Degree of average glycemic control in the 6 months following diagnosis of diabetic foot was a relevant variable. Since patients with optimal glycemic control by quantifying their fasting glucose levels (<130 mg/dl) as well as their glycated hemoglobin values (< 7%), had a less frequency of amputations (p˂0.001; Chi2) compared to those patients without an adequate glycemic control. Conclusions: After the analysis conducted in the population, it was found that being male, having glycated hemoglobin values higher than 7%, and having average fasting glucose values higher than 130 mg/L increase the probability of presenting a lower extremity amputation.  Introducción: La diabetes continua siendo una de las principales causas de discapacidad y  muerte en la población mundial. Se estima que cerca del 25% de las personas que viven con diabetes desarrollarán  una úlcera en alguna  de sus miembros pélvicos inferiores. Objetivo: El presente estudio evalúa los aspectos clínicos relacionados con la amputación del miembro inferior pélvico en una cohorte de pacientes con diabetes mellitus. Métodos: Se realizó un estudio transversal retrospectivo consistente en la revisión del censo total de pacientes con diabetes tipo 2. Recibieron atención médica en el ámbito del estudio en el periodo enero-mayo de 2022. Según la CIE-10 se identificaron a través de sus historias clínicas un total de 2021 pacientes con múltiples complicaciones de la diabetes. Cien pacientes fueron diagnosticados de pie diabético. Resultados: El grado de control glucémico medio en los 6 meses siguientes al diagnóstico de pie diabético fue una variable relevante. Ya que los pacientes con un control glucémico óptimo cuantificando sus niveles de glucosa en ayunas (<130 mg/dl) así como sus valores de hemoglobina glicosilada (< 7%), tuvieron una menor frecuencia de amputaciones (p˂0,001; Chi2) en comparación con aquellos pacientes sin un control glucémico adecuado. Conclusiones: Después del análisis realizado en la población, se encontró que ser hombre, tener valores de hemoglobina glucosilada superiores al 7% y tener valores promedio de glucosa en ayunas superiores a 130 mg/L aumentan la probabilidad de presentar una amputación de extremidad inferior

The Neurotoxic Effect of Ochratoxin-A on the Hippocampal Neurogenic Niche of Adult Mouse Brain

Ochratoxin A (OTA) is a common secondary metabolite of Aspergillus ochraceus, A. carbonarius, and Penicillium verrucosum. This mycotoxin is largely present as a contaminant in several cereal crops and human foodstuffs, including grapes, corn, nuts, and figs, among others. Preclinical studies have reported the involvement of OTA in metabolic, physiologic, and immunologic disturbances as well as in carcinogenesis. More recently, it has also been suggested that OTA may impair hippocampal neurogenesis in vivo and that this might be associated with learning and memory deficits. Furthermore, aside from its widely proven toxicity in tissues other than the brain, there is reason to believe that OTA contributes to neurodegenerative disorders. Thus, in this present in vivo study, we investigated this possibility by intraperitoneally (i.p.) administering 3.5 mg OTA/kg body weight to adult male mice to assess whether chronic exposure to this mycotoxin negatively affects cell viability in the dentate gyrus of the hippocampus. Immunohistochemistry assays showed that doses of 3.5 mg/kg caused a significant and dose-dependent reduction in repetitive cell division and branching (from 12% to 62%). Moreover, the number of countable astrocytes (p < 0.001), young neurons (p < 0.001), and mature neurons (p < 0.001) negatively correlated with the number of i.p. OTA injections administered (one, two, three, or six repeated doses). Our results show that OTA induced adverse effects in the hippocampus cells of adult mice brain tissue when administered in cumulative doses

Additional file 1: of Preclinical evaluation of anti-Helicobacter spp. activity of Hippocratea celastroides Kunth and its acute and sub-acute toxicity

HPTLC profiling of H. celastroides and H. excelsa for alkaloids and triterpenes analysis. (DOCX 239 kb