Hiperglucemia en ayuno, medicamentos y plantas medicinales en un laboratorio clínico de Oaxaca, México / Fasting hyperglycemia, drugs and medicinal plants in a clinical laboratory in Oaxaca, Mexico

Resumen Objetivo: Determinar la prevalencia de hiperglucemia en ayuno y el uso de medicamentos y plantas medicinales en pacientes atendidos en el laboratorio de análisis clínicos HECA LAB de Ciudad Ixtepec, Oaxaca. Materiales y Métodos: Se realizó un estudio descriptivo, transversal durante el cuarto trimestre del año 2021. Se llevó a cabo un muestreo no probabilístico, por conveniencia, incluyendo un total de 205 pacientes. La recolección de los datos se llevó a cabo mediante la aplicación de un cuestionario estructurado que contenía información sociodemográfica. A los pacientes con hiperglucemia se les interrogó sobre el consumo de medicamentos y plantas medicinales. Los niveles de glucosa fueron determinados mediante el analizador semiautomático de química clínica Mindray, modelo BA-88A. Se consideró como diabético un valor de glucosa mayor o igual a 126 mg/dl de acuerdo a los criterios de la Organización Mundial de la Salud. Se calculó la prevalencia por fórmula, según Aschengrau & Seage. Resultados: De los 205 pacientes, el 29% presentó diabetes. El 8% se clasificó como prediabético con predominio del sexo femenino. El rango de edad más frecuente fue de 54-65 años con el 32%. El 82% refirió llevar tratamiento farmacológico y el 63% de los encuestados manifestó consumir plantas medicinales para el tratamiento de la diabetes. Conclusiónes: La prevalencia de diabetes en la muestra estudiada fue del 29%. La monoterapia oral con metformina, fue el tratamiento más frecuente, seguido por el uso de insulina vía subcutánea. Fueron referidas 11 plantas medicinales, siendo la moringa (Moringa oleifera) la más empleada. El 18% de los pacientes con diabetes no recibió tratamiento farmacológico. Por ello, es necesario establecer intervenciones oportunas de educación en salud, sobre cambios en el estilo de vida y adherencia al tratamiento farmacológico de los pacientes con diabetes mellitus; para lograr un manejo adecuado de la enfermedad y evitar complicaciones.

Abstract Objetive: To determinate the prevalence of fasting hyperglycemia and the use of drugs and medicinal plants in patients trated at the HECA LAB clinical analysis laboratory in Ciudad Ixtepec, Oaxaca. Materials and Methods: A descriptive, cross-sectional study was carried out during the fourth quarter of 2021. A non-probabilistic sampling was carried out, for convenience, incluiding a total of 205 patients. Data collection was carried out by applying a structured questionnaire containing sociodemographic information such as age and sex. Patients with hyperglycemia were questioned about the consumption of medicines and medicinal plants. Glucose levels were determined using the Mindray semi-automatic clinical chemistry analyzer, model BA-88A. A glucose value greater than or equal to 126 mg/dl according to the WHO criteria was considered diabetic. Prevalence was calculated by formula, according to Aschengrau & Seage. Results: Of the 205 patients, 29% had diabetes. 8% were classified as prediabetic with a predominance of females. The most frequent age range was 54-65 years with 32%. 82% reported taking pharmacological treatment and 63% of those surveyed reported consuming medicinal plantas for the treatment of diabetes. Conclusions: The prevalence of diabetes in the studied patients was 29%. Oral monotherapy with metformin was the most frequent treatment, followed by the use of subcutaneous insulin. Eleven medicinal plants were referred, being moringa (Moringa oleifera), the most used. 18% did not receive pharmacological treatment. For this reason, it is necessary to establish timely health education interventions on lifestyle changes and adherence to pharmacological treatment of patients with diabetes mellitus; to achieve adequate management of the disease and avoid complications.

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.

Hydrolysis of chiral organophosphorus compounds by phosphotriesterases and mammalian paraoxonase-1.

Some organophosphorus compounds (OPs), which are used in the manufacturing of insecticides and nerve agents, are racemic mixtures with at least one chiral center with a phosphorus atom. Acute exposure of humans to these mixtures induces the covalent modification of acetylcholinesterase (AChE) and neuropathy target esterase (NTE) and causes a cholinergic syndrome or organophosphate-induced delayed polyneuropathy syndrome (OPIDP). These irreversible neurological effects are due to the stereoselective interaction of the racemic OPs with these B-esterases (AChE and NTE) and such interactions have been studied in vivo, ex vivo and in vitro, using stereoselective hydrolysis by A-esterases or phosphotriesterases (PTEs) and the PTE from Pseudomonas diminuta, and paraoxonase-1 (PON1) from mammalian serum. PON1 has a limited hydrolytic potential of the racemic OPs, while the bacterial PTE exhibits a significant catalytic activity on the less toxic isomers P(+) of the nerve agents. Avian serum albumin also shows a hydrolyzing capacity of chiral OPs with oxo and thio forms. There are ongoing environmental and bioremediation efforts to design and produce recombinants as bio-scavengers of OPs.

Copper(II)-dependent hydrolysis of trichloronate by turkey serum albumin.

Trichloronate is a racemic organophosphonothioate insecticide that induced delayed neuropathic in hens and human. The avian are species with great susceptibility to organophosphorus poisoning due to their low levels of A-esterases. However, a significant copper-dependent A-esterase activity has been identified in chicken and turkey serum. This study aims at characterizing the trichloronate enantiomers hydrolysis by serum and albumin from chicken (CSA) and turkey (TSA) in the presence of copper by chiral chromatography. A significant Cu2+-dependent hydrolysis for both trichloronate enantiomers (38%) was observed in turkey serum and TSA (p < 0.05). The hydrolysis is stereoselective for (-)-trichloronate (p < 0.05). TSA incubation (200 µg) showed residual values of 56 µM and 20 µM of (+)-trichloronate and (-)-trichloronate, respectively; while the chicken serum and CSA presented a slight hydrolysis (1-7%) of both enantiomers. This copper-dependent hydrolysis and stereoselectivity of trichloronate by TSA was proportional to the incubation time. The increase of TSA in the assay (200-3000 µg) in the presence of 100 µM copper did not significantly increase the levels of hydrolysis and stereoselectivity, an opposite effect was observed for turkey serum (100-200 mL), which totally inhibited this copper-dependent activity of both isomers. The present study evidences an A-esterase activity of TSA on a thio form OP compound, which is stereoselective and activated by copper.

Copper-dependent hydrolysis of trichloronate by turkey serum studied with use of new analytical procedure based on application of chiral chromatography and UV/Vis spectrophotometry.

Trichloronate is a racemic organophosphate, which has been used for the manufacture of insecticides. This compound induces delayed neuropathy in hen and humans. This study shows the Cu2+-dependent hydrolysis of trichloronate by turkey serum using UV/Vis spectrophotometry and chiral chromatography. The CHIRALCEL OD column and mobile phase of heptane allowed a resolution of 1.15 of its two enantiomers, while the liquid-liquid extraction showed a recovery of 95-98%. The optimum linear response was of 50 to 800 µM with a detection and quantification limit of 0.6 and 2 µM for (+)-trichloronate, and 0.7 and 2.3 µM for (-)-trichloronate. The levels of Cu2+-dependent hydrolysis (µM remaining concentration) quantified for 60 min at 37 °C and pH 7.4 were statistically higher (p ˂ 0.05) for (-)-trichloronate (65%) than (+)-trichloronate (32%). This stereoselective hydrolysis was confirmed by UV/Vis spectrophotometry using 2,4,5­trichlorophenol as standard, each of the enantiomers (93-95% purity) collected by HPLC, as well as aminoantipyrine and ferricyanide reagents to yield a colored product. This method exhibited an optimal linearity (r > 0.99) and a higher Cu2+-dependent hydrolysis (p < 0.05) to (-)-trichloronate (47%) than its corresponding (+)-form (31%). This results shows the Cu2+-dependent stereoselective hydrolysis of a racemic OP in its thio form (P = S) by an A-esterase of the turkey serum through the development of a colorimetric method and optimization of an existing chiral chromatographic method.

Paraoxonase-1 polymorphisms and cerebral ischemic stroke: a pilot study in mexican patients.

BACKGROUND: The serum paraoxonase-1 (PON1) associated to HDL presents two common polymorphisms in the positions 192 and 55. These polymorphisms are considered determinant of the capacity of HDL to protect LDL from their oxidative modification. In this context, the PON1 genotype has been associated with cardiovascular diseases, including stroke. OBJECTIVE: To determine the allelic and genotypic frequencies of PON1 L55M and Q192R as well as the enzymatic activities of PON1 in subjects with and without atherothrombotic stroke. METHODS: There were included 28 people with atherothrombotic stroke and 29 without stroke. The genotyping was carried out by PCR-RFLP and the phenotyping by measurement of the activities of paraoxonase and arylesterase in serum. RESULTS: For the polymorphism Q192R, the allelic frequencies (Q/R) were 0.46/0.54 and 0.48/0.52 (p= 0.843) for the control group and the group with stroke, respectively. While for the polymorphism L55M, the allelic frequencies (L/M) were 0.81/0.19 for the control group, and 0.78/0.22 for the group with stroke (p= 0.610). The activity levels of paraoxonase were not significantly different between the control and stroke groups (450 vs. 348 UI/mL, p= 0.093) While the activity levels of arylesterase were significantly different between the studied groups (90 vs. 70 UI/mL, p= 0.001); however, upon adjustment by multiple linear regression, it was not longer significant. CONCLUSION: The polymorphisms Q192R and L55M, and the paraoxonase activity of PON1 are not risk factors for atherothrombotic stroke according to the results of this study.

INTRODUCCIÓN: La paraoxonasa-1 (PON1) sérica asociada a las HDL presenta dos polimorfismos comunes en las posiciones 192 y 55. Estos polimorfismos se consideran determinantes para la capacidad de las HDL de proteger a las LDL de su modificación oxidativa. En este contexto, el genotipo de PON1 se ha asociado con enfermedades cerebrovasculares, que incluyen el infarto cerebral. OBJETIVO: Determinar las frecuencias alélicas y genotípicas de PON1-L55M y PON1- Q192R, así como las actividades enzimáticas de PON1 en sujetos con y sin infarto cerebral aterotrombótico. MÉTODOS: Se incluyeron 28 personas con infarto cerebral aterotrombótico y 29 sin infarto. Las genotipificaciones se realizaron mediante PCR-RFLP y las fenotipificaciones mediante la medición de las actividades paraoxonasa y arilesterasa en suero. RESULTADOS: Para el polimorfismo Q192R, las frecuencias alélicas (Q/R) fueron 0.46/0.54 y 0.48/0.52 (p= 0.843) para el grupo control y el grupo con infarto, respectivamente. Mientras que para el polimorfismo L55M, las frecuencias alélicas (L/M) fueron 0.81/0.19 para el grupo control y 0.78/0.22 para el grupo con infarto (p= 0.610). Los niveles de actividad paraoxonasa no fueron significativamente diferentes entre los grupos control y con infarto (450 vs. 348 Ul/mL, p= 0.093). Mientras que los niveles de actividad arilesterasa fueron significativamente diferentes entre los grupos estudiados (90 vs. 70 Ul/mL, p= 0.001), sin embargo, al ajustarla por regresión lineal múltiple, dejo de ser significativa. CONCLUSIÓN: Los polimorfismos Q192R y L55M, y la actividad paraoxonasa de la PON1 no son factores de riesgo para el infarto cerebral aterotrombótico en este estudio.

Paraoxonase-1 polymorphisms and cerebral ischemic stroke: a pilot study in mexican patients / Caracterización polimórfica de la paraoxonasa-1 en el infarto cerebral aterotrombótico de una poblacion mexicana: estudio piloto

Abstract Background: The serum paraoxonase-1 (PON1) associated to HDL presents two common polymorphisms in the positions 192 and 55. These polymorphisms are considered determinant of the capacity of HDL to protect LDL from their oxidative modification. In this context, the PON1 genotype has been associated with cardiovascular diseases, including stroke. Objective: To determine the allelic and genotypic frequencies of PON1 L55M and Q192R as well as the enzymatic activities of PON1 in subjects with and without atherothrombotic stroke. Methods: There were included 28 people with atherothrombotic stroke and 29 without stroke. The genotyping was carried out by PCR-RFLP and the phenotyping by measurement of the activities of paraoxonase and arylesterase in serum. Results: For the polymorphism Q192R, the allelic frequencies (Q/R) were 0.46/0.54 and 0.48/0.52 (p= 0.843) for the control group and the group with stroke, respectively. While for the polymorphism L55M, the allelic frequencies (L/M) were 0.81/0.19 for the control group, and 0.78/0.22 for the group with stroke (p= 0.610). The activity levels of paraoxonase were not significantly different between the control and stroke groups (450 vs. 348 UI/mL, p= 0.093) While the activity levels of arylesterase were significantly different between the studied groups (90 vs. 70 UI/mL, p= 0.001); however, upon adjustment by multiple linear regression, it was not longer significant. Conclusion: The polymorphisms Q192R and L55M, and the paraoxonase activity of PON1 are not risk factors for atherothrombotic stroke according to the results of this study.

Resumen Introducción: La paraoxonasa-1 (PON1) sérica asociada a las HDL presenta dos polimorfismos comunes en las posiciones 192 y 55. Estos polimorfismos se consideran determinantes para la capacidad de las HDL de proteger a las LDL de su modificación oxidativa. En este contexto, el genotipo de PON1 se ha asociado con enfermedades cerebrovasculares, que incluyen el infarto cerebral. Objetivo: Determinar las frecuencias alélicas y genotípicas de PON1-L55M y PON1- Q192R, así como las actividades enzimáticas de PON1 en sujetos con y sin infarto cerebral aterotrombótico. Métodos: Se incluyeron 28 personas con infarto cerebral aterotrombótico y 29 sin infarto. Las genotipificaciones se realizaron mediante PCR-RFLP y las fenotipificaciones mediante la medición de las actividades paraoxonasa y arilesterasa en suero. Resultados: Para el polimorfismo Q192R, las frecuencias alélicas (Q/R) fueron 0.46/0.54 y 0.48/0.52 (p= 0.843) para el grupo control y el grupo con infarto, respectivamente. Mientras que para el polimorfismo L55M, las frecuencias alélicas (L/M) fueron 0.81/0.19 para el grupo control y 0.78/0.22 para el grupo con infarto (p= 0.610). Los niveles de actividad paraoxonasa no fueron significativamente diferentes entre los grupos control y con infarto (450 vs. 348 Ul/mL, p= 0.093). Mientras que los niveles de actividad arilesterasa fueron significativamente diferentes entre los grupos estudiados (90 vs. 70 Ul/mL, p= 0.001), sin embargo, al ajustarla por regresión lineal múltiple, dejo de ser significativa. Conclusión: Los polimorfismos Q192R y L55M, y la actividad paraoxonasa de la PON1 no son factores de riesgo para el infarto cerebral aterotrombótico en este estudio.

Relationship Between Paraoxonase-1 and Butyrylcholinesterase Activities and Nutritional Status in Mexican Children.

BACKGROUND: The enzymes butyrylcholinesterase (BuChE) and paraoxonase-1 (PON1) are the primary bioscavenging enzymes in serum and exhibit antioxidant and anti-inflammatory activities. PON1 has been associated with diseases caused by high oxidative stress, whereas BuChE appears to be involved in the pathophysiology of the metabolic syndrome and related disorders. It has been suggested that children from rural communities in Mexico may have a predisposition to develop obesity or type 2 diabetes during adolescence or adulthood. The objective of this study was to determine whether associations exist between the paraoxonase (PONase)/arylesterase (AREase) activity of PON1, its PON1-Q192R and PON1-L55M polymorphisms, and BuChE activity with the nutritional status and lipid profiles in a group of children from rural communities in Mexico. METHODS: A group of 97 boys and girls from a rural community in Mexico were assessed for body mass index, the enzymatic activities of BuChE, PONase, and AREase were measured in serum, and their lipid profiles were determined. Genetic polymorphisms of PON1-L55M and PON1-Q192R were determined by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). RESULTS: The children were classified into four groups: thinness, normal weight, overweight, and obese. Of the children studied, 34.4% were overweight and obese. The mean age of the participants was 9.5 years (standard deviation = 1.8). The L allele of the PON1-L55M genotype was the most frequent (83.3%), and the R allele of the PON1-Q192R genotype was the most frequent (61.8%). Overweight and obese children had higher values of BuChE, total cholesterol, triglycerides (TG), and lower high-density lipoprotein (HDL-C) values than children with thinness or normal weight (P = 0.028, P = 0.019, P = 0.004, P = 0.069 and P = 0.021, respectively). The levels of AREase and PONase and the prevalence of PON1-L55M and PON1-Q192R genotypes were similar between groups (P = 0.484 and P = 0.380, respectively). CONCLUSIONS: This study establishes a positive association of BuChE activity with nutritional status and serum TG.

Relationship between the paraoxonase (PON1) L55M and Q192R polymorphisms and obesity in a Mexican population: a pilot study.

The aim of this study was to examine the relationship between the L55M and Q192R paraoxonase (PON1) polymorphisms and obesity in a population of adult Mexican workers. The study population included 127 adult individuals from the Universidad Autónoma del Estado de Morelos, ranging in age from 20 to 56 years and representing both sexes. Based on body mass index, 63 individuals were classified as obese and 64 as normal weight. The PON1-Q192R and PON1-L55M polymorphisms were determined by restriction fragment length polymorphism PCR analysis. Both arylesterase and paraoxonase activity levels were similar in both groups, whereas systolic pressure, triglyceride, total cholesterol, low-density lipoprotein cholesterol, very-low-density lipoprotein cholesterol, glucose, and insulin levels were higher in the obese group than in the normal-weight group (P < 0.05). An exception was the high-density lipoprotein cholesterol (HDL-C) levels, which were lower in the obese group (P < 0.05). Although the PON1-Q192R polymorphism was not associated with either group, the frequency of the homozygous L genotype for the PON1-L55M polymorphism was higher in the obese group than in the normal-weight group (P < 0.05). In conclusion, this study established a positive association between the PON1-L55M homozygous L genotype and obesity.