Investigation of the cytotoxicity, genotoxicity and antioxidant prospects of JM-20 on human blood cells: A multi-target compound with potential therapeutic applications.

JM-20 is a 1,5-benzodiazepine compound fused to a dihydropyridine fraction with different pharmacological properties. However, its potential toxic effects on blood cells have not yet been reported. Thus, the present study aimed to investigate, for the first time, the possible cytotoxicity of JM-20 through cell viability, cell cycle, morphology changes, reactive species (RS) to DCFH-DA, and lipid peroxidation in human leukocytes, its hemolytic effect on human erythrocytes, and its potential DNA genotoxicity using plasmid DNA in vitro. Furthermore, the compound's ability to reduce the DPPH radical was also measured. Human blood was obtained from healthy volunteers (30 ± 10 years old), and the leukocytes or erythrocytes were immediately isolated and treated with different concentrations of JM-20. A cytoprotective effect was exhibited by 10 µM JM-20 against 1 mM tert-butyl hydroperoxide (t-but-OOH) in the leukocytes. However, the highest tested concentrations of the compound (20 and 50 µM) changed the morphology and caused a significant decrease in the cell viability of leukocytes (p < 0.05, in comparison with Control). All tested concentrations of JM-20 also resulted in a significant increase in intracellular RS as measured by DCFH-DA in these cells (p < 0.05, in comparison with Control). On the other hand, the results point out a potent antioxidant effect of JM-20, which was similar to the classical antioxidant α-tocopherol. The IC50 value of JM-20 against the lipid peroxidation induced by (FeII) was 1.051 µM ± 0.21, while the IC50 value of α-tocopherol in this parameter was 1.065 µM ± 0.34. Additionally, 50 and 100 µM JM-20 reduced the DPPH radical in a statistically similar way to the 100 µM α-tocopherol (p < 0.05, in comparison with the control). No significant hemolysis in erythrocytes, no cell cycle changes in leukocytes, and no genotoxic effects in plasmid DNA were induced by JM-20 at any tested concentration. The in silico pharmacokinetic and toxicological properties of JM-20, derivatives, and nifedipine were also studied. Here, our findings demonstrate that JM-20 and its putative metabolites exhibit similar characteristics to nifedipine, and the in vitro and in silico data support the low toxicity of JM-20 to mammals.

Evidence of acrylamide-induced behavioral deficit, mitochondrial dysfunction and cell death in Drosophila melanogaster.

Acrylamide (ACR), a ubiquitous compound with diverse route of exposure, has been demonstrated to have detrimental effects on human and animal health. The mechanisms underlying its toxicity is multifaceted and not fully elucidated. This study aims to provide further insight into novel pathways underlying ACR toxicity by leveraging on Drosophila melanogaster as a model organism. The concentrations of acrylamide (25, 50 and 100 mg/kg) and period of exposure (7-days) used in this study was established through a concentration response curve. ACR exposure demonstrably reduced organismal viability, evidenced by decline in survival rate, offspring emergence and deficits in activity, sleep and locomotory behaviors. Using a high-resolution respirometry assay, the role of mitochondria respiratory system in ACR-mediated toxicity in the flies was investigated. Acrylamide caused dysregulation in mitochondrial bioenergetics and respiratory capacity leading to an impaired OXPHOS activity and electron transport, ultimately contributing to the pathological process of ACR-toxicity. Furthermore, ACR exacerbated apoptosis and induced oxidative stress in D. melanogaster. The up-regulation of mRNA transcription of Reaper, Debcl and Dark genes and down-regulation of DIAP1, an ubiquitylation catalyzing enzyme, suggests that ACR promotes apoptosis through disruption of caspase and pro-apoptotic protein ubiquitination and a mitochondria-dependent pathway in Drosophila melanogaster. Conclusively, this study provides valuable insights into the cellular mechanism underlying ACR-mediated toxicity. Additionally, our study reinforces the utility of D. melanogaster as a translational tool for elucidating the complex mechanisms of ACR toxicity.

Effect of Prepartum Maternal Supplementation with Diphenyl Diselenide on Biochemical, Immunological, and Oxidative Parameters of the Offspring.

This study aimed to assess the impact of prepartum maternal diphenyl diselenide (PhSe)2 supplementation on the development, biochemical, immune, and antioxidant parameters of calves. Eighteen Holstein breed calves were used, born to females who were or were not subjected to supplementation, at 42, 28, and 14 days prior to calving. The (PhSe)2 group (DDG) was administered 3 µmol/kg of (PhSe)2 in 4 mL of dimethyl sulfoxide (DMSO), while the DMSO and NaCl groups were administered 4 mL of DMSO and 0.9% NaCl, subcutaneously. The calves were evaluated based on their weight, withers height, body condition score 24 h post-birth (0), as well on days 14, 28, 42, 56, 70. Blood samples were also taken to determine serum variables. Calves on the DDG showed higher average levels of total protein, albumin, and globulins on day 0, and the immunoglobulin G level was significantly higher than the other groups on days 0, 14, 56, 70. Maternal supplementation showed immunomodulatory effect on calves, evidenced by the exceptional rates of passive immunity transfer, as well as the enhancement of humoral immunity. Our research offers fresh insights into the immunomodulatory potential of (PhSe)2, making it a viable alternative in facing this challenging phase, rearing dairy calves.

Biochemical and nutritional profile of patients with exclusive enteral nutrition during hospitalization / Perfil bioquímico y nutricional de pacientes con nutrición enteral exclusiva durante hospitalización

BACKGROUND: Enteral nutrition therapy (ENT) is intended to restore the nutritional status of patients. OBJECTIVE: The objective of this study was to evaluate the biochemical and nutritional profile of hospitalized patients with exclusive enteral nutrition. METHODS: It is a longitudinal study, with a sample of 42 hospitalized young and elder adults, with exclusive ENT, for at least seven days. The patients were submitted to nutritional, anthropometric (Body Mass Index, corrected arm muscle area and arm muscle circumference) and biochemical evaluation as albumin, hemoglobin, C-reactive protein, vitamin C, Iron, Zinc and Copper serum. Results and DISCUSSION: It was observed that anthropometric parameters such as weight, BMI, muscle area and circumference increased during hospitalization time only in the elderly (P= 0.016; P=0.018; P = 0.021; P = 0.020). The percentage of adequacy in energy, protein and micronutrients with vitamin C, iron, zinc and copper were adequate during hospitalization for both age groups, according to the estimated average needs. Serum levels of these micronutrients were within normal values for both age groups, with the exception of zinc, which decreased during hospitalization in the elderly. This may be associated with the greater need for this mineral in this age group or with a implicate in its absorption. CONCLUSION: The ENT influence the weight and muscle mass gain in hospitalized elderly patients and, although the appropriate administration of micronutrients, the absorption of zinc was affected. Therefore, monitoring of enteral nutrition is essential in order to avoid worsening nutritional status during hospitalization

INTRODUCCIÓN: La terapia de nutrición enteral (TNE) tiene la finalidad de recuperar el estado nutricional de los pacientes. Objectivo: Se evaluó el perfil bioquímico y nutricional de pacientes hospitalizados con nutrición enteral exclusiva. MÉTODOS: Estudio longitudinal, con muestra compuesta por 42 adultos y ancianos hospitalizados, con TNE exclusiva, por lo menos siete días. Los pacientes fueron sometidos a evaluación nutricional, antropométrica (Índice de Masa Corporal, área muscular del brazo corregida y circunferencia del brazo) y bioquímica como albúmina, proteína C-reactiva, vitamina C, hierro zinc y cobre sérico. Resultados y DISCUSIÓN: Se observó que los parámetros antropométricos como el peso, IMC, área y circunferencia muscular del brazo aumentaron durante el tiempo de internación solo en los ancianos (P= 0.016; P=0.018; P = 0.021; P = 0.020). El porcentaje de adecuación de energía, proteica y micronutrientes como vitamina C, hierro, zinc y cobre fueron adecuados durante el tiempo de internación para ambos grupos de edad, de acuerdo con las necesidades medias estimadas. Los niveles séricos de estos micronutrientes se mantuvieron dentro de los valores normales para ambos grupos de edad, a excepción del zinc, que disminuyó durante la hospitalización en ancianos. Esto puede asociarse a la mayor necesidad de este mineral en este grupo de edad o a un deterioro en su absorción. CONCLUSIÓN: La TNE influye en el aumento de peso y la masa muscular en ancianos y, apesar de la administración adecuada de micronutrientes, se observó un deterioro en la absorción de zinc. Por lo tanto, el monitoreo de la nutrición enteral es esencial para evitar el empeoramiento del estado nutricional durante la hospitalización

Effect of antioxidant potential on severity of cirrhosis in humans / Efecto del potencial antioxidante para las complicaciones de la cirrosis en los seres humanos

Background/Aims: to examine the relationship between the antioxidant potential and severity parameters of cirrhosis in humans. Methods: fifteen patients with hepatic cirrhosis (nine subjects - Child group B, and six subjects - Child group C) and nine control subjects were enrolled in the study. The main criteria taken into account to characterize the diagnosis of cirrhosis and its complications were the AST: ALT ratio, AST to platelet ratio index, Bonacini score, Meld score and Child classification. Those parameters were determined based on laboratory results and patient’s clinical records. Se, Zn, ascorbic acid (AA) levels and oxidative stress parameters were measured in blood samples of cirrhotic patients. Results: the analysis of plasma levels of Se and AA showed low concentrations in cirrhotic patients compared with control subjects (P < 0.05). Though, there was a positive correlation between plasma of Se and severity parameters of cirrhosis in patients of Child group B and C. In the activity of the antioxidant enzymes only catalase was lower in patients of Child group C compared with control group. Conclusion: we found low plasma levels of Se and AA among cirrhotic patients. However, is not clear why selenium levels tend to increase with the severity of liver cirrhosis (AU)

Introducción/Objetivos: examinar la relación entre los potenciales antioxidantes y los parámetros de gravedad de la cirrosis en los seres humanos. Métodos: quince pacientes con cirrosis hepática (nueve sujetos - grupo Child B, y seis sujetos - grupo Child C) y nueve sujetos control fueron incluidos en el estudio. Los principales criterios que se tuvieron en cuenta para caracterizar el diagnóstico de la cirrosis y sus complicaciones fueron la AST: relación de ALT, AST índice de la relación de plaquetas, clasificación Bonacini, clasificación MELD y clasificación de Child. Estos parámetros fueron determinados con base en los resultados de laboratorio y los registros clínicos del paciente. Se midieron los niveles de Zn, ácido ascórbico (AA) y los parámetros de estrés oxidativo en muestras de sangre de pacientes cirróticos. Resultados: el análisis de los niveles plasmáticos de Se y AA mostraron bajas concentraciones en los pacientes cirróticos en comparación con los sujetos control (P < 0,05); sin embargo, hubo una correlación positiva entre el plasma de Se y los parámetros de gravedad de la cirrosis en pacientes del grupo Child B y C. En la actividad de las enzimas antioxidantes catalasa solamente fue menor en los pacientes del grupo Child C, en comparación con el grupo control. Conclusión: se encontraron niveles bajos en plasma de Se y AA en pacientes cirróticos. Sin embargo, no está claro por qué los niveles de selenio tienden a aumentar con la gravedad de la cirrosis hepática (AU)

In vivo and in vitro genotoxicity studies of aqueous extract of Xanthium spinosum

The use of plants as a source of palliative or cure for pathological conditions is quite common worldwide. Xanthium spinosum (Asteraceae), popularly known in Brazil as 'espinho de carneiro', is an annual weed from South America, which has been used by empiric medicine to treat neoplasias. Owing to the extensive use of the above-mentioned plant and to the lack of reports about the real effects of its infusion, current study evaluated the genotoxic potential of its aqueous extract at concentrations 0.02 g L-1, 0.1 g L-1 and 0.2 g L-1 by fish micronucleus test and by comet human leukocytes assay. The micronucleus test featured at least 50 cells with micronuclei to every 2,000 cells scored, as a mutagenic parameter. The comet assay was used as a parameter for assessing the level of cell damage and the damage index. Since no significant changes in strain cells exposed to the aqueous extract in the comet and micronucleus assays were reported, it seems that no genotoxicity evidence is extant at the concentrations and in the assays performed.

Em diversos lugares do mundo a utilização de plantas como fonte paliativa ou de cura para determinadas condições patológicas é bastante comum. No Brasil, essa prática não se torna diferente devido à ampla biodiversidade da fauna e flora presentes no País. Nesse contexto, surge a Xanthium spinosum (Asteraceae), conhecida popularmente como "espinho-de-carneiro", um arbusto anual introduzido na América do Sul, o qual tem sido utilizado empiricamente no tratamento de neoplasias. Sabendo do extensivo uso dessa planta em contrapartida com nenhum estudo reportando os reais efeitos de sua infusão, o objetivo do estudo foi avaliar a genotoxicidade do extrato aquoso nas concentrações de 0,02 g L-1, 0,1 g L-1 e 0,2 g L-1, através do ensaio do micronúcleo písceo e do ensaio cometa em leucócitos de sangue humano. O ensaio do micronúcleo tem como parâmetro mutagênico a presença de no mínimo 50 células com micronúcleo em uma contagem de 2.000 células por amostra, enquanto o ensaio cometa utiliza como parâmetro o nível de dano e o índice de dano. Os resultados mostram que não foi possível observar mudanças significativas nas células expostas ao extrato aquoso, em ambos os testes, o que sugere não existir evidência de genotoxicidade nas concentrações utilizadas no ensaio.

Over-activation of the Drosophila melanogaster hsp83 gene by selenium intoxication

Selenium is an important dietary micronutrient and an essential component of selenoproteins and the active sites of some enzymes, although at high concentrations it is toxic. We investigated diphenyl diselenide ((C6H5)2Se2 ) for its effects on the developmental stages of Drosophila melanogaster and found that in the larval and pupae stages the toxic threshold for this compound when added to the banana-agar medium on which the larva were fed was 350 µmol. In adult flies, fed on the same media, there were no observable toxic effects below 500 µmol but there were toxic effects above 600 µmol, indicating that adult flies were more resistant to selenium intoxication. In larvae, a when diphenyl diselenide was present above the toxic threshold there was increased activation of the hsp83 heat shock protein gene. Selenium promotes oxidation of sulfhydryl groups and affects the folding of proteins and this could explain the over-expression of hsp83 because the product of this gene is involved in protein folding and defense responses, including the response to heat shock.