Asociación entre enfermedad de hígado graso no alcohólico y aterosclerosis subclínica

Introduction: non-alcoholic fatty lever disease is linked to atherosclerosis due to the latest´s risk factors. There are also tendencies that affirm that there is a different factor on its pathogeny. The subclinical atherosclerosis is a preventive medicine concept, and looking for it can help finding clues for future irreversible damage.Objective: to determine the association between subclinical atherosclerosis in dyslipidemic patients with non-alcoholic fatty lever disease. Method: a descriptive, cross-sectional study was carried out in a population of 114 dyslipidemic patients with non-alcoholic fatty lever disease in the dyslipoproteinemias consultation at the Surgical Clinical Hospital ¨Hermanos Amejeiras¨ from 2016 to 2019.Results:  mean age was 52.9 ± 12.4 years, with male patients’ predominance. 62 patients showed thickening in the intima-media wider than 1mm in echocardiography, and just 8 patients had atheroma plaque in the carotid artery. The frequency of the altered lipidic profile was due to hypertriglyceridaemia and HDL cholesterol. Conclusions: no link was found between non-alcoholic fatty lever disease and subclinical atherosclerosis. Through the analysis, atherosclerosis was linked to variables as age and family history of ischemic heart disease. There is no association between subclinical atherosclerosis and the degree of progression in the non-alcoholic fatty lever disease given by the APRI index.Introducción: la enfermedad de hígado graso no alcohólico está asociada con la aterosclerosis como consecuencia de los factores de riesgo que la acompañan. Hay tendencias que apoyan que es un factor independiente en su patogenia. La aterosclerosis subclínica es un concepto de medicina preventiva y su búsqueda permite marcar la aparición futura de daños irreversibles. Objetivo: determinar la asociación de aterosclerosis subclínica en pacientes dislipidémicos con enfermedad de hígado graso no alcohólico. Método: se realizó un estudio descriptivo, transversal en 114 pacientes dislipidémicos con enfermedad de hígado graso no alcohólico de la consulta de dislipoproteinemias en el Hospital Clínico Quirúrgico “Hermanos Ameijeiras” en el período comprendido entre los años 2016 al 2019.Resultados: la media de edad fue de 52,9 ± 12,4 años con predominio del sexo masculino. En ecocardiografía, 62 pacientes con enfermedad de hígado graso no alcohólico presentaron aumento del grosor de la íntima media mayor de 1mm y solo 8 pacientes tenían placa de ateroma de la arteria carotídea. La frecuencia del perfil lipídico alterado fue a expensas de hipertrigliceridemia y de la HDL colesterol.Conclusiones: no existió asociación entre la enfermedad de hígado graso no alcohólico con aterosclerosis subclínica; mediante el análisis multivariado se encontró asociación franca de la aterosclerosis subclínica con variables como la edad y el antecedente familiar de cardiopatía isquémica. No hay asociación de aterosclerosis subclínica con el grado de progresión de enfermedad de hígado graso no alcohólico dado por el índice de APRI

Links between non-alcoholic fatty lever disease and subclinical atherosclerosis

Introduction: non-alcoholic fatty lever disease is linked to atherosclerosis due to the latest´s risk factors. There are also tendencies that affirm that there is a different factor on its pathogeny. The subclinical atherosclerosis is a preventive medicine concept, and looking for it can help finding clues for future irreversible damage. Objective: to determine the association between subclinical atherosclerosis in dyslipidemic patients with non-alcoholic fatty lever disease. Method: a descriptive, cross-sectional study was carried out in a population of 114 dyslipidemic patients with non-alcoholic fatty lever disease in the dyslipoproteinemias consultation at the Surgical Clinical Hospital ¨Hermanos Amejeiras¨ from 2016 to 2019. Results:  mean age was 52.9 ± 12.4 years, with male patients’ predominance. 62 patients showed thickening in the intima-media wider than 1mm in echocardiography, and just 8 patients had atheroma plaque in the carotid artery. The frequency of the altered lipidic profile was due to hypertriglyceridaemia and HDL cholesterol. Conclusions: no link was found between non-alcoholic fatty lever disease and subclinical atherosclerosis. Through the analysis, atherosclerosis was linked to variables as age and family history of ischemic heart disease. There is no association between subclinical atherosclerosis and the degree of progression in the non-alcoholic fatty lever disease given by the APRI index

Benzimidazole derivatives as new and selective inhibitors of arginase from leishmania mexicana with biological activity against promastigotes and amastigotes

17 pags, 6 figs, 3 tabs, 2 schs. -- Supplementary materials are available online at https://www.mdpi.com/article/10 .3390/ijms222413613/s1.Leishmaniasis is a disease caused by parasites of the Leishmania genus that affects 98 countries worldwide, 2 million of new cases occur each year and more than 350 million people are at risk. The use of the actual treatments is limited due to toxicity concerns and the apparition of resistance strains. Therefore, there is an urgent necessity to find new drugs for the treatment of this disease. In this context, enzymes from the polyamine biosynthesis pathway, such as arginase, have been considered a good target. In the present work, a chemical library of benzimidazole derivatives was studied performing computational, enzyme kinetics, biological activity, and cytotoxic effect characterization, as well as in silico ADME-Tox predictions, to find new inhibitors for arginase from Leishmania mexicana (LmARG). The results show that the two most potent inhibitors (compounds 1 and 2) have an I50 values of 52 μM and 82 μM, respectively. Moreover, assays with human arginase 1 (HsARG) show that both compounds are selective for LmARG. According to molecular dynamics simulation studies these inhibitors interact with important residues for enzyme catalysis. Biological activity assays demonstrate that both compounds have activity against promastigote and amastigote, and low cytotoxic effect in murine macrophages. Finally, in silico prediction of their ADME-Tox properties suggest that these inhibitors support the characteristics to be considered drug candidates. Altogether, the results reported in our study suggest that the benzimidazole derivatives are an excellent starting point for design new drugs against leishmanisis.This research was funded by the Consejo Nacional de Ciencia y Tecnología (CONACyT), grants 257848 (A.T.-V.) and 258694 (C.A.-D.). The work in Spain was funded by a grant from the Spanish Ministry of Science, Innovation and Competitiveness PID2020-115331GB-100 to J.A.-H.Peer reviewe

Disulfiram as a novel inactivator of Giardia lamblia triosephosphate isomerase with antigiardial potential

Giardiasis, the infestation of the intestinal tract by Giardia lamblia, is one of the most prevalent parasitosis worldwide. Even though effective therapies exist for it, the problems associated with its use indicate that new therapeutic options are needed. It has been shown that disulfiram eradicates trophozoites in vitro and is effective in vivo in a murine model of giardiasis; disulfiram inactivation of carbamate kinase by chemical modification of an active site cysteine has been proposed as the drug mechanism of action. The triosephosphate isomerase from G. lamblia (GlTIM) has been proposed as a plausible target for the development of novel antigiardial pharmacotherapies, and chemical modification of its cysteine 222 (C222) by thiol-reactive compounds is evidenced to inactivate the enzyme. Since disulfiram is a cysteine modifying agent and GlTIM can be inactivated by modification of C222, in this work we tested the effect of disulfiram over the recombinant and trophozoite-endogenous GlTIM. The results show that disulfiram inactivates GlTIM by modification of its C222. The inactivation is species-specific since disulfiram does not affect the human homologue enzyme. Disulfiram inactivation induces only minor conformational changes in the enzyme, but substantially decreases its stability. Recombinant and endogenous GlTIM inactivates similarly, indicating that the recombinant protein resembles the natural enzyme. Disulfiram induces loss of trophozoites viability and inactivation of intracellular GlTIM at similar rates, suggesting that both processes may be related. It is plausible that the giardicidal effect of disulfiram involves the inactivation of more than a single enzyme, thus increasing its potential for repurposing it as an antigiardial drug. Keywords: Giardiasis, Drug repurposing, Neglected disease, Recombinant protein, Enzyme inactivatio

Species-Specific Inactivation of Triosephosphate Isomerase from Trypanosoma brucei: Kinetic and Molecular Dynamics Studies

Human African Trypanosomiasis (HAT), a disease that provokes 2184 new cases a year in Sub-Saharan Africa, is caused by Trypanosoma brucei. Current treatments are limited, highly toxic, and parasite strains resistant to them are emerging. Therefore, there is an urgency to find new drugs against HAT. In this context, T. brucei depends on glycolysis as the unique source for ATP supply; therefore, the enzyme triosephosphate isomerase (TIM) is an attractive target for drug design. In the present work, three new benzimidazole derivatives were found as TbTIM inactivators (compounds 1, 2 and 3) with an I50 value of 84, 82 and 73 µM, respectively. Kinetic analyses indicated that the three molecules were selective when tested against human TIM (HsTIM) activity. Additionally, to study their binding mode in TbTIM, we performed a 100 ns molecular dynamics simulation of TbTIM-inactivator complexes. Simulations showed that the binding of compounds disturbs the structure of the protein, affecting the conformations of important domains such as loop 6 and loop 8. In addition, the physicochemical and drug-like parameters showed by the three compounds suggest a good oral absorption. In conclusion, these molecules will serve as a guide to design more potent inactivators that could be used to obtain new drugs against HAT

Biochemical Analysis of Two Single Mutants that Give Rise to a Polymorphic G6PD A-Double Mutant

Glucose-6-phosphate dehydrogenase (G6PD) is a key regulatory enzyme that plays a crucial role in the regulation of cellular energy and redox balance. Mutations in the gene encoding G6PD cause the most common enzymopathy that drives hereditary nonspherocytic hemolytic anemia. To gain insights into the effects of mutations in G6PD enzyme efficiency, we have investigated the biochemical, kinetic, and structural changes of three clinical G6PD variants, the single mutations G6PD A+ (Asn126AspD) and G6PD Nefza (Leu323Pro), and the double mutant G6PD A− (Asn126Asp + Leu323Pro). The mutants showed lower residual activity (≤50% of WT G6PD) and displayed important kinetic changes. Although all Class III mutants were located in different regions of the three-dimensional structure of the enzyme and were not close to the active site, these mutants had a deleterious effect over catalytic activity and structural stability. The results indicated that the G6PD Nefza mutation was mainly responsible for the functional and structural alterations observed in the double mutant G6PD A−. Moreover, our study suggests that the G6PD Nefza and G6PD A− mutations affect enzyme functions in a similar fashion to those reported for Class I mutations

RNAi-Mediated Specific Gene Silencing as a Tool for the Discovery of New Drug Targets in Giardia lamblia; Evaluation Using the NADH Oxidase Gene

The microaerophilic protozoan Giardia lamblia is the agent causing giardiasis, an intestinal parasitosis of worldwide distribution. Different pharmacotherapies have been employed against giardiasis; however, side effects in the host and reports of drug resistant strains generate the need to develop new strategies that identify novel biological targets for drug design. To support this requirement, we have designed and evaluated a vector containing a cassette for the synthesis of double-stranded RNA (dsRNA), which can silence expression of a target gene through the RNA interference (RNAi) pathway. Small silencing RNAs were detected and quantified in transformants expressing dsRNA by a stem-loop RT-qPCR approach. The results showed that, in transformants expressing dsRNA of 100–200 base pairs, the level of NADHox mRNA was reduced by around 30%, concomitant with a decrease in enzyme activity and a reduction in the number of trophozoites with respect to the wild type strain, indicating that NADHox is indeed an important enzyme for Giardia viability. These results suggest that it is possible to induce the G. lamblia RNAi machinery for attenuating the expression of genes encoding proteins of interest. We propose that our silencing strategy can be used to identify new potential drug targets, knocking down genes encoding different structural proteins and enzymes from a wide variety of metabolic pathways

The nuclear receptor FXR, but not LXR, up-regulates bile acid transporter expression in non-alcoholic fatty liver disease

Background. Non-alcoholic fatty liver disease (NAFLD) is the most common cause of chronic liver disease. Patients with non-alcoholic steatohepatitis (NASH) have increased plasmatic and hepatic concentrations of bile acids (BA), suggesting that they can be associated with the progression of the disease. Hepatic nuclear receptors are known to modulate genes controlling BA metabolism; thus, in this work we aimed to compare the expression of liver nuclear receptors -farnesoid X (FXR), small heterodimer partner (SHP) and liver X alpha (LXRα) receptors- and BA transporters -sodium+/taurocholate cotransporting polypeptide (NTCP) and bile salt export pump (BSEP)- in liver biopsy samples of patients with simple steatosis (SS) and NASH.Material and methods. Forty patients with biopsy-proven NALFD were enrolled between 2009 and 2012; liver biopsies were classified as SS (N = 20) or NASH (N = 20) according to the NAFLD activity score. Gene expression of nuclear FXR, LXRa, SHP, NTCP and BSEP was analyzed by real-time reverse transcription polymerase chain reaction and protein level was quantified by western blot.Results. Gene expression of FXR, SHP, NTCP and BSEP was significantly up-regulated in the NASH group in comparison with SS patients (P < 0.05). In contrast, protein level for FXR, SHP and NTCP was decreased in the NASH patients vs. the SS group (P < 0.05). Gene and protein profile of LXRa did not show differences between groups.Conclusions. The results suggest that liver nuclear receptors (FXR and SHP) and BA transporters (NTCP and BSEP) are associated with the progression of NAFLD