Antinociceptive effect of LMH-2, a new sigma-1 receptor antagonist analog of haloperidol, on the neuropathic pain of diabetic mice.

This study evaluates the antiallodynic and antihyperalgesic effects of LMH-2, a new haloperidol (HAL) analog that acts as sigma-1 receptor (σ1 R) antagonist, in diabetic mice using a model of neuropathic pain induced by chronic hyperglycemia. Additionally, we compared its effects with those of HAL. Hyperglycemia was induced in mice by nicotinamide-streptozotocin administration (NA-STZ, 50-130 mg/kg). Four weeks later, mechanical allodynia was assessed using the up-down method, and hyperalgesia was evoked with formalin 0.5%. We evaluated antiallodynic and antihyperalgesic effects of LMH-2 (5.6-56.2 mg/kg), HAL (0.018-0.18 mg/kg) and gabapentin (GBP, 5.6-56.2 mg/kg). The results showed that LMH-2 had a more significant antiallodynic effect compared to HAL and GBP (90.4±8.7 vs 75.1±3.1 and 41.9±2.3%, respectively; P<0.05), as well as an antihyperalgesic effect (96.3±1.2 vs 86.9±7.41 and 86.9±4.8%, respectively; P<0.05). Moreover, the antiallodynic and antihyperalgesic effect of both LMH-2 and HAL were completely abolished by PRE-084 (σ1 R agonist); and partially by pramipexole (a D2-like receptor agonist). Finally, the effect of all treatments on the rotarod test, barra, open field and exploratory behaviors showed that LMH-2 did not alter the animals' balance or the exploratory behavior, unlike as HAL or GBP. The molecular docking included indicate that LMH-2 has lower affinity to the D2R than HAL. These results provide evidence that LMH-2 exerts its antinociceptive effects as a σ1 R antagonist without the adverse effects induced by HAL or GBP. Consequently, LMH-2 can be considered a good and safe strategy for treating neuropathic pain caused by hyperglycemia in patients with diabetes.

Design of a multi-target focused library for antidiabetic targets using a comprehensive set of chemical transformation rules.

Virtual small molecule libraries are valuable resources for identifying bioactive compounds in virtual screening campaigns and improving the quality of libraries in terms of physicochemical properties, complexity, and structural diversity. In this context, the computational-aided design of libraries focused against antidiabetic targets can provide novel alternatives for treating type II diabetes mellitus (T2DM). In this work, we integrated the information generated to date on compounds with antidiabetic activity, advances in computational methods, and knowledge of chemical transformations available in the literature to design multi-target compound libraries focused on T2DM. We evaluated the novelty and diversity of the newly generated library by comparing it with antidiabetic compounds approved for clinical use, natural products, and multi-target compounds tested in vivo in experimental antidiabetic models. The designed libraries are freely available and are a valuable starting point for drug design, chemical synthesis, and biological evaluation or further computational filtering. Also, the compendium of 280 transformation rules identified in a medicinal chemistry context is made available in the linear notation SMIRKS for use in other chemical library enumeration or hit optimization approaches.

Enhancing Giardicidal Activity and Aqueous Solubility through the Development of "RetroABZ", a Regioisomer of Albendazole: In Vitro, In Vivo, and In Silico Studies.

Parasitic diseases, including giardiasis caused by Giardia lamblia (G. lamblia), present a considerable global health burden. The limited effectiveness and adverse effects of current treatment options underscore the necessity for novel therapeutic compounds. In this study, we employed a rational design strategy to synthesize retroalbendazole (RetroABZ), aiming to address the limitations associated with albendazole, a commonly used drug for giardiasis treatment. RetroABZ exhibited enhanced in vitro activity against G. lamblia trophozoites, demonstrating nanomolar potency (IC50 = 83 nM), outperforming albendazole (189 nM). Moreover, our in vivo murine model of giardiasis displayed a strong correlation, supporting the efficacy of RetroABZ, which exhibited an eleven-fold increase in potency compared to albendazole, with median effective dose (ED50) values of 5 µg/kg and 55 µg/kg, respectively. A notable finding was RetroABZ's significantly improved water solubility (245.74 µg/mL), representing a 23-fold increase compared to albendazole, thereby offering potential opportunities for developing derivatives that effectively target invasive parasites. The molecular docking study revealed that RetroABZ displays an interaction profile with tubulin similar to albendazole, forming hydrogen bonds with Glu198 and Cys236 of the ß-tubulin. Additionally, molecular dynamics studies demonstrated that RetroABZ has a greater number of hydrophobic interactions with the binding site in the ß-tubulin, due to the orientation of the propylthio substituent. Consequently, RetroABZ exhibited a higher affinity compared to albendazole. Overall, our findings underscore RetroABZ's potential as a promising therapeutic candidate not only for giardiasis but also for other parasitic diseases.

Sociodemographic and clinical characteristics and access to health care in patients with spinal muscular atrophy in Argentina.

The FAME registry gathers the majority of patients with SMA in Argentina. From it, the clinical, sociodemographic and access to treatment characteristics were analyzed in 322 patients (range 8 months-61 years) included from 2008 to 2021. Important data were obtained for the planning of medical care of these patients such as: similar distribution of patient care in public and private hospitals, time gap between onset of symptoms and diagnoses, low level of completion of SMN2 copy count, estimate of 16 new diagnoses per year between 2014 and 2018, and 68% of patient in specific pharmacological treatment.

LC-MS Fingerprinting Development for Standardized Precipitate from Agastache mexicana, Which Induces Antihypertensive Effect through NO Production and Calcium Channel Blockade.

The aim of this work was to evaluate the vasorelaxant and antihypertensive effects of a standardized precipitate of the hydroalcoholic extract from Agastache mexicana (PPAm), comprising ursolic acid, oleanolic acid, acacetin, luteolin and tilianin, among others. In the ex vivo experiments, preincubation with L-NAME (nonspecific inhibitor of nitric oxide synthases) reduced the relaxation induced by PPAm; nevertheless, preincubation with indomethacin (nonspecific inhibitor of cyclooxygenases) did not generate any change in the vasorelaxation, and an opposed effect was observed to the contraction generated by CaCl2 addition. Oral administration of 100 mg/kg of PPAm induced a significant acute decrease in diastolic (DBP) and systolic (SBP) blood pressure in spontaneously hypertensive rats, without changes in heart rate. Additionally, PPAm showed a sustained antihypertensive subacute effect on both DBP and SBP for 10 days compared to the control group. On the other hand, human umbilical vein cells treated with 10 µg/mL of PPAm showed a significant reduction (p < 0.05) in intracellular adhesion molecule-1, compared to the control, but not on vascular cell adhesion molecule-1. In conclusion, PPAm induces a significant antihypertensive effect in acute- and subacute-period treatments, due to its direct vasorelaxant action on rat aortic rings through NO production and Ca2+ channel blockade.

Synergistic interactions of cytarabine-adavosertib in leukemic cell lines proliferation and metabolomic endpoints.

Drug synergy allows reduced dosing, side effects and tolerance. Optimization of drug synergy chemotherapy is fundamental in acute lymphocytic leukemia and other cancers. This study aimed to analyze the pharmacodynamic synergy between the anti-metabolite cytarabine and WEE1 inhibitor adavosertib on acute leukemia cell lines CCRF-CEM and Jurkat. In both cell lines analysis of concentration-inhibition curves of adavosertib-cytarabine combinations and synergy matrixes supported mutually synergistic drug interactions. Overall mean ( ± SD) synergy scores were higher in Jurkat than CCRF-CEM: Jurkat, ZIP 22.51 ± 1.1, Bliss 22.49 ± 1.1, HSA 23.44 ± 1.0, Loewe 14.16 ± 1.2; and, CCRF-CEM, ZIP 9.17 ± 1.9, Bliss 8.13 ± 2.1, HSA 11.48 ± 1.9 and Loewe 4.99 ± 1.8. Jurkat also surpassed CCRF-CEM in high-degree synergistic adavosertib-cytarabine interactions with mean across-models synergy values of ∼89.1% ± 2.9 for 63 nM cytarabine-97 nM adavosertib (91.4% inhibition synergy barometer). Combination sensitivity scores scatter plots confirmed combination's synergy efficacy. This combined approach permitted identification and prioritization of 63 nM cytarabine-97 nM adavosertib for multiple endpoints analysis. This combination did not affect PBMC viability, while exhibiting Jurkat selective synergy. Immunoblots also revealed Jurkat selective synergistically increased γH2AX phosphorylation, while CDC2 phosphorylation effects were attributed to adavosertib's WEE1 inhibition. In conclusion, the high synergistic efficacy combination of cytarabine (63 nM) and adavosertib (97 nM) was associated with remarkable alterations in metabolites related to the Krebs cycle in Jurkat. The metabolic pathways and processes are related to gluconeogenesis, amino acids, nucleotides, glutathione, electron transport and Warburg effect. All above relate to cell survival, apoptosis, and cancer progression. Our findings could pave the way for novel biomarkers in treatment, diagnosis, and prognosis of leukemia and other cancers.

Insulin Sensitization by PPARγ and GLUT-4 Overexpression/Translocation Mediates the Antidiabetic Effect of Plantago australis.

Plantago australis Lam. Subsp. hirtella (Kunth) Rahn is a medicinal plant used as a diuretic, anti-inflammatory, antibacterial, throat cancer treatment and for the control of diabetes. P. australis was collected in the state of Morelos, México. The hydroalcoholic extract (HAEPa) of P. australis was obtained by maceration and concentrated in vacuo. Once dry, it was evaluated through an oral glucose tolerance test (OGTT) in normoglycemic mice and in a non-insulin-dependent diabetic mice model. The expression of PPARγ and GLUT-4 mRNA was determined by rt-PCR, and GLUT-4 translocation was confirmed by confocal microscopy. The toxicological studies were conducted in accordance with the guidelines suggested by the OECD, sections 423 and 407, with some modifications. HAEPa significantly decreased glycemia in OGTT curves, as well as in the experimental diabetes model compared to the vehicle group. In vitro tests showed that HAEPa induced an α-glucosidase inhibition and increased PPARγ and GLUT-4 expression in cell culture. The LD50 of HAEPa was greater than 2000 mg/kg, and sub-chronic toxicity studies revealed that 100 mg/kg/day for 28 days did not generate toxicity. Finally, LC-MS analysis led to the identification of verbascoside, caffeic acid and geniposidic acid, and phytochemical approaches allowed for the isolation of ursolic acid, which showed significant PPARγ overexpression and augmented GLUT-4 translocation. In conclusion, HAEPa induced significant antidiabetic action by insulin sensitization through PPARγ/GLUT-4 overexpression.

El difícil camino al diagnóstico del paciente con atrofia muscular espinal / The difficult path to diagnosis of the patient with spinal muscular atrophy

Introducción. Con las nuevas terapias, el diagnóstico temprano de la atrofia muscular espinal (AME) es esencial. El objetivo de este estudio es analizar los distintos componentes que influyen en el retraso diagnóstico. Población y métodos. Se incluyeron pacientes con un diagnóstico molecular de AME tipo I, II y III. Se estudiaron varios parámetros, como la edad al momento de la aparición del primer signo, qué signo fue y el intervalo entre este y el diagnóstico confirmado. Neurólogos especialistas realizaron entrevistas que se complementaron con la revisión de historias clínicas cuando fue necesario. Resultados. Se entrevistaron 112 pacientes. AME I n = 40, AME II n = 48, AME III n = 24. La mediana de edad en meses al momento del reporte del primer signo fue AME I: 1,5 (R 0-7), AME II: 9 (R 2-20), AME III: 18 (R 8-180). Los primeros signos fueron reconocidos por los padres en el 75 % al 85 % de las veces en todos los subtipos. La mediana del tiempo transcurrido entre el primer signo y la primera consulta médica fue menor a un mes en los tres tipos. La mediana de tiempo transcurrido en meses entre el primer signo y el diagnóstico molecular confirmado fue en AME I: 2 (R 0-11), en AME II: 10 (3-46) y en AME III: 31,5 (R 4-288). Conclusiones. Existe un significativo retraso en el diagnóstico de la AME relacionado fundamentalmente a la falta de sospecha clínica. La demora es menor en AME I y mayor en AME III. Otros factores incluyen deficiencias en el sistema de salud.

Introduction. News treatments, make early diagnosis of spinal muscular atrophy (SMA) critical. The objective of this study is to analyze the different factors that influence delay in diagnosis. Population and methods. Patients with a molecular diagnosis of types I, II, and III SMA were included. Several parameters were studied, such as age at onset of first sign, what sign it was, and the time from recognition of first sign to confirmed diagnosis. Neurologists specialized in SMA conducted interviews, supported by the review of medical records when deemed necessary. Results. A total of 112 patients were interviewed. SMA I n = 40, SMA II n = 48, SMA III n = 24. The median age in months at the time of reporting the first sign was SMA I: 1.5 (R: 0­7), SMA II: 9 (R: 2­20), SMA III: 18 (R: 8­180). In all subtypes, first signs were identified by parents from 75% to 85% of the times. The median time from first sign to first medical consultation was less than a month in all 3 types. The median time in months, from first sign to confirmed molecular diagnosis in SMA I was: 2 (R: 0­11), in SMA II: 10 (R: 3­46), in SMA III: 31.5 (R: 4­288). Conclusions. There is a significant delay in SMA diagnosis mainly related to the absence of clinical suspicion. The delay is shorter in SMA I and longer in SMA III. Other factors include deficiencies in the health care system.

The difficult path to diagnosis of the patient with spinal muscular atrophy. / El difícil camino al diagnóstico del paciente con atrofia muscular espinal.

Introduction. News treatments, make early diagnosis of spinal muscular atrophy (SMA) critical. The objective of this study is to analyze the different factors that influence delay in diagnosis. Population and methods. Patients with a molecular diagnosis of types I, II, and III SMA were included. Several parameters were studied, such as age at onset of first sign, what sign it was, and the time from recognition of first sign to confirmed diagnosis. Neurologists specialized in SMA conducted interviews, supported by the review of medical records when deemed necessary. Results. A total of 112 patients were interviewed. SMA I n = 40, SMA II n = 48, SMA III n = 24. The median age in months at the time of reporting the first sign was SMA I: 1.5 (R: 0-7), SMA II: 9 (R: 2-20), SMA III: 18 (R: 8-180). In all subtypes, first signs were identified by parents from 75% to 85% of the times. The median time from first sign to first medical consultation was less than a month in all 3 types. The median time in months, from first sign to confirmed molecular diagnosis in SMA I was: 2 (R: 0-11), in SMA II: 10 (R: 3-46), in SMA III: 31.5 (R: 4-288). Conclusions. There is a significant delay in SMA diagnosis mainly related to the absence of clinical suspicion. The delay is shorter in SMA I and longer in SMA III. Other factors include deficiencies in the health care system.

Introducción. Con las nuevas terapias, el diagnóstico temprano de la atrofia muscular espinal (AME) es esencial. El objetivo de este estudio es analizar los distintos componentes que influyen en el retraso diagnóstico. Población y métodos. Se incluyeron pacientes con un diagnóstico molecular de AME tipo I, II y III. Se estudiaron varios parámetros, como la edad al momento de la aparición del primer signo, qué signo fue y el intervalo entre este y el diagnóstico confirmado. Neurólogos especialistas realizaron entrevistas que se complementaron con la revisión de historias clínicas cuando fue necesario. Resultados. Se entrevistaron 112 pacientes. AME I n = 40, AME II n = 48, AME III n = 24. La mediana de edad en meses al momento del reporte del primer signo fue AME I: 1,5 (R 0-7), AME II: 9 (R 2-20), AME III: 18 (R 8-180). Los primeros signos fueron reconocidos por los padres en el 75 % al 85 % de las veces en todos los subtipos. La mediana del tiempo transcurrido entre el primer signo y la primera consulta médica fue menor a un mes en los tres tipos. La mediana de tiempo transcurrido en meses entre el primer signo y el diagnóstico molecular confirmado fue en AME I: 2 (R 0-11), en AME II: 10 (346) y en AME III: 31,5 (R 4-288). Conclusiones. Existe un significativo retraso en el diagnóstico de la AME relacionado fundamentalmente a la falta de sospecha clínica. La demora es menor en AME I y mayor en AME III. Otros factores incluyen deficiencias en el sistema de salud.

Giardia lamblia G6PD::6PGL Fused Protein Inhibitors Decrease Trophozoite Viability: A New Alternative against Giardiasis.

Treatments to combat giardiasis have been reported to have several drawbacks, partly due to the drug resistance and toxicity of current antiparasitic agents. These constraints have prompted many researchers to investigate new drugs that act against protozoan parasites. Enzyme inhibition is an important means of regulating pathogen metabolism and has recently been identified as a significant alternative target in the search for new treatments. Glucose-6-phosphate dehydrogenase and 6-phosphogluconolactonase (G6PD::6PGL) is a bifunctional enzyme involved in the pentose phosphate pathway (PPP) in Giardia lamblia (G. lamblia). The G. lamblia enzyme is unusual since, unlike the human enzyme, it is a fused enzyme. Here, we show, through inhibition assays, that an in-house chemical library of 120 compounds and four target compounds, named CNZ-7, CNZ-8, CMC-1, and FLP-2, are potent inhibitors of the G. lamblia G6PD::6PGL fused enzyme. With a constant (k2) of 2.3, 3.2, and 2.8 M−1 s−1, respectively, they provoke alterations in the secondary and tertiary protein structure and global stability. As a novel approach, target compounds show antigiardial activity, with IC50 values of 8.7, 15.2, 15.3, and 24.1 µM in trophozoites from G. lamblia. Moreover, these compounds show selectivity against G. lamblia, since, through counter-screening in Caco-2 and HT29 human cells, they were found to have low toxicity. This finding positions these compounds as a potential and attractive starting point for new antigiardial drugs.

[Chronic degenerative conditions as risk factors for lethal COVID-19Doenças crônicas degenerativas como fator de risco para letalidade por COVID-19]. / Enfermedades crónicas degenerativas como factor de riesgo de letalidad por COVID-19 en México.

Objective: To determine the relative risk of a lethal outcome associated with chronic degenerative conditions in patients with COVID-19. Methods: A cohort study was conducted using electronic medical records belonging to patients who tested positive for COVID-19 on RT-PCR while receiving care as outpatients or inpatients in a social security system facility between March 2020 and March 2021. Two study groups were formed. The exposed group was divided into four subgroups, each of which was diagnosed with one and only one chronic condition (diabetes, hypertension, obesity, or chronic kidney disease); the unexposed group was obtained from the medical records of patients without comorbidities. A total of 1 114 medical records were examined using simple random sampling. Once the minimum sample size was reached, the relative risk was calculated for each chronic condition. Combinations of two, three, and four conditions were created, and each of them was included in the analysis. Results: In the absence of a chronic degenerative condition, the prevalence of a lethal outcome from COVID-19 is 3.8%; in the presence of type 2 diabetes mellitus, 15.8%; in the presence of arterial hypertension, 15.6%; and in the presence of obesity, 15.0%. For diabetes and hypertension combined, the prevalence of a lethal outcome is 54.1%; for diabetes and obesity combined, 36.8%, and for obesity and hypertension combined, 28.1%. Conclusion: In patients with COVID-19, the relative risk of a lethal outcome is 4.17 for those with diabetes, 4.13 for those with hypertension, and 3.96 for those with obesity. For two chronic conditions combined, the relative risk doubles or triples. The relative risk of a lethal outcome is 14.27 for diabetes plus hypertension; 9.73 for diabetes plus obesity, and 7.43 for obesity plus hypertension. Chronic conditions do not present alone; they generally occur together, hence the significance of the relative risks for lethal outcomes presented in this paper.

Objetivo: Determinar o risco de letalidade conferido por doenças crônicas degenerativas em pacientes com COVID-19. Métodos: Foi realizado um estudo de coorte em prontuários eletrônicos de pacientes com RT-PCR positivo para COVID-19 em atendimento ambulatorial ou hospitalar em uma instituição de previdência social, no período de março de 2020 a março de 2021. Foram constituídos dois grupos de estudo. O grupo exposto foi dividido em quatro subgrupos, cada um com diagnóstico único e exclusivo de uma doença crônica (diabetes, hipertensão, obesidade ou doença renal crônica). O grupo não exposto foi constituído por prontuários de pacientes sem comorbidades. Foram revisados 1.114 prontuários no total, utilizando técnica de amostragem aleatória simples. Uma vez obtido o tamanho mínimo da amostra, foi calculado o risco relativo para cada doença crônica. Foram realizadas combinações de 2, 3 e 4, tendo sido feita a análise com cada uma delas. Resultados: Na ausência de doença crônica degenerativa, a prevalência de letalidade na COVID-19 é de 3,8%; na presença de diabetes mellitus tipo 2, a letalidade é de 15,8%; na presença de hipertensão arterial, 15,6%; e na presença de obesidade, 15%. Quando tanto diabetes como hipertensão estão presentes, a letalidade é de 54,1%; com diabetes e obesidade, 36,8%; e obesidade com hipertensão, 28,1%. Conclusões: Em pacientes com COVID-19, o risco relativo de letalidade é de 4,17 naqueles com diabetes; 4,13 naqueles com hipertensão; e 3,96 naqueles com obesidade. Quando duas doenças crônicas são combinadas, o risco relativo dobra ou triplica. Para diabetes e hipertensão, o risco relativo de letalidade é 14,27; para diabetes e obesidade, 9,73; e para obesidade e hipertensão, 7,43. As doenças crônicas não ocorrem sozinhas (geralmente estão associadas), e nessa perspectiva os riscos relativos de letalidade apresentados neste artigo tornam-se relevantes.

Synthesis, in vitro, in silico and in vivo hypoglycemic and lipid-lowering effects of 4-benzyloxy-5-benzylidene-1,3-thiazolidine-2,4-diones mediated by dual PPAR α/γ modulation.

In current work, we prepared a series of nine 4-benzyloxy-5-benzylidene-1,3-thiazolidine-2,4-diones using a two-step pathway. Compounds 1-9 were tested in vitro using a set of three proteins recognized as important targets in diabetes and related diseases: PPARα, PPARγ, and GLUT-4. Compounds 1-3, 5, and 7 showed significant increases in the mRNA expression of PPARγ and GLUT-4, whereas compounds 1-3 did it over PPARα. Compounds 1-3 were identified as a dual PPAR α/γ modulators and were selected for evaluating the in vivo antidiabetic action at 100 mg/kg dose, being orally actives and decreasing blood glucose concentration in a hyperglycemic mice model, as well as reducing the triacylglycerides levels in normolipidemic rats. Docking and molecular dynamics studies were conducted to clarify the dual effect and binding mode of compounds 1-3 on both PPARs. Compounds 2 and 3 exhibited robust in vitro and in vivo efficacy and could be considered dual PPAR modulators with antidiabetic and antidyslipidemic effects.

Enfermedades crónicas degenerativas como factor de riesgo de letalidad por COVID-19 en México

[RESUMEN]. Objetivo. Determinar el riesgo de letalidad de las enfermedades crónicas degenerativas en pacientes con COVID-19. Métodos. Se realizo un estudio de cohorte, en expedientes clínicos electrónicos de pacientes con RT-PCR positiva para COVID-19 en atención ambulatoria o intrahospitalaria en una Institución de Seguridad Social de marzo 2020 a marzo 2021. Se integraron 2 grupos de estudio, el grupo expuesto se dividió en cuatro subgrupos, cada uno con diagnóstico único y exclusivo de una patología crónica (diabetes, hipertensión, obesidad o enfermedad renal crónica); el grupo no expuesto lo integraron expedientes de pacientes sin comorbilidades. Se revisaron 1 114 expedientes en total utilizando técnica muestral aleatoria simple, una vez obtenido el tamaño mínimo de muestra se calculó el riesgo relativo para cada enfermedad crónica, se realizaron combinaciones de 2, 3 y 4, con cada uno de ellos se realizó el análisis. Resultados. En ausencia de enfermedad crónica degenerativa la prevalencia de letalidad en COVID-19 es 3,8%; en presencia de diabetes mellitus tipo 2 la letalidad es 15,8; en hipertensión arterial de 15,6%; y en obesidad 15,0%. Cuando se combinan diabetes e hipertensión la letalidad es 54,1%; en diabetes y obesidad 36,8%; y en obesidad e hipertensión 28,1%. Conclusiones. En pacientes con COVID-19 el riesgo relativo para letalidad de letalidad en diabetes es 4,17; en hipertensión 4,13; y en obesidad 3,96. Cuando se combinan dos enfermedades crónicas el riesgo relativo se duplica o triplica, para diabetes e hipertensión el riesgo relativo para letalidad es 14,2; para diabetes y obesidad 9,73; y para obesidad e hipertensión 7,43. Es verdad que las enfermedades crónicas no se presentan solas, generalmente se encuentra asociadas, y desde esa perspectiva los riesgos relativos para letalidad ofrecidos en este artículo adquieren relevancia.

[ABSTRACT]. Objective. To determine the relative risk of a lethal outcome associated with chronic degenerative conditions in patients with COVID-19. Methods. A cohort study was conducted using electronic medical records belonging to patients who tested positive for COVID-19 on RT-PCR while receiving care as outpatients or inpatients in a social security system facility between March 2020 and March 2021. Two study groups were formed. The exposed group was divided into four subgroups, each of which was diagnosed with one and only one chronic condition (diabetes, hypertension, obesity, or chronic kidney disease); the unexposed group was obtained from the medical records of patients without comorbidities. A total of 1 114 medical records were examined using simple random sampling. Once the minimum sample size was reached, the relative risk was calculated for each chronic condition. Combinations of two, three, and four conditions were created, and each of them was included in the analysis. Results. In the absence of a chronic degenerative condition, the prevalence of a lethal outcome from COVID-19 is 3.8%; in the presence of type 2 diabetes mellitus, 15.8%; in the presence of arterial hypertension, 15.6%; and in the presence of obesity, 15.0%. For diabetes and hypertension combined, the prevalence of a lethal outcome is 54.1%; for diabetes and obesity combined, 36.8%, and for obesity and hypertension combined, 28.1%. Conclusion. In patients with COVID-19, the relative risk of a lethal outcome is 4.17 for those with diabetes, 4.13 for those with hypertension, and 3.96 for those with obesity. For two chronic conditions combined, the relative risk doubles or triples. The relative risk of a lethal outcome is 14.27 for diabetes plus hypertension; 9.73 for diabetes plus obesity, and 7.43 for obesity plus hypertension. Chronic conditions do not present alone; they generally occur together, hence the significance of the relative risks for lethal outcomes presented in this paper.

[RESUMO]. Objetivo. Determinar o risco de letalidade conferido por doenças crônicas degenerativas em pacientes com COVID-19. Métodos. Foi realizado um estudo de coorte em prontuários eletrônicos de pacientes com RT-PCR positivo para COVID-19 em atendimento ambulatorial ou hospitalar em uma instituição de previdência social, no período de março de 2020 a março de 2021. Foram constituídos dois grupos de estudo. O grupo exposto foi dividido em quatro subgrupos, cada um com diagnóstico único e exclusivo de uma doença crônica (diabetes, hipertensão, obesidade ou doença renal crônica). O grupo não exposto foi constituído por prontuários de pacientes sem comorbidades. Foram revisados 1.114 prontuários no total, utilizando técnica de amostragem aleatória simples. Uma vez obtido o tamanho mínimo da amostra, foi calculado o risco relativo para cada doença crônica. Foram realizadas combinações de 2, 3 e 4, tendo sido feita a análise com cada uma delas. Resultados. Na ausência de doença crônica degenerativa, a prevalência de letalidade na COVID-19 é de 3,8%; na presença de diabetes mellitus tipo 2, a letalidade é de 15,8%; na presença de hipertensão arterial, 15,6%; e na presença de obesidade, 15%. Quando tanto diabetes como hipertensão estão presentes, a letalidade é de 54,1%; com diabetes e obesidade, 36,8%; e obesidade com hipertensão, 28,1%. Conclusões. Em pacientes com COVID-19, o risco relativo de letalidade é de 4,17 naqueles com diabetes; 4,13 naqueles com hipertensão; e 3,96 naqueles com obesidade. Quando duas doenças crônicas são combinadas, o risco relativo dobra ou triplica. Para diabetes e hipertensão, o risco relativo de letalidade é 14,27; para diabetes e obesidade, 9,73; e para obesidade e hipertensão, 7,43. As doenças crônicas não ocorrem sozinhas (geralmente estão associadas), e nessa perspectiva os riscos relativos de letalidade apresentados neste artigo tornam-se relevantes.

Kinetic and Molecular Docking Studies to Determine the Effect of Inhibitors on the Activity and Structure of Fused G6PD::6PGL Protein from Trichomonas vaginalis.

Trichomoniasis is a sexually transmitted disease with a high incidence worldwide, affecting 270 million people. Despite the existence of a catalog of available drugs to combat this infection, their extensive use promotes the appearance of resistant Trichomonas vaginalis (T. vaginalis), and some side effects in treated people, which are reasons why it is necessary to find new alternatives to combat this infection. In this study, we investigated the impact of an in-house library comprising 55 compounds on the activity of the fused T. vaginalis G6PD::6PGL (TvG6PD::6PGL) protein, a protein mediating the first reaction step of the pentose phosphate pathway (PPP), a crucial pathway involved in the parasite's energy production. We found four compounds: JMM-3, CNZ-3, CNZ-17, and MCC-7, which inhibited the TvG6PD::6PGL protein by more than 50%. Furthermore, we determined the IC50, the inactivation constants, and the type of inhibition. Our results showed that these inhibitors induced catalytic function loss of the TvG6PD::6PGL enzyme by altering its secondary and tertiary structures. Finally, molecular docking was performed for the best inhibitors, JMM-3 and MCC-7. All our findings demonstrate the potential role of these selected hit compounds as TvG6PD::6PGL enzyme selective inhibitors.

Synthesis, In Vitro, In Vivo and In Silico Antidiabetic Bioassays of 4-Nitro(thio)phenoxyisobutyric Acids Acting as Unexpected PPARγ Modulators: An In Combo Study.

Four isobutyric acids (two nitro and two acetamido derivatives) were prepared in two steps and characterized using spectral analysis. The mRNA concentrations of PPARγ and GLUT-4 (two proteins documented as key diabetes targets) were increased by 3T3-L1 adipocytes treated with compounds 1-4, but an absence of in vitro expression of PPARα was observed. Docking and molecular dynamics studies revealed the plausible interaction between the synthesized compounds and PPARγ. In vivo studies established that compounds 1-4 have antihyperglycemic modes of action associated with insulin sensitization. Nitrocompound 2 was the most promising of the series, being orally active, and one of multiple modes of action could be selective PPARγ modulation due to its extra anchoring with Gln-286. In conclusion, we demonstrated that nitrocompound 2 showed strong in vitro and in vivo effects and can be considered as an experimental antidiabetic candidate.

PPARα/γ, adiponectin, and GLUT4 overexpression induced by moronic acid methyl ester influenced glucose and triglyceride levels of experimental diabetic mice.

The current study aimed to determine the antidiabetic and antidyslipidemic activities of moronic acid methyl ester (1) (compound 1) by in vivo, in vitro, in silico, and molecular biology studies. Compound 1 was evaluated to establish its dose-dependent antidiabetic and antihyperglycemic (50 mg/kg) activities, in diabetic and normoglycemic male CD1 mice, respectively. Also, compound 1 was subjected to a subacute study (50 mg/kg per day for 8 days) to determine blood biochemical profiles and the expression of protein tyrosine phosphatase 1B (PTP-1B), glucose transporter type 4 (GLUT4), peroxisome proliferator-activated receptor α (PPAR-α), PPAR-γ, adiponectin, interleukin-1ß (IL-1ß), and monocyte chemoattractant protein 1 (MCP-1) in adipose tissue of animals after treatment. Different doses in acute administration of compound 1 decreased glycemia (p < 0.05) compared with vehicle, showing greater effectiveness in the range 50-160 mg/kg. Also, the oral glucose tolerance test showed that compound 1 induced a significant antihyperglycemic action by opposing the hyperglycemic peak (p < 0.05). Moreover, compound 1 subacute administration decreased glucose and triglyceride levels after treatment (p < 0.05); while the expression of PPAR-α and PPAR-γ, adiponectin, and GLUT4 displayed an increase (p < 0.05) compared with the diabetic control group. In conclusion, compound 1 showed antihyperglycemic, antidiabetic, and antidyslipidemic effects in normal and diabetic mice, probably due to insulin sensitization through increased mRNA expression of GLUT4, PPAR-α, PPAR-γ, and adiponectin genes.

Enfermedades crónicas degenerativas como factor de riesgo de letalidad por COVID-19 en México / Chronic degenerative conditions as risk factors for lethal COVID-19 / Doenças crônicas degenerativas como fator de risco para letalidade por COVID-19

RESUMEN Objetivo. Determinar el riesgo de letalidad de las enfermedades crónicas degenerativas en pacientes con COVID-19. Métodos. Se realizo un estudio de cohorte, en expedientes clínicos electrónicos de pacientes con RT-PCR positiva para COVID-19 en atención ambulatoria o intrahospitalaria en una Institución de Seguridad Social de marzo 2020 a marzo 2021. Se integraron 2 grupos de estudio, el grupo expuesto se dividió en cuatro subgrupos, cada uno con diagnóstico único y exclusivo de una patología crónica (diabetes, hipertensión, obesidad o enfermedad renal crónica); el grupo no expuesto lo integraron expedientes de pacientes sin comorbilidades. Se revisaron 1 114 expedientes en total utilizando técnica muestral aleatoria simple, una vez obtenido el tamaño mínimo de muestra se calculó el riesgo relativo para cada enfermedad crónica, se realizaron combinaciones de 2, 3 y 4, con cada uno de ellos se realizó el análisis. Resultados. En ausencia de enfermedad crónica degenerativa la prevalencia de letalidad en COVID-19 es 3,8%; en presencia de diabetes mellitus tipo 2 la letalidad es 15,8; en hipertensión arterial de 15,6%; y en obesidad 15,0%. Cuando se combinan diabetes e hipertensión la letalidad es 54,1%; en diabetes y obesidad 36,8%; y en obesidad e hipertensión 28,1%. Conclusiones. En pacientes con COVID-19 el riesgo relativo para letalidad de letalidad en diabetes es 4,17; en hipertensión 4,13; y en obesidad 3,96. Cuando se combinan dos enfermedades crónicas el riesgo relativo se duplica o triplica, para diabetes e hipertensión el riesgo relativo para letalidad es 14,2; para diabetes y obesidad 9,73; y para obesidad e hipertensión 7,43. Es verdad que las enfermedades crónicas no se presentan solas, generalmente se encuentra asociadas, y desde esa perspectiva los riesgos relativos para letalidad ofrecidos en este artículo adquieren relevancia.

ABSTRACT Objective. To determine the relative risk of a lethal outcome associated with chronic degenerative conditions in patients with COVID-19. Methods. A cohort study was conducted using electronic medical records belonging to patients who tested positive for COVID-19 on RT-PCR while receiving care as outpatients or inpatients in a social security system facility between March 2020 and March 2021. Two study groups were formed. The exposed group was divided into four subgroups, each of which was diagnosed with one and only one chronic condition (diabetes, hypertension, obesity, or chronic kidney disease); the unexposed group was obtained from the medical records of patients without comorbidities. A total of 1 114 medical records were examined using simple random sampling. Once the minimum sample size was reached, the relative risk was calculated for each chronic condition. Combinations of two, three, and four conditions were created, and each of them was included in the analysis. Results. In the absence of a chronic degenerative condition, the prevalence of a lethal outcome from COVID-19 is 3.8%; in the presence of type 2 diabetes mellitus, 15.8%; in the presence of arterial hypertension, 15.6%; and in the presence of obesity, 15.0%. For diabetes and hypertension combined, the prevalence of a lethal outcome is 54.1%; for diabetes and obesity combined, 36.8%, and for obesity and hypertension combined, 28.1%. Conclusion. In patients with COVID-19, the relative risk of a lethal outcome is 4.17 for those with diabetes, 4.13 for those with hypertension, and 3.96 for those with obesity. For two chronic conditions combined, the relative risk doubles or triples. The relative risk of a lethal outcome is 14.27 for diabetes plus hypertension; 9.73 for diabetes plus obesity, and 7.43 for obesity plus hypertension. Chronic conditions do not present alone; they generally occur together, hence the significance of the relative risks for lethal outcomes presented in this paper.

RESUMO Objetivo. Determinar o risco de letalidade conferido por doenças crônicas degenerativas em pacientes com COVID-19. Métodos. Foi realizado um estudo de coorte em prontuários eletrônicos de pacientes com RT-PCR positivo para COVID-19 em atendimento ambulatorial ou hospitalar em uma instituição de previdência social, no período de março de 2020 a março de 2021. Foram constituídos dois grupos de estudo. O grupo exposto foi dividido em quatro subgrupos, cada um com diagnóstico único e exclusivo de uma doença crônica (diabetes, hipertensão, obesidade ou doença renal crônica). O grupo não exposto foi constituído por prontuários de pacientes sem comorbidades. Foram revisados 1.114 prontuários no total, utilizando técnica de amostragem aleatória simples. Uma vez obtido o tamanho mínimo da amostra, foi calculado o risco relativo para cada doença crônica. Foram realizadas combinações de 2, 3 e 4, tendo sido feita a análise com cada uma delas. Resultados. Na ausência de doença crônica degenerativa, a prevalência de letalidade na COVID-19 é de 3,8%; na presença de diabetes mellitus tipo 2, a letalidade é de 15,8%; na presença de hipertensão arterial, 15,6%; e na presença de obesidade, 15%. Quando tanto diabetes como hipertensão estão presentes, a letalidade é de 54,1%; com diabetes e obesidade, 36,8%; e obesidade com hipertensão, 28,1%. Conclusões. Em pacientes com COVID-19, o risco relativo de letalidade é de 4,17 naqueles com diabetes; 4,13 naqueles com hipertensão; e 3,96 naqueles com obesidade. Quando duas doenças crônicas são combinadas, o risco relativo dobra ou triplica. Para diabetes e hipertensão, o risco relativo de letalidade é 14,27; para diabetes e obesidade, 9,73; e para obesidade e hipertensão, 7,43. As doenças crônicas não ocorrem sozinhas (geralmente estão associadas), e nessa perspectiva os riscos relativos de letalidade apresentados neste artigo tornam-se relevantes.

Nitazoxanide Exerts Immunomodulatory Effects on Peripheral Blood Mononuclear Cells from Type 2 Diabetes Patients.

BACKGROUND: Type 2 diabetes (T2D) is a low-grade inflammatory condition with abnormalities in the immune response mediated by T lymphocytes and macrophages. Drug repositioning for immunomodulatory molecules is an attractive proposal for treating T2D. Nitazoxanide (NTZ) is a broad-spectrum drug with promising immunomodulatory effects. Thus, we investigated the immunomodulatory effect of NTZ on peripheral blood mononuclear cells (PBMCs) from patients with T2D. METHODS: Fifty patients with T2D were selected, and the proliferative response of T lymphocytes and the M1/M2 ratio of macrophages post cell culture were evaluated by flow cytometry, as well as measuring the concentration of cytokines by ELISA and the relative expression of microRNAs (miRNAs) related to the immune response by real-time PCR. RESULTS: NTZ exerts an inhibitory effect on the cell proliferation of T lymphocytes stimulated with anti-CD3 and anti-CD28 antibodies without modifying cell viability, and significant decreases in the supernatant concentrations of interleukin (IL)-1ß, IL-2, IL-6, IL-10, and IL-12. Furthermore, NTZ negatively regulates the relative expression of miR-155-5p without changes in miR-146a-5p. The M1/M2 ratio of monocytes/macrophages decreased the M1 and increased the M2 subpopulation by NTZ. CONCLUSIONS: Our results suggest that NTZ exerts immunomodulatory effects on PBMCs from T2D patients, and shows potential alternative therapeutic benefits.

Breast Cancer Antiestrogen Resistance 3 (BCAR3) promotes tumor growth and progression in triple-negative breast cancer.

Triple-Negative Breast Cancers (TNBCs) constitute roughly 10-20% of breast cancers and are associated with poor clinical outcomes. Previous work from our laboratory and others has determined that the cytoplasmic adaptor protein Breast Cancer Antiestrogen Resistance 3 (BCAR3) is an important promoter of cell motility and invasion of breast cancer cells. In this study, we use both in vivo and in vitro approaches to extend our understanding of BCAR3 function in TNBC. We show that BCAR3 is upregulated in ductal carcinoma in situ (DCIS) and invasive carcinomas compared to normal mammary tissue, and that survival of TNBC patients whose tumors contained elevated BCAR3 mRNA is reduced relative to individuals whose tumors had less BCAR3 mRNA. Using mouse orthotopic tumor models, we further show that BCAR3 is required for efficient TNBC tumor growth. Analysis of publicly available RNA expression databases revealed that MET receptor signaling is strongly correlated with BCAR3 mRNA expression. A functional role for BCAR3-MET coupling is supported by data showing that both proteins participate in a single pathway to control proliferation and migration of TNBC cells. Interestingly, the mechanism through which this functional interaction operates appears to differ in different genetic backgrounds of TNBC, stemming in one case from potential differences in the strength of downstream signaling by the MET receptor and in another from BCAR3-dependent activation of an autocrine loop involving the production of HGF mRNA. Together, these data open the possibility for new approaches to personalized therapy for individuals with TNBCs.

Identification and In Silico Characterization of Novel Helicobacter pylori Glucose-6-Phosphate Dehydrogenase Inhibitors.

Helicobacter pylori (H. pylori) is a pathogen that can remain in the stomach of an infected person for their entire life. As a result, this leads to the development of severe gastric diseases such as gastric cancer. In addition, current therapies have several problems including antibiotics resistance. Therefore, new practical options to eliminate this bacterium, and its induced affections, are required to avoid morbidity and mortality worldwide. One strategy in the search for new drugs is to detect compounds that inhibit a limiting step in a central metabolic pathway of the pathogen of interest. In this work, we tested 55 compounds to gain insights into their possible use as new inhibitory drugs of H. pylori glucose-6-phosphate dehydrogenase (HpG6PD) activity. The compounds YGC-1; MGD-1, MGD-2; TDA-1; and JMM-3 with their respective scaffold 1,3-thiazolidine-2,4-dione; 1H-benzimidazole; 1,3-benzoxazole, morpholine, and biphenylcarbonitrile showed the best inhibitory activity (IC50 = 310, 465, 340, 204 and 304 µM, respectively). We then modeled the HpG6PD protein by homology modeling to conduct an in silico study of the chemical compounds and discovers its possible interactions with the HpG6PD enzyme. We found that compounds can be internalized at the NADP+ catalytic binding site. Hence, they probably exert a competitive inhibitory effect with NADP+ and a non-competitive or uncompetitive effect with G6P, that of the compounds binding far from the enzyme's active site. Based on these findings, the tested compounds inhibiting HpG6PD represent promising novel drug candidates against H. pylori.