RESUMO
Introduction: Serine proteases play a critical role during SARS-CoV-2 infection. Therefore, polymorphisms of transmembrane protease serine 2 (TMPRSS2) and serpine family E member 1 (SERPINE1) could help to elucidate the contribution of variability to COVID-19 outcomes. Methods: To evaluate the genetic variants of the genes previously associated with COVID-19 outcomes, we performed a cross-sectional study in which 1536 SARS-CoV-2-positive participants were enrolled. TMPRSS2 (rs2070788, rs75603675, rs12329760) and SERPINE1 (rs2227631, rs2227667, rs2070682, rs2227692) were genotyped using the Open Array Platform. The association of polymorphisms with disease outcomes was determined by logistic regression analysis adjusted for covariates (age, sex, hypertension, type 2 diabetes, and obesity). Results: According to our codominant model, the GA genotype of rs2227667 (OR=0.55; 95% CI = 0.36-0.84; p=0.006) and the AG genotype of rs2227667 (OR=0.59; 95% CI = 0.38-0.91; p=0.02) of SERPINE1 played a protective role against disease. However, the rs2227692 T allele and TT genotype SERPINE1 (OR=1.45; 95% CI = 1.11-1.91; p=0.006; OR=2.08; 95% CI = 1.22-3.57; p=0.007; respectively) were associated with a decreased risk of death. Similarly, the rs75603675 AA genotype TMPRSS2 had an OR of 1.97 (95% CI = 1.07-3.6; p=0.03) for deceased patients. Finally, the rs2227692 T allele SERPINE1 was associated with increased D-dimer levels (OR=1.24; 95% CI = 1.03-1.48; p=0.02). Discussion: Our data suggest that the rs75603675 TMPRSS2 and rs2227692 SERPINE1 polymorphisms are associated with a poor outcome. Additionally, rs2227692 SERPINE1 could participate in hypercoagulable conditions in critical COVID-19 patients, and this genetic variant could contribute to the identification of new pharmacological targets and treatment strategies to block the inhibition of TMPRSS2 entry into SARS-CoV-2.
Assuntos
COVID-19 , Diabetes Mellitus Tipo 2 , Humanos , COVID-19/genética , Serina Proteases , SARS-CoV-2 , Estudos TransversaisRESUMO
Bacterioplankton communities play a crucial role in global biogeochemical processes and are highly sensitive to changes induced by natural and anthropogenic stressors in aquatic ecosystems. We assessed the influence of Land Use Land Cover (LULC), environmental, and geographic changes on the bacterioplankton structure in highly connected and impacted shallow lakes within the Salado River basin, Buenos Aires, Argentina. Additionally, we investigated how changes in LULC affected the limnological characteristics of these lakes at a regional scale. Our analysis revealed that the lakes were ordinated by sub-basins (upper and lower) depending on their LULC characteristics and limnological properties. In coincidence, the same ordination was observed when considering the Bacterioplankton Community Composition (BCC). Spatial and environmental predictors significantly explained the variation in BCC, although when combined with LULC the effect was also important. While the pure LULC effect did not explain a significant percentage of BCC variation, the presence of atrazine in water, an anthropogenic variable linked to LULC, directly influenced both the BCC and some Amplicon Sequence Variants (ASVs) in particular. Our regional-scale approach contributes to understanding the complexity of factors driving bacterioplankton structure and how LULC pervasively affect these communities in highly impacted shallow lake ecosystems from the understudied Southern Hemisphere.
Assuntos
Ecossistema , Lagos , Organismos Aquáticos , Argentina , RiosRESUMO
Neurotensin (NTS)-polyplex is a multicomponent nonviral vector that enables gene delivery via internalization of the neurotensin type 1 receptor (NTSR1) to dopaminergic neurons and cancer cells. An approach to improving its therapeutic safety is replacing the viral karyophilic component (peptide KPSV40; MAPTKRKGSCPGAAPNKPK), which performs the nuclear import activity, by a shorter synthetic peptide (KPRa; KMAPKKRK). We explored this issue and the mechanism of plasmid DNA translocation through the expression of the green fluorescent protein or red fluorescent protein fused with KPRa and internalization assays and whole-cell patch-clamp configuration experiments in a single cell together with importin α/ß pathway blockers. We showed that KPRa electrostatically bound to plasmid DNA increased the transgene expression compared with KPSV40 and enabled nuclear translocation of KPRa-fused red fluorescent proteins and plasmid DNA. Such translocation was blocked with ivermectin or mifepristone, suggesting importin α/ß pathway mediation. KPRa also enabled NTS-polyplex-mediated expression of reporter or physiological genes such as human mesencephalic-derived neurotrophic factor (hMANF) in dopaminergic neurons in vivo. KPRa is a synthetic monopartite peptide that showed nuclear import activity in NTS-polyplex vector-mediated gene delivery. KPRa could also improve the transfection of other nonviral vectors used in gene therapy.
Assuntos
Portadores de Fármacos/síntese química , Técnicas de Transferência de Genes , Vetores Genéticos/administração & dosagem , Neurotensina/administração & dosagem , Fragmentos de Peptídeos/síntese química , Transporte Ativo do Núcleo Celular , Animais , Linhagem Celular , Núcleo Celular/metabolismo , Neurônios Dopaminérgicos/citologia , Neurônios Dopaminérgicos/metabolismo , Terapia Genética/métodos , Vetores Genéticos/genética , Masculino , Camundongos , Modelos Animais , Nanopartículas/química , Neurotensina/genética , Neurotensina/farmacocinética , Técnicas de Patch-Clamp , Plasmídeos/genética , Ratos , Receptores de Neurotensina/metabolismo , Análise de Célula Única , Técnicas EstereotáxicasRESUMO
Dyslipidemias are common risk factors for the development of chronic disorders including type 2 diabetes (T2D). Over 100 associated loci have been identified but few reports have evaluated the population attributable fraction captured by them in population-based nationwide surveys. Therefore, we determined the population contribution of a set of known genetic risk variants to the development of dyslipidemias and T2D in Mexico. This study included 1665 participants from a Mexican National Health Survey carried out in the year 2000. It is a probabilistic complex sample survey of households, which comprises representative data at a national level. 103 previously reported SNPs associated with different dyslipidemias or T2D were genotyped and used to compute polygenic risk scores. We found that the previously known variants associated with dyslipidemias explain at most 7% of the total risk variance of lipid levels. In contrast, the known genetic risk component for T2D explained a negligible amount of variance (0.1%). Notably, variants derived from the Native-American ancestry have the strongest effect and contribute with a high proportion of the variance. These results support the need for additional studies aimed to identify specific genetic risk variants for Mexican population.
Assuntos
Diabetes Mellitus Tipo 2 , Dislipidemias , Variação Genética , Genótipo , Adulto , Estudos Transversais , Diabetes Mellitus Tipo 2/epidemiologia , Diabetes Mellitus Tipo 2/genética , Dislipidemias/epidemiologia , Dislipidemias/genética , Feminino , Humanos , Masculino , México , Pessoa de Meia-Idade , Fatores de RiscoRESUMO
Identification of a new class of antitumor agent capable to induce apoptosis without triggering necrotic cell death event is challenging. The present communication describes the multicomponent synthesis of seven new (1S,4S)-2,5-diazabicyclo[2.2.1]heptane-dithiocarbamates and their in vitro antiproliferative activity on cervical cancer cell line (CaSki), breast cancer cell line (MDA-MB231), lung cancer cell line (SK-Lu-1) and human lymphocytes. Among the synthesized dithiocarbamates, compound 9e displayed significant antiproliferative activity without inducing any necrotic cell death (both on tumour cells and lymphocytes) and induced apoptosis in tumor cells by the caspase dependent apoptotic pathway. The compound 9e also exhibited greater tumor selectivity than human lymphocytes. In silico ADME predictions revealed that compound 9e has the potential to be developed as a drug candidate. Rapid chemical modifications of this lead are thus highly necessary for further investigation as a drug like safer antitumor candidate and also to achieve compounds with better activity profile.
Assuntos
Antineoplásicos/farmacologia , Apoptose/efeitos dos fármacos , Compostos Azabicíclicos/farmacologia , Tiocarbamatos/farmacologia , Antineoplásicos/síntese química , Antineoplásicos/química , Compostos Azabicíclicos/síntese química , Compostos Azabicíclicos/química , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Relação Dose-Resposta a Droga , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Estrutura Molecular , Relação Estrutura-Atividade , Tiocarbamatos/síntese química , Tiocarbamatos/químicaRESUMO
This paper summarises results of zinc content and its speciation in human milk from mothers of preterm and full-term infants at different stages of lactation and from synthetic formula milks. Human milk samples (colostrum, 7th, 14th, and 28th day after delivery) from Spanish and Brazilian mothers of preterm and full-term infants (and also formula milks) were collected. After adequate treatment of the sample, total Zn was determined, while speciation analysis of the Zn was accomplished by size exclusion chromatography coupled online with the ICP-MS. It is observed that total zinc content in human milk decreases continuously during the first month of lactation, both for preterm and full term gestations. All infant formulas analysed for total Zn were within the currently legislated levels. For Zn speciation analysis, there were no differences between preterm and full term human milk samples. Moreover Zn species elute mainly associated with immunoglobulins and citrate in human milk whey. Interestingly the speciation in formula milk whey turned out to be completely different as the observed Zn(2+) was bound almost exclusively to low molecular weight ligands (citrate) and only comparatively very low amounts of the metal appeared to be associated with higher mass biomolecules (e.g. proteins).
Assuntos
Cromatografia em Gel , Análise de Alimentos/métodos , Fórmulas Infantis/química , Espectrometria de Massas , Leite Humano/química , Zinco/análise , Brasil , Citratos/química , Humanos , Lactente , Peso MolecularRESUMO
IMPORTANCE: Latino populations have one of the highest prevalences of type 2 diabetes worldwide. OBJECTIVES: To investigate the association between rare protein-coding genetic variants and prevalence of type 2 diabetes in a large Latino population and to explore potential molecular and physiological mechanisms for the observed relationships. DESIGN, SETTING, AND PARTICIPANTS: Whole-exome sequencing was performed on DNA samples from 3756 Mexican and US Latino individuals (1794 with type 2 diabetes and 1962 without diabetes) recruited from 1993 to 2013. One variant was further tested for allele frequency and association with type 2 diabetes in large multiethnic data sets of 14,276 participants and characterized in experimental assays. MAIN OUTCOME AND MEASURES: Prevalence of type 2 diabetes. Secondary outcomes included age of onset, body mass index, and effect on protein function. RESULTS: A single rare missense variant (c.1522G>A [p.E508K]) was associated with type 2 diabetes prevalence (odds ratio [OR], 5.48; 95% CI, 2.83-10.61; P = 4.4 × 10(-7)) in hepatocyte nuclear factor 1-α (HNF1A), the gene responsible for maturity onset diabetes of the young type 3 (MODY3). This variant was observed in 0.36% of participants without type 2 diabetes and 2.1% of participants with it. In multiethnic replication data sets, the p.E508K variant was seen only in Latino patients (n = 1443 with type 2 diabetes and 1673 without it) and was associated with type 2 diabetes (OR, 4.16; 95% CI, 1.75-9.92; P = .0013). In experimental assays, HNF-1A protein encoding the p.E508K mutant demonstrated reduced transactivation activity of its target promoter compared with a wild-type protein. In our data, carriers and noncarriers of the p.E508K mutation with type 2 diabetes had no significant differences in compared clinical characteristics, including age at onset. The mean (SD) age for carriers was 45.3 years (11.2) vs 47.5 years (11.5) for noncarriers (P = .49) and the mean (SD) BMI for carriers was 28.2 (5.5) vs 29.3 (5.3) for noncarriers (P = .19). CONCLUSIONS AND RELEVANCE: Using whole-exome sequencing, we identified a single low-frequency variant in the MODY3-causing gene HNF1A that is associated with type 2 diabetes in Latino populations and may affect protein function. This finding may have implications for screening and therapeutic modification in this population, but additional studies are required.
Assuntos
Diabetes Mellitus Tipo 2/genética , Fator 1-alfa Nuclear de Hepatócito/genética , Adulto , Idade de Início , Idoso , Feminino , Genótipo , Hispânico ou Latino/genética , Humanos , Masculino , México , Pessoa de Meia-Idade , Mutação de Sentido Incorreto , Análise de Sequência de DNA , Estados UnidosRESUMO
Certain steroidal compounds have demonstrated an antiproliferative effect against several tumor cell lines; however, their complete role on cancer cells is not currently established. Herein, we report the synthesis and evaluation of two new 26-hydroxy-22-oxocholestanic steroids on cervical cancer CaSki cells. The title compounds were prepared from diosgenin and hecogenin in excellent yields. We determined their effect on cell proliferation, cell cycle, and cell death. The cytotoxic effect of the title compounds on CaSki and human lymphocytes was also evaluated, indicating that the main cell death process is not necrosis; the null effect on lymphocytes implies that they are not cytotoxic. The observation of apoptotic bodies as well as the increase in the expression of active caspase-3 along with the fragmentation of DNA confirmed that such new cholestanic frameworks induced apoptosis in tumor cells. Significantly, their antiproliferative activity on tumor cells did not affect the proliferative potential of normal fibroblasts from cervix and peripheral blood lymphocytes. The title compounds show selective antitumor activity and therefore serve as promising lead candidates for further optimization.
Assuntos
Antineoplásicos Fitogênicos/síntese química , Antineoplásicos Fitogênicos/farmacologia , Apoptose/efeitos dos fármacos , Colestanóis/química , Colestanóis/síntese química , Colestanóis/farmacologia , Diosgenina/química , Sapogeninas/química , Esteroides/química , Esteroides/síntese química , Esteroides/farmacologia , Sequência de Carboidratos , Caspase 3/metabolismo , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Fragmentação do DNA/efeitos dos fármacos , Feminino , Fibroblastos/efeitos dos fármacos , Corantes Fluorescentes , Humanos , Marcação In Situ das Extremidades Cortadas , Indóis , Linfócitos/efeitos dos fármacos , Espectroscopia de Ressonância Magnética , Dados de Sequência Molecular , Relação Estrutura-Atividade , Neoplasias do Colo do Útero/tratamento farmacológico , Neoplasias do Colo do Útero/patologiaRESUMO
BACKGROUND: Although epidemiological studies have demonstrated an increased predisposition to low high-density lipoprotein cholesterol and high triglyceride levels in the Mexican population, Mexicans have not been included in any of the previously reported genome-wide association studies for lipids. METHODS AND RESULTS: We investigated 6 single-nucleotide polymorphisms associated with triglycerides, 7 with high-density lipoprotein cholesterol, and 1 with both triglycerides and high-density lipoprotein cholesterol in recent Caucasian genome-wide association studies in Mexican familial combined hyperlipidemia families and hypertriglyceridemia case-control study samples. These variants were within or near the genes ABCA1, ANGPTL3, APOA5, APOB, CETP, GALNT2, GCKR, LCAT, LIPC, LPL (2), MMAB-MVK, TRIB1, and XKR6-AMAC1L2. We performed a combined analysis of the family-based and case-control studies (n=2298) using the Z method to combine statistics. Ten of the single-nucleotide polymorphisms were nominally significant and 5 were significant after Bonferroni correction (P=2.20 x 10(-3) to 2.6 x 10(-11)) for the number of tests performed (APOA5, CETP, GCKR, and GALNT2). Interestingly, our strongest signal was obtained for triglycerides with the minor allele of rs964184 (P=2.6 x 10(-11)) in the APOA1/C3/A4/A5 gene cluster region that is significantly more common in Mexicans (27%) than in whites (12%). CONCLUSIONS: It is important to confirm whether known loci have a consistent effect across ethnic groups. We show replication of 5 Caucasian genome-wide association studies lipid associations in Mexicans. The remaining loci will require a comprehensive investigation to exclude or verify their significance in Mexicans. We also demonstrate that rs964184 has a large effect (odds ratio, 1.74) and is more frequent in the Mexican population, and thus it may contribute to the high predisposition to dyslipidemias in Mexicans.
Assuntos
HDL-Colesterol/genética , Dislipidemias/genética , Estudo de Associação Genômica Ampla , Triglicerídeos/genética , População Branca/genética , Alelos , Estudos de Casos e Controles , HDL-Colesterol/sangue , Família , Frequência do Gene , Variação Genética , Genética Populacional , Genótipo , Humanos , Hipertrigliceridemia/genética , Americanos Mexicanos/genética , México , Razão de Chances , Polimorfismo de Nucleotídeo Único , Triglicerídeos/sangueRESUMO
Antecedentes: El quilotórax es el acumulo de quilo en la cavidad pleural, secundario a una pérdida desde el conducto torácico, produciendo alteraciones cardiorrespiratorias, metabólicas y nutritivas. Objetivo: Analizar la metodología diagnóstica y terapéutica aplicada en pacientes con quilotórax. Diseño: Análisis retrospectivo de historias clínicas. Método: Desde enero de 1994 hasta mayo de 2004 se incluyeron 7 pacientes, 6 varones y 1 mujer, con una edad promedio de 40,4 años (rango 24-56). Todos los pacientes presentaron disnea y tos seca. A todos los enfermos se les realizó radiografía y TAC de tórax, estudio de triglicéridos (TG), colesterol y láctico deshidrogenasa (LDH) del líquido pleural, y en un paciente se indicó la linfografía radioisotópica. Todos los enfermos recibieron tratamiento inicial con avenamiento pleural, alimentación oral selectiva y parenteral parcial o total, y en dos casos se indicó tratamiento quirúrgico (Operación de Lampson) ante el fracaso del tratamiento inicial. Resultados: Los dos pacientes a quienes se les realizó la ligadura del conducto torácico tuvieron buena evolución. De los cinco restantes, 3 evolucionaron favorablemente, 1 paciente no concurrió a los controles y otro falleció sin resolver el quilotórax. La mortalidad fue del 14,2 por ciento (1 de 7 pacientes). Conclusiones: El quilotórax es de presentación infrecuente. Se sospecha por la clínica, la radiografía simple de tórax, la TAC, y se confirma por la medición de TG, colesterol y LDH en líquido pleural. Todos los pacientes son tratados inicialmente con drenaje pleural, dieta selectiva y/o parenteral, reservando la conducta quirúrgica ante el fracaso de éstos
Assuntos
Humanos , Masculino , Adulto , Pessoa de Meia-Idade , Feminino , Algoritmos , Derrame Pleural , Quilotórax/diagnóstico , Quilo , Esofagectomia , Neoplasias do Mediastino , Procedimentos Cirúrgicos Cardíacos/efeitos adversos , Procedimentos Cirúrgicos Torácicos/efeitos adversos , Quilotórax/cirurgia , Quilotórax/dietoterapia , Estudos RetrospectivosRESUMO
Antecedentes: El quilotórax es el acumulo de quilo en la cavidad pleural, secundario a una pérdida desde el conducto torácico, produciendo alteraciones cardiorrespiratorias, metabólicas y nutritivas. Objetivo: Analizar la metodología diagnóstica y terapéutica aplicada en pacientes con quilotórax. Diseño: Análisis retrospectivo de historias clínicas. Método: Desde enero de 1994 hasta mayo de 2004 se incluyeron 7 pacientes, 6 varones y 1 mujer, con una edad promedio de 40,4 años (rango 24-56). Todos los pacientes presentaron disnea y tos seca. A todos los enfermos se les realizó radiografía y TAC de tórax, estudio de triglicéridos (TG), colesterol y láctico deshidrogenasa (LDH) del líquido pleural, y en un paciente se indicó la linfografía radioisotópica. Todos los enfermos recibieron tratamiento inicial con avenamiento pleural, alimentación oral selectiva y parenteral parcial o total, y en dos casos se indicó tratamiento quirúrgico (Operación de Lampson) ante el fracaso del tratamiento inicial. Resultados: Los dos pacientes a quienes se les realizó la ligadura del conducto torácico tuvieron buena evolución. De los cinco restantes, 3 evolucionaron favorablemente, 1 paciente no concurrió a los controles y otro falleció sin resolver el quilotórax. La mortalidad fue del 14,2 por ciento (1 de 7 pacientes). Conclusiones: El quilotórax es de presentación infrecuente. Se sospecha por la clínica, la radiografía simple de tórax, la TAC, y se confirma por la medición de TG, colesterol y LDH en líquido pleural. Todos los pacientes son tratados inicialmente con drenaje pleural, dieta selectiva y/o parenteral, reservando la conducta quirúrgica ante el fracaso de éstos (AU)
Assuntos
Humanos , Masculino , Adulto , Pessoa de Meia-Idade , Feminino , Quilotórax/diagnóstico , Derrame Pleural/etiologia , Algoritmos , Quilotórax/cirurgia , Quilotórax/dietoterapia , Quilo , Esofagectomia/efeitos adversos , Procedimentos Cirúrgicos Cardíacos/efeitos adversos , Procedimentos Cirúrgicos Torácicos/efeitos adversos , Neoplasias do Mediastino/complicações , Estudos RetrospectivosRESUMO
Autosomal recessive hypercholesterolemia (ARH) is characterized by elevated LDL serum levels, xanthomatosis, and premature coronary artery disease. Three loci have been described for this condition (1p35, 15q25-q26 and 13q). Recently, the responsible gene at the 1p35 locus, encoding an LDL receptor adaptor protein (ARH) has been identified. We studied a Mexican ARH family with two affected siblings. Sequence analysis of the ARH gene (1p35 locus) revealed that the affected siblings are homozygous for a novel mutation (IVS4+2T>G) affecting the donor splice site in intron 4, whereas both the parents and an unaffected sister are heterozygous for this mutation. The IVS4+2T>G mutation results in a major alternative transcript derived from a cryptic splice site, which carries an in-frame deletion of 78 nucleotides in the mature mRNA. The translation of this mRNA yields a mutant protein product (ARH-26) lacking 26 amino acids, resulting in the loss of beta-strands beta6 and beta7 from the PTB domain. This is the first case where a naturally occurring mutant with an altered PTB domain has been identified.