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1.
J Med Virol ; 95(8): e29015, 2023 08.
Artigo em Inglês | MEDLINE | ID: mdl-37539979

RESUMO

It has been proposed that infection by adipogenic viruses constitutes a "low risk" factor for obesity. Here, we report the presence of adenovirus 36 (Ad36) and its viral load copy number in fat tissue of participants with obesity and normal weight; phylogenetic analysis was performed to describe their relationship and genetic variability among viral haplotypes. Adipose tissue obtained from 105 adult patients with obesity (cases) and 26 normal-weight adult participants as controls were analyzed by quantitative polymerase chain reaction (qPCR) amplifying the partial Ad36 E1a gene. The amplicons were examined by melting curves and submitted to sequencing. Then, genetic diversity and phylogenetic inferences were performed. Ad36 was identified at rates of 82% and 46% in the case and control groups, respectively (p = 1.1 × 10-4 , odds ratio = 5.28); viral load copies were also significantly different between both groups, being 25% higher in the case group. Melting curve analysis showed clear amplification among positive samples. Phylogenetic inferences and genetic diversity analyses showed that the Ad36 E1a gene exhibits low genetic variability and differentiation with strong gene flow due to an expanding process. Our results suggest that the phenomenon of infectobesity by Ad36 might not be a low-risk factor, as has been previously argued by other authors.


Assuntos
Infecções por Adenoviridae , Adenovírus Humanos , Adulto , Humanos , Adenovírus Humanos/genética , Gordura Intra-Abdominal , Filogenia , Carga Viral , Adenoviridae/genética , Obesidade/genética
2.
Mar Drugs ; 21(12)2023 Nov 28.
Artigo em Inglês | MEDLINE | ID: mdl-38132936

RESUMO

The majority of natural products utilized to treat a diverse array of human conditions and diseases are derived from terrestrial sources. In recent years, marine ecosystems have proven to be a valuable resource of diverse natural products that are generated to defend and support their growth. Such marine sources offer a large opportunity for the identification of novel compounds that may guide the future development of new drugs and therapies. Using the National Oceanic and Atmospheric Administration (NOAA) portal, we explore deep-sea coral and sponge species inhabiting a segment of the U.S. Exclusive Economic Zone, specifically off the western coast of Florida. This area spans ~100,000 km2, containing coral and sponge species at sea depths up to 3000 m. Utilizing PubMed, we uncovered current knowledge on and gaps across a subset of these sessile organisms with regards to their natural products and mechanisms of altering cytoskeleton, protein trafficking, and signaling pathways. Since the exploitation of such marine organisms could disrupt the marine ecosystem leading to supply issues that would limit the quantities of bioactive compounds, we surveyed methods and technological advances that are necessary for sustaining the drug discovery pipeline including in vitro aquaculture systems and preserving our natural ecological community in the future. Collectively, our efforts establish the foundation for supporting future research on the identification of marine-based natural products and their mechanism of action to develop novel drugs and therapies for improving treatment regimens of human conditions and diseases.


Assuntos
Antozoários , Produtos Biológicos , Poríferos , Animais , Produtos Biológicos/farmacologia , Produtos Biológicos/uso terapêutico , Descoberta de Drogas , Ecossistema , Florida
3.
Parasitol Res ; 122(7): 1701-1707, 2023 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-37191687

RESUMO

Blastocystis sp. is a common eukaryotic microorganism that colonizes the intestinal tract of several animals, including humans, although its role as a pathogen is still unclear. In the present study, we report the prevalence and risk factors associated with Blastocystis infection in scholars from a rural community in Mexico. A cross-sectional observational study was carried out on schoolchildren aged 3 to 15 years old; fecal samples were analyzed by culture, Faust technique, and molecular analysis. In addition, a structured questionnaire was applied to identify possible risk factors. Of the 177 samples obtained, Blastocystis sp. was the microorganism that presented the highest frequency (n=78, 44%), and included the following subtypes (STs): ST1 (n=43, 56.5%), ST2 (n=18, 23.6%), and ST3 (n=15, 19.7%); Blastocystis STs were not identified in two cases. No associating factors were found between Blastocystis infection or among STs vs. symptoms. During bivariate analysis, no statistically significant risk factors were found, except for the variable of "eating sweets, snacks, and handmade food on the way home" (p=0.04). Therefore, it is plausible to conclude that schoolchildren become infected with Blastocystis sp. mainly outside their homes, perhaps by eating contaminated handmade food on their way to or from school; however, this variable should be evaluated in detail in future studies.


Assuntos
Infecções por Blastocystis , Blastocystis , Animais , Humanos , Criança , Pré-Escolar , Adolescente , Blastocystis/genética , Infecções por Blastocystis/epidemiologia , População Rural , México/epidemiologia , Estudos Transversais , Fezes , Prevalência , Fatores de Risco , Filogenia , Variação Genética
4.
J Med Virol ; 93(10): 5969-5976, 2021 10.
Artigo em Inglês | MEDLINE | ID: mdl-34196423

RESUMO

In-house assays for the diagnosis of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) by quantitative reverse-transcription polymerase chain reaction (qRT-PCR), are feasible alternatives, particularly in developing countries. Cycle threshold (Ct ) values obtained by qRT-PCR were compared with clinical and laboratory data from saliva of inpatients with COVID-19 and asymptomatic health workers (AHW) were studied. Saliva specimens from 58 inpatients confirmed by qRT-PCR for SARS-CoV-2 using nasopharyngeal specimens, and 105 AHW were studied by qRT-PCR using three sets of primers for the N (N1, N2, and N3) gene of SARS-CoV-2, according to the CDC Diagnostic Panel protocol, showing a positivity of 88% for inpatients and 8% for AHW. Bivariate analysis revealed an association between Ct < 38.0 values for N2 and mechanical ventilation assistance among patients (p = .013). In addition, values of aspartate-transaminase, lactate dehydrogenase, and ferritin showed significant correlations with Ct values of N1 and N3 genes in inpatients. Therefore, our results show that Ct values correlate with some relevant clinical data for inpatients with COVID-19.


Assuntos
Teste de Ácido Nucleico para COVID-19/estatística & dados numéricos , COVID-19/diagnóstico , Pessoal de Saúde/estatística & dados numéricos , Pacientes Internados/estatística & dados numéricos , Adulto , Idoso , Infecções Assintomáticas , Biomarcadores/sangue , Proteínas do Nucleocapsídeo de Coronavírus/genética , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Fosfoproteínas/genética , SARS-CoV-2/genética , SARS-CoV-2/isolamento & purificação , Saliva/virologia , Índice de Gravidade de Doença
5.
New Microbiol ; 44(1): 24-32, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33582825

RESUMO

Cervical lymph node tuberculosis (LNTB) is the most common manifestation of extrapulmonary tuberculosis, resulting from the interaction of environmental and genetic factors. The immune response against TB is regulated by several cytokines, which have single nucleotide polymorphisms (SNPs), leading to different levels of expression. The aim of this study was to evaluate the association of LNTB with the TNF, IL8, IL10, IL12B and IFNG gene polymorphisms in Mexican patients. We investigated the association of ten SNPs in 14 patients with LNTB and 138 healthy controls. Significant differences were found for the allele TNF-238A (P=0.03) and the genotypes TNF-238GA (P=0.03), IL8+396GG (P=0.01) and IL12B+1188CC (P=0.04). Allele IL8+781C showed some association trend (P=0.08). Haplotypes TNF-AA and IL10-GTA were of susceptibility, whereas haplotype IL8-ATT was of protection. No association was found with IFNG. The association of these polymorphisms with extrapulmonary TB was compared in different populations. Our results suggest that these cytokine SNPs may influence the manifestation of LNTB in Mexican patients; however, we are aware of the limitations of our study, so it is necessary to make a replica using a larger sample of patients, as well as an increased number of cytokines with SNPs.


Assuntos
Interleucina-10 , Tuberculose dos Linfonodos , Estudos de Casos e Controles , Predisposição Genética para Doença , Humanos , Interferon gama/genética , Interleucina-10/genética , Subunidade p40 da Interleucina-12/genética , Interleucina-8 , Polimorfismo de Nucleotídeo Único , Tuberculose dos Linfonodos/genética
6.
J Sport Rehabil ; 29(5): 533-540, 2020 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-31034307

RESUMO

CONTEXT: Previous research has analyzed how the sport influences sagittal spinal curvatures in young athletes and has found that spinal curves may be modified as a consequence of repeated movement patterns and postures of each discipline. OBJECTIVE: To analyze sagittal spinal alignment by equestrian discipline and its relation to training load, and to describe "sagittal integrative morphotype" in young riders. DESIGN: Observational descriptive study. SETTING: Training room. PARTICIPANTS: A total of 23 riders (aged 9-17 y)-13 dressage riders (3 males and 10 females) and 10 show jumping riders (5 males and 5 females)-participated voluntarily. MAIN OUTCOME MEASURES: Mann-Whitney U test was applied to determine differences between riders' characteristics (gender, discipline, and training load) and spine variables. RESULTS: According to normality ranges for spinal curves, females showed an increase for lumbar curvature in standing position. It was found that show jumping riders manifested an increment in thoracic and lumbar curves while standing and an increase in the thoracic curvature in slump sitting. Statistically significant differences were found when lumbar curvature, "sit and reach" distance, and lumbo-horizontal angle in flexion were analyzed by gender in "sit and reach" test. No statistical significant differences were found when spinal curves in each position were analyzed depending on the training load. With regard to "sagittal integrative morphotype," all riders presented a hyperkyphotic dorsal morphotype no matter what their discipline. As for the lumbar curve, dressage and show jumping riders presented a functional hyperkyphotic morphotype. CONCLUSIONS: It is important to note that many riders presented a sagittal imbalance for the thoracic and lumbar curves. Therefore, as the sagittal spinal misalignments persist and worsen over time, exercise programs to prevent or rehabilitate these imbalances in young riders will be needed. The "sagittal integrative morphotype" assessment is an essential tool in order to identify the spinal misalignment.


Assuntos
Postura/fisiologia , Coluna Vertebral/anatomia & histologia , Esportes Juvenis/fisiologia , Adolescente , Animais , Criança , Feminino , Cavalos , Humanos , Região Lombossacral/anatomia & histologia , Masculino , Movimento/fisiologia , Avaliação de Resultados em Cuidados de Saúde , Fatores Sexuais , Postura Sentada , Curvaturas da Coluna Vertebral/diagnóstico , Posição Ortostática , Estatísticas não Paramétricas , Vértebras Torácicas/anatomia & histologia
7.
Langmuir ; 34(50): 15551-15559, 2018 12 18.
Artigo em Inglês | MEDLINE | ID: mdl-30453736

RESUMO

The self-assembly behavior of a diphenylalanine amphiphile blocked at the C-terminus with a 9-fluorenylmethyl ester and stabilized at the N-terminus with a trifluoroacetate (TFA) anion, TFA·FF-OFm, has been examined. At low peptide concentration (0.5 mg/mL), long amyloid-like fibrils, which come from the fusion of two or more helical ribbons and/or thinner fibrils, organized in bundles or as individual entities are detected. Microbeam synchrotron radiation infrared spectroscopy has shown that TFA·FF-OFm molecules in amyloid-like fibrils arrange, forming antiparallel ß-sheets. Alteration of the experimental conditions to prioritize the thermodynamic contribution with respect to the kinetic one in the self-assembly process inhibits the organization of amyloid-like structures in favor of the formation of conventional fibrous structures. On the basis of experimental observations, a structural model where the individual antiparallel ß-sheets are oriented in parallel has been proposed for TFA·FF-OFm amyloid-like fibrils.


Assuntos
Fluorenos/química , Fenilalanina/análogos & derivados , Dipeptídeos , Estrutura Molecular , Peptídeos/química , Fenilalanina/síntese química , Fenilalanina/química
8.
Parasitol Res ; 117(12): 3935-3943, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30298236

RESUMO

The potential role of Blastocystis as a pathogen is controversial because it is found in both symptomatic and asymptomatic carriers. Since Cathepsin B has been identified as a main virulence factor that contributes to the pathogenesis of this parasite, the purpose of this study was to analyze the genetic polymorphisms of cathepsin B from Blastocystis from patients with irritable bowel syndrome and from asymptomatic carriers. DNA from fecal samples of both groups, which were previously genotyped by 18S sequencing, was used to amplify a fragment of the cathepsin B gene. Phylogenetic reconstructions were performed and some genetic population indexes were obtained. Amplicons of 27 samples (15 cases, 10 controls, and two commercial ATCC strains) were obtained and analyzed. Phylogenetic reconstructions using nucleotides or inferred amino acid sequences did not separate between cases or controls or among subtypes. Regarding the values of genetic variability, we found that the haplotype and nucleotide diversity indexes of cathepsin B from cases and controls were similar to the values of 18S from controls. By contrast, 18S from cases showed low variability, suggesting that the genetic variability of cathepsin B was not related to the symptomatology of Blastocystis carriers. However, since no polymorphisms related to cases or controls were found, it is logical to assume that the potential damage caused by Blastocystis in situ may be due to unclear mechanisms of Cathepsin B regulation and expression that should be studied in future studies.


Assuntos
Infecções por Blastocystis/parasitologia , Blastocystis/genética , Blastocystis/patogenicidade , Catepsina B/genética , Síndrome do Intestino Irritável/parasitologia , Adulto , Sequência de Aminoácidos/genética , Blastocystis/classificação , Fezes/parasitologia , Feminino , Genética Populacional , Genótipo , Haplótipos , Humanos , Masculino , Pessoa de Meia-Idade , Filogenia , Polimorfismo Genético , Fatores de Virulência/genética
9.
Rev Med Chil ; 146(3): 399-402, 2018 Mar.
Artigo em Espanhol | MEDLINE | ID: mdl-29999113

RESUMO

Aplastic anemia (AA) or acquired aplastic anemia is an uncommon and potentially fatal disease. It is defined as reduction of at least two peripheral blood series, associated with persistent bone marrow hypocellularity. It's association with pregnancy is even more uncommon, and it may result in high mortality for the mother and the child. We report an adolescent female with aplastic anemia, which worsened during pregnancy. Her hemoglobin was 5.2 g/dl, her leukocytes were 1,833/ul and her platelets were 19,000/ul. She was initially treated with cyclosporine and horse antithymocyte globulin without hematologic improvement. At 36 weeks of pregnancy methylprednisolone and platelet transfusions were used and a cesarean section was performed. Six month later she continues to require red blood cell and platelet transfusions.


Assuntos
Anemia Aplástica/terapia , Complicações Hematológicas na Gravidez/terapia , Adolescente , Feminino , Humanos , Recém-Nascido , Masculino , Gravidez , Resultado da Gravidez , Índice de Gravidade de Doença
10.
Phys Chem Chem Phys ; 19(39): 27038-27051, 2017 Oct 11.
Artigo em Inglês | MEDLINE | ID: mdl-28959820

RESUMO

Alkyne and azide, which are commonly used in the cycloaddition reaction recognized as "click chemistry", have been used as capping groups of two engineered diphenylalanine (FF) derivatives due to their ability to form weak intermolecular interactions (i.e. dipole-π and π-π stacking). In Poc-FF-N3, alkyne and azide act as N- and C-terminal capping groups, respectively, while such positions are exchanged in N3-FF-OPrp. The self-assembly of such two synthesized peptides has been extensively studied in their "pre-click" state, considering the influence of three different factors: the peptide concentration, the polarity of the medium, and the nature of the substrate. Poc-FF-N3 assembles into microfibers that, depending on the medium and the substrate, can aggregate hierarchically in supramolecular structures with different morphologies. The most distinctive one corresponds to very stable birefringent dendritic-like microstructures, which are derived from the ordered agglomeration of microfibers. These branched supramolecular structures, which are observed under a variety of conditions, are relatively uncommon in short FF sequences. At the molecular level, Poc-FF-N3 organizes in antiparallel ß-sheets stabilized by N-HO intermolecular hydrogen bonds and re-enforced by weak interactions between the azide and alkyne groups of neighbouring molecules. In contrast, N3-FF-OPrp exhibits a very poor tendency to organize into structures with a well-defined morphology. Theoretical calculations on model complexes indicate that the tendency of the latter peptide to organize into small amorphous agglomerates is due to its poor ability to form specific intermolecular interactions in comparison with Poc-FF-N3. The implications of the weak interactions induced by the alkyne and azide groups, which strengthen peptidepeptide hydrogen bonds and π-ladders due to the stacked aromatic phenyl side groups, are discussed.

11.
Am J Pathol ; 185(8): 2076-82, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-26055209

RESUMO

Amniocentesis in rats is associated with different malformations, such as cleft palate and limb deformation, resembling the human congenital amniotic band syndrome (ABS). Despite many human cases reported in the literature, little is known about the mechanisms involved in ABS. This study addressed if the activation of the transforming growth factor-ß1 (TGF-ß1) pathway is, in part, associated with amniotic band formation and growth restriction induced in rats by amniocentesis, as by a previously published model. For this purpose, quantification of TGF-ß1, α-smooth muscle actin, and collagen type I mRNA and protein levels were determined by quantitative PCR and Western blot analysis, respectively, in the fetus, its amniotic membrane, and the uterus of experimental and control rats. We found that TGF-ß1 mRNA levels are increased in the fetus and the amniotic membrane at 6 hours, whereas α-smooth muscle actin, phosphorylated Smad3, and collagen type I increased at 48 hours, suggesting that a fibrotic response is induced after the amniotic sac puncture. Furthermore, fetuses had hemorrhages, syndactyly, and amputation of limbs, similar to human ABS.


Assuntos
Síndrome de Bandas Amnióticas/metabolismo , Modelos Animais de Doenças , Fator de Crescimento Transformador beta1/metabolismo , Actinas/genética , Actinas/metabolismo , Amniocentese , Síndrome de Bandas Amnióticas/genética , Animais , Colágeno Tipo I/genética , Colágeno Tipo I/metabolismo , Feminino , Retardo do Crescimento Fetal/genética , Retardo do Crescimento Fetal/metabolismo , Ratos , Fator de Crescimento Transformador beta1/genética
12.
Hereditas ; 153: 2, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-28096764

RESUMO

BACKGROUND: Human obesity is due to a complex interaction among environmental, behavioral, developmental and genetic factors, including the interaction of leptin (LEP) and leptin receptor (LEPR). Several LEPR mutations and polymorphisms have been described in patients with early onset severe obesity and hyperphagic eating behavior; however, some contradictory findings have also been reported. In the present study we explored the association of six LEPR gene polymorphisms in patients with morbid obesity. FINDINGS: Twenty eight patients with morbid obesity and 56 non-obese Mexican Mestizo individuals were included. Typing of rs1137100, rs1137101, rs1805134, Ser492Thr, rs1805094 and rs1805096 LEPR polymorphisms was performed by PCR and allele specific hybridization. The LEPR Ser492Thr polymorphism was monomorphic with the presence of only the Ser492Thr-G allele. Allele C and genotype T/C for rs1805134 polymorphism were associated with susceptibility to morbid obesity (p = 0.02 and p = 0.03, respectively). No association was observed with any haplotype. Linkage disequilibrium (LD) showed that five polymorphisms (rs1137100, rs1137101, rs1805134, rs1805094 and rs1805096) were in absolute (D' = 1) but none in perfect (r2 = 1) LD. CONCLUSIONS: Our results suggest that rs1805134 polymorphism could be involved in the development of morbid obesity, whilst none of the alleles of the LEPR gene, rs1137100, rs1137101, rs1805094 and rs1805096 were associated as risk factors. However, more studies are necessary to confirm or reject this hypothesis.


Assuntos
Obesidade Mórbida/genética , Polimorfismo de Nucleotídeo Único , Receptores para Leptina/genética , Adulto , Alelos , Estudos de Casos e Controles , Feminino , Frequência do Gene , Genótipo , Haplótipos , Humanos , Desequilíbrio de Ligação , Masculino , México
13.
Chemistry ; 21(47): 16895-905, 2015 Nov 16.
Artigo em Inglês | MEDLINE | ID: mdl-26419936

RESUMO

Three different tetraphenylalanine (FFFF) based peptides that differ at the N- and C-termini have been synthesized by using standard procedures to study their ability to form different nanoassemblies under a variety of conditions. The FFFF peptide assembles into nanotubes that show more structural imperfections at the surface than those formed by the diphenylalanine (FF) peptide under the same conditions. Periodic DFT calculations (M06L functional) were used to propose a model that consists of three FFFF molecules defining a ring through head-to-tail NH3(+)⋅⋅⋅(-)OOC interactions, which in turn stack to produce deformed channels with internal diameters between 12 and 16 Å. Depending on the experimental conditions used for the peptide incubation, N-fluorenylmethoxycarbonyl (Fmoc) protected FFFF self-assembles into a variety of polymorphs: ultra-thin nanoplates, fibrils, and star-like submicrometric aggregates. DFT calculations indicate that Fmoc-FFFF prefers a parallel rather than an antiparallel ß-sheet assembly. Finally, coexisting multiple assemblies (up to three) were observed for Fmoc-FFFF-OBzl (OBzl = benzyl ester), which incorporates aromatic protecting groups at the two peptide terminals. This unusual and noticeable feature is attributed to the fact that the assemblies obtained by combining the Fmoc and OBzl groups contained in the peptide are isoenergetic.


Assuntos
Fluorenos/química , Nanotubos/química , Peptídeos/química , Peptídeos/síntese química , Fenilalanina/análogos & derivados , Fenilalanina/química , Fenilalanina/síntese química , Dipeptídeos , Simulação de Dinâmica Molecular
14.
Environ Microbiol ; 16(9): 2791-803, 2014 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-24707812

RESUMO

This is the first study using molecular and culture-based methods aimed at investigating the composition of the intestinal yeast microbiota of wild and reared carnivorous salmonids, croaker and yellowtail, to characterize their cores and to evaluate the enzymatic activities of the cultivated yeast. Among 103 samples from salmonids, croaker and yellowtail, yeast were detected in 85.4%, with 43 species identified. The core of reared fish was composed of eight species, in contrast to the wild fish core, which consisted of two species: Debaryomyces hansenii and Rhodotorula mucilaginosa. Despite the smaller diversity of the wild fish core, similar enzymatic profiles were detected for the species from the wild and reared cores. For principal component analysis, samples grouped together independently of host species, domestication status and location. A high proportion of yeast produced aminopeptidases and lipases, which may be explained by the high proportion of protein and lipids in the carnivorous diet. This study reveals the presence of a yeast community in the fish gut that appears to be strongly shaped by a carnivorous diet. Yeast in the gut increases the repertoire of microorganisms interacting with the host intestine, which could influence health and disease.


Assuntos
Intestinos/microbiologia , Microbiota , Perciformes/microbiologia , Rhodotorula/isolamento & purificação , Salmonidae/microbiologia , Leveduras/isolamento & purificação , Animais , DNA Fúngico/genética
15.
Int J Mol Sci ; 15(5): 7064-123, 2014 Apr 25.
Artigo em Inglês | MEDLINE | ID: mdl-24776758

RESUMO

Poly(alkylene dicarboxylate)s constitute a family of biodegradable polymers with increasing interest for both commodity and speciality applications. Most of these polymers can be prepared from biobased diols and dicarboxylic acids such as 1,4-butanediol, succinic acid and carbohydrates. This review provides a current status report concerning synthesis, biodegradation and applications of a series of polymers that cover a wide range of properties, namely, materials from elastomeric to rigid characteristics that are suitable for applications such as hydrogels, soft tissue engineering, drug delivery systems and liquid crystals. Finally, the incorporation of aromatic units and α-amino acids is considered since stiffness of molecular chains and intermolecular interactions can be drastically changed. In fact, poly(ester amide)s derived from naturally occurring amino acids offer great possibilities as biodegradable materials for biomedical applications which are also extensively discussed.


Assuntos
Materiais Biocompatíveis/química , Ácidos Dicarboxílicos/química , Glicóis/química , Nylons/química , Poliésteres/química , Animais , Materiais Biocompatíveis/síntese química , Materiais Biocompatíveis/metabolismo , Ácidos Dicarboxílicos/síntese química , Ácidos Dicarboxílicos/metabolismo , Sistemas de Liberação de Medicamentos , Glicóis/síntese química , Glicóis/metabolismo , Humanos , Modelos Moleculares , Nylons/síntese química , Nylons/metabolismo , Poliésteres/síntese química , Poliésteres/metabolismo , Polimerização , Engenharia Tecidual
16.
J Vis Exp ; (208)2024 Jun 14.
Artigo em Inglês | MEDLINE | ID: mdl-38949304

RESUMO

The dot-blot is a simple, fast, sensitive, and versatile technique that enables the identification of minimal quantities of DNA specifically targeted by probe hybridization in the presence of carrier DNA. It is based on the transfer of a known amount of DNA onto an inert solid support, such as a nylon membrane, utilizing the dot-blot apparatus and without electrophoretic separation. Nylon membranes have the advantage of high nucleic acid binding capacity (400 µg/cm2), high strength, and are positively or neutrally charged. The probe used is a highly specific ssDNA fragment of 18 to 20 bases long labeled with digoxigenin (DIG). The probe will conjugate with the Leptospira DNA. Once the probe has hybridized with the target DNA, it is detected by an anti-digoxigenin antibody, allowing its easy detection through its emissions revealed in an X-ray film. The dots with an emission will correspond to the DNA fragments of interest. This method employs the non-isotopic labeling of the probe, which may have a very long half-life. The drawback of this standard immuno-label is a lower sensitivity than isotopic probes. Nevertheless, it is mitigated by coupling polymerase chain reaction (PCR) and dot-blot assays. This approach enables the enrichment of the target sequence and its detection. Additionally, it may be used as a quantitative application when compared against a serial dilution of a well-known standard. A dot-blot application to detect Leptospira from the three main clades in water samples is presented here. This methodology can be applied to large amounts of water once they have been concentrated by centrifugation to provide evidence of the presence of Leptospiral DNA. This is a valuable and satisfactory tool for general screening purposes, and may be used for other non-culturable bacteria that may be present in water, enhancing the comprehension of the ecosystem.


Assuntos
Leptospira , Reação em Cadeia da Polimerase , Leptospira/genética , Leptospira/isolamento & purificação , Reação em Cadeia da Polimerase/métodos , DNA Bacteriano/genética , DNA Bacteriano/análise , Hibridização de Ácido Nucleico/métodos , Microbiologia da Água
17.
Microorganisms ; 12(6)2024 Jun 05.
Artigo em Inglês | MEDLINE | ID: mdl-38930533

RESUMO

In the present study, we compared the genetic variability of fragments from the internal transcribed spacer region (ITS) and the small subunit ribosomal DNA (SSUrDNA) as nuclear markers, in contrast with the ribosomal protein large two (rpl2) loci, placed in the mitochondrion-related organelles (MROs) within and among human fecal samples with Blastocystis. Samples were analyzed using polymerase chain reaction (PCR)-sequencing, phylogenies, and genetics of population structure analyses were performed. In total, 96 sequences were analyzed, i.e., 33 of SSUrDNA, 35 of rpl2, and 28 of ITS. Only three subtypes (STs) were identified, i.e., ST1 (11.4%), ST2 (28.6%), and ST3 (60%); in all cases, kappa indexes were 1, meaning a perfect agreement among ST assignations. The topologies of phylogenetic inferences were similar among them, clustering to each ST in its specific cluster; discrepancies between phylogeny and assignment of STs were not observed. The STRUCTURE v2.3.4 software assigned three subpopulations corresponding to the STs 1-3, respectively. The population indices were consistent with those previously reported by other groups. Our results suggest the potential use of the ITS and rpl2 genes as molecular markers for Blastocystis subtyping as an alternative approach for the study of the genetic diversity observed within and between human isolates of this microorganism.

18.
Artigo em Inglês | MEDLINE | ID: mdl-38324874

RESUMO

Currently, there are some concerns about the situation and, in particular, about the future of the COVID-19 pandemic and the new emerging variants of SARS-CoV-2. Rodents are an example of synanthropic animals in urban environments that harbor important zoonoses. Although the molecular identification of SARS-CoV-2 in Rattus norvegicus from New York City had been reported, in other studies, urban wild rodents infected with this virus have not been found. This study aimed to molecularly identify the presence of SARS-CoV-2 in urban wild rodents from Mexico City, trapped along a water channel of a public park as part of a pest control program, at the beginning of the COVID-19 pandemic, during the fall and winter of 2020. Up to 33 Mus musculus and 52 R. norvegicus were captured and euthanized, large intestine samples with feces from the animals were obtained. RNAs were obtained and subjected to qRT-PCR for SARS-CoV-2 identification and threshold cycle (Ct) values were obtained. Four mice (12.1%) and three rats (5.8%) were positive, three rodents exhibited Ct<30. Our results on the frequency of SARS-CoV-2 in urban rats are in line with other previous reports. Thus, similar to other authors, we suggest that surveillance for the detection of SARS-CoV-2 in urban wild rodents, as sentinel animals, should be maintained.


Assuntos
COVID-19 , Roedores , Ratos , Camundongos , Animais , Humanos , COVID-19/epidemiologia , SARS-CoV-2 , México/epidemiologia , Pandemias
19.
PLoS One ; 18(5): e0285752, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37192191

RESUMO

COVID-19 exposed and exacerbated health disparities, and a core challenge has been how to adapt pandemic response and public health in light of these disproportionate health burdens. Responding to this challenge, the County of Santa Clara Public Health Department designed a model of "high-touch" contact tracing that integrated social services with disease investigation, providing continued support and resource linkage for clients from structurally vulnerable communities. We report results from a cluster randomized trial of 5,430 cases from February to May 2021 to assess the ability of high-touch contact tracing to aid with isolation and quarantine. Using individual-level data on resource referral and uptake outcomes, we find that the intervention, randomized assignment to the high-touch program, increased the referral rate to social services by 8.4% (95% confidence interval, 0.8%-15.9%) and the uptake rate by 4.9% (-0.2%-10.0%), with the most pronounced increases in referrals and uptake of food assistance. These findings demonstrate that social services can be effectively combined with contact tracing to better promote health equity, demonstrating a novel path for the future of public health.


Assuntos
COVID-19 , Humanos , COVID-19/epidemiologia , COVID-19/prevenção & controle , Busca de Comunicante/métodos , Tato , Promoção da Saúde , SARS-CoV-2 , Serviço Social
20.
Mol Biol Rep ; 39(9): 8837-43, 2012 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-22740130

RESUMO

Irritable bowel syndrome (IBS) is one of the most common gastrointestinal diagnoses seen by primary care providers and gastroenterologists. Proinflammatory cytokines interleukin (IL)-6 and IL-8 have been found increased in IBS patients. Cytokine gene single nucleotide polymorphisms (SNPs) of IL-8 and IL-10 have not been assessed in Mexican IBS patients. DNA was extracted from peripheral blood leucocytes of 45 IBS unrelated patients and 137 controls. Allele, genotype, and haplotype frequencies were determined by analyzing SNPs of IL-8 and IL-10 genes. IL-8 + 396 G allele (P = 0.02), IL-8 + 396(G/G) and IL-8 + 781(C/T) genotypes (P < 0.001), IL-10 - 1082A allele and IL-10 - 1082(A/A) genotype (P < 0.001) were significantly increased in the IBS group. Haplotypes IL-8 ATCC (P = 0.03) and IL-10 ACC (P < 0.001) were associated with susceptibility to develop IBS. An association of certain polymorphisms of IL-8 and IL-10 in IBS patients compared to controls was demonstrated, suggesting a role of these cytokine SNPs in the pathophysiology of IBS.


Assuntos
Interleucina-10/genética , Interleucina-8/genética , Síndrome do Intestino Irritável/genética , Polimorfismo de Nucleotídeo Único , Adulto , Alelos , Estudos de Casos e Controles , Feminino , Frequência do Gene , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade
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