RESUMEN
Histoplasmosis is caused by the fungus Histoplasma capsulatum and is often fatal for individuals with acquired immunodeficiency syndrome (AIDS). Delayed diagnosis is a major factor in worsening coinfection, as it can be mistaken for other diseases. Thus, rapid identification of Histoplasma in immunocompromised patients is essential. Molecular techniques, particularly polymerase chain reaction (PCR), were used in this study to identify H. capsulatum in patients coinfected with histoplasmosis and AIDS. Blood samples from 14 individuals with AIDS and disseminated histoplasmosis were collected and analyzed. The PCR method successfully amplified the fungal region in whole blood samples, while PCR-RFLP analysis confirmed a consistent profile in the samples. Genetic sequencing further confirmed the fungal species. Compared to clinical tests such as fungal culture and urinary antigen detection, molecular analysis proved faster, more sensitive, and cost-effective. These molecular markers can potentially be incorporated into routine diagnostics in the future. Further studies are needed to expand and enhance this diagnostic approach, particularly in patients with nonprogressive clinical forms of histoplasmosis.
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Infecciones Oportunistas Relacionadas con el SIDA , Histoplasma , Histoplasmosis , Reacción en Cadena de la Polimerasa , Humanos , Histoplasmosis/diagnóstico , Histoplasmosis/microbiología , Histoplasma/genética , Histoplasma/aislamiento & purificación , Infecciones Oportunistas Relacionadas con el SIDA/microbiología , Infecciones Oportunistas Relacionadas con el SIDA/diagnóstico , Masculino , Femenino , ADN de Hongos/análisis , ADN de Hongos/genética , ADN de Hongos/sangre , Adulto , Polimorfismo de Longitud del Fragmento de Restricción , Síndrome de Inmunodeficiencia Adquirida/complicaciones , Síndrome de Inmunodeficiencia Adquirida/microbiología , Persona de Mediana EdadRESUMEN
The aim of this study was to investigate the DGAT1 gene polymorphism and its effects on lamb weight in kazakh and tajik sheep breeds. A total of 97 blood samples were collected from purebred (еdilbay Ñ Ðµdilbay) and crossbred lambs (еdilbay x gissar) breеd by the Baiserke Agro Scientific and Production Center in the Talgar District of the Almaty Region of Kazakhstan. Animals were genotyped for DGAT1-AluI polymorphism using the polymerase chain reaction-restriction length polymorphism (PCR-RFLP) method. The result of PCR-RFLP showed that purebred (еdilbay Ñ Ðµdilbay) sheep had three genotypes (CC, CT and TT) and crossbred sheep had two genotypes (CC and CT). The predominant genotype was CC with a frequency of 0.70 and 0.58 in purebred sheep and crossbred sheep breeds, respectively. The DGAT1 gene showed no significant association with live weight of lambs at different times in both breeds studied. However, the study showed that the CC genotype produced higher live weight at day 60 in purebred sheep (CC: 33,668 kg and CT: 32,444) and at day 120 (CC: 41,487 and CT: 40,929) in crossbred lambs. The present study was the first to investigate the polymorphism and relationships between genotypes and lamb live weights for DGAT1 gene in sheep breeds, purebred and crossbred. We conclude that further comprehensive investigations should be done for the exact evidence of the effects of DGAT1/Alui polymorphism on lamb live weights.
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Peso Corporal , Diacilglicerol O-Acetiltransferasa , Genotipo , Reacción en Cadena de la Polimerasa , Polimorfismo Genético , Polimorfismo de Longitud del Fragmento de Restricción , Animales , Diacilglicerol O-Acetiltransferasa/genética , Ovinos/genética , Reacción en Cadena de la Polimerasa/veterinaria , Peso Corporal/genética , Frecuencia de los Genes , Kazajstán , MasculinoRESUMEN
The work aims to analyze the associations of polymorphic variants of the PRL and BLG genes with resistance and susceptibility to mastitis in Holstein cows. The experimental study consisted of the selection of biomaterial samples from 250 heads of Holstein cows aged 3 years divided into two groups (healthy and with a confirmed diagnosis of mastitis). The determination of animal genotypes was carried out using polymerase chain reaction and restriction fragment length polymorphism. The study of the nature of the association of polymorphic variants of the PRL and BLG gene with resistance/increased risk of mastitis established a significant deviation from the theoretically expected distribution of bBLG-HaeIII genotypes in the group of animals suffering from mastitis (the value of χ2 was 0.24). The bBLG-HaeIIIBB genotype can act as a marker of an increased risk of developing mastitis in Holstein cows; its frequency in the group of sick animals exceeds the frequency in the control group by more than 2 times (44.0 compared to 17.0%, respectively). The bBLG-HaeIIIAB genotype is significantly associated with mastitis resistance in Holstein cows; its frequency is 2 times lower than in the control group (28.0 compared to 54.0%).
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Predisposición Genética a la Enfermedad , Genotipo , Lactoglobulinas , Mastitis Bovina , Reacción en Cadena de la Polimerasa , Polimorfismo Genético , Prolactina , Animales , Bovinos , Femenino , Mastitis Bovina/genética , Prolactina/genética , Reacción en Cadena de la Polimerasa/veterinaria , Lactoglobulinas/genética , Polimorfismo de Longitud del Fragmento de Restricción , Frecuencia de los GenesRESUMEN
Genetic improvement of local rabbit breeds using modern approaches such as marker-assisted selection requires accurate and precise information about markerâtrait associations in animals with different genetic backgrounds. Therefore, this study was designed to estimate the association between two mutations located in the Neuropeptide Y (NPY, g.1778G > C) and Phosphoglycerate Mutase 2 (PGAM2, c.195 C > T) genes in New Zealand White (NZW), Baladi (BR), and V-line rabbits. The first mutation was genotyped using high-resolution melting, and the second mutation was genotyped using the PCR-RFLP method. The results revealed significant associations between the NPY mutation and body weight at 10 (V-line) and 12 weeks of age (NZW, BR, and V-line), body weight gain (BWG) from 10 to 12 weeks of age (BR), BWG from 6 to 12 weeks of age (NZW, BR, and V-line), average daily gain (NZW, BR, and V-line, and BR), growth rate (GR) from 8 to10 weeks (V-line), 10 to 12 weeks (BR), and GR from 6 to 12 weeks of age (BR, and V-line). The PGAM2 mutation was associated with body weight at 10 (V-line) and 12 (NZW, and V-line) weeks of age, with significant positive additive effects at 12 weeks of age in all breeds, and was associated with BWG from 8 to 10 and 10 to 12 in BR, and BWG from 6 to 12 weeks of age (NZW, and BR), and average daily gain (NZW, and BR), and was associated with GR form 8 to 10 weeks (BR), from10 to 12 weeks (BR, and V-line) and from 6 to 12 weeks (BR). The results highlighted the importance of the two mutations in growth development, and the possibility of considering them as candidate genes for late growth in rabbits.
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Neuropéptido Y , Fosfoglicerato Mutasa , Polimorfismo de Nucleótido Simple , Animales , Conejos/crecimiento & desarrollo , Conejos/genética , Fosfoglicerato Mutasa/genética , Fosfoglicerato Mutasa/metabolismo , Neuropéptido Y/genética , Neuropéptido Y/metabolismo , Masculino , Femenino , Genotipo , Peso Corporal/genética , Polimorfismo de Longitud del Fragmento de Restricción , Aumento de Peso/genéticaRESUMEN
BACKGROUND: Opisthorchis-like eggs are a public health problem in northern and northeastern Thailand. However, the genetic epidemiology and structure of these parasites in northern Thailand are unknown. Thus, this study investigated their population genetic structure using cytochrome c oxidase subunit 1 (cox1) and NADH dehydrogenase subunit 1 (nad1) nucleotide sequences. METHODOLOGY/PRINCIPAL FINDINGS: A study was conducted in the hill tribe regions of Chiang Mai Province, northern Thailand. Internal transcribed spacer 2 polymerase chain reaction and restriction fragment length polymorphism were used to distinguish 205 positive feces samples for Opisthorchis-like eggs. The results showed that the prevalence of O. viverrini and Haplorchis taichui was 10.5% and 38.2%, respectively, and the co-infection rate was 37.2%. To determine the genetic structure of O. viverrini and H. taichui using cox1 and nad1 genes, genetic analysis was performed using 30 randomly chosen fecal samples for Opisthorchis-like eggs. Pairwise FST analysis indicated that O. viverrini and H. taichui displayed nonsignificant genetic differentiation within Chiang Mai Province and between interpopulations from different geographic areas. Moreover, within the intrapopulation in Chiang Mai Province, cox1 presented higher gene flow than nad1 in O. viverrini, while nad1 demonstrated higher gene flow than cox1 in H. taichui. The neutrality tests based on Fu's Fs indicated population expansion and selective sweep from bottleneck or hitchhiking in O. viverrini and H. taichui populations, supported by haplotype network patterns. Phylogenetic tree analysis based on cox1 and nad1 revealed the monophyly of O. viverrini and H. taichui and genetic relationships with other isolates collected from Thailand, Lao People's Democratic Republic (PDR), and Vietnam. CONCLUSIONS/SIGNIFICANCE: This study investigated the molecular discrimination and genetic structure of Opisthorchis-like eggs in northern Thailand. The genetic information derived from this study could be associated with the background, molecular epidemiology, and disease severity of these parasites.
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Complejo IV de Transporte de Electrones , Heces , Opistorquiasis , Opisthorchis , Animales , Tailandia/epidemiología , Opistorquiasis/parasitología , Opistorquiasis/epidemiología , Opisthorchis/genética , Opisthorchis/clasificación , Opisthorchis/aislamiento & purificación , Heces/parasitología , Humanos , Complejo IV de Transporte de Electrones/genética , NADH Deshidrogenasa/genética , Genes Mitocondriales/genética , Variación Genética , Polimorfismo de Longitud del Fragmento de Restricción , Filogenia , PrevalenciaRESUMEN
PURPOSE: The present study was conducted to determine the presence of Toxoplasma gondii in donkeys by molecular tests and genetic diversity analysis of the obtained DNA samples from central Kenya. METHOD: A total of 363 blood samples were collected from donkeys in Meru and Kirinyaga Counties, and 96 samples that were previously seropositive for T. gondii using indirect ELISA were subjected to nested PCR based on the amplification of the internal transcribed spacer 1 (ITS-1) gene followed by DNA sequencing and phylogenetic analysis. Genotyping was performed on 15 selected positive samples using multilocus nested polymerase chain reaction restriction fragment length polymorphism (Mn-PCR-RFLP) with eight genetic markers ('SAG 2, 5'SAG 2, Alt. SAG 2, SAG 3, GRA 6, C29-2, BTUB and L358). RESULTS: Toxoplasma gondii DNA was detected in 36.5% (35/96) of the blood samples. The sequences obtained exhibited 98.2-99.5% homology with those deposited in GenBank. Phylogenetic analysis demonstrated that the obtained sequences are conserved and clustered with those of infecting animals from other regions of the world. Eighteen distinct T. gondii haplotypes were identified to be circulating in donkeys from central Kenya. The T. gondii DNA samples exhibited high haplotype diversity (Hd: 0.915) and limited genetic diversity (π = 0.01027). PCR-RFLP of T. gondii DNA-positive samples revealed three different genetic combinations that consisted of alleles I, II and III, indicating the dissemination of atypical genotypes. CONCLUSION: This study demonstrated that T. gondii is widespread in donkeys from Kenya and could be a possible source of infection in humans. These findings are important for designing control strategies for this parasite to improve the livestock sector, which is one of the main sources of livelihood for farmers in Kenya.
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ADN Protozoario , Equidae , Variación Genética , Filogenia , Polimorfismo de Longitud del Fragmento de Restricción , Toxoplasma , Toxoplasmosis Animal , Animales , Toxoplasma/genética , Toxoplasma/aislamiento & purificación , Toxoplasma/clasificación , Kenia/epidemiología , Equidae/parasitología , Toxoplasmosis Animal/parasitología , Toxoplasmosis Animal/epidemiología , ADN Protozoario/genética , Reacción en Cadena de la Polimerasa/veterinaria , Genotipo , Análisis de Secuencia de ADN , HaplotiposRESUMEN
BACKGROUND/OBJECTIVE: The cestode Echinococcus granulosus causes cystic echinococcosis, a zoonotic parasitic infection that constitutes a significant public health risk. This parasite has been documented to have potential reservoirs and carriers among wild canids, namely wolves, foxes and jackals. This study aimed to determine the prevalence and molecular characteristics of E. granulosus sensu lato species/genotypes among wild canids in three northern, northeastern and north-western Iran regions. METHODS: From 2019 to 2022, 93 wild canid carcasses (69 jackals), (22 foxes) and (2 wolves) were collected that were killed in car accidents or illnesses. Analyses of morphology and morphometry were performed to verify the presence of E. granulosus. To determine E. granulosus s.l. species/genotypes, polymerase chain reaction (PCR)-RFLP (ITS1) was performed utilizing the Bsh1236I (BstUI) restriction enzyme. COX1, NADH1 and ITS1 gene sequencing were also performed to confirm the PCR-RFLP results. RESULTS: During this study, 93 wild canids were examined, and 3.2% (95% CI: 0%-7%) of the 93 were infected with Echinococcus. The north-western region of Iran showed two out of 30 jackals (6.6%) infected with adult Echinococcus compared to one out of 35 jackals (2.8%) in the northern region. DNA from Echinococcus was detected in these individuals by PCR. Based on PCR-RFLP analysis of the ITS1 gene and sequencing of COX1, NADH1 and ITS1 gene, E. granulosus sensu stricto genotype was confirmed in the jackals that had been infected. CONCLUSION: Evidence shows that E. granulosus occurs in jackals in Iran, with the E. granulosus s.s. genotype being the most common. This parasite has been identified as a zoonotic parasite with a genotype that can be transmitted to livestock and humans. Establishing effective control measures to prevent the spread of echinococcosis and ensure public health is crucial.
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Equinococosis , Echinococcus granulosus , Genotipo , Animales , Echinococcus granulosus/genética , Irán/epidemiología , Equinococosis/veterinaria , Equinococosis/epidemiología , Equinococosis/parasitología , Chacales/parasitología , Zorros/parasitología , Lobos/parasitología , Reacción en Cadena de la Polimerasa/veterinaria , Prevalencia , Polimorfismo de Longitud del Fragmento de RestricciónRESUMEN
Bacterial Artificial chromosome (BAC) recombineering is a powerful genetic manipulation tool for the efficient development of recombinant genetic resources. Long homology arms of more than 150 kb composed of BAC constructs not only substantially enhance genetic recombination events, but also provide a variety of single nucleotide polymorphisms (SNPs) that are useful markers for accurately docking BAC constructs at target sites. Even if the BAC construct is homologous to the sequences of the target region, different variations may be distributed between various SNPs within the region and those within the BAC construct. Once the BAC construct carrying these variations was precisely replaced in the target region, the SNP profiles within the target genomic locus were directly replaced with those in the BAC. This alteration in SNP profiles ensured that the BAC construct accurately targeted the designated site. In this study, we introduced restriction fragment length polymorphism or single-strand conformation polymorphism analyses to validate and evaluate BAC recombination based on changes in SNP patterns. These methods provide a simple and economical solution to validation steps that can be cumbersome with large homologous sequences, facilitating access to the production of therapeutic resources or disease models based on BAC-mediated homologous recombination.
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Cromosomas Artificiales Bacterianos , Recombinación Homóloga , Polimorfismo de Nucleótido Simple , Cromosomas Artificiales Bacterianos/genética , Polimorfismo de Longitud del Fragmento de Restricción , HumanosRESUMEN
BACKGROUND: The insulin-like growth factor (IGF-I) and growth hormone (GH) genes have been identified as major regulators of milk yield and composition, and reproductive performance in cattle. Genetic variations/polymorphism in these genes have been found to influence milk production, yield and quality. This investigation aimed to explore the association between IGF-I and GH polymorphisms and milk yield and composition, and reproductive performance in a herd consisting of 1000 Holstein-Friesian (HF) dairy cattle from El-Alamia farm. The experimental animals were 76 ± 7.25 months in age, with an average live weight of 750 ± 50.49 kg, and raised under the same conditions of feeding and weather. The studied animals were divided into three categories; high producers (n = 280), medium producers (n = 318) and low producers (n = 402). RESULTS: The digestion of 249 bp for IGF-I-SnaBI using the Restriction-fragment-length-polymorphism (RFLP) technique yielded two alleles; T (0.59) and C (0.41) and three genotypes; TT (0.52), TC (0.39) and CC (0.09) and this agrees with the results of DNA/gene sequencing technique. The sequencing analysis of the IGF-I gene revealed polymorphism in position 472 (C > T). Nucleotide sequencing of the amplified fragment of the IGF-I gene of different genotypes was done and submitted to the NCBI GenBank with Accession no. MH156812.1 and MH156811.1. While the digestion of 432 bp for GH-AluI using the RFLP technique yielded two alleles; A (0.81) and G (0.19) and two genotypes; AA (0.77) and AG (0.23) and this agrees with the results of DNA/gene sequencing technique. The sequencing analysis of the GH gene revealed polymorphism in the position 1758 C > G and in turn led to changes in amino acid sequence as Alanine for (A) compared to Glycine for (G). Nucleotide sequencing of the amplified fragment of the GH gene was done and submitted to the NCBI GenBank with Accession no. MH156810.1. The results of this study demonstrate the effects of variants of the GH-IGF-I somatotrophic axis on milk production and composition traits in commercial HF cattle. The greatest values of milk yield and reproductive performance were observed on IGF-I-SnaBI-TC and GH-AluI-AG genotypes. While the greatest % fat and % protein values were observed on IGF-I-SnaBI-CC and GH-AluI-AA genotyped individuals. CONCLUSION: The genetic variation of the studied genes can be utilized in selecting animals with superior milk yield, composition and reproductive performance in Holstein-Friesian Dairy Cattle under subtropical conditions.
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Hormona del Crecimiento , Factor I del Crecimiento Similar a la Insulina , Lactancia , Leche , Reproducción , Animales , Bovinos/genética , Bovinos/fisiología , Factor I del Crecimiento Similar a la Insulina/genética , Factor I del Crecimiento Similar a la Insulina/metabolismo , Leche/química , Leche/metabolismo , Hormona del Crecimiento/genética , Femenino , Reproducción/genética , Lactancia/genética , Polimorfismo Genético , Genotipo , Polimorfismo de Longitud del Fragmento de RestricciónRESUMEN
BACKGROUND: MiR-34b/c takes an important role in various aspects of carcinogenesis. Notably, pri miR34b/c (rs4938723) T>C polymorphism has been identified as a significant biomarker in various kinds of cancer. The objective of this study was to explore whether pri-miR34b/c rs4938723) T>C was associated with breast cancer susceptibility. Moreover, the association of pri-miR34b/c (rs4938723) T>C and clinicopathologic data, including survival outcomes, were studied in Thai breast cancer patients. METHODS: DNA extracted from the blood of 100 Thai female breast cancer patients and 100 Thai healthy women were investigated for pri-miR34b/c (rs4938723) T>C polymorphism using polymerase chain reaction-restriction fragment-length polymorphism (PCR-RFLP). RESULTS: There was no statistically significant difference between the frequency of pri miR34b/c (rs4938723) T>C genotype between Thai breast cancer patients and normal subjects. This study showed that there is no association between pri-miR34b/c (rs4938723) genotypes and breast cancer susceptibility, clinicopathologic parameters, and survival time. However, age greater than 50 and histologic grade III were the prognostic factors affecting survival in breast cancer patients (p=0.017, p=0.010, respectively). CONCLUSION: The pri-miR34b/c (rs4938723) genotypes had no association with cancer susceptibility and clinicopathologic parameters in Thai breast cancer patients. Patients with older age and patients with higher histologic grade, but not the pri miR34b/c (rs4938723) genotype, affected survival time among breast cancer patients.
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Biomarcadores de Tumor , Neoplasias de la Mama , Predisposición Genética a la Enfermedad , Genotipo , MicroARNs , Humanos , Femenino , MicroARNs/genética , Neoplasias de la Mama/genética , Neoplasias de la Mama/patología , Neoplasias de la Mama/mortalidad , Persona de Mediana Edad , Pronóstico , Estudios de Casos y Controles , Biomarcadores de Tumor/genética , Tasa de Supervivencia , Polimorfismo de Nucleótido Simple , Adulto , Tailandia/epidemiología , Estudios de Seguimiento , Anciano , Polimorfismo de Longitud del Fragmento de RestricciónRESUMEN
Toxoplasma gondii (T. gondii) is an intracellular protozoan that can cause toxoplasmosis in all warm-blooded hosts. This study focused on the prevalence and genetic characterize of T. gondii in ducks from Fujian province, China. Genomic DNA was extracted from duck tissue samples (heart, liver, lung, and muscle). To assess the genetic diversity of the T. gondii isolates, it was determined by using multilocus polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technology. A total of 586 ducks from 5 cities in Fujian province were tested, and 35 (6.0%) of which were found to be positive for the T. gondii B1 gene. Further genotyping of these positive samples at 10 genetic markers (SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, and Apico) using PCR-RFLP revealed that one tissue samples (heart samples from Fuzhou ducks) were identified as Type I (ToxoDB#10). This study is the first report on the prevalence and genetic characterization of T. gondii in ducks in Fujian province, and Type I (ToxoDB#10) is found in ducks in China for the first time. The findings document the genetic characterization of T. gondii in free-range ducks from Fujian Province, thereby enriching the understanding of T. gondii genetic diversity in China. Moreover, these results provide essential data support for further prospective studies and underscores the "One Health" concept, emphasizing the integral link among human, animal, and environmental health.
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Patos , Enfermedades de las Aves de Corral , Toxoplasma , Toxoplasmosis Animal , Animales , Toxoplasma/genética , Toxoplasma/aislamiento & purificación , Toxoplasmosis Animal/epidemiología , Toxoplasmosis Animal/parasitología , China/epidemiología , Enfermedades de las Aves de Corral/parasitología , Enfermedades de las Aves de Corral/epidemiología , Prevalencia , Polimorfismo de Longitud del Fragmento de Restricción , Salud Pública , Reacción en Cadena de la Polimerasa/veterinaria , Genotipo , Variación GenéticaRESUMEN
OBJECTIVE: Nonalcoholic fatty liver disease is a chronic liver disease and a growing global epidemic. The aim of this study was to investigate the association between a visfatin gene (NAMPT) variant and nonalcoholic fatty liver disease, owing to the connection between this disease and insulin resistance, obesity, inflammation, and oxidative stress, and the role of visfatin in these metabolic disorders. METHODS: In the present case-control study, we enrolled 312 genetically unrelated individuals, including 154 patients with biopsy-proven nonalcoholic fatty liver disease and 158 controls. The rs2058539 polymorphism of NAMPT gene was genotyped using the PCR-RFLP method. RESULTS: Genotype and allele distributions of NAMPT gene rs2058539 polymorphism conformed to the Hardy-Weinberg equilibrium both in the case and control groups (p>0.05). The distribution of NAMPT rs2058539 genotypes and alleles differed significantly between the cases with nonalcoholic fatty liver disease and controls. The "CC" genotype of the NAMPT rs2058539 compared with "AA" genotype was associated with a 2.5-fold increased risk of nonalcoholic fatty liver disease after adjustment for confounding factors [p=0.034; odds ratio (OR)=2.52, 95% confidence interval (CI)=1.36-4.37]. Moreover, the NAMPT rs2058539 "C" allele was significantly overrepresented in the nonalcoholic fatty liver disease patients than controls (p=0.022; OR=1.77, 95%CI=1.14-2.31). CONCLUSION: Our findings indicated for the first time that the NAMPT rs2058539 "CC" genotype is a marker of increased nonalcoholic fatty liver disease susceptibility; however, it needs to be supported by further investigations in other populations.
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Citocinas , Predisposición Genética a la Enfermedad , Genotipo , Nicotinamida Fosforribosiltransferasa , Enfermedad del Hígado Graso no Alcohólico , Polimorfismo de Nucleótido Simple , Humanos , Nicotinamida Fosforribosiltransferasa/genética , Enfermedad del Hígado Graso no Alcohólico/genética , Femenino , Masculino , Estudios de Casos y Controles , Persona de Mediana Edad , Factores de Riesgo , Adulto , Predisposición Genética a la Enfermedad/genética , Citocinas/genética , Frecuencia de los Genes/genética , Alelos , Polimorfismo de Longitud del Fragmento de Restricción , Reacción en Cadena de la PolimerasaRESUMEN
OBJECTIVES: Nonalcoholic fatty liver disease is the term used for a range of conditions in which fat builds up in the liver and exceeds 5% of hepatocytes without inordinate alcohol intake or other causes of lipid accumulation. Regarding the fact that insulin resistance and obesity play key roles in the pathogenesis of nonalcoholic fatty liver disease, as well as the connection between resistin and these metabolic diseases, the association between nonalcoholic fatty liver disease and a resistin gene (RETN) polymorphism was examined. METHODS: In this genetic case-control association study, 150 biopsy-proven nonalcoholic fatty liver disease patients and 154 controls were enrolled and genotyped for the RETN rs1862513 (-420C>G) gene polymorphism using PCR-RFLP method. RESULTS: The -420C>G genotype frequency distributions in both groups were consistent with Hardy-Weinberg equilibrium (HWE; p>0.05). The carriers of the RETN -420C>G "CC" genotype compared with the "GG" genotype occurred less frequently in the cases with nonalcoholic fatty liver disease than in the controls, and the difference remained significant even after adjustment for confounding factors (p=0.030; OR=0.47, 95%CI=0.36-0.93). Interestingly, the RETN -420C>G "C" allele was also associated with a decreased risk for nonalcoholic fatty liver disease too (p=0.042; OR=0.72, 95%CI=0.53-0.95). CONCLUSION: We found for the first time an association between biopsy-proven nonalcoholic fatty liver disease and RETN -420C>G promoter polymorphism. The carriers of the RETN -420C>G "CC" genotype had a 53% decreased risk for nonalcoholic fatty liver disease. Our findings, however, need to be corroborated by further studies.
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Predisposición Genética a la Enfermedad , Genotipo , Enfermedad del Hígado Graso no Alcohólico , Regiones Promotoras Genéticas , Resistina , Humanos , Enfermedad del Hígado Graso no Alcohólico/genética , Resistina/genética , Femenino , Masculino , Predisposición Genética a la Enfermedad/genética , Estudios de Casos y Controles , Persona de Mediana Edad , Regiones Promotoras Genéticas/genética , Adulto , Polimorfismo de Nucleótido Simple/genética , Frecuencia de los Genes/genética , Reacción en Cadena de la Polimerasa , Factores de Riesgo , Polimorfismo de Longitud del Fragmento de RestricciónRESUMEN
OBJECTIVE: The aim of this study was to evaluate the vitamin D receptor (VDR) BsmI variant in morbidly obese patients compared with healthy normal controls. METHODS: The study included 103 patients with morbid obesity and 120 healthy individuals serving as normal controls. The DNA samples obtained from blood were genotyped using the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique. The gender, age, smoking status, triglycerides, total cholesterol, insulin, mean body mass index, and frequency of allele and genotype of the BsmI variant in the VDR gene in morbidly obese patients were evaluated. RESULTS: The body mass index of the patients was 47.14 ± 7.19. The VDR B/B, B/b, and b/b genotype frequencies were 27.2% versus 28.3%; 54.4% versus 50%; and 18.4% versus 21.7% in the morbidly obese patients and the control group, respectively. There was no statistically significant difference between patients and control subjects in the genotype and allele distribution of the VDR BsmI variant (p>0.05). Both patients and control genotype frequencies are consistent with Hardy-Weinberg equilibrium. CONCLUSION: The BsmI variant in the VDR gene may not seem to predispose to morbid obesity in our study population. Further studies with a larger number of subjects are needed to make a more precise evaluation of this relationship.
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Índice de Masa Corporal , Frecuencia de los Genes , Predisposición Genética a la Enfermedad , Genotipo , Obesidad Mórbida , Polimorfismo de Longitud del Fragmento de Restricción , Receptores de Calcitriol , Humanos , Receptores de Calcitriol/genética , Obesidad Mórbida/genética , Masculino , Femenino , Adulto , Estudios de Casos y Controles , Persona de Mediana Edad , Frecuencia de los Genes/genética , Polimorfismo de Longitud del Fragmento de Restricción/genética , Predisposición Genética a la Enfermedad/genética , Reacción en Cadena de la Polimerasa , Factores de Riesgo , AlelosRESUMEN
Sauroleishmania spp. comprises one of the four Leishmania subgenera, which has been historically considered a non-pathogenic protozoan of reptiles. However, some strains appear to be transiently infective to mammals, and recent findings have detected these parasites in dogs and humans in areas where leishmaniasis is endemic. Herein, the digestion pattern of PCR-RFLP of the 234 bp-hsp70 fragment was evaluated as a simpler and cheaper tool to distinguish the Sauroleishmania species from the other Leishmania subgenera. As a result, the digestion of the 234 bp-hsp70 fragments with HaeIII produced a banding pattern specific to the four Sauroleishmania strains assessed. This technique could contribute to the identification of Leishmania parasites isolated from sandflies, reptiles, or even mammals in fieldworks as an alternative to the use of laborious and expensive methodologies.
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Proteínas HSP70 de Choque Térmico , Leishmania , Reacción en Cadena de la Polimerasa , Polimorfismo de Longitud del Fragmento de Restricción , Animales , Proteínas HSP70 de Choque Térmico/genética , Reacción en Cadena de la Polimerasa/métodos , Leishmania/genética , Leishmania/clasificación , Leishmania/aislamiento & purificación , Perros , Humanos , ADN Protozoario/genética , Parasitología/métodos , Leishmaniasis/parasitología , Leishmaniasis/veterinaria , Reptiles/parasitologíaRESUMEN
Human ascariasis is a soil-transmitted helminthiasis and remains a neglected tropical disease. Ascaris suum has the potential to cause cross-infections between humans and pigs. In this study, we present a rare case of a patient with asymptomatic infection by Ascaris suum. A 66-year-old male underwent colonoscopy, and a white linear worm body was found in the hepatic curvature. The worm was collected by aspiration and submitted to the laboratory for parasite identification. The patient had no symptoms related to parasitic infection. The worm was highly suspected to be of the genus Ascaris. Because of the difficulty of morphological classification, genetic analysis was performed. From PCR-restriction fragment length polymorphism results and sequence analysis of the internal transcribed spacer-1 region, it was determined to be A. suum. The experience with rapid differentiation of A. suum by performing genetic analysis will be useful for future examinations of parasitic infections.
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Ascariasis , Ascaris suum , ADN de Helmintos , Reacción en Cadena de la Polimerasa , Polimorfismo de Longitud del Fragmento de Restricción , Análisis de Secuencia de ADN , Humanos , Ascariasis/parasitología , Ascariasis/diagnóstico , Masculino , Animales , Anciano , Ascaris suum/genética , Ascaris suum/aislamiento & purificación , Reacción en Cadena de la Polimerasa/métodos , ADN de Helmintos/genética , Infecciones Asintomáticas , ADN Espaciador Ribosómico/genéticaRESUMEN
INTRODUCTION: Studies in different populations have shown that single-nucleotide polymorphisms (SNPs) of tumor necrosis factor-alpha (TNFα) and TNF receptors 1 and 2 (TNFR1 and TNFR2) may be involved in the pathogenesis of lepromatous leprosy (LL). To further explore the results in a Mexican population, we compared the frequencies of the polymorphisms in - 308 G>A TNFA (rs1800629), - 383 A>C TNFRS1A (rs2234649), and + 196 T >G TNFSR1B (rs1061622) genes in LL patients (n = 133) and healthy subjects (n = 198). METHODOLOGY: The genotyping was performed with the polymerase chain reaction-based restriction fragment length polymorphism (PCR-RFLP) technique. Statistical analysis was performed using the χ2 test, within the 95% confidence interval. Odds ratios (OR) were calculated and Hardy-Weinberg equilibrium was verified for all control subjects and patients. RESULTS: We found an association between the TNFSR1 -383 A>C genotype and the risk of lepromatous leprosy when leprosy patients were compared to controls (OR = 1.71, CI: 1.08-2.69, p = 0.02). Furthermore, it was also associated with the risk of LL in a dominant model (AC + CC vs AA, OR: 1.65, 95% CI: 1.05-2.057, p = 0.02). Similar genotype and allele frequencies for the SNPs TNFA - 308 G>A and TNFSR2 + 196 T>G were observed between leprosy patients and healthy subjects. CONCLUSIONS: The TNFSR1 -383 A>C could be a potential marker for the identification of high-risk populations. However, additional studies, using larger samples of different ethnic populations, are required.
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Predisposición Genética a la Enfermedad , Lepra Lepromatosa , Polimorfismo de Nucleótido Simple , Receptores Tipo II del Factor de Necrosis Tumoral , Receptores Tipo I de Factores de Necrosis Tumoral , Factor de Necrosis Tumoral alfa , Humanos , México , Masculino , Femenino , Adulto , Persona de Mediana Edad , Lepra Lepromatosa/genética , Receptores Tipo I de Factores de Necrosis Tumoral/genética , Factor de Necrosis Tumoral alfa/genética , Receptores Tipo II del Factor de Necrosis Tumoral/genética , Adulto Joven , Anciano , Frecuencia de los Genes , Polimorfismo de Longitud del Fragmento de Restricción , Estudios de Casos y Controles , Genotipo , Adolescente , Reacción en Cadena de la PolimerasaRESUMEN
Patients with systemic sclerosis (SSc) have a disproportionately high prevalence of reduced bone mineral density (BMD). Polymorphisms of the vitamin D receptor (VDR) gene have been associated with osteoporosis in patients with autoimmune diseases. The aim of this study was to investigate the prevalence and possible effects of VDR polymorphism on BMD and bone metabolism in patients with SSc. In patients with SSc measurement of BMD was performed using dual-energy X-ray absorptiometry. VDR polymorphisms (FokI, BsmI) were genotyped using restriction fragment length polymorphism analysis. Markers of bone metabolism (calcium, osteocalcin, ß-crosslaps) were determined. Primary endpoint was the prevalence of VDR gene polymorphisms and the association with reduced BMD. Secondary endpoints included associations between bone metabolism and VDR gene polymorphism. 79 Caucasian patients with SSc were included. Overall, 83.5% had reduced BMD (51.9% osteopenia, 31.6% osteoporosis). The prevalence of VDR gene polymorphism (73% BsmI, 77% FokI) was comparable to studies in healthy and rheumatic populations. The homozygous presence of FokI polymorphism, but not BsmI, was significantly associated with reduced axial BMD. Fokl polymorphism was significantly associated with reduced CTX levels, although changes remained within the reference limits. VDR polymorphisms can frequently be found in patients with SSc in comparable prevalence to healthy and rheumatic populations. The homozygous presence of FokI polymorphism, but not BsmI, was significantly associated with reduced axial BMD. This could be a possible contributor for the high prevalence of reduced BMD in 83.5% of patients with SSc in this study.Trial registration. DRKS00032768, date: 05.10.2023, retrospectively registered.
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Densidad Ósea , Receptores de Calcitriol , Esclerodermia Sistémica , Humanos , Receptores de Calcitriol/genética , Esclerodermia Sistémica/genética , Femenino , Densidad Ósea/genética , Masculino , Persona de Mediana Edad , Anciano , Adulto , Prevalencia , Osteoporosis/genética , Absorciometría de Fotón , Polimorfismo Genético , Polimorfismo de Longitud del Fragmento de Restricción , Enfermedades Óseas Metabólicas/genética , Enfermedades Óseas Metabólicas/epidemiología , GenotipoRESUMEN
AIM: Bacteria that promote plant growth, such as diazotrophs, are valuable tools for achieving a more sustainable production of important non-legume crops like rice. Different strategies have been used to discover new bacteria capable of promoting plant growth. This work evaluated the contribution of soil diazotrophs to the endophytic communities established in the roots of rice seedlings cultivated on seven representative soils from Uruguay. METHODS AND RESULTS: The soils were classified into two groups according to the C and clay content. qPCR, terminal restriction fragment length polymorphism (T-RFLP), and 454-pyrosequencing of the nifH gene were used for analyzing diazotrophs in soil and plantlets' roots grown from seeds of the same genotype for 25 days under controlled conditions. A similar nifH abundance was found among the seven soils, roots, or leaves. The distribution of diazotrophs was more uneven in roots than in soils, with dominance indices significantly higher than in soils (nifH T-RFLP). Dominant soils' diazotrophs were mainly affiliated to Alphaproteobacteria and Planctomycetota. Conversely, Alpha, Beta, Gammaproteobacteria, and Bacillota were predominant in different roots, though undetectable in soils. Almost no nifH sequences were shared between soils and roots. CONCLUSIONS: Root endophytic diazotrophs comprised a broader taxonomic range of microorganisms than diazotrophs found in soils from which the plantlets were grown and showed strong colonization patterns.
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Endófitos , Oryza , Raíces de Plantas , Microbiología del Suelo , Suelo , Oryza/microbiología , Oryza/crecimiento & desarrollo , Raíces de Plantas/microbiología , Endófitos/genética , Endófitos/aislamiento & purificación , Endófitos/clasificación , Suelo/química , Polimorfismo de Longitud del Fragmento de Restricción , Bacterias/genética , Bacterias/aislamiento & purificación , Bacterias/clasificación , Fijación del Nitrógeno , Oxidorreductasas/genéticaRESUMEN
BACKGROUND: Snails of the Lymnaeidae family are the intermediate hosts of Fasciola species, the causative agents of fascioliasis. The purpose of this study was to determine the prevalence of Fasciola species in lymnaeid snails and to investigate the association of geoclimatic factors and Fasciola species distribution in northwestern provinces of Iran using geographical information system (GIS) data. METHODS: A total of 2000 lymnaeid snails were collected from 33 permanent and seasonal habitats in northwestern Iran during the period from June to November 2021. After identification by standard morphological keys, they were subjected to shedding and crushing methods. Different stages of Fasciola obtained from these snails were subjected to the ITS1 polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method for species identification. The associations of weather temperature, rainfall, humidity, evaporation, air pressure, wind speed, elevation, and land cover with the distribution of Fasciola species were investigated. Geographical and statistical analysis was performed using ArcMap and SPSS software, respectively, to determine factors related to Fasciola species distribution. RESULTS: Of the 2000 snails collected, 19 were infected with Fasciola hepatica (0.09%), six with F. gigantica (0.03%), and 13 with other trematodes. Among geoclimatic and environmental factors, mean humidity, maximum humidity, and wind speed were significantly higher in areas where F. hepatica was more common than F. gigantica. The altitude of F. hepatica-prevalent areas was generally lower than F. gigantica areas. No significant relationship was observed between other investigated geoclimatic factors and the distribution of infected snails. CONCLUSIONS: The present study showed the relationship of humidity and wind speed with the distribution of snails infected with F. hepatica or F. gigantica in the northwestern regions of Iran. In contrast to F. gigantica, F. hepatica was more prevalent in low-altitude areas. Further research is recommended to elucidate the relationship between geoclimatic factors and the presence of intermediate hosts of the two Fasciola species.