Vitamin D, together with its nuclear receptor (VDR), plays an important role in modulating the immune response, decreasing the inflammatory process. Some polymorphisms of the VDR gene, such as BsmI (G>A rs1544410), ApaI (G>T rs7975232), and TaqI (T>C rs731236) could affect its stability and mRNA transcription activity, while FokI T>C (rs2228570) gives a truncated protein with three fewer amino acids and more efficiency in binding vitamin D. This study evaluated these four polymorphisms in the immunopathogenesis of leprosy in 404 patients and 432 control individuals without chronic or infectious disease in southern Brazil. When analyzing differences in the allele and genotype frequency of polymorphisms between patients (leprosy per se, multibacillary, and paucibacillary clinical forms) and controls, we found no statistically significant association. Regarding haplotype analysis, the bAt haplotype was associated with protection from leprosy per se (P = 0.004, OR = 0.34, CI = 0.16–0.71) and from the multibacillary clinical form (P = 0.005, OR = 0.30, CI = 0.13–0.70). In individuals aged 40 or more years, this haplotype has also showed protection against leprosy per se and multibacillary (OR = 0.26, CI = 0.09–0.76; OR = 0.26, CI = 0.07–0.78, respectively), while the BAt haplotype was a risk factor for leprosy per se in the same age group (OR = 1.34, CI = 1.04–1.73). In conclusion, despite having found no associations between the VDR gene polymorphisms with the development of leprosy, the haplotypes formed by the BsmI, ApaI, and TaqI polymorphisms were associated with leprosy per se and the multibacillary clinical form.
BackgroundKiller-cell immunoglobulin-like receptors (KIRs) are a group of regulatory molecules able to activate or inhibit natural killer cells upon interaction with human leukocyte antigen (HLA) class I molecules. Combinations of KIR and HLA may contribute to the occurrence of different immunological and clinical responses to infectious diseases. Leprosy is a chronic neglected disease, both disabling and disfiguring, caused mainly by Mycobacterium leprae. In this case–control study, we examined the influence of KIRs and HLA ligands on the development of multibacillary leprosy.Methodology/Principal findingsGenotyping of KIR and HLA genes was performed in 264 multibacillary leprosy patients and 518 healthy unrelated controls (238 healthy household contacts and 280 healthy subjects). These are unprecedented results in which KIR2DL2/KIR2DL2/C1/C2 and KIR2DL3/2DL3/C1/C1 indicated a risk for developing lepromatous and borderline leprosy, respectively. Concerning to 3DL2/A3/A11+, our study demonstrated that independent of control group (contacts or healthy subjects), this KIR receptor and its ligand act as a risk factor for the borderline clinical form.Conclusions/SignificanceOur finding suggests that synergetic associations of activating and inhibitory KIR genes may alter the balance between these receptors and thus interfere in the progression of multibacillary leprosy.
RESUMONo presente estudo o objetivo foi selecionar primers ISSR (Inter-Simple Sequence Repeats), adequados para futuros estudos de variabilidade genética em plantas da cultivar 'Itália' (Vitis vinifera L.). Além disso, padronizar o método de quantificação de ADN (Ácido Desoxirribonucleico) e a reação de PCR para os primers selecionados. Para extração do ADN foram utilizadas amostras de folhas jovens de videira coletadas na região de Marialva, PR, e o método de quantificação do ADN escolhido foi o espectrofotômetro Picodrop®. Os primers selecionados por apresentarem um número satisfatório de bandas nítidas (106 no total) em gel de agarose 2% quando submetidos à eletroforese, foram: ISSR-1, ISSR-2, ISSR-5, ISSR-6, ISSR-7, ISSR-8, ISSR-9, ISSR-11, ISSR-12, ISSR-13, ISSR-14 e ISSR-15, usando uma temperatura para ligação dos primers de 50 °C na PCR. O número médio de bandas por primer foi de 8,84, sendo o ISSR-5 o que gerou uma maior quantidade (13) de regiões ISSR amplificadas. SUMMARYThe objective of present study was to select ISSR (Inter-Simple Sequence Repeats) primers suitable for future genetic variability studies of the grape cultivar 'Itália' (Vitis vinifera L.) plants. Besides, it is aimed to standardize the method for quantifying DNA and PCR reaction for the selected primers. For DNA extraction, samples from young vine leaves collected in the region of Marialva-PR were used, and the chosen DNA quantification method was Picodrop® spectrophotometer. The primers selected for presenting a satisfactory number of sharp bands (106 in total) in 2% agarose gel when subjected to electrophoresis were: ISSR-1, ISSR-2, ISSR-5, ISSR-6, ISSR-7, ISSR-8, ISSR-9, ISSR-11, ISSR-12, ISSR-13, ISSR-14 and ISSR-15 using an annealing temperature of 50°C at the PCR. The average number of bands per primer was 8.84, whereas ISSR-5 primer produced the highest number (13) of amplified ISSR regions.Palavras-chave: eletroforese, biologia vegetal, ISSR, uva.
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