Genetic polymorphisms of toll-like receptors in leprosy patients from southern Brazil.

Leprosy is a chronic disease and also a global health issue, with a high number of new cases per year. Toll-like receptors can respond to mycobacterial molecules in the early stage of infection. As important components of the innate immune response, alterations in genes coding for these receptors may contribute to susceptibility/protection against diseases. In this context, we used a case-control study model (183 leprosy cases vs. 185 controls) to investigate whether leprosy patients and the control group, in southern Brazil, have different frequencies in TLR1 (TLR1 G>T; rs5743618), TLR2 (TLR2 T>C, rs1816702 and rs4696483), and TLR4 (TLR4 A>G, rs1927911) polymorphisms. Analysis of the TLR1 1805G>T polymorphism presented the G/G genotype more frequently in the control group. TLR2 T>C rs1816702 and TLR2 T>C rs4696483, the T/T and C/T genotype, respectively, were more frequent in the control group than in leprosy patients, suggesting protection from leprosy when the T allele is present (rs4696483). Haplotype analyses between TLR1 (rs5743618) and TLR2 (rs1816702 and rs4696483) polymorphisms suggest risk for the presence of the TCC haplotype and protection in the presence of the TCT haplotype. This study suggests that polymorphisms in TLR1 and TLR2 are factors that may contribute to development/resistance of leprosy.

Rapid determination of L-ascorbic acid content in vitamin C serums by ultra-high-performance liquid chromatography-tandem mass spectrometry.

OBJECTIVE: This study aimed to develop and validate a rapid, simple, accurate and precise analytical method for the quantification of L-AA in vitamin C serums. Moreover, the developed method was further applied to determine L-AA in eight different brands of vitamin C serums. A complementary study was also carried out to evaluate the stability of L-AA in the vitamin C serum samples after 15, 30, 45 and 60 days of storage at ambient temperature (15-35°C). METHODS: Ultra-high-performance liquid chromatography-tandem mass spectrometry was applied. RESULTS: Quantitative analyses were performed with a total chromatographic run time of 1.5 min by matrix-matched calibration, and the analytical curve was linear over the range of 1-1700 µg L-1 with a correlation coefficient of 0.9998. The limits of detection (LOD) and quantification (LOQ) were 0.3 and 1.0 µg L-1 , respectively. Intra- and inter-assay precisions, expressed in terms of relative standard deviation, ranged from 0.3% and 2.2%, respectively, and recoveries in concentration levels of 1 and 5 µg L-1 were 103.9% and 101.2%, respectively. The proposed analytical method was successfully applied to determine the L-AA content in eight commercial vitamin C serum samples. The stability of the target analyte in samples stored at ambient temperature (15-35°C) was evaluated throughout 60 days with a 15-day interval between analyses. At 0 days, L-AA content in samples ranged from 1.05 to 169.91 mg L-1 , which decreases over time. CONCLUSION: The proposed method could be powerful in routine analyses to ensure the quantification of L-AA in vitamin C serums since it proved to be a simple, reliable, fast, precise, accurate and sensitive analytical method.

OBJECTIF: Cette étude visait à développer et valider une méthode analytique rapide, simple, exacte et précise pour la quantification de l'acide L-ascorbique dans les sérums à la vitamine C. De plus, la méthode développée a été appliquée pour déterminer l'acide L-ascorbique dans huit différentes marques de sérums à la vitamine C. Une étude complémentaire a également été réalisée pour évaluer la stabilité de l'acide L-ascorbique dans les échantillons de sérum à la vitamine C après 15, 30, 45 et 60 jours de conservation à température ambiante (15 à 35 °C). MÉTHODES: La chromatographie en phase liquide à haute performance avec spectrométrie de masse en tandem a été employée. RÉSULTATS: Des analyses quantitatives ont été réalisées avec une durée totale d'exécution chromatographique de 1,5 minute par calibration matricielle appariée, et la courbe analytique était linéaire sur la plage de 1 à 1700 µg L-1 avec un coefficient de corrélation de 0,9998. La limite de détection (LD) et la limite de quantification (LQ) ont été déterminées à 0,3 et 1,0 µg L−1 , respectivement. Les précisions intra- et inter-essais, exprimées en termes d'écart-type relatif, étaient de 0,3 % et 2,2 %, respectivement, et les récupérations aux niveaux de concentration de 1 et 5 µg L-1 étaient de 103,9 % et 101,2 %, respectivement. La méthode analytique proposée a été employée avec succès pour déterminer la teneur en acide L-ascorbique de huit échantillons de sérum à la vitamine C commerciaux. La stabilité de l'analyte cible dans les échantillons conservés à température ambiante (15 à 35 °C) a été évaluée sur 60 jours avec un intervalle de 15 jours entre les analyses. À 0 jour, la teneur en acide L-ascorbique dans les échantillons était comprise entre 1,05 et 169,91 µg L-1 , ce qui diminue au fil du temps. CONCLUSION: La méthode proposée pourrait être puissante dans les analyses de routine pour assurer la quantification de l'acide L-ascorbique dans les sérums à la vitamine C puisqu'elle s'est avérée être une méthode analytique simple, fiable, rapide, précise, exacte et sensible.

Vitamin D Receptor Gene Polymorphisms Are Associated With Leprosy in Southern Brazil.

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.

Bartonella henselae bacteremia diagnosed post-mortem in a myelodysplastic syndrome patient.

This study involves a 49-year-old male, who for three years suffered with a myelodysplastic syndrome and who needed frequent blood transfusions. One day following a transfusion, he presented fever and abdominal pain. The fever became persistent and only improved temporarily with two cycles of intravenous ciprofloxacin. Nearly 120 days after beginning the second cycle of treatment, he had experienced a weight loss of 16 kg and recurring fever. Screening for fever of unknown origin was conducted, including Bartonella infection. No etiology could be found. The patient improved with an antimicrobial regimen composed of oral doxycycline and intravenous ciprofloxacin. After 15 days afebrile, the patient was discharged with a four-month oral prescription of doxycycline and ciprofloxacin. Eight months following the antibiotic treatment, the patient received an allogeneic bone marrow transplant. Five days following the transplant, the patient initiated a febrile neutropenia and died. From a blood sample collected and stored at the time of hospitalization, a microbiological and molecular study was performed again. Blood- and liquid culture-PCRs from the same blood sample were all negative, but an isolate from solid subculture was found. The molecular reactions from this isolate were all positive and the sequence was 100% homologous to Bartonella henselae . The present report points to the limitations of laboratory techniques currently available for investigation of possible cases of bartonellosis in clinical practice, and the potential risk of Bartonella spp. transmission through blood transfusions.

Vancomycin-induced linear IgA bullous dermatosis (LABD)-an atypical presentation.

We report an uncommon presentation of bullous dermatosis by linear IgA. There are few cases reported in the literature with this form of presentation starting with mucosal lesions and then evolving into a similar bullous pemphigoid pattern. In addition, we emphasize the importance of direct immunofluorescence for the definitive diagnosis.

IL8 and IL17A polymorphisms associated with multibacillary leprosy and reaction type 1 in a mixed population from southern Brazil.

We evaluated the influence of the IL8 T-738A (nonidentified rs), IL8 T-353A (rs4073), IL17A G197A (rs2275913), and IL17F T7488C (rs763780) single-nucleotide polymorphisms on leprosy. The AA genotype of IL8 T-353A was observed as a risk factor for multibacillary leprosy, regardless of gender and age-of-onset of disease, considering the recessive model (OR, 3.8; 95% CI, 1.1-13.5; P, 0.023). Furthermore, the AA genotype of IL17A G197A was associated with leprosy type 1 reaction (OR, 2.4; 95% CI, 1.1-5.1; P, 0.026) when compared to the group without reaction, which was adjusted for gender and age-of-onset of disease by the model log additive. These results indicate association of IL8 and IL17A polymorphisms with the progression to multibacillary leprosy and with the type 1 reaction, respectively.