Correlation between clinical efficacy on pruritus and serum interleukin-31 levels in dogs with atopic dermatitis treated with lokivetmab.

Studies on serum interleukin (IL)-31 levels in dogs with atopic dermatitis (AD) and their correlation with disease severity are limited. To the author's knowledge, there are no studies that measured serum IL-31 in dogs treated with lokivetmab injections, a selective inhibitor of this key cytokine in pruritus. The aim of the study was to evaluate serum IL-31 levels in dogs treated with lokivetmab and correlate it with the severity of canine atopic dermatitis using the pruritus visual analog scale (pVAS) and canine atopic dermatitis extent and severity index (CADESI-04). Ten client-owned dogs diagnosed with AD received two injections of lokivetmab four weeks apart. Disease severity was assessed using the pVAS and CADESI-04 scores before and after both injections. In addition, canine serum IL-31 levels were measured at the same moments. Serum IL-31 was detected in all dogs in the study. There was a significant reduction in pVAS scores and serum IL-31 after administrations. However, there was no difference in CADESI-04 scores, and there was no significant correlation between CADESI-04 scores and serum IL-31 in dogs diagnosed with AD. Nonetheless, a significant positive correlation was observed between the pVAS scores and serum IL-31 levels with lokivetmab therapy, which reinforces the role of IL-31 in the pathogenesis of pruritus in dogs with AD. The data presented here provide further evidence that IL-31 is directly involved in pruritus pathogenesis in dogs with AD. In addition, blocking IL-31 has a significant antipruritic effect, but has no influence on skin lesion severity and extension.

A robust wavelet-based approach for dominant frequency analysis of atrial fibrillation in body surface signals.

OBJECTIVE: Atrial dominant frequency (DF) maps undergoing atrial fibrillation (AF) presented good spatial correlation with those obtained with the non-invasive body surface potential mapping (BSPM). In this study, a robust BSPM-DF calculation method based on wavelet analysis is proposed. APPROACH: Continuous wavelet transform along 40 scales in the pseudo-frequency range of 3-30 Hz is performed in each BSPM signal using a Gaussian mother wavelet. DFs are estimated from the intervals between the peaks, representing the activation times, in the maximum energy scale. The results are compared with the traditionally widely applied Welch periodogram and the robustness was tested on different protocols: increasing levels of white Gaussian noise, artificial DF harmonics presence and reduction in the number of leads. A total of 11 AF simulations and 12 AF patients are considered in the analysis. For each patient, intracardiac electrograms were acquired in 15 locations from both atria. The accuracy of both methods was assessed by calculating the absolute errors of the highest DF BSPM (HDF BSPM ) with respect to the atrial HDF, either simulated or intracardially measured, and assumed correct if ≤1 Hz. The spatial distribution of the errors between torso DFs and atrial HDFs were compared with atria driving mechanism locations. Torso HDF regions, defined as portions of the maps with [Formula: see text] Hz were identified and the percentage of the torso occuping these regions was compared between methods. The robustness of both methods to white Gaussian noise, ventricular influence and harmonics, and to lower spatial resolution BSPM lead layouts was analyzed: computer AF models (567 leads vs 256 leads down to 16 leads) and patient data (67 leads vs 32 and 16 leads). MAIN RESULTS: The proposed method allowed an improvement in non-invasive estimation of the atria HDF. For the models the median relative errors were 7.14% for the wavelet-based algorithm vs 60.00% for the Welch method; in patients, the errors were 10.03% vs 12.66%, respectively. The wavelet method outperformed the Welch approach in correct estimations of atrial HDFs in models (81.82% vs 45.45%, respectively) and patients (66.67% vs 41.67%). A low positive BSPM-DF map correlation was seen between the techniques (0.47 for models and 0.63 for patients), highlighting the overall differences in DF distributions. The wavelet-based algorithm was more robust to white Gaussian noise, residual ventricular activity and harmonics, and presented more consistent results in lead layouts with low spatial resolution. SIGNIFICANCE: Estimation of atrial HDFs using BSPM is improved by the proposed wavelet-based algorithm, helping to increase the non-invasive diagnostic ability in AF.

High glucose level as a modifier factor in CMT1A patients.

Charcot-Marie-Tooth disease type 1A (CMT1A) is the most common type of hereditary neuropathy worldwide and diabetes mellitus (DM) is the most frequent cause of peripheral neuropathy in the Western world. CMT1A typically manifest as a predominant motor neuropathy, while, DM-related neuropathy often manifests as a predominant sensory disorder. There are some evidences that CMT1A patients that also had DM had a more severe neuropathy. Although the real frequency and the underlying mechanisms related to this association has not yet been addressed in the literature. We sought to characterize the phenotypic variability of CMT1A patients with persistent high glucose levels (DM or impaired glucose tolerance [IGT]). Nineteen patients with CMT1A and DM (CMTdiab), seven with CMT1A and IGT (CMTintol) and 27 with CMT1A without comorbidities were analyzed. They were evaluated through clinical assessment, application of the following scales: visual analogue scale, McGill, CMTNS, SF-36 and COMPASS 31 and electrophysiological studies. Patients CMTdiab had a more severe motor and sensory neuropathy, more intense autonomic symptoms and worse quality of life. Surprisingly, proximal weakness and temporal dispersion on nerve conduction studies are frequently observed in this group. Patients CMTintol also had a more severe neuropathy. Curiously, we observed that the association of CMT1A and glucose metabolism disorders (CMTglic) clustered in some families. Patients CMTglic develop a more severe neuropathy. As there is yet no cure to CMT1A, a strict blood sugar control may be a useful measure.

Chronic aerobic exercise associated to low-dose L-NAME improves contractility without changing calcium handling in rat cardiomyocytes.

Nitric oxide (NO) inhibition by high-dose NG-nitro-L-arginine methyl ester (L-NAME) is associated with several detrimental effects on the cardiovascular system. However, low-dose L-NAME increases NO synthesis, which in turn induces physiological cardiovascular benefits, probably by activating a protective negative feedback mechanism. Aerobic exercise, likewise, improves several cardiovascular functions in healthy hearts, but its effects are not known when chronically associated with low-dose L-NAME. Thus, we tested whether the association between low-dose L-NAME administration and chronic aerobic exercise promotes beneficial effects to the cardiovascular system, evaluating the cardiac remodeling process. Male Wistar rats were randomly assigned to control (C), L-NAME (L), chronic aerobic exercise (Ex), and chronic aerobic exercise associated to L-NAME (ExL). Aerobic training was performed with progressive intensity for 12 weeks; L-NAME (1.5 mg·kg-1·day-1) was administered by orogastric gavage. Low-dose L-NAME alone did not change systolic blood pressure (SBP), but ExL significantly increased SBP at week 8 with normalization after 12 weeks. Furthermore, ExL promoted the elevation of left ventricle (LV) end-diastolic pressure without the presence of cardiac hypertrophy and fibrosis. Time to 50% shortening and relaxation were reduced in ExL, suggesting a cardiomyocyte contractile improvement. In addition, the time to 50% Ca2+ peak was increased without alterations in Ca2+ amplitude and time to 50% Ca2+ decay. In conclusion, the association of chronic aerobic exercise and low-dose L-NAME prevented cardiac pathological remodeling and induced cardiomyocyte contractile function improvement; however, it did not alter myocyte affinity and sensitivity to intracellular Ca2+ handling.

Influence of tropical reservoirs eutrophication on the polyunsaturated fatty acid profile in Astyanax altiparanae (Pisces: Characidae).

The eutrophication increases the quantity of algae that are deficient in highly unsaturated fatty acids (HUFA) n3, as EPA (eicosapentaenoic acid) and DHA (docosahexaenoic acid), altering the bottom-up transfer of the entire food chain. Due to the importance of the fatty acids (FA) in physiological processes related to the main role of the membrane phospholipids (PL) and precursors of eicosanoids, and also the little knowledge about the influence of eutrophication in tropical ecosystems, it is important to understand how anthropogenic changes in the aquatic ecosystem affect the lipid quality, specifically n3 HUFAs. This study aimed to investigate the influence of eutrophication on the FA profile of the hepatic PL, as well as prostaglandin (PG) levels in Astyanax altiparanae females. Fish were collected in reservoirs with different eutrophication degrees: Jundiaí (Ju) reservoir and Embu-Guaçu (EG) branch, considered as mesotrophic points, and Barragem (Ba) and Aracati (Ar), branches from Guarapiranga reservoir, considered as supereutrophic and hypereutrophic points, respectively. The FA profile of the liver PL was analyzed by gas chromatography, and the concentration of liver PGE2 was analyzed using ELISA immunoassay kits. The degree of eutrophication did not alter the PG concentration and produced few modifications in the FA profile of hepatic PL. Fish collected at Ba and EG presented similar FA profiles, both with high percentages of n3 HUFA, especially DHA, in comparison to fish from Ju. However, no change was observed in the n3 HUFA profile of the fish from Ar. These data demonstrated that the FA profile in A. altiparanae seems to be more related to different physiological requirements of n3 HUFA of the species than eutrophication. So, this study contributed to findings on the trophic transfer dynamics in tropical freshwater reservoirs, where a direct effect of eutrophication on the availability of HUFA n3 to animals is not suggested.

Chronic aerobic exercise associated to low-dose L-NAME improves contractility without changing calcium handling in rat cardiomyocytes

Nitric oxide (NO) inhibition by high-dose NG-nitro-L-arginine methyl ester (L-NAME) is associated with several detrimental effects on the cardiovascular system. However, low-dose L-NAME increases NO synthesis, which in turn induces physiological cardiovascular benefits, probably by activating a protective negative feedback mechanism. Aerobic exercise, likewise, improves several cardiovascular functions in healthy hearts, but its effects are not known when chronically associated with low-dose L-NAME. Thus, we tested whether the association between low-dose L-NAME administration and chronic aerobic exercise promotes beneficial effects to the cardiovascular system, evaluating the cardiac remodeling process. Male Wistar rats were randomly assigned to control (C), L-NAME (L), chronic aerobic exercise (Ex), and chronic aerobic exercise associated to L-NAME (ExL). Aerobic training was performed with progressive intensity for 12 weeks; L-NAME (1.5 mg·kg-1·day-1) was administered by orogastric gavage. Low-dose L-NAME alone did not change systolic blood pressure (SBP), but ExL significantly increased SBP at week 8 with normalization after 12 weeks. Furthermore, ExL promoted the elevation of left ventricle (LV) end-diastolic pressure without the presence of cardiac hypertrophy and fibrosis. Time to 50% shortening and relaxation were reduced in ExL, suggesting a cardiomyocyte contractile improvement. In addition, the time to 50% Ca2+ peak was increased without alterations in Ca2+ amplitude and time to 50% Ca2+ decay. In conclusion, the association of chronic aerobic exercise and low-dose L-NAME prevented cardiac pathological remodeling and induced cardiomyocyte contractile function improvement; however, it did not alter myocyte affinity and sensitivity to intracellular Ca2+ handling.

The ability of three nickel-titanium mechanized systems to negotiate and shape MB2 canals in extracted maxillary first molars: a micro-computed tomographic study.

AIM: To evaluate the influence of three engine-driven NiTi file systems manufactured from different NiTi alloys for the preparation of second mesiobuccal (MB2) canals in extracted maxillary first molars using a micro-computed tomographic (micro-CT) scanner. METHODOLOGY: Thirty maxillary molars with two canals in the mesiobuccal root were selected and randomly divided into three groups (n = 10): Reciproc [REC; size 25, .08 taper; VDW, Munich, Germany], ProDesign R [PDR; size 25, .06 taper; Easy, Belo Horizonte, Brazil] and Mtwo [MO; size 25, .06 taper; VDW, Munich, Germany]. Before and after root canal preparation of the MB2 canal, the teeth were scanned using a micro-CT to evaluate canal transportation, centring ability, dentine thickness and volume change. The working time to achieve working length was also evaluated. All parameters were compared statistically using the Kruskal-Wallis and Dunn test for multiple comparisons, with a significance level of 5%. RESULTS: There was no significant difference amongst the groups regarding canal transportation and centring ability (P > 0.05). However, the PDR size 25, .06 taper group had significantly lower canal volume and volume of dentine removal compared with a MO size 25, .06 taper and REC size 25, .08 taper (P < 0.05). A root perforation was detected in MO size 25, .06 taper and REC size 25, .08 taper groups, respectively. Regarding the working time, the PDR size 25, .06 taper required a significantly longer time to achieve working length than MO size 25, .06 taper and REC size 25, .08 taper (P < 0.05). CONCLUSIONS: All NiTi systems had similar canal transportation, centring ability and increase in apical volume after preparation of MB2 canals. However, the PDR size 25, .06 taper had less volume of dentine removal, absence of root canal perforation and required a longer time to accomplish the root canal preparation.

Application of the 2015 diagnostic criteria for neuromyelitis optica spectrum disorders in a cohort of Latin American patients.

BACKGROUND: The 2015 International Panel for neuromyelitis optica (NMO) spectrum disorders (NMOSD) diagnosis (IPND) criteria was recently proposed. However, because there are no studies evaluating application of the IPND criteria in Latin American populations, we aimed to assess whether these new criteria improve the diagnostic rate and reduce the time taken to make the diagnosis in a cohort of Latin American patients. METHODS: We reviewed medical records and applied both the 2006 and 2015 diagnostic criteria to all patients seen in four centers in Argentina, Brazil and Venezuela. Patients with multiple sclerosis (MS, n = 915) or other well-established central nervous system (CNS) inflammatory diseases were excluded. AQP4-ab status was measured using indirect immunofluorescence (23%) and cell-based assay (CBA, 77%). In addition, data on gender, ethnicity, age and symptoms at onset, relapses, neuroimaging and immunosuppressive therapy were collected. RESULTS: A total of 104 patients were classified as presenting NMOSD (2015 IPND). Of these, 64 patients (61.5%) fulfilled the 2006 NMO criteria (32 AQP4-ab positive, 17 AQP4-ab negative and 15 unknown). Thus, 40 new patients (38.5%) were classified as presenting NMOSD using the 2015 IPND criteria (33 AQP4-ab positive, 5 AQP4-ab negative and 2 unknown AQP4-ab status), with a median time taken to fulfill the 2015 NMOSD criteria (n = 104) of 1 month (95% CI: 0.6-1.3) and a median time taken to fulfill the 2006 NMO criteria (n = 64) of 18 months (95% CI: 9-26) (log-rank test: p < 0.0001). Females, with median age of 37 years, white ethnicity and recurrent course, predominated in all samples. Ninety-nine patients (95.1%) had at least 1 of the 3 major core clinical characteristics, of which optic neuritis (56.7%) was the most frequent symptom at disease onset. CONCLUSION: This study showed that there was a 62.5% increase in the rate of diagnosing NMOSD through the 2015 IPND criteria, in comparison with the 2006 NMO criteria, with a shorter median time to diagnosis.

Overweight Risk and Food Habits in Portuguese Pre-school Children.

The relationship between dietary intake and overweight-risk was assessed in 4349 children aged 3-5 years. Eating and sedentary behaviours were assessed by questionnaire. Logistic regressions were used. Children who consumed daily soft-drinks were 1.52 times more likely to be obese, and 72% more likely to be classified as overweight children.

Characterization of virulence and antibiotic profile and agr typing of Staphylococcus aureus from milk of subclinical mastitis bovine in State of Rio de Janeiro / Caracterização do perfil de virulência e resistência antimicrobiana e tipagem agr de Staphylococcus aureus oriundos do leite bovino de mastite subclinica no estado do Rio de Janeiro

This study aims to detect the main virulence and antimicrobial resistance genes in Stapylococcus aureus from bovine mastitic milk as well as classifying them according to agr typing. A total of 55 strains from six dairy unities in the state of Rio de Janeiro were selected, of these 27.3% presented fbnA and 78,2% for fbnB genes, respectively. None of the strains tested were positive for cap5 gene, 3.6% were positive for cap8 gene. Additionally, 94.5% of strains had hlA gene and 89.1% had hlB gene while 67.3% of the strains had icaA gene and 87.3% had icaD gene. From these results it was possible to establish 12 different virulence profiles. Prevalence of agrII type was detected in 81.8% of the isolates. Concerning antimicrobial resistance evaluation, the studied strains were susceptible to all antibiotics tested except penicillin, 83.6% being resistant strains. None of the strains had mecA gene, however, 40% of the strains had blaZ gene. Associating virulence and resistance data made it possible to obtain 23 different profiles. This great diversity of strains shows wide array of bacterial strategies and the challenge of mastitis prevention in cattle. Despite antimicrobial susceptibility, these strains presented certain genes that allow its persistence in the herd.(AU)

O presente estudo teve como objetivo detectar os principais genes de virulência e resistência antimicrobiana em Staphylococcus aureus oriundos de leite bovino mastítico e classificá-los de acordo com a tipagem do gene agr. Foram selecionados 55 isolados de seis unidades produtores no estado do Rio de Janeiro. Destas, o gene fbnA foi encontrado em 27,3% das cepas e 78,2% possuíam o gene fbnB. Em nenhuma cepa foi encontrado o gene cap5 e 3,6% possuíam o gene cap8. O gene hlA foi encontrado em 94,5% das cepas e 89,1% possuíam o gene hlB. O gene icaA foi encontrado em 67,3% das cepas e 87,3% possuíam o gene icaD. Com base nesses resultados, foi possível estabelecer 12 diferentes perfis de virulência. Prevalência do agr tipo II foi detectada em 81,8% dos isolados. Considerando-se a avaliação da resistência antimicrobiana, as cepas estudadas foram suscetíveis a todos os antibióticos exceto penicilina, sendo detectado um percentual de 83,6% de cepas resistentes. Nenhuma das cepas apresentou o gene mecA, contudo 40% das cepas apresentaram o gene blaZ. Vinte e três perfis diferentes foram estabelecidos por associação de dados de virulência e resistência. Essa grande diversidade de cepas mostra a ampla gama de estratégias bacterianas e o desafio da prevenção à mastite no gado bovino, considerando-se que, a despeito da suscetibilidade antimicrobiana, essas cepas apresentam genes que permitem sua persistência no rebanho.(AU)

Comparison of oral fluid collection methods for the molecular detection of hepatitis B virus.

BACKGROUND: This study aims to compare the efficiency of four oral fluid collection methods (Salivette, FTA Card, spitting and DNA-Sal) to detect HBV DNA by qualitative PCR. MATERIALS AND METHODS: Seventy-four individuals (32 HBV reactive and 42 with no HBV markers) donated serum and oral fluid. In-house qualitative PCR to detect HBV was used for both samples and commercial quantitative PCR for serum. RESULTS: HBV DNA was detected in all serum samples from HBV-infected individuals, and it was not detected in control group. HBV DNA from HBV group was detected in 17 samples collected with Salivette device, 16 samples collected by FTA Card device, 16 samples collected from spitting and 13 samples collected by DNA-Sal device. Samples that corresponded to a higher viral load in their paired serum sample could be detected using all oral fluid collection methods, but Salivette collection device yielded the largest numbers of positive samples and had a wide range of viral load that was detected. CONCLUSION: It was possible to detect HBV DNA using all devices tested, but higher number of positive samples was observed when samples were collected using Salivette device, which shows high concordance to viral load observed in the paired serum samples.

GROWING UP IN PORTUGAL: CAPE VERDEAN ANCESTRY CHILDREN EXHIBIT LOW OVERWEIGHT AND OBESITY COMPARED WITH PORTUGUESE IN URBAN LISBON.

Portugal has one of the highest rates of childhood overweight and obesity in Europe. However, little is known about the health of ethnic minorities living in its capital city, Lisbon. The Cape Verdean community in Lisbon tend to have low educational levels, material deprivation and struggle with discrimination and racism, factors that would probably be associated with a higher prevalence of overweight and obesity. Data for the Cape Verdean population were collected in three different time periods by three different research teams in 1993, 2009 and 2013 and included children aged 6-12 years living in the Cova da Moura neighbourhood of the Greater Lisbon Metro Area. The Portuguese national survey was conducted between 2009 and 2010 at public and private schools in mainland Portugal and included height, weight, skinfolds and arm and waist circumferences. From these survey data body mass index (BMI) and prevalence of stunting (chronic malnutrition - low height-for-age) and underweight (low weight-for-age) were calculated according to reference values proposed by Frisancho (2008). Overweight and obesity prevalence values were defined based on the references established by the International Obesity Task Force. The results show significant differences in height between Cape Verdean and Portuguese boys and girls. Generally, Cape Verdeans' growth falls within the healthy range of international growth references across all of the survey data collected. Cape Verdean rates for combined overnutrition (overweight and obesity) in 2013 (9.8% for boys and 16.7% for girls) were lower than those of the Portuguese (33% for boys and 31.7% for girls). Logistic regression models showed that Cape Verdean children had a lower risk of being overweight or obese when accounting for breast-feeding, birth weight, maternal education and occupation. Despite living in a deprived neighbourhood these Cape Verdean children seemed to have grown more healthily than Portuguese ancestry children. The challenge for policymakers will be to support improvement of the poverty-related living conditions of this community without creating a risky environment for increasing prevalence of overweight and obesity.

Maternal smoking in pregnancy association with childhood adiposity and blood pressure.

BACKGROUND: Maternal smoking during pregnancy has been associated with increased risk of childhood overweight/obesity defined by body mass index (BMI). We examined its association with a range of adiposity measures and cardiovascular indicators in children aged 3-10 years. METHODS: We used data from a cross-sectional study of schoolchildren across mainland Portuguese districts (2009-2010). We applied quantile regressions to examine maternal smoking associations with adiposity (n = 17 286), blood pressure (BP) and resting pulse rate (RPR) (n ≈ 2500) measures across the age range, adjusting for prenatal and early life factors. RESULTS: Maternal smoking during pregnancy was associated with increases in offspring adiposity levels. The difference in median BMI between children of smokers and non-smokers was 0.39 kg m(-2) (95% confidence interval: 0.25, 0.53) in boys and 0.46 kg m(-2) (0.31, 0.62) in girls; 0.55 cm (0.24, 0.87) and 0.82 cm (0.45, 1.19), respectively, in median waist circumference; and 0.94 mm (0.49, 1.40) and 1.47 mm (0.87, 2.07) in median sum of (triceps, subscapular, suprailiac) skin-folds. The associations appeared to be stronger with increasing age. The differences in the 90th centile tended to be greater than those in median. There was no consistent association of maternal smoking with BP and RPR. CONCLUSIONS: Children whose mother smoked during pregnancy had higher adiposity levels than children of non-smokers, across several measures, particularly among older children. Although there was no consistent association with cardiovascular indicators, maternal smoking association with childhood obesity may have implications for cardiovascular risk factors over the life course.

Pretransplant biopsy in expanded criteria donors: do we really need it?

INTRODUCTION: Renal transplantation is the best treatment for end-stage renal disease, including when using expanded criteria donors (ECD) kidneys. However, these suboptimal kidneys should be evaluated rigorously to meet their usefulness. Opinions differ about the best way to evaluate them. MATERIALS AND METHODS: We retrospectively reviewed kidneys from ECD harvested by a single academic institution between January 2008 and September 2013. Needle biopsies were performed at the time of the harvest when considered relevant by the transplant team. Two pathologists where responsible for their analysis; the Remuzzi classification has been used in all cases. RESULTS: We evaluated 560 ECD kidneys. Biopsies were made in 197 (35.2%) organs, 20 of which were considered not usable and 36 good only for double transplantation. Sixty-three kidneys (11.3%) were discarded by the transplant team based on the biopsy result and clinical criteria. Donors who underwent a biopsy were older (P < .001) and had a worse glomerular filtration rate (GFR; P = .001). Comparing donors approved and rejected by the biopsy, the rejected donors were heavier (P = .003) and had a lower GFR (P = .002). Cold ischemia time was longer for the biopsy group (P < .001). Regarding graft function, the biopsy overall score correlated with the transplant outcome in the short and long term. Separately, glomeruli and interstitium scores were correlated with recipient's GFR in the earlier periods (3 months; P = .025 and .037), and the arteries and tubules correlated with GFR in the longer term (at 3 years P = .004 and .010). CONCLUSION: The decision on the usability of ECD grafts is complex. At our center, we chose a mixed approach based on donor risk. Low-risk ECD do not require biopsy. In more complex situations, especially older donors or those with a lower GFR, prompted a pretransplant biopsy. The biopsy results proved to be useful as they relate to subsequent transplant outcomes, thereby allowing us to exclude grafts whose function would most probably be less than optimal.

Infective endocarditis in Rio de Janeiro, Brazil: a 5-year experience at two teaching hospitals.

INTRODUCTION: Despite the recent advances in diagnosis and treatment, mortality rates due to infective endocarditis (IE) remain high if not aggressively treated with antibiotics, whether or not associated with surgery. Data on the prevalence, epidemiology and etiology of IE from developing countries remain scarce. The aim of this observational, prospective cohort study was to report a 5-year experience of IE at two teaching hospitals in Rio de Janeiro, Brazil. MATERIAL AND METHODS: Demographical, anamnestic and microbiological characteristics of 71 IE patients were evaluated during the period of January 2009 to March 2013. RESULTS: The mean age of the IE patients was 49.8 ± 2.4 years, of which 41 (57.7%) were males. The median time between the onset of symptoms and diagnosis of IE was 35.8 ± 4.8 days. A total of 31 (43.6%) cases of community-acquired infective endocarditis (CAIE) and 40 (56.3%) cases of healthcare-acquired infective endocarditis (HAIE) were observed. Staphylococcus aureus (30%) was the predominant cause of IE. Streptococcus spp. (45.1 %) was the predominant cause of the CAIE while S. aureus (32.5%) and Enterococcus spp. (27.2 %) were the main etiological agents of HAIE. For 64 (90.1 %) patients with native valve endocarditis, the mitral valve was the most commonly affected (48.3%). The main source of IE in this cohort was intravascular catheter. The tricuspid valve and renal chronic insufficiency were more frequent in patients with HAIE than CAIE (p = 0.001). The risk factors associated with in-hospital mortality rate (46.4%) in IE patients were: age over 45 (OR 3.4; 95% CI 1.03-11.24; p = 0.04) and chronic renal insufficiency (OR 38.3; 95% CI 3.2-449.4; p = 0.004). CONCLUSIONS: At two main teaching hospitals in Brazil, Streptococcus spp. was the principal pathogen of CAIE while S. aureus and Enterococcus spp. were the most frequent causes of HAIE. IE remains a serious disease associated with high in-hospital mortality rate (46.6%); especially, in individuals over 45 years of age and with renal failure. Data suggest that early surgery may improve the outcome of IE patients.

Pulmonary cryptococcosis in childhood systemic lupus erythematosus and Sjögren syndrome overlap: a rare opportunistic infection.

Meningitis is the main manifestation of cryptococcosis in adult systemic lupus erythematosus (SLE) patients, and other organs and systems, such as the lungs, are rarely affected in this fungal infection. To our knowledge, no case of pulmonary cryptococcosis has been described in the pediatric lupus population. Therefore, we report herein one patient with childhood SLE (C-SLE) and Sjögren's syndrome overlap that presented encapsulated Cryptococcus yeast cells in lung tissue. A 14-year-old girl was diagnosed with C-SLE. At the age of 16 years and 5 months, she presented with fever, cough and dyspnea, without headache, vomiting, and also without signs of meningeal irritation or other clinical manifestations. She was being treated with mycophenolate mofetil, hydroxychloroquine and prednisone. Chest radiography and chest computer tomography showed a single nodule in the left posterior apex and three nodular lesions in the left hemithorax respectively. Bronchoalveolar lavage and transbronchial biopsy were normal and without isolation of bacteria or fungi. Voriconazole was empirically introduced for 21 days. Fifteen days after the first biopsy, she underwent open thoracotomy with surgical left lung biopsy and was diagnosed with pulmonary cryptococcosis. Voriconazole was replaced with oral fluconazole and this antifungal therapy was maintained with improvement of clinical manifestations and without marked alteration of radiological images. In conclusion, we report the first case of pulmonary cryptococcosis in Sjögren's and C-SLE patient with a satisfactory clinical response to antifungal therapy. Fungal infections should be excluded in the presence of lung nodules and etiological identification is required for proper treatment.

The first case report of non-nosocomial healthcare-associated infective endocarditis due to methicillin-resistant Staphylococcus aureus USA400 in Rio de Janeiro, Brazil.

Staphylococcus aureus is the main causal pathogen of infective endocarditis (IE), which may have distinct origins, namely, community, nosocomial, or non-nosocomial healthcare-associated (NNHCA). We report the first case of NNHCA-IE caused by methicillin-resistant S. aureus strain USA400/SCCmec IV in which the combination therapy of rifampin and vancomycin had a favorable outcome for the patient.

Simultaneous detection of hepatitis C virus antigen and antibodies in dried blood spots.

BACKGROUND: Enzyme immunoassays (EIA) designed to detect hepatitis C virus (HCV) core antigen and anti-HCV antibodies (HCV AgAb) simultaneously can improve the early detection of HCV infection when molecular diagnostic methods are not widely available. OBJECTIVES: To evaluate the suitability of dried blood spot (DBS) samples for detecting HCV AgAb using commercial EIAs. STUDY DESIGN: Paired serum and DBS samples were assayed using two commercial EIAs for HCV AgAb (Monolisa™ HCV AgAb ULTRA and Murex HCV AgAb). Manufacturer's recommendations were followed for sera while sample volume, incubation time and cut-off (CO) determination were evaluated for the DBS samples. The values of sensitivity, specificity, inter-rater agreement, detection limit, assay precision and stability of DBS samples at different conditions (22-26°C, 2-8°C and -20°C) were determined. RESULTS: It was necessary to increase the DBS sample volume fourfold compared to the sera samples to approximate the DBS Optical Density (OD) values to the sera OD values. Using ROC curve to recalculate CO values for the DBS samples, sensitivity was 97.5% for both EIAs, while the specificity was 99.71% for Monolisa™ HCV AgAb ULTRA and 95.95% for Murex HCV AgAb. Accurate testing results were obtained with DBS samples for 60 days at all conditions evaluated; storage at -20°C resulted in low OD variation. Both EIAs demonstrated the same limit of detection among DBS samples [estimated viral load of 3.1 International Units per millilitre (IU/mL)] and low OD value variability in repetitivity and reproducibility studies. CONCLUSION: DBS samples can be used for the detection of HCV AgAb by EIA as they present comparable performance characteristics and excellent stability among various storage conditions.

Dried blood spot samples: optimization of commercial EIAs for hepatitis C antibody detection and stability under different storage conditions.

This study was undertaken to optimize and compare the efficiency of two commercial EIAs for anti-HCV detection (HCV Ab Radim, Pomezzia, Italy and ETI-AB-HCVK-4 DiaSorin, Vercelli, Italy), in dried blood spot (DBS) samples. The long-term stability of anti-HCV on DBS samples stored at three environmental conditions was also evaluated at: 2-8 °C, 20-25 °C, and -20 °C. Paired DBS and serum samples were obtained from individuals with or without anti-HCV. The type of elution buffer, sample and conjugate volume, sample incubation time and cut-off values were evaluated. For both EIAs, a larger sample volume was used, and the cut-off value determined by the manufacturer was employed for Radim EIA; however, ROC curve analysis was used for the DiaSorin EIA. The sensitivity and specificity of Radim EIA on DBS were 97.5% and 99.5%, respectively, and of DiaSorin EIA were 88.9% and 98.9%, respectively. Accurate results were obtained for a period of 117 days using DBS samples stored at all storage conditions, but storage at -20 °C resulted in the lowest variation among the absorbance values. Both EIAs demonstrated the same limit of detection (until dilution of 1:10(4) with estimated viral load of 3.1 × 10(-1) UI/ml), but the Radim EIA was associated with the best performance because a low coefficient of variation was observed in the repetition and reproducibility studies. In conclusion, commercial EIAs can be optimized for anti-HCV detection in DBS samples that are extremely stable at different conditions for more than 100 days.