Thiamine Deficiency Modulates p38MAPK and Heme Oxygenase-1 in Mouse Brain: Association with Early Tissue and Behavioral Changes.

Thiamine deficiency (TD) produces severe neurodegenerative lesions. Studies have suggested that primary neurodegenerative events are associated with both oxidative stress and inflammation. Very little is known about the downstream effects on intracellular signaling pathways involved in neuronal death. The primary aim of this work was to evaluate the modulation of p38MAPK and the expression of heme oxygenase 1 (HO-1) in the central nervous system (CNS). Behavioral, metabolic, and morphological parameters were assessed. Mice were separated into six groups: control (Cont), TD with pyrithiamine (Ptd), TD with pyrithiamine and Trolox (Ptd + Tr), TD with pyrithiamine and dimethyl sulfoxide (Ptd + Dmso), Trolox (Tr) and DMSO (Dmso) control groups and treated for 9 days. Control groups received standard feed (AIN-93M), while TD groups received thiamine deficient feed (AIN-93DT). All the groups were subjected to behavioral tests, and CNS samples were collected for cell viability, histopathology and western blot analyses. The Ptd group showed a reduction in weight gain and feed intake, as well as a reduction in locomotor, grooming, and motor coordination activities. Also, Ptd group showed a robust increase in p38MAPK phosphorylation and mild HO-1 expression in the cerebral cortex and thalamus. The Ptd group showed a decreased cell viability, hemorrhage, spongiosis, and astrocytic swelling in the thalamus. Groups treated with Trolox and DMSO displayed diminished p38MAPK phosphorylation in both the structures, as well as attenuated thalamic lesions and behavioral activities. These data suggest that p38MAPK and HO-1 are involved in the TD-induced neurodegeneration in vivo, possibly modulated by oxidative stress and neuroinflammation.

Incomplete myelopathy and human T cell lymphotropic virus type-1 (HTLV-1).

This was a cross-sectional prospective study. We performed a multivariate statistical analysis of the neurological signs and symptoms of patients infected with human T cell lymphotropic virus type 1 (HTLV-1) in an attempt to separate them into distinct groups and identify clinical-neurological manifestations that could differentiate the various profiles. The study was performed in the city of Belém (state of Pará), located in the Amazon region of Brazil, from 2014 to 2016. We determined muscle strength and tone, reflexes, sensations, sphincter function, gait, and the Expanded Disability Status Scale score among individuals with HTLV-I. We then used exploratory statistical methods in an attempt to find different profiles and establish distinct groups. We analyzed 60 patients with HTLV-1. The filtering of the data, performed with mixed PCA, gave rise to a streamlined database with the most informative data and suggested the formation of three statistically distinct groups: asymptomatic carriers (AC), mono/oligosymptomatic (MOS), and HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSPd), AC and MOS (p = 0.002), AC and HAM/TSPd (p < 0.001), and HAM/TSPd and MOS (p = 0.001). The subsequent cluster analysis confirmed the formation of three clusters. The classification and regression tree demonstrated that altered gait was the most important variable for the classification of an individual with HAM/TSPd and that, in the absence of this impairment, hyperreflexia characterized MOS. The present study was able to separate patients infected by HTLV-1 into three clinical groups (AC, HAM/TSPd, and MOS) and identify clinical manifestations that could differentiate the various patient groups.

Multivariate analysis and multiple linear regression as a tool to estimate the behavior of hexazinone in Brazilian soils.

Weed control efficiency and the environmental contamination potential of herbicides depend on soil sorption and desorption. Among the indexes that evaluate the soil adsorption processes, the coefficients sorption (Kfs) and desorption (Kfd) obtained by Freundlich isotherms can provide accurate information about the behavior of an herbicide in the soil. The values of Kfs and Kfd of an herbicide vary according to the physicochemical characteristics of the soil, so it is possible to estimate these coefficients with high precision if good predictive mathematical models are constructed. Therefore, our objective aimed to evaluate the use of multiple regression models (MLR) associated with multivariate techniques to estimate the coefficient Kfs and Kfd for the hexazinone based on the chemical and physical attributes of soils. The correlation analyses, principal components, and clustering analysis allowed the multiple linear regression technique to generate models with higher adjustment coefficient (R2) for Kfs (0.73 to 0.99) and Kfd (0.94 to 0.99), and lower root mean squared error (RMSE) for Kfs (0.003 to 0.065) and Kfd (0.018 to 0.120). Regression models created from groups of soils showed greater prediction performance for Kfs and Kfd. The organic matter followed by the cation exchange capacity was the most important attributes of soils in sorption and desorption processes of hexazinone.

Late-onset congenital central hypoventilation syndrome and a rare PHOX2B gene mutation.

INTRODUCTION: Congenital central hypoventilation syndrome (CCHS) is a rare lifelong disorder characterized by an abnormal ventilatory response with persistent hypercapnia and hypoxia, which worsen during sleep. About 90 % of CCHS individuals are heterozygous for a mutation in the exon 3 of the PHOX2B gene. With higher awareness and better diagnostic tools, cases are identified in late childhood and adulthood, often with distinct mutations. CLINICAL CASE: The authors present a 4-year-old girl admitted to the intensive care unit at 9, 11 and 13 months suffering from severe hypercapnic respiratory failure during viral respiratory infections. Hypercapnia during sleep improved with wakefulness. CCHS was confirmed genetically (heterozygous insertion of an adenine at position 23, leading to a premature stop codon in exon 1 of the PHOX2B gene). The parents' DNA showed no PHOX2B mutations. Hypoventilation was observed by polysomnography, with no autonomic response to declining oxygen or increasing carbon dioxide values. A subsequent sleep study showed less hypoxia and hypercapnia. The patient has been on non-invasive ventilation during sleep, showing good growth and neurocognitive development. DISCUSSION: A greater awareness is required to diagnose late-onset CCHS. A respiratory infection can trigger the disease, with a significant difference in CO2 between sleep and wakefulness as the warning signal. Given the clinical suspicion, a genetic study should be performed. Polysomnography is essential for patient characterization. Follow-up and ventilator support adjustment prevent serious hypoxia and hypercapnia, which impair cardiovascular and neurocognitive functions. This patient's mutation has not been previously described; hence, clinical evolution cannot be predicted.

Herbicide Leaching in Soil with Different Properties: Perspectives from Commercial Formulations and Analytical Standards.

The leaching of herbicides into the soil is essential to control germinating seeds and parts of vegetative weeds. However, herbicide transportation to deeper soil layers can result in groundwater contamination and, consequently, environmental issues. In this research, our objective was to investigate differences in herbicide leaching between commercial formulations and analytical standards using three different soils. Leaching experiments were carried out for diuron, hexazinone, and sulfometuron-methyl herbicides isolated and in binary and ternary mixtures. The herbicide residue quantification was performed by ultra-high-performance liquid chromatography coupled to a mass spectrometer (LC-MS/MS). Diuron had less mobility in soils and was retained in the most superficial layers. Hexazinone and sulfometuron-methyl were more mobile and leached into deeper layers. The leaching process was more intense for hexazinone and sulfometuron-methyl. The additives present in the commercial formulation favored the leaching in soils of diuron, hexazinone, and sulfometuron-methyl herbicides isolated and mixture compared to the analytical standard. This fact highlights the importance of considering these effects for the positioning of herbicides in the field to increase the efficiency of weed control and minimize the potential for environmental contamination.

Streptococcus suis meningitis in a COVID-19 patient: A case report.

Streptococcus suis is a Gram-positive, facultative anaerobic bacterium and has its natural reservoir in pigs. Infection by this microorganism usually manifests in humans as meningitis, endocarditis, sepsis, and/or arthritis after contact with pigs or pork. Meningitis is a very common manifestation and Streptococcus suis may be considered its second most common cause. A clinical course may manifest acutely or with a more chronic course. Streptococcus suis is endemic in pork-consuming and pig-rearing countries, but may occur all over the world, especially in individuals with occupational exposure to pigs and/or pork, such as abattoir workers, butchers, and farmers. Most infections are observed in adults without preexisting diseases. We report a case of Streptococcus suis meningitis in a healthy patient with occupational exposure and with an asymptomatic infection by severe acute respiratory syndrome coronavirus 2, who was admitted to an intensive care unit in a tertiary hospital in the North of Portugal.

The Caloric Necessities of Critical Care Patients During the First Week of Admission.

INTRODUCTION:  The nutritional needs of critically ill patients have been the subject of intense controversy. In accordance with international guidelines, it is advocated to optimize a nutritional intake based on the following recommendation: 25-30 kcal/kg body weight per day. However, there still are authors who recommend permissive underfeeding in the first week of hospitalization. Nevertheless, energy expenditure (EE) and necessity are influenced by the catabolic phase of critical illness, which may vary over time on a patient and from patient to patient. OBJECTIVE: The objective of this study is to assess if the energy needs of critically ill patients admitted in our intensive care unit (ICU) in the first week of hospitalization are in line with those recommended by the European Society for Clinical Nutrition and Metabolism (ESPEN) international guidelines. METHODS: A prospective cross-sectional study was carried out from September to December 2019. The energy needs were evaluated by indirect calorimetry and by the Harris-Benedict equation. Stress variables were evaluated, namely, the type of pathology, hemodynamic support, sedation, temperature, sequential organ failure assessment (SOFA) score, and state at discharge. RESULTS: Forty-six patients were included in this study, with an average energy expenditure by indirect calorimetry of 19.22 ± 4.67 kcal/kg/day. The energy expenditure was less than 20 kcal/kg/day in 63% of the measurements. The concordance rate did not show the relationship between the Harris-Benedict equation and the values of indirect calorimetry. Stress variables were analyzed, with the SOFA score as the only variable with values close to statistical significance. CONCLUSION: In our ICU, the energy needs of critically ill patients in the first week of hospitalization are lower than the intake recommended by the guidelines.

Analyzing giant pituitary adenomas: An 8-year review (2012-2020) at a reference center in Brazil.

PURPOSE: This study aimed to review the cases of giant adenomas that were operated on from 2012 to 2020 and to analyze the epidemiological profile, complications, and resection rates using the transseptal/transnasal endoscopy technique. We also compared the results with data from revised literature. METHODS: This is an observational, retrospective study includes a review of the medical records of 26 patients diagnosed with giant adenomas (diameter > 4 cm or volume >10 cm3) who underwent 28 surgical procedures. RESULTS: Of the 57 patients operated for pituitary adenoma, 26 (50.8%) had giant adenomas. The mean volume was 17.28 cm3 (95% confidence interval [CI], 8.3359-26.2241). Moreover, 64.28% of the patients were graded Knosp 3 (p < 0.0001). Most cases were of nonsecretory adenomas (88.46%). Visual impairment was present in 89.2% of the patients and hormonal deficit and headache affected 53.5% of them. The visual impairment improved in 60% of the patients. The most common surgical complication was cerebrospinal fluid fistula (10.71%). Tumor resection > 90% was achieved in 53.56% of the cases, and the mean resection rate was 78.36% (95% CI, 71.316-87.956). CONCLUSIONS: The endoscopic endonasal technique is a good treatment option for giant adenomas, showing satisfactory optic apparatus decompression rates and a low incidence of complications when performed by surgeons with expertise in this approach.

Interaction between herbicides applied in mixtures alters the conception of its environmental impact.

Herbicide mixtures have often been used to control weeds in crops worldwide, but the behavior of these mixtures in the environment is still poorly understood. Laboratory and greenhouse tests have been conducted to study the interaction of the herbicides diuron, hexazinone, and sulfometuron-methyl which have been applied alone and in binary and ternary mixtures in the processes of sorption, desorption, half-life, and leaching in the soil. A new index of the risk of leaching of these herbicides has also been proposed. The sorption and desorption study has been carried out by the batch equilibrium method. The dissipation of the herbicides has been evaluated for 180 days to determine the half-life (t1/2). The leaching tests have been carried out on soil columns. The herbicides isolated and in mixtures have been quantified using ultra-high performance liquid chromatography coupled to the mass spectrometer. Diuron, hexazinone, and sulfometuron-methyl in binary and ternary mixtures have less sorption capacity and greater desorption when compared to these isolated herbicides. Dissipation of diuron alone is slower, with a half-life (t1/2) = 101 days compared to mixtures (t1/2 between 44 and 66 days). For hexazinone and sulfometuron-methyl, the dissipation rate is lower in mixtures (t1/2 over 26 and 16 days), with a more pronounced effect in mixtures with the presence of diuron (t1/2 = 47 and 56 and 17 and 22 days). The binary and ternary mixtures of diuron, hexazinone, and sulfometuron-methyl promoted more significant transport in depth (with the three herbicides quantified to depth P4, P7, and P7, respectively) compared to the application of these isolated herbicides (quantified to depth P2, P4, and P5). Considering the herbicides' desorption and solubility, the new index proposed to estimate the leaching potential allowed a more rigorous assessment concerning the risk of leaching these pesticides, with hexazinone and sulfometuron-methyl presenting a higher risk of contamination of groundwater.

[Pregnancy and Hemodialysis: About a Successful Twin Pregnancy]. / Gravidez e Hemodiálise: A Propósito de uma Gravidez Gemelar Bem Sucedida.

Pregnancy is a rare condition in women with chronic kidney disease on hemodialysis, but its incidence has increased in recent years. Despite being a high-risk condition for both mother and fetus, there has been an improvement in care due to the increased number of cases described and the experience gained in dialysis therapy. We report the case of a multiparous 32-years-old women with chronic hypertension and chronic kidney disease with 10 years of evolution and difficult control, with a diagnosis of dichorionic/diamniotic twin pregnancy, 4 months after beginning hemodialysis. Pregnancy was managed by a multidisciplinary team, hemodialysis sessions were clinically and hemodynamically well tolerated, with good blood pressure and analytical control, as well as adequate development of both fetuses. She went into labor at 34 weeks and 6 days, so a caesarean section was performed, with 2 boys born with adequate weight for gestational age.

A gravidez é uma situação rara em mulheres com doença renal crónica sob hemodiálise. Contudo, a sua incidência tem aumentado nos últimos anos. Apesar de ser uma situação de elevado risco materno e fetal, tem havido uma melhoria na assistência, devido ao aumento do número de casos descritos e da experiência adquirida na terapia dialítica. Relatamos o caso de uma multípara de 32 anos com hipertensão crónica e doença renal crónica com 10 anos de evolução e difícil controlo, com diagnóstico de gravidez gemelar bicoriónica/biamniótica, quatro meses após início da hemodiálise. A gestação foi vigiada por uma equipa multidisciplinar, as sessões de hemodiálise bem toleradas clínica e hemodinamicamente, com bom controlo tensional e analítico seriado, assim como adequado desenvolvimento de ambos os fetos. Foi submetida a cesariana às 34 semanas e seis dias por início de trabalho de parto, tendo nascido dois meninos com peso adequado para a idade gestacional.

Prevention of Human Papillomavirus Infection. Beyond Cervical Cancer: A Brief Review.

INTRODUCTION: Human papillomavirus is responsible for almost all cases of cervical cancer, an important portion of anogenital and oropharyngeal invasive and preinvasive lesions, as well as genital warts (condyloma acuminatum) and recurrent respiratory papillomatosis. Currently, three prophylactic vaccines against high-risk Human papillomavirus are commercialized in many countries worldwide. METHODS: To this non-systematic review the authors searched in MEDLINE/PubMed for systematic reviews, meta-analysis and randomized controlled trials, published in the last six years, using the terms "HPV", "non-cervical cancer" and "vaccine". Non-cervical cancers caused by human papillomavirus are less common lesions. However, its incidence has been increasing, while cervical cancer has declined, due mainly to highly effective screening programs. There are no formal screening programs for non-cervical cancers, so universal vaccination could have an important impact. The preventive effect of the vaccine is mainly studied and established in relation to cervical cancer, although it has also been demonstrated in the development of vulvar and vaginal lesions. To date, the efficacy in preventing anal and oropharyngeal diseases related with human papillomavirus is uncertain due to scarce supporting data and low vaccination coverage in men. The prevalence of injuries and subsequent absolute benefit of vaccination is lower in men, but it provides an additional benefit to the herd immunity achieved with the vaccination of women. CONCLUSION: The total fraction of malignant and pre-malignant lesions attributed to Human papillomavirus genotypes contained in the nonavalent vaccine is significant in both women and men, which turns this vaccine into a great asset in terms of Public Health.

Introdução: O vírus do papiloma humano é responsável por quase todos os casos de cancro do colo do útero, de uma importante fração de lesões anogenitais e orofaríngeas pré-invasivas e invasivas bem como de condilomas genitais e da papilomatose respiratória recorrente. Atualmente existem três vacinas profiláticas contra o vírus do papiloma humano de alto risco comercializadas em vários países do mundo. Métodos: Para esta revisão não-sistemática, os autores pesquisaram na MEDLINE/PubMed revisões sistemáticas, metanálises e ensaios clínicos randomizados, publicados nos últimos seis anos, utilizando os termos "HPV", "cancro não cervical" e "vacina". Os cancros não cervicais causados pelo vírus do papiloma humano são lesões menos comuns. Contudo, a sua incidência tem aumentado, a par de uma diminuição do cancro do colo do útero, devido principalmente à implementação de programas de rastreio altamente eficazes. Uma vez que não existem programas oficiais de rastreio para cancros não cervicais, a vacinação universal pode ter um impacto importante. O efeito preventivo da vacina é principalmente estudado e estabelecido em relação ao cancro do colo do útero, embora também tenha sido demonstrado no desenvolvimento de lesões vulvares e vaginais. Até ao momento, a eficácia na prevenção de doenças anais e orofaríngeas relacionadas com o vírus do papiloma humano é incerta, devido à escassez de dados na literatura e baixa cobertura de vacinação em homens. A prevalência de lesões e o consequente benefício absoluto da vacinação é inferior nos homens, porém proporciona um benefício adicional à imunidade de grupo alcançada com a vacinação de mulheres. Conclusão: A fração total de lesões malignas e pré-malignas atribuídas aos genótipos de vírus do papiloma humano contidos na vacina nonavalente é significativa tanto em mulheres quanto em homens, o que confere a essa vacina um grande potencial em termos de Saúde Pública.

Are cloth masks a substitute to medical masks in reducing transmission and contamination? A systematic review.

During the COVID-19 pandemic the use of cloth masks has increased dramatically due to the shortage of medical masks. However, the efficiency of this material is controversial. We aimed to investigate the efficiency of cloth masks in reducing transmission and contamination by droplets and aerosols for the general population and healthcare workers. Electronic databases were searched without year or language restrictions. Clinical and laboratorial studies were included. The risk of bias (RoB) was assessed using an adapted quality checklist for laboratory-based studies. ROBINS-I tool and Cochrane RoB 2.0 were used to evaluate non-randomized (n-RCT) and randomized clinical trials (RCT), respectively. The quality of the evidence was assessed through GRADE tool. From the eleven studies selected, eight were laboratory-based studies, one non-randomized and one RCT supported by laboratory data. Between the evaluated fabrics only three presented a filtration efficiency > 90%. Hybrid of cotton/chiffon (95%CI 95.2 to 98.8), hybrid of cotton/silk (95%CI 92.2 to 95.8) and cotton quilt (95%CI 94.2 to 97.8). However, cloth masks are not recommended for healthcare workers. A meta-analysis was not feasible due to a high methodological heterogeneity. The overall quality of evidence ranged from very low to moderate. Despite the lower efficiency compared to medical masks, laboratorial results may underestimate the efficiency of cloth masks in real life. Cloth mask efficiency is higher when made of hybrid fabrics (cotton/chiffon, cotton/silk) and cotton quilt, mainly with multiple layers.

Primary endemic Cryptococcosis gattii by molecular type VGII in the state of Pará, Brazil.

In order to study the infectious agents causing human disseminated cryptococcosis in the state of Pará, North Brazil, 56 isolates of Cryptococcusspp. (54 isolated from cerebral spinal fluid and two from blood cultures) from 43 cases diagnosed between 2003-2007 were analysed. The species were determined through morphological and physiological tests and genotypes were determined by URA5-RFLP and PCR-fingerprinting (wild-type phage M13). The following species and genotypes were identified: Cryptococcus neoformans VNI (28/56, 50%), Cryptococcus gattii VGII (25/56, 44.64%) and C. gattii VGI (3/56, 5.26%). The genotype VNI occurred in 12 out of 14 HIV-positive adults, whereas the genotype VGII occurred in 11 out of 21 HIV-negative adults (p < 0.02, OR = 6.6 IC95% 0.98-56.0). All patients less than 12 years old were HIV negative and six cases were caused by the VGII genotype, one by the VGI and one by VNI. Therefore, endemic primary mycosis in HIV-negative individuals, including an unexpectedly high number of children, caused by the VGII genotype deserves further study and suggests the need for surveillance on cryptococcal infection in the state of Pará, Eastern Amazon.

Amprolium exposure alters mice behavior and metabolism in vivo.

BACKGROUND: Thiamine deficiency (TD) models have been developed, mainly using the thiamine analog pyrithiamine. Other analogs have not been used in rodents. We aimed to evaluate the effects and mechanisms of intraperitoneal (ip) amprolium-induced TD in mice. We also evaluated the associated pathogenesis using antioxidant and anti-inflammatory compounds (Trolox, dimethyl sulfoxide). METHODS: Male mice were separated into two groups, one receiving a standard diet (control animals), and the other a TD diet (deficient groups) for 20 days. Control mice were further subdivided into three groups receiving daily ip injections of saline (NaCl 0.9%; Cont group), Tolox (Tr group) or dimethyl sulfoxide (DMSO; Dmso group). The three TD groups received amprolium (Amp group), amprolium and Trolox (Amp+Tr group), or amprolium and DMSO (Amp+Dmso group). The animals were subjected to behavioral tests and then euthanized. The brain and viscera were analyzed. RESULTS: Amprolium exposure induced weight loss with hyporexia, reduced the behavioral parameters (locomotion, exploratory activity, and motor coordination), and induced changes in the brain (lower cortical cell viability) and liver (steatosis). Trolox co-treatment partially improved these conditions, but to a lesser extent than DMSO. CONCLUSIONS: Amprolium-induced TD may be an interesting model, allowing the deficiency to develop more slowly and to a lesser extent. Amprolium exposure also seems to involve oxidative stress and inflammation, suggested as the main mechanisms of cell dysfunction in TD.

Failure of zanamivir therapy for pneumonia in a bone-marrow transplant recipient infected by a zanamivir-sensitive influenza A (H1N1) virus.

Influenza A viruses are responsible for significant morbidity and mortality after bone marrow transplantation. Here we report failure of inhaled zanamivir treatment in a bone-marrow transplant recipient with pneumonia caused by an influenza A (H1N1) virus, although the influenza viruses isolated from bronchoalveolar lavages before and after treatment were clearly found to be sensitive to zanamivir using cell-based and enzymatic assays. Subsequent oral treatment with oseltamivir allowed complete recovery. Poor bioavailability of zanamivir in the peripheral lungs might have been limiting treatment efficacy in such an immunocompromised patient.

Deep Infiltrating Endometriosis-Urinary Tract Involvement and Predictive Factors for Major Surgery.

OBJECTIVE: To evaluate urinary tract involvement by deep infiltrating endometriosis as well as the surgical treatment and existence of predictive factors for major urologic surgery. METHODS: We conducted a retrospective analysis of 656 women submitted to surgery for endometriosis, of which 28 patients underwent minor or major surgery for deep infiltrating endometriosis involving the urinary tract, with a mean age of 38 ± 6.9 years (27-50) at diagnosis. Clinical data, surgeries performed, and complications were analyzed. Minor surgery was defined by endoscopic surgery or insertion of a percutaneous nephrostomy catheter, and major surgery included open or laparoscopic procedures. RESULTS: Endometriomas affected the ureter in 13 (46.4%), the bladder in 11 (39.3%), and both structures in 4 (14.3%) patients. Twelve (42.9%) patients had decreased renal function, and ureteral involvement was predictive of renal function loss (P = .034). Minor surgeries were performed in most women with isolated bladder involvement and in 12 (42.9%) patients with ureteral infiltration. Patients with ureteric involvement underwent major surgeries more often (n = 12 vs n = 3; P = .025) and had longer hospitalization (8.2 vs 3.1 days, P = .05). After a mean follow-up of 36.3 (1-102) months, there was no bladder involvement recurrence. The most common complication was ureteral stenosis (Clavien-Dindo grade IIIb) in 3 (10.7%) patients. CONCLUSION: Surgery is highly successful in most cases. Patients with ureteric involvement are more likely to lose kidney function, undergo major surgery, and have longer hospitalization.

Diagnosis of human rabies cases by polymerase chain reaction of neck-skin samples.

Rapid diagnosis of rabies in suspected human cases influences post-exposure prophylaxis for potential contacts of the patient and ensures appropriate patient management. Apart from the central nervous system (CNS), rabies virus (RABV) is usually present in small sensory nerves adjacent to hair follicles of infected humans. We used an RT-PCR, with primers targeted to the 3' terminal portion of the nucleoprotein gene (N), to test neck-skin samples of nine patients who had rabies in order to validate a diagnostic method that could serve as an additional tool for rabies diagnosis, particularly in antemortem samples. Six of eight postmortem samples were found to be positive for rabies by RT-PCR, and one of two samples collected antemortem was positive with this same technique. Results were confirmed by DNA sequencing; this validates RT-PCR and neck-skin as a suitable technique and type of sample, respectively, for use in the diagnosis of human rabies. RT-PCR applied to neck-skin biopsies could allow early diagnosis and lead to more effective rabies treatment.

Persistence of Rabies Virus-Neutralizing Antibodies after Vaccination of Rural Population following Vampire Bat Rabies Outbreak in Brazil.

BACKGROUND: Animal control measures in Latin America have decreased the incidence of urban human rabies transmitted by dogs and cats; currently most cases of human rabies are transmitted by bats. In 2004-2005, rabies outbreaks in populations living in rural Brazil prompted widespread vaccination of exposed and at-risk populations. More than 3,500 inhabitants of Augusto Correa (Pará State) received either post-exposure (PEP) or pre-exposure (PrEP) prophylaxis. This study evaluated the persistence of rabies virus-neutralizing antibodies (RVNA) annually for 4 years post-vaccination. The aim was to evaluate the impact of rabies PrEP and PEP in a population at risk living in a rural setting to help improve management of vampire bat exposure and provide additional data on the need for booster vaccination against rabies. METHODOLOGY/PRINCIPAL FINDINGS: This prospective study was conducted in 2007 through 2009 in a population previously vaccinated in 2005; study participants were followed-up annually. An RVNA titer >0.5 International Units (IU)/mL was chosen as the threshold of seroconversion. Participants with titers ≤0.5 IU/mL or Equivalent Units (EU)/mL at enrollment or at subsequent annual visits received booster doses of purified Vero cell rabies vaccine (PVRV). Adherence of the participants from this Amazonian community to the study protocol was excellent, with 428 of the 509 (84%) who attended the first interview in 2007 returning for the final visit in 2009. The long-term RVNA persistence was good, with 85-88.0% of the non-boosted participants evaluated at each yearly follow-up visit remaining seroconverted. Similar RVNA persistence profiles were observed in participants originally given PEP or PrEP in 2005, and the GMT of the study population remained >1 IU/mL 4 years after vaccination. At the end of the study, 51 subjects (11.9% of the interviewed population) had received at least one dose of booster since their vaccination in 2005. CONCLUSIONS/SIGNIFICANCE: This study and the events preceding it underscore the need for the health authorities in rabies enzootic countries to decide on the best strategies and timing for the introduction of routine rabies PrEP vaccination in affected areas.

Zika virus epidemic in Brazil. I. Fatal disease in adults: Clinical and laboratorial aspects.

BACKGROUND: Zika virus (ZIKV) was first detected in Brazil in May 2015 and the country experienced an explosive epidemic. However, recent studies indicate that the introduction of ZIKV occurred in late 2013. Cases of microcephaly and deaths associated with ZIKV infection were identified in Brazil in November, 2015. OBJECTIVES: To determine the etiology of three fatal adult cases. STUDY DESIGN: Here we report three fatal adult cases of ZIKV disease. ZIKV infection in these patients was confirmed by cells culture and/or real-time reverse transcriptase polymerase chain reaction (RT-qPCR) and by antigen detection using immunohistochemical assay. Samples of brain and other selected organs taken at autopsy from three patients were also analyzed by histopathological and immunohistological examination. RESULTS: The first patient, a 36-year-old man with lupus and receiving prednisone therapy, developed a fulminant ZIKV infection. At autopsy, RT-qPCR of blood and tissues was positive for ZIKV RNA, and the virus was cultured from an organ homogenate. The second patient, a previously healthy female, 16 years of age, presented classic symptoms of Zika fever, but later developed severe thrombocytopenia, anemia and hemorrhagic manifestations and died. A blood sample taken on the seventh day of her illness was positive RT-PCR for ZIKV RNA and research in the serum was positive for antinuclear factor fine speckled (1/640), suggesting Evans syndrome (hemolytic anemia an autoimmune disorder with immune thrombocytopenic purpura) secondary to ZIKV infection. The third patient was a 20-year-old woman hospitalized with fever, pneumonia and hemorrhages, who died on 13days after admission. Histopathological changes were observed in all viscera examined. ZIKV antigens were detected by immunohistochemistry in viscera specimens of patients 1 and 3. These three cases demonstrate other potential complications of ZIKV infection, in addition to microcephaly and Guillain-Barre syndrome (GBS), and they suggest that individuals with immune suppression and/or autoimmune disorders may be at higher risk of developing severe disease, if infected with ZIKV.

Using the International Classification of Functioning, Disability and Health as a tool for analysis of the effect of physical therapy on spasticity in HAM/TSP patients.

INTRODUCTION: This study aimed to evaluate spasticity in human T-lymphotropic virus type 1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) patients before and after physical therapy using the International Classification of Functioning, Disability and Health (ICF). METHODS: Nine subjects underwent physical therapy. Spasticity was evaluated using the Modified Ashworth Scale. The obtained scores were converted into ICF body functions scores. RESULTS: The majority of subjects had a high degree of spasticity in the quadriceps muscles. According to the ICF codes, the spasticity decreased after 20 sessions of physical therapy. CONCLUSIONS: The ICF was effective in evaluating spasticity in HAM/TSP patients.