Expression in Escherichia coli, purification, refolding and antifungal activity of an osmotin from Solanum nigrum

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Evaluation of the genetic polymorphism of Plasmodium falciparum P126 protein (SERA or SERP) and its influence on naturally acquired specific antibody responses in malaria-infected individuals living in the Brazilian Amazon

<p>Abstract</p> <p>Background</p> <p>The <it>Plasmodium falciparum </it>P126 protein is an asexual blood-stage malaria vaccine candidate antigen. Antibodies against P126 are able to inhibit parasite growth <it>in vitro</it>, and a major parasite-inhibitory epitope has been recently mapped to its 47 kDa N-terminal extremity (octamer repeat domain – OR domain). The OR domain basically consists of six octamer units, but variation in the sequence and number of repeat units may appear in different alleles. The aim of the present study was to investigate the polymorphism of P126 N-terminal region OR domain in <it>P. falciparum </it>isolates from two Brazilian malaria endemic areas and its impact on anti-OR naturally acquired antibodies.</p> <p>Methods</p> <p>The study was carried out in two villages, Candeias do Jamari (Rondonia state) and Peixoto de Azevedo (Mato Grosso state), both located in the south-western part of the Amazon region. The repetitive region of the gene encoding the P126 antigen was PCR amplified and sequenced with the di-deoxy chain termination procedure. The antibody response was evaluated by ELISA with the Nt47 synthetic peptide corresponding to the P126 OR-II domain.</p> <p>Results</p> <p>Only two types of OR fragments were identified in the studied areas, one of 175 bp (OR-I) and other of 199 bp (OR-II). A predominance of the OR-II fragment was observed in Candeias do Jamari whereas in Peixoto de Azevedo both fragments OR-I and OR-II were frequent as well as mixed infection (both fragments simultaneously) reported here for the first time. Comparing the DNA sequencing of OR-I and OR-II fragments, there was a high conservation among predicted amino acid sequences of the P126 N-terminal extremity. Data of immune response demonstrated that the OR domain is highly immunogenic in natural conditions of exposure and that the polymorphism of the OR domain does not apparently influence the specific immune response.</p> <p>Conclusion</p> <p>These findings confirm a limited genetic polymorphism of the P126 OR domain in <it>P. falciparum </it>isolates and that this limited genetic polymorphism does not seem to influence the development of a specific humoral immune response to P126 and its immunogenicity in the studied population.</p

SAÚDE E TRABALHO DOS PROFESSORES DE EDUCAÇÃO FÍSICA QUE ATUAM COM ATIVIDADES AQUÁTICAS

Objetivo: O objetivo do presente estudo foi identificar a relação entre o processo de trabalho e a saúde dos professores de educação física que atuam com atividades aquáticas, bem como, verificar as diferenças entre os gêneros. Método: Foram pesquisados 184 professores de educação física, através de um levantamento realizado com questionários anônimos. Resultados: Foi observada desigualdade entre os gêneros no que se refere à distribuição da remuneração, embora não significativa. O estudo também permitiu observar uma excessiva queixa de dores (n=97; 52,72%). A percepção sobre o desgaste revelou um valor médio de 14,18 para escala de Borg e o sofrimento psíquico, medido pelo SRQ-20, apresentou valores de 2,58; 2,04 e 3,03 para todos, homens e mulheres, respectivamente, com diferença significativa entre os gêneros (

Identification of the Begomoviruses Squash Leaf Curl Virus and Watermelon Chlorotic Stunt Virus in Various Plant Samples in North America

Geminiviruses are a group of plant-infecting viruses with single-stranded DNA genomes. Within this family, viruses in the genus Begomovirus are known to have a worldwide distribution causing a range of severe diseases in a multitude of dicotyledonous plant species. Begomoviruses are transmitted by the whitefly Bemisia tabaci, and their ssDNA genomes can be either monopartite or bipartite. As part of a viral survey, various plants including those in the families Alliaceae, Amaranthaceae, Apiaceae, Asteraceae, Brassicaceae, Cactaceae, Cucurbitaceae, Lamiaceae, Lauraceae, Malvaceae, Oleaceae and Solanaceae were sampled and screened for begomoviruses using both a high-throughput sequencing and a begomovirus-specific primer pair approach. Based on the sequences derived using these approaches, the full-length genome of various begomoviruses were amplified from plants using abutting primers. Squash leaf curl virus (SLCV) and watermelon chlorotic stunt virus (WCSV) were identified in Cactaceae (n = 25), Solanaceae (n = 7), Cucurbitaceae (n = 2) and Lamiaceae (n = 1) samples. WCSV is an Old World bipartite begomovirus that has only recently been discovered infecting watermelons in the Americas. Our discovery of WCSV in the USA is the first indication that it has reached this country and indicates that this virus might be widespread throughout North America. Phylogenetic analysis suggests WCSV was introduced to the New World twice. The detection of begomoviruses in cactus plants suggests possible spillover events from agricultural areas into native vegetation. Since WCSV and SLCV have previously been found in mixed infections, pseudo-recombination infection experiments were conducted. We demonstrate that WCSV DNA-B is successfully trans-replicated by SLCV DNA-A despite very low degree of similarity between the replication-associated iterative sequences present in their common region, an essential feature for binding of the replication associated protein. This study highlights the importance of viral surveys for the detection of spillover events into native vegetation, but also suggests the need for more surveillance of WCSV in the USA, as this virus is a serious threat to watermelon cultivation in the Middle East

PHYTOPLANKTON BIOMASS INCREASES IN A SILT-IMPACTED AREA IN AN AMAZONIAN FLOOD-PLAIN LAKE OVER 15 YEARS

Funding Information: We thank Mineração Rio do Norte S.A. and Limnologia/UFRJ for fieldwork support, Dr. Janet W. Reid (JWR Associates) for language revision, and Leonardo Preza Rodrigues for map charting. VLMH, JCN, FAE, RLB, and FR are partially supported by the National Council for Scientific and Technological Development (CNPq), Brazil, RLB, and FAE by FAPERJ, Brazil, and CGR financially supported by Sakari Alhopuro Foundation, Finland. Publisher Copyright: © 2022, Universidade Federal do Rio de Janeiro. All rights reserved.Tailings from bauxite mining in Porto Trombetas (Pará state, Central Amazonia, Brazil) was discharged (1979–1989) into Batata Lake affecting about 30% of its area. The lake belongs to a clear-water flood-plain system along the Trombetas River, a tributary of the Amazon River. Siltation is the main perceived factor impacting aquatic and flooded communities. Besides natural regeneration, a program to restore a section of igapó forest in the impacted area (IA) has been conducted since 1991. Decreased light is the main factor reducing total phytoplankton biomass (PhyBM) in IA. We hypothesized that PhyBM in IA increases over time because of the improvement of the underwater light conditions due to the natural regeneration and restoration. We sampled quarterly PhyBM and limnological variables (depth, transparency, temperature, pH, conductivity, dissolved oxygen, turbidity, suspended solids, total Kjeldahl nitrogen, and total phosphorus), over 15 years (2005–2019) at eight sampling sites in the two areas (N = 349). We also obtained daily climatic and hydrologic data. PhyBM was higher in NIA than in IA. The temporal trend in the annual mean of PhyBM increased significantly over time only in the IA, approximating the NIA values, confirming our general hypothesis. The increase of PhyBM in the IA was negatively related to the residual light attenuation caused by non-phytoplankton turbidity and to total phosphorus, and positively to air temperature and site depth (p < 0.05; Marginal r2 = 0.18; Conditional r2 = 0.29). Instead, in NIA, PhyBM was explained only by the increase in air temperature (p < 0.05; Marginal r2 = 0.15; Conditional r2 = 0.34). We concluded that the PhyBM in the IA positively responds to the synergy between increasing light availability, air temperature, and site depth, and decreasing total phosphorus concentrations, regardless of hydrologic phase.Peer reviewe

An optimized nanoparticle delivery system based on chitosan and chondroitin sulfate molecules reduces the toxicity of amphotericin B and is effective in treating tegumentary leishmaniasis

Amphotericin B (AmpB) is active against leishmaniasis, but its use is hampered due to its high toxicity observed in patients. In this study, a nanoparticles-delivery system for AmpB (NQC-AmpB), containing chitosan and chondroitin sulfate molecules, was evaluated in BALB/c mice against Leishmania amazonensis. An in vivo biodistribution study, including biochemical and toxicological evaluations, was performed to evaluate the toxicity of AmpB. Nanoparticles were radiolabeled with technetium-99m and injected in mice. The products presented a similar biodistribution in the liver, spleen, and kidneys of the animals. Free AmpB induced alterations in the body weight of the mice, which, in the biochemical analysis, indicated hepatic and renal injury, as well as morphological damage to the kidneys of the animals. In general, no significant organic alteration was observed in the animals treated with NQC-AmpB. Mice were infected with L. amazonensis and treated with the nanoparticles or free AmpB; then, parasitological and immunological analyses were performed. The NQC-AmpB group, as compared to the control groups, presented significant reductions in the lesion size and in the parasite burden in all evaluated organs. These animals presented significantly higher levels of IFN-γ and IL-12, and low levels of IL-4 and IL-10, when compared to the control groups. The NQC-AmpB system was effective in reducing the infection in the animals, and proved to be effective in diminishing the toxicity evoked by AmpB, which was observed when it was administered alone. In conclusion, NQC-AmpB could be considered a viable possibility for future studies in the treatment of leishmaniasisThis work was supported by grants from Pró-Reitoria de Pesquisa from UFMG (Edital 01/2014), Instituto Nacional de Ciência e Tecnologia em Nano-biofarmacêutica (INCT-Nanobiofar), FAPEMIG (CBB-APQ-00496-11 and CBB-APQ-00819-12), and CNPq (APQ-472090/2011-9 and APQ-482976/2012-8). MACF is a grant recipient of FAPEMIG/CAPES. EAFC, VNC, and AAGF are grant recipients of CNPq. Eduardo AF Coelho and André AG Faraco are co-senior authors of this stud

Estratégias de coping utilizadas por familiares de indivíduos em tratamento hemodialítico

This descriptive, qualitative study aims to identify the coping strategies used most by the family members of patients with chronic kidney disease receiving hemodialysis. The data were collected through interviews, with the Coping Strategies Inventory questionnaire administered to ten family members, and were submitted to descriptive statistics and to thematic content analysis. The strategy used most was escape-avoidance, demonstrating that the family members prefer not to face the situation. The second most-used strategy was social support, ratified by the fact that they stated that they talk with other persons and professionals, seeking information about the problem, and because they accept the support received positively. The third was problem resolution, represented by the attitude of accepting responsibility for the care, either through strong affective bonds or through a feeling of responsibility established prior to the disease. The families use coping strategies of different intensities, while, nevertheless, indicating involvement and major participation in the care for the patient.Estudio cualitativo, que trató de identificar las estrategias de afrontamiento empleadas por las familias de los pacientes con insuficiencia renal crónica en hemodiálisis. Se aplicó el Inventario de Estrategias de Afrontamiento y una questión abierta con diez familiares. Los datos cuantitativos fueron sometidos a procedimientos estatísticos y análisis de contenido cualitativo. La estrategia de mayor utilización fue el escape y evitación (oito), lo que demuestra que los miembros de las familisa preferían no tener que enfrentar esta situación. La siguiente estrategia fue utilizada más apoyo social, ya hablé con otras personas y profesionales que buscan información sobre el problema, y aceptar el apoyo positivo recibido. A terceira foi resolução de problemas, representada pela atitude em assumir o cuidado, seja pelo forte vínculo afetivo ou pelo sentimento de responsabilidade já estabelecido antes da doença. Familias de las estrategias de afrontamiento empleadas con diferentes intensidades, pero mostrando una gran implicación y la participación en el cuidado del paciente crónico.Estudo descritivo, de natureza quali-quantitativa, que objetivou identificar as estratégias de enfrentamento mais utilizadas por familiares de pacientes com doença renal crônica em hemodiálise. Os dados foram coletados por meio de entrevista, com aplicação do Inventário de Estratégias de Coping a dez familiares, e submetidos à estatística descritiva e à análise de conteúdo temática. A estratégia mais utilizada foi fuga e esquiva, demonstrando que os familiares preferiam não enfrentar a situação. A segunda estratégia mais usada foi suporte social, ratificada ao afirmarem que conversaram com outras pessoas e profissionais buscando informações sobre o problema, e por aceitarem positivamente o apoio recebido. A terceira foi resolução de problemas, representada pela atitude em assumir o cuidado, seja pelo forte vínculo afetivo ou pelo sentimento de responsabilidade já estabelecido antes da doença. As famílias empregaram estratégias de enfrentamento com intensidades diferentes, porém indicando envolvimento e grande participação no cuidado ao doente.Universidade Federal de São Paulo (UNIFESP)aff2Universidade Estadual de MaringáUEMUNIFESPSciEL

PCR colorimetric dot-blot assay and clinical pretest probability for diagnosis of Pulmonary Tuberculosis in Smear-Negative patients

<p>Abstract</p> <p>Background</p> <p>Smear-negative pulmonary tuberculosis (SNPTB) accounts for 30% of Pulmonary Tuberculosis (PTB) cases reported annually in developing nations. Polymerase chain reaction (PCR) may provide an alternative for the rapid detection of <it>Mycobacterium tuberculosis </it>(MTB); however little data are available regarding the clinical utility of PCR in SNPTB, in a setting with a high burden of TB/HIV co-infection.</p> <p>Methods</p> <p>To evaluate the performance of the PCR dot-blot in parallel with pretest probability (Clinical Suspicion) in patients suspected of having SNPTB, a prospective study of 213 individuals with clinical and radiological suspicion of SNPTB was carried out from May 2003 to May 2004, in a TB/HIV reference hospital. Respiratory specialists estimated the pretest probability of active disease into high, intermediate, low categories. Expectorated sputum was examined by direct microscopy (Ziehl-Neelsen staining), culture (Lowenstein Jensen) and PCR dot-blot. Gold standard was based on culture positivity combined with the clinical definition of PTB.</p> <p>Results</p> <p>In smear-negative and HIV subjects, active PTB was diagnosed in 28.4% (43/151) and 42.2% (19/45), respectively. In the high, intermediate and low pretest probability categories active PTB was diagnosed in 67.4% (31/46), 24% (6/25), 7.5% (6/80), respectively. PCR had sensitivity of 65% (CI 95%: 50%–78%) and specificity of 83% (CI 95%: 75%–89%). There was no difference in the sensitivity of PCR in relation to HIV status. PCR sensitivity and specificity among non-previously TB treated and those treated in the past were, respectively: 69%, 43%, 85% and 80%. The high pretest probability, when used as a diagnostic test, had sensitivity of 72% (CI 95%:57%–84%) and specificity of 86% (CI 95%:78%–92%). Using the PCR dot-blot in parallel with high pretest probability as a diagnostic test, sensitivity, specificity, positive and negative predictive values were: 90%, 71%, 75%, and 88%, respectively. Among non-previously TB treated and HIV subjects, this approach had sensitivity, specificity, positive and negative predictive values of 91%, 79%, 81%, 90%, and 90%, 65%, 72%, 88%, respectively.</p> <p>Conclusion</p> <p>PCR dot-blot associated with a high clinical suspicion may provide an important contribution to the diagnosis of SNPTB mainly in patients that have not been previously treated attended at a TB/HIV reference hospital.</p

Linfadenites tuberculóides em suínos abatidos no Estado de São Paulo, Brasil: aspectos macroscópicos histopatológicos e pesquisa de micobactérias

INTRODUÇÃO: Como a ocorrência de linfadenites tuberculóides nos suínos oferece riscos à saúde pública, sobretudo em indivíduos imunocomprometidos, foi estudada a distribuição das lesões tuberculóides e a presença de micobactérias em linfonodos, tecido hepático e muscular de suínos de abate, no Estado de São Paulo, SP (Brasil), no período de 1993-1994. MATERIAL E MÉTODO: Foram estudadas 60 carcaças de suínos abatidos, sendo que, trinta apresentavam lesões tuberculóides macroscópicas (grupo A) e trinta estavam livres de tais lesões (grupo B ou controle). Foram analisadas seis localizações: linfonodos (retrofaríngeos, jejunais e mediastínicos), tecido hepático e tecido muscular (masséter e diafragma). Os exames executados foram: histopatologia e o cultivo para micobactérias. RESULTADOS: No grupo A, 14 em 30 carcaças apresentaram granuloma na histopatologia e, em 14, houve o isolamento de representantes do Complexo MAC (Mycobacterium avium-intracellulare). No grupo B não foram observadas lesões na histopatologia, e de quatro carcaças houve o isolamento de micobactérias de rápido crescimento. As lesões macroscópicas foram encontradas predominantemente nos linfonodos mesentéricos. O Complexo MAC foi isolado apenas em linfonodos do grupo com lesão, não sendo isolado em tecido hepático e muscular. CONCLUSÃO: Não foi possível o estabelecimento de relação entre o tipo de micobactéria isolada e as características macroscópicas das lesões.INTRODUCTION: As the occurrence of tuberculosis lymphadenitis in swine constitutes a public health risk, especially in immunosuppressed individuals, the distribution of tuberculoid lesions and the presence of micobacteria in lymphonodes and hepatic and muscular tissue in swine for slaughter, in the State of S. Paulo, SP (Brazil), in the period from 1993 to 1994, were studied. MATERIAL E MÉTODO: Tuberculous lesions in 60 carcasses of swine, slaughtered at abattoirs during the period 1993-1994, were studied. When macroscopic lesions were observed, a representative sampling of lymphnodes, hepatic and muscular tissues (masseter and diaphragm) were examined using histophatological and micobacterial isolation techniques. RESULTS: The macroscopic lesions were found predominantly in mesenteric lymphnodes. The MAI Complex (M.avium-intracellulare) was isolated mainly in the carcass group that showed lesions and was not found in the control group. Microorganisms of the MAI Complex were not isolated from hepatic and muscular tissues. CONCLUSION: No clear relationship between the type of mycobacteria isolated and the macroscopic lesions observed during the carcass inspection was found