Toll-like receptor 4 (TLR4) is the major pattern recognition receptor triggering the protective effect of a Candida albicans extracellular vesicle-based vaccine prototype in murine systemic candidiasis.

Systemic candidiasis remains a significant public health concern worldwide, with high mortality rates despite available antifungal drugs. Drug-resistant strains add to the urgency for alternative therapies. In this context, vaccination has reemerged as a prominent immune-based strategy. Extracellular vesicles (EVs), nanosized lipid bilayer particles, carry a diverse array of native fungal antigens, including proteins, nucleic acids, lipids, and glycans. Previous studies from our laboratory demonstrated that Candida albicans EVs triggered the innate immune response, activating bone marrow-derived dendritic cells (BMDCs) and potentially acting as a bridge between innate and adaptive immunity. Vaccination with C. albicans EVs induced the production of specific antibodies, modulated cytokine production, and provided protection in immunosuppressed mice infected with lethal C. albicans inoculum. To elucidate the mechanisms underlying EV-induced immune activation, our study investigated pathogen-associated molecular patterns (PAMPs) and pattern recognition receptors (PRRs) involved in EVs-phagocyte engagement. EVs from wild-type and mutant C. albicans strains with truncated mannoproteins were compared for their ability to stimulate BMDCs. Our findings revealed that EV decoration with O- and N-linked mannans and the presence of ß-1,3-glucans and chitin oligomers may modulate the activation of specific PRRs, in particular Toll-like receptor 4 (TLR4) and dectin-1. The protective effect of vaccination with wild-type EVs was found to be dependent on TLR4. These results suggest that fungal EVs can be harnessed in vaccine formulations to selectively activate PRRs in phagocytes, offering potential avenues for combating or preventing candidiasis.IMPORTANCESystemic candidiasis is a serious global health concern with high mortality rates and growing drug resistance. Vaccination offers a promising solution. A unique approach involves using tiny lipid-coated particles called extracellular vesicles (EVs), which carry various fungal components. Previous studies found that Candida albicans EVs activate the immune response and may bridge the gap between innate and adaptive immunity. To understand this better, we investigated how these EVs activate immune cells. We demonstrated that specific components on EV surfaces, such as mannans and glucans, interact with receptors on immune cells, including Toll-like receptor 4 (TLR4) and dectin-1. Moreover, vaccinating with these EVs led to strong immune responses and full protection in mice infected with Candida. This work shows how harnessing fungal EVs might lead to effective vaccines against candidiasis.

Assessment of description and implementation fidelity of clinical trials involving exercise-based treatment in individuals with rotator cuff tears: a scoping review.

BACKGROUND: For evidence-based practice, clinicians and researchers can rely on well-conducted randomized clinical trials that exhibit good methodological quality, provide adequate intervention descriptions, and implementation fidelity. OBJECTIVE: To assess the description and implementation fidelity of exercise-based interventions in clinical trials for individuals with rotator cuff tears. METHODS: A systematic search was conducted in PubMed, Embase, CINAHL, LILACS, Cochrane Library, Web of Science, SCOPUS and SciELO. Randomized clinical trials that assessed individuals with rotator cuff tears confirmed by imaging exam were included. All individuals must have received an exercise-based treatment. The methodological quality was scored with the Physiotherapy Evidence Database (PEDro) scale. The Template for Intervention Description and Replication (TIDieR) checklist and the National Institutes of Health Behaviour Change Consortium (NIHBCC) were used to assess intervention description and implementation fidelity, respectively. RESULTS: A total of 13 studies were included. Despite their adequate methodological quality, the description of the intervention was poor with TIDieR scores ranging from 6 to 15 out of 24 total points. The TIDieR highest-scoring item was item 1 (brief name) that was reported in all studies. Considering fidelity, only one of the five domains of NIHBCC (i.e., treatment design) reached just over 50%. CONCLUSION: Exercise-based interventions used in studies for individuals with rotator cuff tears are poorly reported. The description and fidelity of the intervention need to be better reported to assist clinical decision-making and support evidence-based practice.

Agar dehydration: a simple method for long-term storage of Acanthamoeba spp. collection at room temperature.

This study describes dehydration of agar containing cysts as a novel and inexpensive method for long-term storage of Acanthamoeba spp. collections at room temperature. Five hundred microliters of axenically cultured Acanthamoeba spp. trophozoites (106 cells/mL) in PYG media or 150 µl of amoeba suspension (106 cells or cysts/mL) from monoxenic plate culture was spread onto the surface of non-nutritive agar (NNA, 2-3-mm thick) without or with a layer of heat-inactivated Escherichia coli, respectively. The plates were sealed and incubated at 30 °C. After the encystment, the Parafilm® was removed, and the plates were kept at the same temperature until the NNA was completely dehydrated. The dehydrated cyst-containing NNA was cut in rectangles and stored in airtight tubes at room temperature for up to 3 years. Cyst viability was assessed by inoculating them in fresh NNA with a layer of E. coli and in PYG followed by incubation at 30 °C. One hundred percent of samples from all specimens (19) stored over the 3 years allowed new cultures to be re-established; however, two strains showed reduced viability, at 66.7% and 62.5%, after 2 years of room temperature storage. One hundred percent of the cyst samples produced axenically and maintained in dry NNA allowed the re-establishment of axenic cultures through direct incubation in PYG, with excystment occurring within 24 or 48 h. For the first time, we report the dehydration of cyst-containing agar as an economical and effective method for the long-term storage of Acanthamoeba spp. collections at room temperature. It enables the creation of large collections using reduced space and economical transport of Acanthamoeba strains, in addition to allowing better organization of the collection.

COVID-19 is more lethal in hemodialysis chronic kidney disease patients with low physical capacity.

INTRODUCTION: The mortality rate in patients on maintenance hemodialysis (HD) is high and is associated with low functional capacity (FC). We analyzed the association of FC and COVID-19 lethality among HD patients. METHODS: A cohort study that included evaluations of FC in patients on HD was underway, and the association between FC and lethality was included as an ad-hoc outcome. RESULTS: One hundred and twelve patients were submitted to a physical, 31 patients contracted symptomatic COVID-19, 20 recovered and 11 died (lethality rate of 35.5%). There was a difference between survivors and non-survivors on six-minutes-walking-test (6MWT) 386.1 ± 112.8 versus 296.9 ± 103.3 meters (p = 0.04), 30 s sit-to-stand test (30CST) score 11.7 ± 3.1 versus 7.7 ± 4.1 (p = 0.006), and timed up and go test (TUG) 9.4 versus 13.6 s (p = 0.009). There was also an association between percentages of predicted 6MWT, 30CST, and TUG with COVID-19 lethality. CONCLUSION: Patients on maintenance HD with poorer physical performance presented the worst prognosis from COVID-19 pandemic.

Molecular and serological detection of Anaplasma spp. in small ruminants in an area of Cerrado Biome in northeastern Brazil.

Anaplasmosis, caused by bacteria of the genus Anaplasma, is an important tick-borne disease that causes economic losses to livestock farms in many countries. Even though Anaplasma spp. have been detected in goats and sheep worldwide, few studies investigate the occurrence and genetic identity of these agents in small ruminants from Brazil. Thus, this work aimed to detect and determine the genetic identity of Anaplasma spp. in small ruminants from the Baixo Parnaíba region, state of Maranhão, northeastern Brazil. For this purpose, blood samples were collected from 161 animals (91 goats; 70 sheep) from 4 municipalities in the Baixo Parnaíba region. Sheep and goat serum samples were subjected to recombinant membrane surface protein (MSP5)-based iELISA. Whole blood samples were subject to DNA extraction and molecular diagnosis using PCR assays for Anaplasma spp. targeting msp1ß, msp1α, 16S rRNA and msp4 genes. Positive samples were sequenced and then subjected to Anaplasma marginale msp1α genetic diversity analysis and phylogenetic inferences based on the 16S rRNA and msp4 genes. The serological survey detected the presence of anti-A. marginale IgG antibodies in 18 animals (11.1%): 2.9% (2/70) sheep and 17.4% (16/91) goats. Anaplasma marginale DNA was detected in 2 goats (1.2%) using qPCR based on the msp1ß gene. Two distinct A. marginale msp1α strains, namely α ß and α ß ΓγΓγΓγΓγ were found in the infected goats, each one found in a different animal, both belonging to the H genotype. Phylogenetic analysis based on the 16S rRNA gene showed the sequences positioned in three different clades and grouped with sequences from 'Candidatus Anaplasma boleense', A. platys and A. marginale. Phylogenetic inferences based on the msp4 gene positioned the sequence variants in the A. marginale clade. The present work represents the first molecular detection of sequence variants phylogenetic associated to 'Candidatus Anaplasma boleense' and A. platys and α ß and α ß ΓγΓγΓγΓγ in goats from Brazil.

Analysis of the perinatal mortality rate in the metropolitan region of grande Vitória, Espírito Santo, Brazil, between 2008 and 2017

Introduction: perinatal mortality is characterized by fetal deaths that occur after the 22nd week of management and neonatal deaths that precede six full days of life. This indicator has been a matter of concern and discussion on the part of entities and organizations involved in comprehensive health care for women and children.Objective: to characterize perinatal deaths in the Metropolitan Region of Greater Vitória (RMGV) in Espírito Santo and identify associated maternal factors in the period between 2008 and 2017.Methods: ecological and descriptive study with a quantitative approach, carried out in 2019 on perinatal mortality from 2008 to 2017 at RMGV. Data collection was performed by extracting data from the SIM, SINASC, IBGE databases of the Espírito Santo State Health Department, about perinatal deaths and associated maternal factors. The research respects the ethical precepts of resolution 466/12 of the National Health Council.Results: the distribution of deaths did not occur homogeneously in the municipalities in the RMGV. The municipality of Vitória had the lowest perinatal mortality rates during the study period, on the other hand, in the comparative analysis between the different municipalities that make up the RMGV, the municipality of Fundão presents the worst scenario regarding perinatal mortality over the years. Regarding the underlying causes of death, it is noted that in this study, the three causes with the highest number of occurrences are complications of the placenta, umbilical cord and maternal affections, not necessarily related to the current pregnancy and intrauterine hypoxia.Conclusion: there were no significant changes in mortality rates in the Metropolitan Region of Greater Vitória. However, the main deaths occurred in neighborhoods with greater socioeconomic inequalities. Maternal causes were highly representative of deaths, raising issues associated with the improvement of public health policies.

Introdução: a mortalidade perinatal caracteriza-se pelos óbitos fetais que ocorrem a partir da 22ª semana de gestão e os óbitos neonatais que antecedem seis dias completos de vida. Este indicador tem sido motivo de preocupação e discussão por parte de entidades e organizações envolvidas na atenção à saúde integral da mulher e da criança. Objetivo: caracterizar os óbitos perinatais da Região Metropolitana da Grande Vitória (RMGV) no Espírito Santo e identificar fatores maternos associados, no período entre 2008 e 2017. Método: estudo ecológico e descritivo de abordagem quantitativa, realizado no ano de 2019 acerca da mortalidade perinatal entre os anos de 2008 a 2017 na RMGV. A coleta de dados foi realizada através extração dos dados das bases SIM, SINASC, IBGE da Secretaria de Saúde do Estado do Espírito Santo, acerca dos óbitos perinatais e fatores maternos associados. A pesquisa respeita os preceitos éticos da resolução 466/12 do Conselho Nacional de Saúde. Resultados: a distribuição dos óbitos não ocorreu de forma homogênea nos municípios na RMGV. O município de Vitória apresentou os menores índices de mortalidade perinatal durante o período estudado, em contrapartida, na análise comparativa entre os diferentes municípios que compõe a RMGV, o município de Fundão apresenta o pior cenário relativo à mortalidade perinatal ao longo dos anos. Acerca das causas bases de óbitos, nota-se que neste estudo, as três causas com maior número de ocorrência são complicações da placenta, do cordão umbilical e afecções maternas, não obrigatoriamente relacionadas com a gravidez atual e hipóxia intrauterina. Conclusão: não houve mudanças significativas nas taxas de mortalidade na Região Metropolitana da Grande Vitória. Contudo, os principais óbitos ocorreram em bairros com maiores desigualdades socieconomicas. As causas maternas representaram uma grande representatividade frente aos óbitos, levantando questões associadas a melhora de políticas públicas de saúde.

Mesenchymal Stem Cells Combined with a P(VDF-TrFE)/BaTiO3 Scaffold and Photobiomodulation Therapy Enhance Bone Repair in Rat Calvarial Defects.

BACKGROUND: Tissue engineering and cell therapy have been the focus of investigations on how to treat challenging bone defects. This study aimed to produce and characterize a P(VDF-TrFE)/BaTiO3 scaffold and evaluate the effect of mesenchymal stem cells (MSCs) combined with this scaffold and photobiomodulation (PBM) on bone repair. METHODS AND RESULTS: P(VDF-TrFE)/BaTiO3 was synthesized using an electrospinning technique and presented physical and chemical properties suitable for bone tissue engineering. This scaffold was implanted in rat calvarial defects (unilateral, 5 mm in diameter) and, 2 weeks post-implantation, MSCs were locally injected into these defects (n = 12/group). Photobiomodulation was then applied immediately, and again 48 and 96 h post-injection. The µCT and histological analyses showed an increment in bone formation, which exhibited a positive correlation with the treatments combined with the scaffold, with MSCs and PBM inducing more bone repair, followed by the scaffold combined with PBM, the scaffold combined with MSCs, and finally the scaffold alone (ANOVA, p ≤ 0.05). CONCLUSIONS: The P(VDF-TrFE)/BaTiO3 scaffold acted synergistically with MSCs and PBM to induce bone repair in rat calvarial defects. These findings emphasize the need to combine a range of techniques to regenerate large bone defects and provide avenues for further investigations on innovative tissue engineering approaches.

Molecular detection and characterization of vector-borne agents in common opossums (Didelphis marsupialis) from northeastern Brazil.

Opossums are synanthropic marsupials able to interchange among wild, peri­urban and urban environments, playing an epidemiologically important role as hosts for emerging pathogens and ectoparasites of relevance in public health. The present study aimed to detect and molecularly characterize vector-borne agents in a population of common opossums (Didelphis marsupialis) from the Island of São Luís do Maranhão, northeastern Brazil. Of the 45 animals analyzed, one (2.22%) was positive in the nested PCR assay based on the 18S rRNA gene of piroplasmids. The obtained sequence was phylogenetically positioned in a clade containing sequences of Babesia sp. previously detected in Didelphis aurita, Didelphis albiventris and associated ticks from Brazil. Eight (17.77%) samples were positive in PCR for Ehrlichia spp. based on the dsb gene; four samples were sequenced and positioned into a new clade, sister to E. minasensis and Ehrlichia sp. clade detected in Superorder Xenarthra mammals. No samples tested positive in the screening PCR assays based on the 16S rRNA gene of Anaplasma spp. Two samples were positive in the qPCR for Bartonella spp. based on the nuoG gene. Seven animals (15.56%) were positive in the nPCR based on the 16S rRNA gene of hemoplasmas. Of these, three were positive in a PCR based on the 23S rRNA gene. The phylogenies based on both 16S rRNA and 23S rRNA genes corroborated to each other and positioned the sequences in the same clade of hemoplasmas previously detected in D. aurita and D. albiventris sampled in Brazil. Finally, three (6.66%) animals were positive in the PCR for Hepatozoon spp.; the obtained 18S rRNA sequence was positioned into the H. felis clade.The present study showed, for the first time, the circulation of piroplasmids, Hepatozoon spp., Ehrlichia spp., hemoplasmas and Bartonella spp. in D. marsupialis sampled in northeastern Brazil, with description of putative novel genotypes of Ehrlichia and Hepatozoon and copositivity by different vector-borne agents. The present work consolidates the "South American Marsupialia" piroplasmid clade, adding one more genotype of Babesia sp. to this clade.

Incidence of hospital admissions in bioimpedance-guided fluid management among maintenance hemodialysis patients-Results of a randomized controlled trial.

INTRODUCTION: Hemodialysis is life-sustaining in kidney failure. However, proper regulation of body fluids depends on an accurate estimate of target weight. This trial aims to compare clinical endpoints between target weight estimation guided by bioimpedance spectroscopy and usual care in hemodialysis patients. METHODS: This is an open-label, parallel-group, controlled trial that randomized, through a table of random numbers, adult patients on maintenance hemodialysis to target weight estimation based on monthly clinical evaluation alone or added to evaluation by bioimpedance twice a year. The primary outcome was survival, and the secondary outcomes were the rate of hospital admissions, change in blood pressure (BP), and antihypertensive drugs load. Participants were followed for 2 years. Survival analysis was performed using Kaplan-Meier estimator and Log-rank test, and hospital admissions were analyzed by the incidence-rate ratio. FINDINGS: One hundred and ten patients were randomized to the usual care (52) or bioimpedance (58) groups, with a mean age of 57.4 (15.4) years, 64 (58%) males. There was no difference between the groups at baseline. Survival was not significantly different between groups (log-rank test p = 0.68), but the trial was underpowered for this outcome. There was also no difference between the groups in the change in systolic or diastolic BP or in the number of antihypertensive drugs being used. The incidence rate of hospital admissions was 3.1 and 2.1 per person-year in usual care and bioimpedance groups, respectively, with a time-adjusted incidence rate ratio of 1.48 (95% CI: 1.20-1.82, p = 0.0001) and attributable fraction of risk among exposed individuals of 0.32 (95% CI: 0.17-0.45). DISCUSSION: The inclusion of bioimpedance data to guide the estimation of target weight in hemodialysis patients had no detectable impact on survival or BP control, but significantly reduced the incidence rate of hospital admissions. The study was registered at ClinicalTrials.gov Identifier: NCT05272800.

Scapular movement training is not superior to standardized exercises in the treatment of individuals with chronic shoulder pain and scapular dyskinesis: randomized controlled trial.

PURPOSE: To investigate whether scapular movement training (SMT) is superior to standardized exercises in improving scapular biomechanics, behavioral, and clinical aspects of individuals with shoulder pain. METHODS AND METHODS: A total of 64 individuals with chronic shoulder pain were randomly assigned to receive 16 sessions of SMT or SE over 8 weeks. Outcome measures included three-dimensional scapular kinematics, muscle activity of scapulothoracic muscles, pain, disability, fear-avoidance, kinesiophobia, and self-perceived changes. Kinematics and muscle activity were assessed at baseline and after treatment, and self-reported measurements at baseline, 4, 8, and 12 weeks. RESULTS: SMT significantly (p < 0.05) decreased scapular internal rotation during arm elevation and lowering at sagittal and scapular planes (mean difference [MD]: ranged from 2.8 to 4.1°), and at lower angles of arm elevation and lowering at the frontal plane (MD: 3.4° and 2.4°, respectively), increased upper trapezius (UT) activity (MD: 10.3%) and decreased middle trapezius (MT) (MD: 60.4%) and serratus anterior (MD: 9.9%) activity during arm lowering compared to SE. Both groups significantly improved pain, disability, fear-avoidance, kinesiophobia, and self-perceived changes over 4 weeks, which was sustained over the remaining 8 weeks. CONCLUSIONS: SMT is not superior to standardized exercises in improving scapular biomechanics, behavioral, and clinical aspects of individuals with shoulder pain. CLINICAL TRIAL REGISTRATION NUMBER: NCT03528499Implications for rehabilitationScapular movement training (SMT) showed small and likely not clinically relevant changes in scapular kinematics and muscle activity compared to standardized exercises.SMT and standardized exercises presented similar improvements in pain, disability, fear-avoidance beliefs, kinesiophobia, and self-perceived change in health condition immediately following 4-weeks of treatment, which was sustained over the following 8 weeks.The changes in patient-reported outcome measures are unlikely to be associated with changes in scapular kinematics and electromyographic activity.Clinicians should consider other factors than scapular movement during the treatment of patients with shoulder pain.

Real-time PCR quantification and histopathological findings of Mycoplasma hyopneumoniae infection in the lungs of pigs slaughtered in São Luís, Maranhão, Brazil / Quantificação por PCR em tempo real e achados histopatológicos da infecção por Mycoplasma hyopneumoniae em pulmão de suínos abatidos na cidade de São Luís, Maranhão, Brasil

Porcine enzootic pneumonia (PES), mainly caused by the bacteria Mycoplasma hyopneumoniae, is the main cause of respiratory problems in pigs. Infection by M. hyopneumoniae leads to production losses and the predisposition of affected animals to secondary infections, which may result in the condemnation of carcasses and organs due to lung lesions at the time of slaughter. The objective of the research was to evaluate the infection by M. hyopneumoniae in pigs submitted to slaughter in São Luís Island/MA, using molecular and histopathological diagnostic methods. One hundred fifty lung samples were collected from inspected (n=65) and non-inspected (n=85) slaughter pigs on São Luís Island, Maranhão, from July 2019 to August 2021. Of the 150 DNA samples collected, 121 showed an amplified product for Cyt B in the PCR assay. Thus, 121 samples were submitted to qPCR of M. hyopneumoniae, of which 44 (36.36%) showed positive results. The mean amount of bacterial load ranged from 1.20 × 101 to 7.20 × 104, with a mean of 1.73 × 104 copies. Of the reagent samples, 81.81% (36 samples) were obtained from non-inspected slaughter, while 18.18% (8 samples) were obtained from slaughterhouses. In the histopathological analysis, 44 positive qPCR samples were evaluated, of which 28 (63.63%) presented results compatible with the main inflammatory process associated with the presence of M. hyopneumoniae, that is, bronchial-associated lymphoid tissue hyperplasia (BALT). Three samples that showed the highest bacterial load (qPCR: 5.63 × 10³, 2.19 × 104 and 7.23 × 104) showed more evident lesions in this study. The microscopic findings associated with the quantifications indicated a relationship between the amount of bacterial load and the presence of microscopic lesions; higher bacterial load in lung tissue is associated with increased histopathologic staining for BALT hyperplasia. In conclusion, the results point to the circulation of the etiological agent in the sampled animals and the need for preventive measures on pig farms in Maranhão with the involvement of producers, sanitary defense and inspection agencies.

A pneumonia enzoótica suína (PES), causada principalmente pela bactéria Mycoplasma hyopneumoniae, é a principal causa de problemas respiratórios em suínos. A infecção por M. hyopneumoniae leva a perdas produtivas e a predisposição dos animais acometidos a infecções secundárias, o que pode resultar em condenação de carcaças e órgãos por lesões pulmonares no momento do abate. O objetivo da pesquisa foi avaliar a infecção por M. hyopneumoniae em suínos submetidos ao abate na Ilha de São Luís, por meio de métodos diagnósticos moleculares e histopatológicos. Para isso, foram coletadas 150 amostras de pulmão de suínos de abate inspecionado (n=65) e não inspecionado (n=85) na Ilha de São Luís/Maranhão, no período de julho de 2019 a agosto de 2021. Das 150 amostras de DNA coletadas, 121 apresentaram produto amplificado para Cyt B no ensaio de PCR. Assim, 121 amostras foram submetidas à qPCR de M. hyopneumoniae, das quais 44 (36,36%) apresentaram resultados positivos. A quantidade média de carga bacteriana variou de 1,20 × 101 a 7,20 × 104, com média de 1,73 × 104 cópias. Das amostras reagentes, 81,81% (36 amostras) foram obtidas de abate não inspecionado, enquanto 18,18% (8 amostras) foram obtidas em abatedouro. Na análise histopatológica, foram avaliadas 44 amostras positivas para qPCR, das quais 28 (63,63%) apresentaram resultados compatíveis com o principal processo inflamatório associado à presença de M. hyopneumoniae, ou seja, hiperplasia do tecido linfóide associado ao brônquio (BALT). Três amostras que apresentaram maior carga bacteriana (qPCR: 5,63 × 10³, 2,19 × 104 e 7,23 × 104) foram mais evidentes neste estudo. Os achados microscópicos associados às quantificações indicaram uma relação entre a quantidade de carga bacteriana e a presença de lesão microscópica; a maior carga bacteriana no tecido pulmonar está associada a maior alteração histopatológica para hiperplasia BALT. Em conclusão, os resultados obtidos sinalizam para a circulação do agente etiológico nos animais amostrados e a necessidade de medidas preventivas nas criações de suínos do estado do Maranhão com envolvimento dos produtores, órgãos de defesa sanitária e inspeção.

Cryptosporidium sp. in cultivated oysters and the natural oyster stock of the state of Maranhão, Brazil / Detecção de Cryptosporidium sp. em ostras de cultivo e estoque natural no estado do Maranhão, Brasil

This study detected Cryptosporidium spp. in cultivated oysters and the natural oyster stock of the state of Maranhão and determine the elective tissue(s) to examine this protozoan. For this purpose, 200 cultivated oysters were purchased from the municipality of Raposa and another 100 from Paço do Lumiar. Additionally, 100 oysters were extracted from the natural stock of the municipality of Primeira Cruz, thus making up a total of 400 oysters. They were grouped into 80 pools consisting of 5 oysters each. From each pool, the gills and visceral mass were removed to obtain 160 pools, 80 pools for the gill group and another 80 for the visceral mass group. Then, DNA was extracted from each pool using a commercial kit with modifications. Subsequently, the protozoan DNA was detected using nested polymerase chain reaction. With this technique, the DNA of the protozoan under investigation was detected in 2.5% (n = 2/80) of the pools containing gills, with 1.25% of the pools (n = 1/80) belonging to the cultivation group of oysters and the other 1.25% (n = 1/80) to the natural stock. With the results obtained in this study, it was concluded that the analyzed oysters of the genus Crassostrea, from cultivation and natural stock groups, found in the state of Maranhão, were contaminated by Cryptosporidium spp. and may become potential sources of infection in humans and other animals. In addition, the gills are the elective tissue for the study of Cryptosporidium spp. in oysters.

Objetivou-se com o estudo detectar Cryptosporidium sp. em ostras de cultivo e estoque natural no estado do Maranhão e determinar o(s) tecido(s) eletivo(s) para pesquisa desse protozoário. Para a realização do estudo foram adquiridas 200 ostras de cultivo do município de Raposa e 100 de Paço do Lumiar, além de 100 ostras extraídas de estoque natural do município de Primeira Cruz, totalizando 400 ostras. Estas foram agrupadas em 80 pools constituídos por cinco animais. De cada pool, as brânquias e a massa visceral foram removidas totalizando 160 pools, sendo 80 para o grupo das brânquias e 80 para o grupo de massa visceral. Na sequência, procedeu-se à extração de DNA de cada pool com a utilização de kit comercial com modificações. Posteriormente, realizou-se a detecção do DNA do protozoário por meio da técnica de Nested-PCR. Com a técnica utilizada, foi detectado o DNA do protozoário pesquisado em 2,5% (n=2/80) pools apenas de brânquias, sendo 1,25% pools (n=1/80) oriundos de cultivo e os outros 1,25% (n=1/80) de estoque natural. Com os resultados obtidos nesse estudo, conclui-se que as ostras analisadas do gênero Crassostrea sp., oriundas de cultivo e estoque natural no estado do Maranhão, estavam contaminadas por Cryptosporidium sp. e podem se reverter em fontes potenciais para seres humanos e outros animais. Para a pesquisa de Cryptosporidium sp. em ostras, as brânquias são o tecido eletivo.

Cryptosporidium sp. in cultivated oysters and the natural oyster stock of the state of Maranhão, Brazil / Detecção de Cryptosporidium sp. em ostras de cultivo e estoque natural no estado do Maranhão, Brasil

ABSTRACT: This study detected Cryptosporidium spp. in cultivated oysters and the natural oyster stock of the state of Maranhão and determine the elective tissue(s) to examine this protozoan. For this purpose, 200 cultivated oysters were purchased from the municipality of Raposa and another 100 from Paço do Lumiar. Additionally, 100 oysters were extracted from the natural stock of the municipality of Primeira Cruz, thus making up a total of 400 oysters. They were grouped into 80 pools consisting of 5 oysters each. From each pool, the gills and visceral mass were removed to obtain 160 pools, 80 pools for the gill group and another 80 for the visceral mass group. Then, DNA was extracted from each pool using a commercial kit with modifications. Subsequently, the protozoan DNA was detected using nested polymerase chain reaction. With this technique, the DNA of the protozoan under investigation was detected in 2.5% (n = 2/80) of the pools containing gills, with 1.25% of the pools (n = 1/80) belonging to the cultivation group of oysters and the other 1.25% (n = 1/80) to the natural stock. With the results obtained in this study, it was concluded that the analyzed oysters of the genus Crassostrea, from cultivation and natural stock groups, found in the state of Maranhão, were contaminated by Cryptosporidium spp. and may become potential sources of infection in humans and other animals. In addition, the gills are the elective tissue for the study of Cryptosporidium spp. in oysters.

RESUMO: Objetivou-se com o estudo detectar Cryptosporidium sp. em ostras de cultivo e estoque natural no estado do Maranhão e determinar o(s) tecido(s) eletivo(s) para pesquisa desse protozoário. Para a realização do estudo foram adquiridas 200 ostras de cultivo do município de Raposa e 100 de Paço do Lumiar, além de 100 ostras extraídas de estoque natural do município de Primeira Cruz, totalizando 400 ostras. Estas foram agrupadas em 80 pools constituídos por cinco animais. De cada pool, as brânquias e a massa visceral foram removidas totalizando 160 pools, sendo 80 para o grupo das brânquias e 80 para o grupo de massa visceral. Na sequência, procedeu-se à extração de DNA de cada pool com a utilização de kit comercial com modificações. Posteriormente, realizou-se a detecção do DNA do protozoário por meio da técnica de Nested-PCR. Com a técnica utilizada, foi detectado o DNA do protozoário pesquisado em 2,5% (n=2/80) pools apenas de brânquias, sendo 1,25% pools (n=1/80) oriundos de cultivo e os outros 1,25% (n=1/80) de estoque natural. Com os resultados obtidos nesse estudo, conclui-se que as ostras analisadas do gênero Crassostrea sp., oriundas de cultivo e estoque natural no estado do Maranhão, estavam contaminadas por Cryptosporidium sp. e podem se reverter em fontes potenciais para seres humanos e outros animais. Para a pesquisa de Cryptosporidium sp. em ostras, as brânquias são o tecido eletivo.

IMPACTO-MR: um estudo brasileiro de plataforma nacional para avaliar infecções e multirresistência em unidades de terapia intensiva / IMPACTO-MR: a Brazilian nationwide platform study to assess infections and multidrug resistance in intensive care units

RESUMO Objetivo: Descrever o IMPACTO-MR, um estudo brasileiro de plataforma nacional em unidades de terapia intensiva focado no impacto das infecções por bactérias multirresistentes relacionadas à assistência à saúde. Métodos: Descrevemos a plataforma IMPACTO-MR, seu desenvolvimento, critérios para seleção das unidades de terapia intensiva, caracterização da coleta de dados, objetivos e projetos de pesquisa futuros a serem realizados na plataforma. Resultados: Os dados principais foram coletados por meio do Epimed Monitor System® e consistiram em dados demográficos, dados de comorbidades, estado funcional, escores clínicos, diagnóstico de internação e diagnósticos secundários, dados laboratoriais, clínicos e microbiológicos e suporte de órgãos durante a internação na unidade de terapia intensiva, entre outros. De outubro de 2019 a dezembro de 2020, 33.983 pacientes de 51 unidades de terapia intensiva foram incluídos no banco de dados principal. Conclusão: A plataforma IMPACTO-MR é um banco de dados clínico brasileiro de unidades de terapia intensiva focado na pesquisa do impacto das infecções por bactérias multirresistentes relacionadas à assistência à saúde. Essa plataforma fornece dados para o desenvolvimento e pesquisa de unidades de terapia intensiva individuais e ensaios clínicos observacionais e prospectivos multicêntricos.

ABSTRACT Objective: To describe the IMPACTO-MR, a Brazilian nationwide intensive care unit platform study focused on the impact of health care-associated infections due to multidrug-resistant bacteria. Methods: We described the IMPACTO-MR platform, its development, criteria for intensive care unit selection, characterization of core data collection, objectives, and future research projects to be held within the platform. Results: The core data were collected using the Epimed Monitor System® and consisted of demographic data, comorbidity data, functional status, clinical scores, admission diagnosis and secondary diagnoses, laboratory, clinical, and microbiological data, and organ support during intensive care unit stay, among others. From October 2019 to December 2020, 33,983 patients from 51 intensive care units were included in the core database. Conclusion: The IMPACTO-MR platform is a nationwide Brazilian intensive care unit clinical database focused on researching the impact of health care-associated infections due to multidrug-resistant bacteria. This platform provides data for individual intensive care unit development and research and multicenter observational and prospective trials.

What factors contribute to the Scapular Assistance Test result? A classification and regression tree approach.

The aim of this study was to determine predictive factors related to the Scapular Assistance Test in individuals with shoulder pain during arm elevation, and to analyze how these predictors interact in a nonlinear manner to discriminate the result of a positive and negative Scapular Assistance Test. Eighty-four individuals with shoulder pain with positive (n = 47, average age 38.4 years) and negative (n = 37, average age 37.8 years) Scapular Assistance Test completed the study. Demographic data, affected shoulder, pain duration, pain at rest, angular onset of pain, scapular dyskinesis, serratus anterior and lower trapezius muscle strength, Disabilities of Arm, Shoulder and Hand questionnaire and Pain Catastrophizing Scale were assessed in all participants. The Classification and Regression Tree analysis was used to determine which factors would predict the occurrence of a positive or negative Scapular Assistance Test and possible interactions among them. The resulting tree presented seven levels that combine the following variables: angular onset of pain, presence of scapular dyskinesis, pain catastrophizing, serratus anterior and lower trapezius muscle strength. The angular onset of pain during arm elevation was the main predictor of a positive Scapular Assistance Test selected by the Classification and Regression Tree. This study indicates that the Scapular Assistance Test result may be explained not only by biomechanical variables, but also by psychological factors. Disability of the upper limbs does not seem to contribute to the Scapular Assistance Test result.

To what extent do typical components of shoulder clinical evaluation explain upper-extremity disability? A cross-sectional study.

BACKGROUND: Physical therapists use several evaluation measures to identify the most important factors related to disability. However, the degree to which these evaluation components explain shoulder disability is not well known and that may detract clinicians from the best clinical reasoning. OBJECTIVE: To determine how much evaluation components explain shoulder function. METHODS: Eighty-one individuals with unilateral shoulder pain for at least four weeks and meeting clinical exam criteria to exclude cervical referred pain, adhesive capsulitis, and shoulder instability, participated in this study. Several typical clinical evaluation components were assessed as potential independent variables in a regression model using the Disabilities of the Arm, Shoulder, and Hand (DASH) score as a proxy to shoulder function. Two multivariate models were built to include (1) evaluation components from physical exam plus clinical history and (2) a model considering all previous variables and magnetic resonance imaging (MRI) data. RESULTS: Pain catastrophizing was the best variable in the model explaining at least 10% of the DASH variance. Sex and lower trapezius muscle strength explained considerably less of shoulder function. The MRI data did not improve the model performance. CONCLUSION: The complexity of shoulder function is not independently explained by pathoanatomical abnormalities. Psychological aspects may explain more of shoulder function even when combined with physical components in some patients.

Overhydration in acute kidney injury: Is it always a menace to critically ill patients? A survival study using bioimpedance spectroscopy.

INTRODUCTION: The hydration status is a predictor of survival in critically ill patients. However, it is unclear whether this association depends on the patient clinical condition. This study was designed to analyze the impact of hydration status on survival of critically ill patients with acute kidney injury (AKI) with or without sepsis. METHODS: A prospective cohort study following critically ill patients with AKI consecutively admitted over a one-year period to a teaching hospital intensive care unit (ICU). All patients with AKI stage three or higher of the KDIGO score were included. The hydration status was evaluated through the overhydration (OH) parameter of spectroscopy bioimpedance and sepsis was defined according Sepse-3 criteria. The survival analysis used adjusted competing-risks regression. RESULTS: Forty-eight patients were included, 27 (56%) with a sepsis diagnosis. The main negative predictors of survival among sepsis patients, adjusted by SAPS3 score, were higher OH (SHR 1.1, 95% CI 1.0-1.2, p = 0.02), mechanical ventilation (SHR 6.9, 95% CI 1.0-47.8, p = 0.04) and older age (SHR 1.1, 95% CI 1.0-1.1, p = 0.005). The predictors in non-sepsis patients were lower OH (SHR 0.82, 95% CI 0.71-0.95, p = 0.008) and mechanical ventilation (SHR 12, 95% CI 2.4-6.6, p < 0.001). CONCLUSIONS: This finding suggests that extracellular overhydration is an independent predictor of survival in critically ill patients with sepsis, but it seems to have opposite effect in non-sepsis patients.

Is the angular onset of pain during arm elevation associated to functioning in individuals with rotator cuff related shoulder pain?

BACKGROUND: Rotator cuff related shoulder pain (RCRSP) is the main diagnosis for shoulder pain. A painful arc during active arm elevation is a common finding in RCRSP. The angular onset of pain during arm elevation may play an important role on functioning of the upper extremities. OBJECTIVE: This study aimed to: 1) determine and characterize the association between the angular onset of pain during arm elevation and upper-limb self-reported functioning, 2) verify whether demographic and clinical characteristics contribute to this association, and 3) investigate whether these characteristics differ considering distinct ranges of angular onset of pain in individuals with RCRSP. METHODS: 252 individuals with RCRSP were divided in 3 groups based on the angular onset of pain during arm elevation in the sagittal plane: <60°, 60°-120°, and >120°. The Disabilities of the Arm, Shoulder and Hand (DASH) questionnaire was used to assess functioning of the upper limbs. Demographic and clinical characteristics were selected for the multivariable analysis. RESULTS: Individuals with an angular onset of pain above 120° of arm elevation presented lower DASH score (lesser disability) than individuals with an angular onset of pain between 60°- 120°. Male sex, age, dominance of the side affected, duration of symptoms, and the angular onset of pain during arm elevation explained 31% of the DASH score variance. CONCLUSION: Individuals with RCRSP and angular onset of pain above 120° of arm elevation present better functioning than individuals with onset of pain between 60°-120°, and similar functioning as those with pain below 60°. Male sex, lower age, non-dominant side being affected and longer duration of symptoms are also associated to better functioning as assessed by the DASH questionnaire.

Endoparasite diversity and liver alterations in Hoplerythrinus unitaeniatus and Cichlasoma bimaculatum in a quilombola area in Maranhão, Brazil.

Our aim was to assess endoparasite diversity and liver alterations in Hoplerythrinus unitaeniatus (jeju) and Cichlasoma bimaculatum (acará preto) in a quilombola area in Maranhão, Brazil. For this, 21 H. unitaeniatus and 21 C. bimaculatum were caught in a natural environment and transported to a laboratory. After these had been euthanized, endoparasites were collected and identified. Liver alterations were evaluated histological analysis based on the severity of each lesion: stage I, organ functioning not compromised; stage II, more severe lesions that impair normal functioning of the organs; and stage III, very severe and irreversible lesions. Among the fish evaluated, 71.43% H. unitaeniatus and 61.90% C. bimaculatum were parasitized. Contracaecum sp. was found in both species; while acanthocephalans, only in H. unitaeniatus. The alterations were vacuolization, nucleus in the cell periphery, deformation of the cell outline, melanomacrophage center, hyperemia, cytoplasmic degeneration and nuclear vacuolization. Through calculating a histological alteration index, it was found that 26.19% of the specimens presented lesions in stage I; 38.09% lesions in stage II and 9.52% lesions in stage III. It was concluded that there is high prevalence of Contracaecum sp. and that the liver lesions may be adaptive responses by the fish to endoparasitic infection.