Colchicine reduces the activation of NLRP3 inflammasome in COVID-19 patients.

OBJECTIVE: To evaluate whether colchicine treatment was associated with the inhibition of NLRP3 inflammasome activation in patients with COVID-19. METHODS: We present a post hoc analysis from a double-blinded placebo-controlled randomized clinical trial (RCT) on the effect of colchicine for the treatment of COVID-19. Serum levels of NOD-like receptor protein 3 (NLRP3) inflammasome products-active caspase-1 (Casp1p20), IL-1ß, and IL-18-were assessed at enrollment and after 48-72 h of treatment in patients receiving standard-of-care (SOC) plus placebo vs. those receiving SOC plus colchicine. The colchicine regimen was 0.5 mg tid for 5 days, followed by 0.5 mg bid for another 5 days. RESULTS: Thirty-six patients received SOC plus colchicine, and thirty-six received SOC plus placebo. Colchicine reduced the need for supplemental oxygen and the length of hospitalization. On Days 2-3, colchicine lowered the serum levels of Casp1p20 and IL-18, but not IL-1ß. CONCLUSION: Treatment with colchicine inhibited the activation of the NLRP3 inflammasome, an event triggering the 'cytokine storm' in COVID-19. TRIAL REGISTRATION NUMBERS: RBR-8jyhxh.

Fluoroquinolone treatment as a protective factor for 10-day mortality in Streptococcus pneumoniae bacteremia in cancer patients.

To evaluate the prognostic factors in adult cancer patients with pneumococcal bacteremia, describe episode features and the phenotypic characteristics of the isolated strains. We evaluated the episodes in patients admitted to a cancer hospital between 2009 and 2015. The outcomes were defined as 48 h mortality and mortality within 10 days after the episode. The variables evaluated were: age, sex, ethnicity, ECOG, Karnofsky score, SOFA, cancer type, metastasis, chemotherapy, radiotherapy, neutropenia, previous antibiotic therapy, community or healthcare-acquired infection, comorbidities, smoking, pneumococcal vaccination, infection site, presence of fever, polymicrobial infection, antimicrobial susceptibility, serotype and treatment. 165 episodes were detected in 161 patients. The mean age was 61.3 years; solid tumors were the most prevalent (75%). 48 h and 10-day mortality were 21% (34/161) and 43% (70/161) respectively. The 48 h mortality- associated risk factors were SOFA and polymicrobial bacteremia; 10-day mortality-associated risk factors were fever, neutropenia, ECOG 3/4, SOFA and fluoroquinolones as a protective factor. Pneumococcal bacteremia presented high mortality in cancer patients, with prognosis related to intrinsic host factors and infection episodes features. Fluoroquinolone treatment, a protective factor in 10-day mortality, has potential use for IPDs and severe community-acquired pneumonia in cancer patients.

Antinociceptive and anti-inflammatory activities of a triterpene-rich fraction from Himatanthus drasticus.

Himatanthus drasticus (Mart.) Plumel belongs to the Apocynaceae family and the latex from its trunk bark (Hd) is known as "janaguba milk". This latex is widely used in Northeast Brazil, mainly in the Cariri region, for its gastroprotective, anti-inflammatory, and antitumor properties. The objective of this study was to investigate a triterpene-rich fraction (FJNB) from H. drasticus latex on acute models of nociception and inflammation and to clarify its mechanisms of action. Wistar rats or Swiss mice were subjected to the carrageenan-induced paw edema test or the formalin test, respectively, after the acute oral treatment with FJNB. The inflamed paws from the carrageenan-induced paw edema and formalin tests were processed for histological and immunohistochemical assays, respectively. The results were analyzed by ANOVA and considered significant at P<0.05. FJNB (10 mg/kg) decreased the paw edema by 25% at the 3rd h after the carrageenan injection. Indomethacin, used as reference, inhibited the paw edema by 59% at the same time-point. In the formalin test, FJNB inhibited the 1st phase by 27, 49, and 52% and the 2nd phase by 37, 50, and 67%, at the doses of 1, 5, and 10 mg/kg, respectively. In addition, FJNB significantly inhibited the expressions of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and the inflammatory cytokine tumor necrosis factor (TNF)-alpha. The histone deacetylase (HDAC) expression and the transcription factor nuclear factor kappa (NF-kB) were also inhibited at the same doses. In conclusion, the FJNB inhibitory actions on iNOS, COX-2, TNF-α, HDAC, and NF-kB could be involved with the drug anti-inflammatory activity.

Antinociceptive and anti-inflammatory activities of a triterpene-rich fraction from Himatanthus drasticus

Himatanthus drasticus (Mart.) Plumel belongs to the Apocynaceae family and the latex from its trunk bark (Hd) is known as "janaguba milk". This latex is widely used in Northeast Brazil, mainly in the Cariri region, for its gastroprotective, anti-inflammatory, and antitumor properties. The objective of this study was to investigate a triterpene-rich fraction (FJNB) from H. drasticus latex on acute models of nociception and inflammation and to clarify its mechanisms of action. Wistar rats or Swiss mice were subjected to the carrageenan-induced paw edema test or the formalin test, respectively, after the acute oral treatment with FJNB. The inflamed paws from the carrageenan-induced paw edema and formalin tests were processed for histological and immunohistochemical assays, respectively. The results were analyzed by ANOVA and considered significant at P<0.05. FJNB (10 mg/kg) decreased the paw edema by 25% at the 3rd h after the carrageenan injection. Indomethacin, used as reference, inhibited the paw edema by 59% at the same time-point. In the formalin test, FJNB inhibited the 1st phase by 27, 49, and 52% and the 2nd phase by 37, 50, and 67%, at the doses of 1, 5, and 10 mg/kg, respectively. In addition, FJNB significantly inhibited the expressions of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and the inflammatory cytokine tumor necrosis factor (TNF)-alpha. The histone deacetylase (HDAC) expression and the transcription factor nuclear factor kappa (NF-kB) were also inhibited at the same doses. In conclusion, the FJNB inhibitory actions on iNOS, COX-2, TNF-α, HDAC, and NF-kB could be involved with the drug anti-inflammatory activity.

The population density of Lymnaea columella (Say, 1817) (Mollusca, Lymnaeidae) an intermediate host of Fasciola hepatica (Linnaeus, 1758), in the Caparaó microregion, ES, Brazil / Densidade populacional de Lymnaea columella (Say, 1817) (Mollusca, Lymnaeidae) hospedeiro intermediário de Fasciola hepatica (Linnaeus, 1758) na Microrregião do Caparaó, ES, Brasil

Abstract The aim of this study was to monitor the population density of Lymnaea columella, an intermediate host of Fasciola hepatica, in various aquatic habitats and in drinking water in the area of the Instituto Federal de Educação, Ciência e Tecnologia do Espírito Santo, on Caparaó Microregion, municipality of Alegre, state of Espírito Santo, Brazil. Monthly samplings were performed at certain points between drainage areas and drinking water in cattle and goat production systems during the years 2010 to 2013. The mean temperature, precipitation and the frequency of samples of L. columella were analysed graphically according the monthly average during the study period. A total of 2,038 molluscs were collected, 1558 of which were L. columella, that predominated in all sampled points. The highest average of specimens observed for L. columella was in the years 2010 and 2013 (51.0), and occurred decreased in 2011 (19.8). The temperature and precipitation averaged is 23.7 °C and 141 mm/year, respectively. Rainfall peak occurred in March (2011, 2013) and November (2012), during these periods the population of L. columella growth. There was no significant difference in the relationship between the specimens observed with seasons (dry-wet), thus the population of L. columella remained stable and can be found throughout the year.

Resumo O objetivo deste estudo foi acompanhar a variação da densidade populacional de Lymnaea columella, hospedeiro intermediário de Fasciola hepatica, em diferentes tipos de coleções hídricas e bebedouros na área do Instituto Federal de Educação, Ciência e Tecnologia do Espírito Santo, na microrregião do Caparaó, município de Alegre, estado do Espírito Santo, Brasil. Foram realizadas coletas mensais em pontos determinados entre áreas de drenagem e bebedouros de animais nos setores de bovinocultura e caprinocultura durante os anos de 2010 a 2013. As médias de temperatura, precipitação e a frequência de exemplares de L. columella foram analisadas graficamente de acordo com a média mensal ao longo do período avaliado. Foram coletados 2.038 moluscos, sendo 1558 de L. columella, espécie predominante em todos os pontos amostrados. A maior média de espécimes observadas para L. columella foi nos anos de 2010 e 2013 (51,0), ocorrendo um decréscimo em 2011 (19,8). A temperatura apresentou média de 23,7°C e precipitação de 141 mm/ano. Picos de precipitação ocorreram em Março (2011, 2013) e Novembro (2012), nestes períodos foi observado aumento populacional de L. columella. Não houve diferença significativa na relação entre as espécimes observadas com as estações do ano (seca-chuvosa), dessa forma, a população de L. columella se mostrou estável, podendo ser encontrada durante todo ano.

The population density of Lymnaea columella (Say, 1817) (Mollusca, Lymnaeidae) an intermediate host of Fasciola hepatica (Linnaeus, 1758), in the Caparaó microregion, ES, Brazil.

The aim of this study was to monitor the population density of Lymnaea columella, an intermediate host of Fasciola hepatica, in various aquatic habitats and in drinking water in the area of the Instituto Federal de Educação, Ciência e Tecnologia do Espírito Santo, on Caparaó Microregion, municipality of Alegre, state of Espírito Santo, Brazil. Monthly samplings were performed at certain points between drainage areas and drinking water in cattle and goat production systems during the years 2010 to 2013. The mean temperature, precipitation and the frequency of samples of L. columella were analysed graphically according the monthly average during the study period. A total of 2,038 molluscs were collected, 1558 of which were L. columella, that predominated in all sampled points. The highest average of specimens observed for L. columella was in the years 2010 and 2013 (51.0), and occurred decreased in 2011 (19.8). The temperature and precipitation averaged is 23.7 °C and 141 mm/year, respectively. Rainfall peak occurred in March (2011, 2013) and November (2012), during these periods the population of L. columella growth. There was no significant difference in the relationship between the specimens observed with seasons (dry-wet), thus the population of L. columella remained stable and can be found throughout the year.

What Makes Umbilical Cord Tissue-Derived Mesenchymal Stromal Cells Superior Immunomodulators When Compared to Bone Marrow Derived Mesenchymal Stromal Cells?

MSCs derived from the umbilical cord tissue, termed UCX, were investigated for their immunomodulatory properties and compared to bone marrow-derived MSCs (BM-MSCs), the gold-standard in immunotherapy. Immunogenicity and immunosuppression were assessed by mixed lymphocyte reactions, suppression of lymphocyte proliferation and induction of regulatory T cells. Results showed that UCX were less immunogenic and showed higher immunosuppression activity than BM-MSCs. Further, UCX did not need prior activation or priming to exert their immunomodulatory effects. This was further corroborated in vivo in a model of acute inflammation. To elucidate the potency differences observed between UCX and BM-MSCs, gene expression related to immune modulation was analysed in both cell types. Several gene expression profile differences were found between UCX and BM-MSCs, namely decreased expression of HLA-DRA, HO-1, IGFBP1, 4 and 6, ILR1, IL6R and PTGES and increased expression of CD200, CD273, CD274, IL1B, IL-8, LIF and TGFB2. The latter were confirmed at the protein expression level. Overall, these results show that UCX seem to be naturally more potent immunosuppressors and less immunogenic than BM-MSCs. We propose that these differences may be due to increased levels of immunomodulatory surface proteins such as CD200, CD273, CD274 and cytokines such as IL1ß, IL-8, LIF and TGFß2.

Selective B-cell expression of the MHV-68 latency-associated M2 protein regulates T-dependent antibody response and inhibits apoptosis upon viral infection.

To better understand the role of the M2 protein of the murine herpes virus strain 68 (MHV-68) in vivo, B-lymphocyte-restricted, M2-transgenic mice were constructed. The transgenic mice contained normal B-cell subpopulations in bone marrow, lymph nodes and spleen. After immunization with sheep red blood cells, spleens from M2-transgenic mice had increased germinal centres. Transgenic mice responded to the T-cell-dependent antigen keyhole limpet haemocyanin (KLH) with higher levels of secondary IgM and IgG2a antibodies than WT mice. Normal and M2-transgenic mice were infected with WT and M2 frame-shift mutant (M2FS) MHV-68 viruses. The pathogenesis of M2-transgenic mice infected with the M2-deficient mutant virus did not revert to that observed upon infection of normal mice with WT virus. However, the higher reactivation levels late after M2-transgenic mice were infected with WT virus reflected the importance of M2 as a target for the immune response, and thus with an impact on the establishment of latency. Finally, there was markedly less apoptosis in B-cells from M2-transgenic mice infected with either WT or M2FS mutant than from similarly infected WT mice, consistent with the published inhibitory influence of M2 on apoptosis in vitro. Thus, M2 provides a strategy to increase the pool of germinal centre B-cells through inhibition of apoptosis in the infected cell.

A novel TLR3 inhibitor encoded by African swine fever virus (ASFV).

African swine fever virus (ASFV) encodes proteins that manipulate important host antiviral mechanisms. Bioinformatic analysis of the ASFV genome revealed ORF I329L, a gene without any previous functional characterization as a possible inhibitor of TLR signaling. We demonstrate that ORF I329L encodes a highly glycosylated protein expressed in the cell membrane and on its surface. I329L also inhibited dsRNA-stimulated activation of NFκB and IRF3, two key players in innate immunity. Consistent with this, expression of I329L protein also inhibited the activation of interferon-ß and CCL5. Finally, overexpression of TRIF reversed I329L-mediated inhibition of both NFκB and IRF3 activation. Our results suggest that TRIF, a key MyD88-independent adaptor molecule, is a possible target of this viral host modulation gene. The demonstration of an ASFV host evasion molecule inhibiting TLR responses is consistent with the ability of this virus to infect vertebrate and invertebrate hosts, both of which deploy innate immunity controlled by conserved TLR systems.

Atividade antimicrobiana do extrato bruto etanólico de raízes e partes aéreas de Ottonia martiana Miq. (Piperaceae) / Antimicrobial activity of crude ethanolic extracts of Ottonia martiana Miq. (Piperaceae) roots and aerial parts

Ottonia martiana Miq., Piperaceae, conhecida na medicina popular brasileira como "anestésia", é um arbusto característico da Floresta Atlântica. Devido à ação anestésica sobre a mucosa oral, raízes e partes aéreas são popularmente utilizadas em dores de dente, na forma de alcoolaturas (bochechos) ou pela mastigação de fragmentos de partes da planta. Como os microrganismos estão adquirindo resistência aos antibióticos tradicionais, a procura por novos agentes antimicrobianos, a partir de plantas, vem sendo intensificada. Nesta perspectiva, avaliou-se o efeito antimicrobiano do extrato bruto etanólico dos órgãos totais desta espécie (EBEtOH) frente às bactérias Enterococcus faecium (ATCC 6569), Enterobacter aerogenes (ATCC 13048) e Pseudomonas aeruginosa (ATCC 15442), utilizando-se os métodos de difusão em ágar e bioautografia. Os resultados obtidos por difusão em ágar mostraram que o EBEtOH de O. martiana apresenta potencial antibacteriano contra E. faecium, evidenciado pelo aparecimento de zonas de inibição de crescimento no respectivo bioautograma e não demonstraram ação antimicrobiana contra as outras cepas testadas.

Ottonia martiana Miq., Piperaceae, known in Brazilian folk medicine as "anestésia", is an herbaceous shrub found in Atlantic Tropical Forest. Due to its anesthetic action on the mucous membranes of the mouth, roots and aerial parts are commonly used to relief toothache, usually as alcoholic macerates (mouth washing) or by chewing parts of the plant. As most microorganisms are acquiring resistance to traditional antibiotics, the search for new antimicrobials agents, specially from plants, is being intensified. Based on this, the activity of the crude ethanolic extract from roots and aerial parts (EBEtOH) of O. martiana against Enterococcus faecium (ATCC 6569), Enterobacter aerogenes (ATCC 13048) and Pseudomonas aeruginosa (ATCC 15442) was evaluated using the agar difusion and bioautographic methods. Results from the agar diffusion assay showed that O. martiana EBEtOH has a potencial effect against E. faecium, which was corroborated by the presence of inhibition zones in the bioautographic test, whereas no inhibitory action against the other assayed bacteria was observed.

First confirmed case of a vancomycin-resistant Enterococcus faecium with vanA phenotype from Brazil: isolation from a meningitis case in São Paulo.

The importance of enterococci as a nosocomial etiologic agent is well documented; however, enterococci are also capable of causing a variety of community-acquired infections. Vancomycin resistance in a clinical Enterococcus isolate was first reported in 1986, and since then vancomycin-resistant enterococci (VRE) have been reported world-wide. This report describes a case of E. faecium with the VanA phenotype, isolated from meningitis in Sao Paulo, Brazil. Two E. faecium strains were isolated. One strain showed VanA phenotype, and the molecular characterization of the VanA gene was confirmed by polymerase chain reaction. The other strain was susceptible to vancomycin and teicoplanin. The authors would like to call the attention of the scientific community to this first identification of a VRE case in Sao Paulo, Brazil.

Primeiros resultados do tratamento da lepra com a kanamicina

Os autores apresentam seus resultados no tratamento de 10 pacientes lepromatosos com a kanamicina, na dose de 1,0 grama diária por um período de 90 dias. Tendo em vista a toxicidade da droga, realizaram exames hematológicos, de urina, e ORL (otorrinoloringológicos) durante a experimentação e observaram albuminúria transitória em 6 pacientes e comprometimento auditivo em 8, evidenciável pela audiometria. Apesar disto somente um paciente revelou sintomatologia aparente com tonturas, surdez, zumbido, sendo obrigado a suspender o medicamento. Nos demais pacientes, o dano cólceovestibular foi nas frequências mais altas fora dos limites da conversação social, não sendo percebido pelos mesmos. Os resultados cl¡nicos foram idênticos aos observados com outros antibióticos tais como rifamicina e oxitetraciclina, sendo bastante evidentes nos primeiros 30 dias, os casos em franca piora cl¡nica. Os autores chamam a atenção sobre as alterações morfológicas precoces dos bacilos nos casos estudados e negativação baciloscópica em 3 pacientes. Consideram ser a kanamicina dotada de atividade bactericida para o bacilo de Hansen, particularmente evidente nos germes alongados caracter¡sticos dos casos piorados. Os exames histopatológicos confirmaram os resultados cl¡nico-baciloscópicos por eles conseguidos. Os fenômenos reacionais tipo ENL foram descritos e não se manifestaram na maioria dos pacientes. Concluem considerando a droga dotada de alta toxicidade para o ouvido, devendo o seu uso ser sempre acompanhado de vigilância clínica cuidadosa e sempre que poss¡vel com controle audiométrico. Não sendo evidentemente um medicamento que se prestasse para "campanha de massa" o seu emprêgo contudo deveria ser recomendado para os casos que não respondem aos tratamentos clássicos ou se acham em franca piora clínica, até a remissão das lesões reativadas. O período de tratamento não deveria ultrapassar 30 dias quando a droga fosse usada isoladamente, porque foi dentro desse per¡odo que...