CRZ1 regulator and calcium cooperatively modulate holocellulases gene expression in Trichoderma reesei QM6a.

Trichoderma reesei is the main filamentous fungus used in industry to produce cellulases. Here we investigated the role of CRZ1 and Ca2+signaling in the fungus T. reesei QM6a concerning holocellulases production. For this, we first searched for potential CRZ1 binding sites in promoter regions of key genes coding holocellulases, as well as transcriptional regulators and sugar and calcium transporters. Using a nearly constructed T. reeseiAcrz1 strain, we demonstrated that most of the genes expected to be regulated by CRZ1 were affected in the mutant strain induced with sugarcane bagasse (SCB) and cellulose. In particular, our data demonstrate that Ca2+ acts synergistically with CRZ1 to modulate gene expression, but also exerts CRZ1-independent regulatory role in gene expression in T. reesei, highlighting the role of the major regulator Ca2+ on the signaling for holocellulases transcriptional control in the most part of cellulases genes here investigated. This work presents new evidence on the regulatory role of CRZ1 and Ca2+ sensing in the regulation of cellulolytic enzymes in T. reesei, evidencing significant and previously unknown function of this Ca2+sensing system in the control key transcriptional regulators (XYR1 and CRE1) and on the expression of genes related to sugar and Ca2+ transport.

Social characterization of COVID-19 in the state of Amazonas, Brazil.

OBJECTIVES: To compare the number of deaths that occurred in the state of Amazonas and in Brazil, from March 16th to August 20th 2020, using the variables skin color, sex, place of death, age group and association with COVID-19, and secondly, to verify whether between 2019 and 2020, in the period from March 16th to August 20th, there was a significant change in the number of deaths from diseases not associated with COVID-19. METHODS: We searched the databases of the Brazilian public agency "Transparency Portal" for the data on deaths that occurred in the state of Amazonas and Brazil in the period from March 16th to August 20th, 2019 and 2020. The absolute frequencies and percentages of the variables studied were used for statistical analysis. RESULTS: COVID-19 was responsible for an 11.01% increase in deaths in Brazil; however, this rate quadrupled in the state of Amazonas. In relation to age group, there was a similar percentage between Amazonas and the national average. The stratified analysis showed significant differences between genders and races, with higher death rates in men and people of brown/black skin. The number of deaths at home increased significantly, especially those from causes not associated with COVID-19. CONCLUSIONS: The national drop in deaths from diseases not associated with COVID between March and August 2020 in Brazil is misleading and may be due to the result of misreported causes of death.

[Immunoglobulin isotype and IgG subclass profiles in American tegumentary leishmaniasis]. / Perfil de isotipos de imunoglobulinas e subclasses de IgG na leishmaniose tegumentar americana.

The present work investigated the serum antibody profiles in 37 patients with American tegumentary leishmaniasis, who were attended at Hospital de Clinicas - Universidade Federal de Uberlandia, MG, Brazil. The immunoglobulin class and IgG subclass profiles were analyzed by indirect ELISA using Leishmania (Leishmania) amazonensis soluble antigen. The antibody avidity was determined by 6 M urea treatment after incubation with immunoenzymatic conjugate. It was observed that 97% of the serum samples presented anti-Leishmania antibodies for IgE class, 94.6% IgG, 57.5% IgA and 21.5% IgM class. For IgG subclasses the profiles were in the following order of frequency: IgG1>IgG3>IgG2>IgG4. High avidity of anti-Leishmania IgE antibodies was found in 44.4% of the samples. On the other hand, moderate avidity of specific IgG and IgA was observed in 62.8% and 47.8% of samples, respectively. These results indicate a very complex antibody response profile against American tegumentary leishmaniasis.

Immunoblotting analyses using two-dimensional gel electrophoresis of Trypanosoma cruzi excreted-secreted antigens.

Trypanosoma cruzi trypomastigotes excrete-secrete a complex mixture of antigenic molecules. This antigenic mixture denominated trypomastigote excreted-secreted antigens contains a 150-160 kDa band that shows excellent performance in Chagas' disease diagnosis by immunoblotting. The present study partially characterized by two-dimensional gel electrophoresis the immunoreactivity against the 150-160 kDa protein using sera samples from chagasic patients in different phases of the disease. Trypomastigote excreted-secreted antigen preparations were subjected to high-resolution two-dimensional (2D) gel electrophoresis followed by immunoblotting with sera from chagasic and non-chagasic patients. The 150-160 kDa protein presented four isoforms with isoelectric focusing ranging from 6.2 to 6.7. The four isoforms were recognized by IgM from acute phase and IgG from chronic phase sera of chagasic patients. The 150-160 kDa isoform with IF of approximately 6.4 became the immunodominant spot with the progression of the disease. No cross-reactivity was observed with non-chagasic or patients infected with Leishmania sp. In this study we provide basic knowledge that supports the validation of trypomastigote excreted-secreted antigens for serological diagnosis of Chagas' disease.

[Prevalence and clinical significance of interactions drug-enteral nutrition in Intensive Care Units]. / Prevalencia e significancia clinica de interacoes farmaco-nutricao enteral em Unidades de Terapia Intensiva.

This is a multicenter, cross-sectional retrospective study, which aimed to determine the prevalence of interactions drug-enteral nutrition (EN) in Intensive Care Units (ICU) of seven teaching hospitals in Brazil, and to analyze the clinical significance of them. Information on medications and EN administered with 24 hours and 120 hours of hospitalization were collected. For identification of drug-EN interaction was employed software Drug Reax®. It was investigated 1.124 records. Of these, 320 patients, with 24 hours of hospitalization, were on EN, and 20 (6.3%) had drug-EN interaction. Of the 504 patients with 120 hours of hospitalization, 39 (7.7%) had drug-EN interaction. The most frequent drug-EN interactions of clinical significance were phenytoin-EN, warfarin-EN and levothyroxine-EN. Drug-EN interactions may interfere with the quality and cost effectiveness of care in ICU, so it is essential that the health team has knowledge about them.

Pseudoparasitism by Calodium hepaticum (syn. Capillaria hepatica; Hepaticola hepatica) in the Negro River, Brazilian Amazon.

We report the finding of eggs of Calodium spp. (syn. Capillaria spp.; Hepaticola spp.) in a fecal sample from an old woman living in a riverine community in the Negro River Basin and describe the associated epidemiological investigation. The case probably does not represent true parasitism; the eggs, which were compatible with the species Calodium hepaticum, were most likely ingested upon consumption of infected tapir (Tapirus terrestris) liver, subsequently passing through the gut and being eliminated. The evolution of these eggs to infective stages in the environment, given the poor sanitation background, could provide the risk of occurrence of hepatic disease in humans.

Prevalencia e significancia clinica de interacoes farmaco-nutricao enteral em Unidades de Terapia Intensiva / Prevalence and clinical significance of interactions drug-enteral nutrition in Intensive Care Units / Prevalencia y significado clinico de interacciones farmaco- nutricion enteral en Unidades de Cuidados Intensivos

Estudo multicêntrico, transversal e retrospectivo, que objetivou determinar a prevalência de interações fármaco-nutrição enteral (NE) em Unidades de Terapia Intensiva (UTI) de sete hospitais de ensino do Brasil, e analisar a significância clínica das mesmas. Foram coletadas informações sobre medicamentos e NE administrados com 24 horas e 120 horas de internação. Para identificação das interações fármaco-NE empregou-se o software Drug Reax®. Foram investigados 1.124 prontuários. Destes, 320 pacientes, com 24 horas de internação, estavam em uso de NE, sendo que 20 (6,3%) apresentaram interação fármaco-NE. Dos 504 pacientes, com 120 horas de internação, 39 (7,7%) apresentaram interação fármaco-NE. As potenciais interações fármaco-NE mais frequentes foram: fenitoína-NE, levotiroxina-NE e varfarina-NE. As interações fármaco-NE podem interferir na qualidade e no custo da assistência prestada em UTI e, desta forma, é essencial que a equipe de saúde tenha conhecimento sobre as mesmas.

This is a multicenter, cross-sectional retrospective study, which aimed to determine the prevalence of interactions drug-enteral nutrition (EN) in Intensive Care Units (ICU) of seven teaching hospitals in Brazil, and to analyze the clinical significance of them. Information on medications and EN administered with 24 hours and 120 hours of hospitalization were collected. For identification of drug-EN interaction was employed software Drug Reax®. It was investigated 1.124 records. Of these, 320 patients, with 24 hours of hospitalization, were on EN, and 20 (6.3%) had drug-EN interaction. Of the 504 patients with 120 hours of hospitalization, 39 (7.7%) had drug-EN interaction. The most frequent drug-EN interactions of clinical significance were phenytoin-EN, warfarin-EN and levothyroxine-EN. Drug-EN interactions may interfere with the quality and cost effectiveness of care in ICU, so it is essential that the health team has knowledge about them.

Estudio multicéntrico, transversal retrospectivo, que tuvo como objetivo determinar la prevalencia de interacciones drogasnutrición enteral (NE) en Unidades de Cuidados Intensivos (UCI) de siete hospitales de enseñanza en Brasil, y analizar el significado clínico de las mismas. Fueran colectadas informaciones en 24 horas y 120 horas de hospitalización en los registros médicos. Para identificar las interacciones fármaco-NE se utilizó el software de Drug Reax®. Fueran investigados 1.124 registros. De éstos, 320 pacientes, con 24 horas de hospitalización, se encontraban en la NE, 20 (6,3%) presentarán interacción. De los 504 pacientes, con 120 horas de hospitalización, 39 (7,7%) tuvieron la interacción. Las interacciones fármacos-NE más frecuentes fueron: fenitoína-NE, levotiroxina-NE, warfarina-NE. Las interacciones fármacos-NE pueden interferir con la calidad de los cuidados en la UCI, por lo que es esencial que el equipo de salud tenga conocimientos al respecto.

Perfil de isotipos de imunoglobulinas e subclasses de IgG na leishmaniose tegumentar americana / Immunoglobulin isotype and IgG subclass profiles in american tegumentary leishmaniasis

O presente trabalho avaliou o perfil de anticorpos em amostras de soro de 37 pacientes com diagnóstico clínico confirmado ou compatível com leishmaniose tegumentar americana atendidos no Hospital de Clínicas da Universidade Federal de Uberlândia, MG. Os perfis das classes de imunoglobulinas e subclasses de IgG foram analisados pelo teste ELISA indireto, utilizando-se antígeno solúvel de Leishmania (Leishmania) amazonensis. A avidez dos anticorpos foi determinada pelo tratamento com uréia a 6 M, após incubação dos soros com o antígeno. Observou-se que 97 por cento, 94,6 por cento, 57,5 e 21,5 por cento das amostras testadas apresentaram anticorpos anti-Leishmania das classes IgE, IgG, IgA e IgM, respectivamente e, os perfis das subclasses de IgG demonstraram, IgG1>IgG3>IgG2>IgG4. Os anticorpos IgE anti-Leishmania de alta avidez corresponderam a 44,4 por cento. Por outro lado, IgG e IgA anti-Leishmania foram em sua maioria (62,8 e 47,8 por cento, respectivamente), de média avidez. A variação do perfil de isotipos, bem como a avidez das imunoglobulinas refletiu a complexidade da resposta imune humoral contra a leishmaniose tegumentar americana.

Isolation and partial characterization of a D-galactose-binding lectin from the latex of Synadenium carinatum

No presente trabalho, foi purificada por cromatografia de afinidade em D-galactose imobilizada em agarose, uma lectina do latex de Synadenium carinatum (ScLL). Essa lectina é uma potente aglutinina para eritrócitos humanos, cuja atividade hemaglutinante foi inibida com 3,0 mM de N acetil-D-galactopiranosidio, 6,3 mM de metil-b-D-galactopiranosidio ou 50 mM metil-a-D-pironosidio ou D-fucose, porém, nenhuma inibição foi evidenciada por L-fucose, revelando uma especificidade anomérica e conformacional da lectina. A análise por SDS-PAGE dessa lectina pareceu ser uma glicoproteína composta por duas cadeias polipeptídicas de aproximadamente 28 e 30 kDa, porém, em cromatografia de exclusão por tamanho sobre Sephadex G100 e em gel nativo apresentou um peso molecular aparente de 120-130 kDa, a qual mostrou ser composta de uma mistura de subunidades de peptídeos de 28 e 30 kDa. Essa lectina manteve-se estável em pH de 6 a 9 e temperatura de 4 a 56ºC. A seqüência N-terninal contem uma região conservada GPN a qual também é observada em outras lectinas de látex de outras Euphorbiaceas ligante de D-galactose.

Immunoblotting analyses using two-dimensional gel electrophoresis of Trypanosoma cruzi excreted-secreted antigens

Formas tripomastigotas de Trypanosoma cruzi excretam/secretam uma complexa mistura de moléculas antigênicas. Essa mistura é chamada trypomastigote excreted-secreted antigens e contém uma banda de massa molecular em torno de 150-160kDa que possui excelente performance para diagnóstico de doença de Chagas em immunoblotting. No presente estudo foi caracterizado parcialmente, por gel bidimensional, a proteína de 150-160kDa pela análise da reatividade de anticorpos de pacientes chagásicos nas diversas fases da doença. Proteínas do trypomastigote excreted-secreted antigens foram separadas por eletroforese de alta resolução em duas dimensões (2D) e submetidas a immunoblotting com soros de pacientes chagásicos e não chagásicos. A proteína de 150-160kDa foi identificada em quatro isoformas com pontos isoelétricos variando entre 6,2 a 6,7. As quatro isoformas foram reconhecidas por anticorpos IgM na fase aguda e por anticorpos IgG na fase crônica da doença de Chagas. A isoforma de 150-160kDa, com ponto isoelétrico de aproximadamente 6,4 tornou-se imunodominante dentre as demais com a progressão da doença. Não foi detectada reatividade cruzada com os soros de pacientes não chagásicos ou pacientes infectados com Leishmania sp. Os dados obtidos nesse trabalho, reforçam a importância da utilização do trypomastigote excreted-secreted antigens para o diagnóstico sorológico da doença de Chagas.