Estudo da influência dos genótipos 1 e 3 do vírus da hepatite C sobre os indicadores do metabolismo lipídico em hepatopatas crônicos

Os perfis metabólicos correlacionam-se com a infecção pelo VHC e são prognósticos da resposta viral em pacientes crônicos. Porém, pouco se sabe a respeito da associação entre perfis lipídicos e carga viral do VHC entre infecções dos genótipos 1, 2 ou 3. Portanto, o objetivo deste trabalho foi estudar a influência da viremia e dos genótipos do VHC sobre o metabolismo lipídico através das variações de lipoproteínas séricas (colesterol total, LDL, HDL, VLDL, triglicérides) e apolipoproteína B (Apo B) em hepatopatas crônicos, avaliando se o VHC predispõe os indivíduos ao aparecimento de complicações vasculares. O grupo amostral constituiu-se de um total de 150 pacientes crônicos do VHC com genótipos 1, 2 ou 3, e de um grupo controle de 20 indivíduos saudáveis (10 homens e 10 mulheres) em idade adulta (20 à 50 anos). Os níveis séricos de HDL (28%), VLDL (26%) e triglicérides (26%) nos portadores crônicos do VHC se mostraram diminuídos em relação ao grupo controle, enquanto os níveis de LDL (25%) e Apo B (29%) se mostraram elevados, resultados que foram mais importantes nos portadores do genótipo 3a. Observou-se correlação positiva entre a viremia e alterações nos níveis de apo B (r = 0,5763) nos portadores do genótipo 1b. Assim, foi pressuposto que o risco de pacientes portadores do VHC desenvolverem complicações vasculares é elevado, pois 1% de redução nos níveis de LDL está associado com uma redução de 2-3% no risco de desenvolvimento de doenças cardíacas, e como cerca de 90% da proteína na LDL se constitui de apo B, sua concentração plasmática indica o número total de partículas potencialmente aterogênicas. Desta forma, o perfil lipídico auxilia no diagnóstico da severidade da infecção hepática causada pelo VHC e ainda atua como um bom sinal prognóstico.The metabolic profiles correlate with the hepatitis C virus infection and are prognostics for the viral reply in chronic patients. However, little is known regarding the distinguishing association between lipid profiles and hepatitis C viral load in patients carrying genotypes 1, 2 or 3. Therefore, the objective of this work was to study viremia and genotypes on the lipid metabolism through the serum lipoprotein variations (total cholesterol, LDL, HDL, VLDL, triglycerides) and apolipoprotein B (Apo B) in chronic carriers of this infection, evaluating if the HCV premakes the individual to the lipidic disequilibrium and favors the appearance of vascular complications. The amostral group consisted of 150 HCV chronic patients with genotypes 1, 2 or 3, and a control group consisted of 20 healthful individuals (10 men and 10 women) in adult age (20 to 50 years). The levels of HDL (28%), VLDL (26%) and triglycerides (26%) of the HCV chronic patients were lower than the control group, while the LDL levels (25%) and the Apo B levels (29%) were higher. These findings were more significant in the genotype 3a carrying patients. Positive correlation occurred between the viremia and the alterations in the Apo B levels (r = 0.5763) in the genotype 1b carrying patients. Consequently it was inferred that the risk of HCV patients to develop vascular complication is elevated. In general, 1% of reduction in the LDL levels is associated with a reduction of 2-3% in the risk of development of cardiac illnesses, and, as about 90% of the protein in the LDL is constituted of apo B, its plasmatic concentration indicates the total potentially atherogenics particles number. The lipid profile aids in the diagnosis of the severity of the hepatic infection and equally acts as a good signal prognostic, therefore its analysis must be carried through in all the cases of advanced hepatic infection.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES

Busca de antígenos para diagnóstico da leishmaniose visceral canina: potencial uso e aplicação da proteína rLc36

A leishmaniose visceral, forma clínica mais severa das leishmanioses, pode ser fatal se não tratada. Cães infectados são reservatórios potenciais da doença, sendo um diagnóstico específico e cuidadoso requerido para a identificação dos animais infectados, com o objetivo de promover um melhor controle epidemiológico e, consequentemente, controlar a transmissão da doença para o homem. Diversos trabalhos na literatura descrevem vários testes utilizados atualmente para diagnóstico da leishmaniose visceral canina (LVC), porém, apresentam variada especificidade e sensibilidade. Dessa forma, o desenvolvimento de um teste diagnóstico com maior sensibilidade e especificidade se torna importante para correta identificação de cães infectados. O presente trabalho teve como objetivo caracterizar a proteína rLc36, codificada por um gene exclusivo de Leishmania infantum, quanto ao potencial antigênico para desenvolvimento de um teste diagnóstico mais sensível e específico para LVC baseado no ensaio ELISA (Enzyme Linked Immuno Sorbent Assay). Diferentes concentrações desta proteína foram testadas por ELISA usando soros de cães infectados com LV e soros de cães não infectados. A concentração de 1,0?g/mL de rLc36 foi capaz de diferenciar os soros positivos dos soros negativos e apresentou sensibilidade de 85% e especificidade de 71%, em um teste com 95% de confiança, mostrando que a proteína rLc36 é antigênica e apresenta potencial para ser aplicada no desenvolvimento de um kit diagnóstico. A proteína rLc36 foi testada em associação com a proteína A2, o qual já foi previamente avaliado como antígeno no diagnóstico sorológico da leishmaniose, e foi possível observar aumento na sensibilidade do teste (77%), mas não na especificidade (79%). Soros de outras parasitoses foram testados frente à proteína rLc36. Foi possível observar reatividade com os soros positivos para babesiose...Visceral leishmaniasis, the most severe clinical form of the leishmaniasis, can be fatal if untreated. Infected dogs are potential reservoirs of the disease, being a specific and careful diagnosis required for identification of the infected animals, with the aim of promoting a better epidemiological control and consequently control the transmission of the disease to humans. Several works in the literature describe various tests used for diagnosis of canine visceral leishmaniasis (CVL), however, they show varied specificity and sensitivity. In this way, the development of a diagnosis test with greater sensitivity and specificity becomes important for the correct identification of infected dogs. Thus, the development of a more sensitive and specific diagnosis test based on recombinant antigens may contribute in the correct identification of dogs truly infected and thus contribute to the more effective control of the transmission of the disease. The present work had as objective to characterize the protein rLc36, encoded by a gene unique to Leishmania infantum, for the antigenic potential for development of a more sensitive and specific test for CVL based on the ELISA (Enzyme Linked Immuno Sorbent Assay). Different protein concentrations were tested by ELISA using sera from dogs infected with LV and sera from non-infected dogs. The concentration of 1.0?g/mL of rLc36 protein was able to differentiate positive from negative sera and showed sensitivity of 85% and specificity of 71% on a test with 95% of confidence, showing that the protein rLc36 is antigenic and has potential for be applied in a new diagnosis test. The rLc36 protein was tested in association with the A2 protein, which was previously assessed as antigen in serological diagnosis of leishmaniasis, and it was possible to observe an increase in the sensitivity of the test (77%), but no increase was observed in the specificity (79%). Sera from..

Avaliação do perfil lipídico entre indivíduos com hepatite C

Metabolic profiles correlate with hepatitis C virus (HCV) infection and are prognostic for the viral response. However, little is known about the association between lipid profiles and viral load in chronic patients carrying HCV genotypes 1, 2 and 3. The aim of this study was to investigate the influence of the viremia and viral genotype on lipid metabolism by observing the variations in serum lipoprotein and apolipoprotein B, to assess whether HCV predisposes individuals to lipid imbalance and favors the appearance of vascular complications. A sample group of 150 chronic HCV patients with viral genotypes 1, 2 or 3 and a control group of 20 healthy adults (10 men and 10 women), all aged from 20 to 50 years were studied. The serum lipid profile of the chronic patients was analyzed and compared to that of the control group. The high-density lipoprotein (HDL), very low-density lipoprotein (VLDL) and triglyceride levels of the sample group were lower than those of the control group, while the low-density lipoprotein (LDL) and apolipoprotein B levels of the patients were higher. These differences were more significant in patients carrying genotype 3a. There was a positive correlation between the viremia and the changes in apolipoprotein B levels in patients carrying genotype 1b. It was inferred that the risk of developing vascular complications raised in HCV patients. As 90% of LDL protein is composed of apolipoprotein B, the plasmatic concentration of the latter indicates the number of potentially atherogenic particles. Therefore, the lipid profile monitoring may aid in the diagnosis of hepatic infection severity and equally act as a good prognostic marker

Evaluation of the lipid profile between individuals with hepatitis C

Metabolic profiles correlate with hepatitis C virus (HCV) infection and are prognostic for the viral response. However, little is known about the association between lipid profiles and viral load in chronic patients carrying HCV genotypes 1, 2 and 3. The aim of this study was to investigate the influence of the viremia and viral genotype on lipid metabolism by observing the variations in serum lipoprotein and apolipoprotein B, to assess whether HCV predisposes individuals to lipid imbalance and favors the appearance of vascular complications. A sample group of 150 chronic HCV patients with viral genotypes 1, 2 or 3 and a control group of 20 healthy adults (10 men and 10 women), all aged from 20 to 50 years were studied. The serum lipid profile of the chronic patients was analyzed and compared to that of the control group. The high-density lipoprotein (HDL), very low-density lipoprotein (VLDL) and triglyceride levels of the sample group were lower than those of the control group, while the low-density lipoprotein (LDL) and apolipoprotein B levels of the patients were higher. These differences were more significant in patients carrying genotype 3a. There was a positive correlation between the viremia and the changes in apolipoprotein B levels in patients carrying genotype 1b. It was inferred that the risk of developing vascular complications raised in HCV patients. As 90% of LDL protein is composed of apolipoprotein B, the plasmatic concentration of the latter indicates the number of potentially atherogenic particles. Therefore, the lipid profile monitoring may aid in the diagnosis of hepatic infection severity and equally act as a good prognostic marker

Discovery of a Novel Lineage Burkholderia cepacia ST 1870 Endophytically Isolated from Medicinal Polygala paniculata Which Shows Potent In Vitro Antileishmanial and Antimicrobial Effects

In this study, we report the isolation and identification of an endophytic strain of Burkholderia cepacia (COPS strain) associated with Polygala paniculata roots. Polygala plants are rich sources of promising microbiomes, of which the literature reports several pharmacological effects, such as trypanocidal, antinociceptive, anesthetic, anxiolytics, and anticonvulsant activities. B. cepacia COPS belongs to a new sequence type (ST 1870) and harbors a genome estimated in 8.3 Mbp which exhibits the aminoglycosides and beta-lactams resistance genes aph(3′)-IIa and blaTEM-116, respectively. Analysis performed using MLST, average nucleotide identity, and digital DNA-DNA hybridization support its species-level identification and reveals its novel housekeeping genes alleles gyrB, lepA, and phaC. The root endophyte B. cepacia COPS drew our attention from a group of 14 bacterial isolates during the primary screening for being potentially active against Staphylococcus aureus ATCC 29213, Enterococcus faecalis ATCC 29212, Micrococcus luteus ATCC 9341, Escherichia coli ATCC 25922, and Candida albicans ATCC 10231 and exhibited the broad-spectrum activity against phytopathogenic fungi. In addition, COPS strain showed production of protease, lipase, and esterase in solid media, and its natural product extract showed potent inhibition against fungal plant pathogens, such as Moniliophthora perniciosa, whose antagonism index (89.32%) exceeded the positive control (74.17%), whereas Sclerotinia sclerotiorum and Ceratocystis paradoxa showed high percentages of inhibition (85.53% and 82.69%, respectively). COPS crude extract also significantly inhibited S. epidermidis ATCC 35984, E. faecium ATCC 700221 (MIC values of 32 μg/mL for both), E. faecalis ATCC 29212 (64 μg/mL), and S. aureus ATCC 25923 (128 μg/mL). We observed moderate antagonistic activity against A. baumannii ATCC 19606 and E. coli ATCC 25922 (both at 512 μg/mL), as well as potent cytotoxic effects on Leishmania infantum and Leishmania major promastigote forms with 78.25% and 57.30% inhibition. In conclusion, this study presents for the first time the isolation of an endophytic B. cepacia strain associated with P. paniculata and enough evidence that these plants may be considered a rich source of microbes for the fight against neglected diseases

In Vitro Modulator Effect of Total Extract from the Endophytic Paenibacillus polymyxa RNC-D in Leishmania (Leishmania) amazonensis and Macrophages

Leishmaniases are diseases with high epidemiological relevance and wide geographical distribution. In Brazil, Leishmania (Leishmania) amazonensis is related to the tegumentary form of leishmaniasis. The treatment for those diseases is problematic as the available drugs promote adverse effects in patients. Therefore, it is important to find new therapeutic targets. In this regard, one alternative is the study of biomolecules produced by endophytic microorganisms. In this study, the total extract produced by the endophytic Paenibacillus polymyxa RNC-D was used to evaluate the leishmanicidal, nitric oxide, and cytokines production using RAW 264.7 macrophages. The results showed that, in the leishmanicidal assay with L. amazonensis, EC50 values at the periods of 24 and 48 hours were 0.624 mg/mL and 0.547 mg/mL, respectively. Furthermore, the cells treated with the extract presented approximately 25% of infected cells with an average of 3 amastigotes/cell in the periods of 24 and 48 hours. Regarding the production of cytokines in RAW 264.7 macrophages infected/treated with the extract, a significant increase in TNF-α was observed at the periods of 24 and 48 hours in the treated cells. The concentrations of IFN-γ and IL-12 showed significant increase in 48 hours. A significant decrease in IL-4 was observed in all cells treated with the extract in 24 hours. It was observed in the treated cells that the NO production by RAW 264.7 macrophages increased between 48 and 72 hours. The endophytic Paenibacillus polymyxa RNC-D extract modulates the mediators of inflammation produced by RAW 264.7 macrophages promoting L. amazonensis death. The immunomodulatory effects might be a promising target to develop new immunotherapeutic and antileishmanial drugs

Inflammatory response of endothelial cells to hepatitis C virus recombinant envelope glycoprotein 2 protein exposure

The hepatitis C virus (HCV) encodes approximately 10 different structural and non-structural proteins, including the envelope glycoprotein 2 (E2). HCV proteins, especially the envelope proteins, bind to cell receptors and can damage tissues. Endothelial inflammation is the most important determinant of fibrosis progression and, consequently, cirrhosis. The aim of this study was to evaluate and compare the inflammatory response of endothelial cells to two recombinant forms of the HCV E2 protein produced in different expression systems (Escherichia coli and Pichia pastoris). We observed the induction of cell death and the production of nitric oxide, hydrogen peroxide, interleukin-8 and vascular endothelial growth factor A in human umbilical vein endothelial cells (HUVECs) stimulated by the two recombinant E2 proteins. The E2-induced apoptosis of HUVECs was confirmed using the molecular marker PARP. The apoptosis rescue observed when the antioxidant N-acetylcysteine was used suggests that reactive oxygen species are involved in E2-induced apoptosis. We propose that these proteins are involved in the chronic inflammation caused by HCV

Recombinant hepatitis C virus-envelope protein 2 interactions with low-density lipoprotein/CD81 receptors

Hepatitis C virus (HCV) envelope protein 2 (E2) is involved in viral binding to host cells. The aim of this work was to produce recombinant E2B and E2Y HCV proteins in Escherichia coli and Pichia pastoris, respectively, and to study their interactions with low-density lipoprotein receptor (LDLr) and CD81 in human umbilical vein endothelial cells (HUVEC) and the ECV304 bladder carcinoma cell line. To investigate the effects of human LDL and differences in protein structure (glycosylated or not) on binding efficiency, the recombinant proteins were either associated or not associated with lipoproteins before being assayed. The immunoreactivity of the recombinant proteins was analysed using pooled serum samples that were either positive or negative for hepatitis C. The cells were immunophenotyped by LDLr and CD81 using flow cytometry. Binding and binding inhibition assays were performed in the presence of LDL, foetal bovine serum (FCS) and specific antibodies. The results revealed that binding was reduced in the absence of FCS, but that the addition of human LDL rescued and increased binding capacity. In HUVEC cells, the use of antibodies to block LDLr led to a significant reduction in the binding of E2B and E2Y. CD81 antibodies did not affect E2B and E2Y binding. In ECV304 cells, blocking LDLr and CD81 produced similar effects, but they were not as marked as those that were observed in HUVEC cells. In conclusion, recombinant HCV E2 is dependent on LDL for its ability to bind to LDLr in HUVEC and ECV304 cells. These findings are relevant because E2 acts to anchor HCV to host cells; therefore, high blood levels of LDL could enhance viral infectivity in chronic hepatitis C patients

Potential application of rLc36 protein for diagnosis of canine visceral leishmaniasis

Visceral leishmaniasis (VL) is fatal if left untreated. Infected dogs are important reservoirs of the disease, and thus specific identification of infected animals is very important. Several diagnostic tests have been developed for canine VL (CVL); however, these tests show varied specificity and sensitivity. The present study describes the recombinant protein rLc36, expressed by Leishmania infantum, as potential antigen for more sensitive and specific diagnosis of CVL based on an immunoenzymatic assay. The concentration of 1.0 μg/mL of rLc36 enabled differentiation of positive and negative sera and showed a sensitivity of 85% and specificity of 71% (with 95% confidence), with an accuracy of 76%