Rat hepatocyte invasion by Listeria monocytogenes and analysis of TNF-alpha role in apoptosis

Listeria monocytogenes, etiological agent of severe human foodborne infection, uses sophisticated mechanisms of entry into host cytoplasm and manipulation of the cellular cytoskeleton, resulting in cell death. The host cells and bacteria interaction may result in cytokine production as Tumor Necrosis Factor (TNF) alpha. Hepatocytes have potential to produce pro-inflammatory cytokines as TNF-alpha when invaded by bacteria. In the present work we showed the behavior of hepatocytes invaded by L. monocytogenes by microscopic analysis, determination of TNF-alpha production by bioassay and analysis of the apoptosis through TUNEL technique. The presence of bacterium, in ratios that ranged from 5 to 50,000 bacteria per cell, induced the rupture of cellular monolayers. We observed the presence of internalized bacteria in the first hour of incubation by electronic microscopy. The levels of TNF-alpha increased from first hour of incubation to sixth hour, ranging from 0 to 3749 pg/mL. After seven and eight hours of incubation non-significant TNF-alpha levels decrease occurred, indicating possible saturation of cellular receptors. Thus, the quantity of TNF-alpha produced by hepatocytes was dependent of the incubation time, as well as of the proportion between bacteria and cells. The apoptosis rate increased in direct form with the incubation time (1 h to 8 + 24 h), ranging from 0 to 43%, as well as with the bacteria : cells ratio. These results show the ability of hepatocyte invasion by non-hemolytic L. monocytogenes, and the main consequences of this phenomenon were the release of TNF-alpha by hepatocytes and the induction of apoptosis. We speculate that hepatocytes use apoptosis induced by TNF-alpha for release bacteria to extracellular medium. This phenomenon may facilitate the bacteria destruction by the immune system.

The pattern of immune cell infiltration in chromoblastomycosis: involvement of macrophage inflammatory protein-1 alpha/CCL3 and fungi persistence

Chromoblastomycosis (CR) is a subcutaneous chronic mycosis characterized by a granulomatous inflammatory response. However, little is known regarding the pattern of leukocyte subsets in CR and the pathways involved in their recruitment. The objective of this study was to assess the cellular subsets, chemokine, chemokine receptors and enzymes in CR. The inflammatory infiltrate was characterized by immunohistochemistry using antibodies against macrophages (CD68), Langerhans'cells (S100), lymphocytes (CD3, CD4, CD8, CD45RO, CD20 and CD56) and neutrophils (CD15). The expression of MIP-1alpha (Macrophage inflammatory protein-1alpha), chemokine receptors (CXCR3 and CCR1) and enzymes (superoxide dismutase-SOD and nitric oxide synthase-iNOS) was also evaluated by the same method. We observed an increase in all populations evaluated when compared with the controls. Numbers of CD15+ and CD56+ were significantly lower than CD3+, CD4+, CD20+ and CD68+ cells. Statistical analysis revealed an association of fungi numbers with CD3, CD45RO and iNOS-positive cells. Furthermore, MIP-1alpha expression was associated with CD45RO, CD68, iNOS and CXCR3. Our results suggest a possible role of MIP-1alpha and fungi persistence in the cell infiltration in CR sites.

Cryoglobulinemia in chronic hepatitis C: clinical aspects and response to treatment with interferon alpha and ribavirin

INTRODUCTION: The main extra-hepatic manifestation of hepatitis C is mixed cryoglobulinemia (MC). The aim of this study was to evaluate its prevalence among patients with chronic hepatitis C (CHC), to correlate its presence to host and virological variables and to the response to combined therapy with interferon-alpha and ribavirin. CASUISTIC AND METHODS: 202 CHC naive patients (136 with chronic hepatitis and 66 with cirrhosis) were consecutively evaluated for the presence of cryoglobulins. Cryoprecipitates were characterized by immunoelectrophoresis and classified according to the Brouet's criteria. RESULTS: The prevalence of MC was 27% (54/202), and 24% of them (13/54) showed major clinical manifestation of the disease. Even though type III MC was more frequent (78%), symptomatic MC was more common in type II MC. The presence of cirrhosis (RR = 2.073; IC95% = 1.029 - 4.179; p = 0.041), and age of the patients (RR = 1.035; IC95% = 1.008 - 1.062; p = 0.01) were independently associated with the presence of cryoglobulins. No relationship was found with viral load and genotype. 102 patients were treated with interferon alpha and ribavirin. Among these, 31 had MC. Sustained virological response (around 30%) was similar in patients with and without MC (p = 0.971). CONCLUSION: MC represents a prevalent complication in patients with CHC, specially older and cirrhotic patients. Only 24% of these patients show clinical manifestation of the disease, specially those with type II MC. The presence of MC did not affect the response to therapy.

Estudo evolutivo da leishmaniose mucosa (7 a 17 anos de seguimento) causada por Leishmania (Viannia ) braziliensis em Três Braços, Bahia

No período de agosto a dezembro de 1993 realizou-se um levantamento de 77 pacientes da casuística de leishmaniose mucosa do programa de Três Braços, registrada no período de 1976 a 1986. Encontrou-se 65 pacientes vivos e 12 falecidos. Os primeiros foram submetidos a exame clínico geral e otorrinolaringológico e à pesquisa de anticorpos por imunofluorescência indireta. Os pacientes com lesões ativas foram submetidos à biópsia da mucosa nasal para isolar o parasita em meio de cultura NNN a partir da inoculação direta do material ou das lesões desenvolvidas no hamster e para estudo histopatológico. Todos os isolados foram caracterizados como Leishmania (Viannia) braziliensis utilizando anticoipos monoclonais. Cinqüenta e seis (86%) pacientes estavam curados e nove (13%) encontravam-se com lesões ativas. Entre os pacientes falecidos, cinco (41%) estavam curados no momento do óbito. O programa teve 79% de pacientes curados ao longo de 17 anos. O tempo médio de seguimento clínico foi de 10 anos (7-17 anos). A doença como causa-mortis foi sugerida em 3 casos.

Evaluation of IFN-gamma and TNF-alpha as immunological markers of clinical outcome in cutaneous leishmaniasis

To evaluate if IFN-gamma and TNF-alpha levels could be used as markers of therapeutic response in cutaneous leishmaniasis, 54 patients with history of one ulcerated cutaneous lesion, with up to 30 days onset, were enrolled in the study. IFN-gammaand TNF-alpha were measured by ELISA in lymphocyte cultures supernatant before and 60 days after initiating therapy. Cure was considered to be a complete healing of lesion 60 days after treatment. IFN-gamma and TNF-alpha levels were similar in both groups of patients before therapy. There was a tendency to increase IFN-gamma levels in patients that were cured in 60 days, however the values did not reach statistical significance. In both groups of patients, TNF-alpha levels were similar before therapy and fell significantly after treatment, irrespective of cure or maintenance of active lesion.

Autoimmune hemolytic anemia in HCV/HIV coinfected patients during treatment with pegylated alpha-2a interferon plus ribavirin

Two cases of autoimmune hemolytic anemia that occurred during the treatment of chronic hepatitis C with pegylated alpha-2a interferon and ribavirin, in HIV coinfected patients, are presented and described. The late occurrence (after six months of therapy) of this severe hemolytic anemia leads to the recommendation that hemoglobin levels should be monitored throughout the treatment period, even among patients who presented stable hemoglobin levels in the preceding months.

Serum levels of interleukin-6, tumor necrosis factor-alpha and interferon-gama in infants with and without dengue

This study compared the serum levels of IL-6, TNF-alpha and IFN-gamma, in children under 1 year of age with and without dengue. Sera were collected from a total of 41 children living in the Department of Antioquia, Colombia (27 patients with dengue and 14 controls). The results showed higher cytokine levels in children with dengue than without dengue, with statistically significant differences for IL-6 and IFN-gamma. No statistically significant differences were found between clinical forms, although IL-6 and IFN-gamma levels were higher in dengue fever cases than in dengue hemorrhagic fever cases. On the other hand, TNF-alpha levels were higher in dengue hemorrhagic fever than in dengue fever. The levels of IL-6 and TNF-alpha were higher in secondary infection than in primary infection, although IFN-gamma levels were higher in primary infection. These results suggest that IL-6, TNF-alpha and IFN-gamma are involved in dengue infection independently of the clinical form.

Schistosomiasis in Southern Brazil 17 years after the confirmation of the first autochthonous case

INTRODUCTION: Schistosomiasis, a parasitic disease, is an important public health issue in Brazil, particularly Northern Brazil. Since the first recorded occurrence of Biomphalaria glabrata in the States of Santa Catarina and Rio Grande do Sul, it has spread to South Brazil. METHODS: Using the Information System for Notifiable Diseases (SINAN), we assessed the disease spread in Southern Brazil. RESULTS: In Rio Grande do Sul, nine localities had confirmed cases (n = 95, 2001-2013). CONCLUSIONS: We confirmed disease expansion to Southern Brazil. We demonstrated the effectiveness of SINAN to monitor notifiable diseases. Our results are useful to develop preventive actions for schistosomiasis control.

Susceptibility of Triatoma sordida Stal, 1859 (Hemiptera: Reduviidae) to alpha-cypermethrin under natural climatic conditions

Abstract:INTRODUCTION:Despite the recommendations by interpretation of resistance ratios obtained in laboratory bioassays, little is known about the actual impact of these results in the effectiveness of vector control activities in the field. In this context, our objective was to determine the mean value of different resistance ratios obtained by laboratory bioassays performed as part of the chemical control strategies of Triatoma sordida in the field.METHODS:Field bioassays were developed in Monte Azul and Coração de Jesus (Southeast, Brazil). In each location, samples were formed with three domestic units treated with alpha-cypermethrin 20.0% (Alfatek (r) 200 SC). One day after spraying, 10 fifth-instar nymphs remained in contact with the surfaces treated (adobe with plaster, adobe without plaster, or wood) with insecticide in plastic cones for 72h. Three cones were exposed inside the intradomicile and the peridomicile. The insects in the control group were exposed to an insecticide-free piece of cardboard. Mortality was measured 72h after removal of the insects from the treated surfaces. The tests were realized in triplicate.RESULTS:Mortality was 100.0% in all locations, except for Monte Azul; Landinho (96.6%) and Coração de Jesus; Barriguda (96.6%).CONCLUSIONS:Although the resistant populations in laboratory tests proved to be susceptible in the field, this observation is not sufficient to suggest that the cut-off points used to justify the resistance ratio should be changed. In this sense, we recommend that laboratory and field bioassays are carried out with a greater number of Triatominae populations to allow more in-depth consideration of the subject.

Preoperative serum levels of ca 72-4, cea, ca 19-9, and Alpha-fetoprotein in patients with gastric cancer

INTRODUCTION: The clinical importance of preoperative serum levels of CA 72-4, carcinoembryonic antigen (CEA), CA 19-9, and alpha-fetoprotein (AFP) was prospectively evaluated in 44 patients with gastric cancer. METHOD: The serum tumor marker levels were determined by commercial radioimmunoassay kits. Positivity for CA 72-4 (>4 U/mL), CEA (>5 ng/mL), CA 19-9 (>37 U/mL), and AFP (>10 ng/mL) were correlated according to the stage, histology, and lymph node metastasis. RESULTS AND DISCUSSION: CA 72-4 showed a higher positivity rate for gastric cancer (47.7%) than CEA (25%), CA 19-9 (25%), and AFP (0%). The combination of CA 72-4 with CEA and CA 19-9 increased the sensitivity to 61.4%. The positivity rates of CA 72-4 in patients at stages I and II (initial disease) and in patients at stages III and IV (advanced disease) were 9% and 60.6%, respectively (P < 0.005). No correlation was found between CEA and CA 19-9 levels and the stage of gastric cancer. There was a tendency of positivity for CA 72-4 to suggest lymph node involvement, but it was not significant (P = 0.075). Serum levels of tumor markers did not show a correlation with the histological types of gastric cancer. CONCLUSION: Preoperative serum levels of CA 72-4 provided a predictive value in indicating advanced gastric cancer.

Ants obtained from trees of a "Jacareuba" (Calophyllum brasiliense) forest plantation in Central Amazonia by canopy fogging: first results

Canopies of the locally occurring Amazonian tree species Calophyllum brasiliense. Camb. (Guttiferae, height 10 m) were fogged after dawn on the same day at five adjacent localities in a 20-year-old plantation at the Adolpho Ducke Forest Reserve near Manaus/Brazil, using natural pyrethrum (0.5%, 1.0%, 1.5%), Baythroid (0.3%) and diesel oil, respectively. The 1152 ants collected represented 52 species, 17 genera and five subfamilies. The highest number of ant species was obtained with the synthetic pyrethrum Baythroid 0.3% and with natural pyrethrum 1.5%. About 46% of the total ant species were only collected in the first hour after fogging, and 17% only during the second hour, after trees had been heavily shaken. About 73% of the total ant specimens dropped in the first hour. Data on alpha-diversity, eveness and frequency of ants indicated a rapidly decreasing knock-down effect of both the synthetic and natural pyrethrum after application. The ant fauna in the canopy of C. brasiliense is somewhat distinct compared with that of Goupia glabra Aubl. (Celastraceae, height 45 m) from the primary forest of the reserve.

Efeitos de área e de borda sobre a estrutura florestal em fragmentos de floresta de terra-firme após 13-17 anos de isolamento

As estimativas de densidade e biomassa de árvores vivas com DAP > 10 cm e arvoretas 1-9,9 cm de DAP, liteira lenhosa grossa caída (LCG diâmetro > 10 cm), árvores mortas em pé (> 10 de DAP) e liteira lenhosa fina caída (LCF - 2,5 9,9 cm de diâmetro) foram quantificadas em 56 parcelas permanentes de 1 ha, distribuídas em quatro categorias de tamanho de fragmento - fragmentos de 1 ha (4 parcelas), fragmentos de 10 ha (12 parcelas) e fragmentos de 100 ha (14 parcelas) e floresta contínua (19 parcelas) e em duas classes de distância da borda - < 300 m de distância da borda (29 parcelas) e > 300 m (21 parcelas). A densidade e a biomassa de árvores e arvoretas de espécies de estágios sucessionais mais avançados não diferiram significativamente entre as diferentes categorias de tamanho e entre as duas distâncias da borda. Por outro lado, fragmentos florestais e locais < 300 m de distâcia da borda tiveram maior biomassa e densidade de árvores e arvoretas de espécies pioneiras do que floresta contínua e locais > 300 m da borda, respectivamente. Fragmentos florestais apresentaram maior quantidade de LCG e LCF do que a floresta contínua. Houve também diferenças significativas entre ambas as distâncias da borda para a quantidade de LCG e LCF e necromassa total. Uma análise de covariância mostrou que não houve efeito de tamanho do fragmento, mas a distância da borda teve um efeito significativo sobre a quantidade de LCG e LCF. A quantidade de LCG e LCF foi correlacionada negativamente com a distância da borda - locais mais próximos à borda tiveram cerca de 40% e 60% mais LCG do que locais mais distantes.