Dengue infection in mice inoculated by the intracerebral route: neuropathological effects and identification of target cells for virus replication.

Dengue is an important arboviral infection, causing a broad range symptom that varies from life-threatening mild illness to severe clinical manifestations. Recent studies reported the impairment of the central nervous system (CNS) after dengue infection, a characteristic previously considered as atypical and underreported. However, little is known about the neuropathology associated to dengue. Since animal models are important tools for helping to understand the dengue pathogenesis, including neurological damages, the aim of this work was to investigate the effects of intracerebral inoculation of a neuroadapted dengue serotype 2 virus (DENV2) in immunocompetent BALB/c mice, mimicking some aspects of the viral encephalitis. Mice presented neurological morbidity after the 7th day post infection. At the same time, histopathological analysis revealed that DENV2 led to damages in the CNS, such as hemorrhage, reactive gliosis, hyperplastic and hypertrophied microglia, astrocyte proliferation, Purkinje neurons retraction and cellular infiltration around vessels in the pia mater and in neuropil. Viral tropism and replication were detected in resident cells of the brain and cerebellum, such as neurons, astrocyte, microglia and oligodendrocytes. Results suggest that this classical mice model might be useful for analyzing the neurotropic effect of DENV with similarities to what occurs in human.

Evaluation of energy expenditure in forward and backward movements performed by soccer referees.

The aim of this study was to measure the energy expenditure for locomotor activities usually performed by soccer referees during a match (walking, jogging, and running) under laboratory conditions, and to compare forward with backward movements. The sample was composed by 10 male soccer referees, age 29±7.8 years, body mass 77.5±6.2 kg, stature 1.78±0.07 m and professional experience of 7.33±4.92 years. Referees were evaluated on two separate occasions. On the first day, maximal oxygen uptake (VO2max) was determined by a maximal treadmill test, and on the second day, the oxygen consumption was determined in different speeds of forward and backward movements. The mean VO2max was 41.20±3.60 mL·kg-1·min-1 and the mean heart rate achieved in the last stage of the test was 190.5±7.9 bpm. When results of forward and backward movements were compared at 1.62 m/s (walking speed), we found significant differences in VO2, in metabolic equivalents, and in kcal. However, the same parameters in forward and backward movements at jogging velocities (2.46 m/s) were not significantly different, showing that these motor activities have similar intensity. Backward movements at velocities equivalent to walking and jogging are moderate-intensity activities, with energy expenditure less than 9 kcal. Energy expenditure was overestimated by at least 35% when calculated by mathematical equations. In summary, we observed that backward movements are not high-intensity activities as has been commonly reported, and when calculated using equations available in the literature, energy expenditure was overestimated compared to the values obtained by indirect calorimetry.

Peripheral effects induced in BALB/c mice infected with DENV by the intracerebral route.

The lack of an immunocompetent animal model for dengue mimicking the disease in humans is a limitation for advances in this field. Inoculation by intracerebral route of neuroadapted dengue strains in mice is normally lethal and provides a straightforward readout parameter for vaccine testing. However, systemic effects of infection and the immune response elicited in this model remain poorly described. In the present work, BALB/c mice infected by the intracerebral route with neuroadapted DENV2 exhibited several evidences of systemic involvement. DENV-inoculated mice presented virus infective particles in the brain followed by viremia, especially in late stages of infection. Infection induced cellular and humoral responses, with presence of activated T cells in spleen and blood, lymphocyte infiltration and tissue damages in brain and liver, and an increase in serum levels of some pro-inflammatory cytokines. Data highlighted an interplay between the central nervous system commitment and peripheral effects under this experimental condition.

Evaluation of energy expenditure in forward and backward movements performed by soccer referees

The aim of this study was to measure the energy expenditure for locomotor activities usually performed by soccer referees during a match (walking, jogging, and running) under laboratory conditions, and to compare forward with backward movements. The sample was composed by 10 male soccer referees, age 29±7.8 years, body mass 77.5±6.2 kg, stature 1.78±0.07 m and professional experience of 7.33±4.92 years. Referees were evaluated on two separate occasions. On the first day, maximal oxygen uptake (VO2max) was determined by a maximal treadmill test, and on the second day, the oxygen consumption was determined in different speeds of forward and backward movements. The mean VO2max was 41.20±3.60 mL·kg-1·min-1 and the mean heart rate achieved in the last stage of the test was 190.5±7.9 bpm. When results of forward and backward movements were compared at 1.62 m/s (walking speed), we found significant differences in VO2, in metabolic equivalents, and in kcal. However, the same parameters in forward and backward movements at jogging velocities (2.46 m/s) were not significantly different, showing that these motor activities have similar intensity. Backward movements at velocities equivalent to walking and jogging are moderate-intensity activities, with energy expenditure less than 9 kcal. Energy expenditure was overestimated by at least 35% when calculated by mathematical equations. In summary, we observed that backward movements are not high-intensity activities as has been commonly reported, and when calculated using equations available in the literature, energy expenditure was overestimated compared to the values obtained by indirect calorimetry.

Trypanosomatid essential metabolic pathway: new approaches about heme fate in Trypanosoma cruzi.

Trypanosoma cruzi, the causal agent of Chagas disease, has a complex life cycle and depends on hosts for its nutritional needs. Our group has investigated heme (Fe-protoporphyrin IX) internalization and the effects on parasite growth, following the fate of this porphyrin in the parasite. Here, we show that epimastigotes cultivated with heme yielded the compounds α-meso-hydroxyheme, verdoheme and biliverdin (as determined by HPLC), suggesting an active heme degradation pathway in this parasite. Furthermore, through immunoprecipitation and immunoblotting assays of epimastigote extracts, we observed recognition by an antibody against mammalian HO-1. We also detected the localization of the HO-1-like protein in the parasite using immunocytochemistry, with antibody staining primarily in the cytoplasm. Although HO has not been described in the parasite's genome, our results offer new insights into heme metabolism in T. cruzi, revealing potential future therapeutic targets.

A protocol proposition of cell therapy for the treatment of chronic obstructive pulmonary disease.

The main feature of pulmonary emphysema is airflow obstruction resulting from the destruction of the alveolar walls distal to the terminal bronchioles. Existing clinical approaches have improved and extended the quality of life of emphysema patients. However, no treatment currently exists that can change the disease course and cure the patient. The different therapeutic approaches that are available aim to increase survival and/or enhance the quality of life of emphysema patients. In this context, cell therapy is a promising therapeutic approach with great potential for degenerative pulmonary diseases. In this protocol proposition, all patients will be submitted to laboratory tests, such as evaluation of heart and lung function and routine examinations. Stem cells will be harvested by means of 10 punctures on each anterior iliac crest, collecting a total volume of 200mL bone marrow. After preparation, separation, counting and labeling (optional) of the mononuclear cells, the patients will receive an intravenous infusion from the pool of Bone Marrow Mononuclear Cells (BMMC). This article proposes a rational and safe clinical cellular therapy protocol which has the potential for developing new projects and can serve as a methodological reference for formulating clinical application protocols related to the use of cellular therapy in COPD. This study protocol was submitted and approved by the Brazilian National Committee of Ethics in Research (CONEP - Brazil) registration number 14764. It is also registered in ClinicalTrials.gov (NCT01110252).

Melatonin affects conjugation of 4-hydroxynonenal with glutathione in liver of pacu, a hypoxia-tolerant fish.

In cytosol from liver of pacu, Piaractus mesopotamicus, a hypoxia-tolerant fish that dwells in Pantanal, we found an enzyme activity capable of modulating the alkenal 4-hydroxy-2-nonenal (HNE) by conjugating it with glutathione (GST-HNE activity). HNE is a downstream metabolite from the oxidation of polyunsaturated fatty acids by reactive oxygen species arisen from mitochondria of animal cells. HNE production may increase more intensively under oxidative stress. Harmful effects to cell survival may occur when HNE increases over 10(-4) M. Pacus submitted to hypoxia in July (cold season in Pantanal) showed 40% less of this GST-HNE conjugating activity in their liver cytosol. Injecting pacus subjected to hypoxia during the cold season with a summer physiological dose of melatonin caused their liver cytosolic GST-HNE activity to increase up to the levels found in the warm season. From October to March (warm season in Pantanal), pacus are prone to oxidative stress particularly during potamodromous active oxygen-demanding swimming, when they migrate up rivers to spawn. Thus, our findings point out that the higher levels of melatonin in circulation during the summer are important to avoid the increase of 4-HNE inside liver cells of this fish species.

Behavior of Enterobacter pulveris in amorphous and crystalline powder matrices treated with supercritical carbon dioxide.

The resistance of an Enterobacter pulveris strain to combined heat and supercritical carbon dioxide (scCO(2)) treatments in different powder matrices was examined. The strain proved resistant to scCO(2) treatment up to 50 MPa pressure at temperatures >73 °C for at least 20 min in a commercial infant formula. Water availability was shown to be important for the observed thermotolerance, because introduction of water in the scCO(2) gas flow during treatment resulted in a 1 log(10) cfu/g reduction of the initial inoculum. Interestingly, similar tolerance to heat and scCO(2) treatment was observed in a less complex matrix, a maltodextrin powder. In contrast, the bacterial strain proved sensitive to lower temperatures (55-65 °C) over shorter times (≤10 min) in a dextrose powder composed of crystalline particles. Therefore, the microorganism demonstrates heat sensitivity in the crystalline powder matrix closer to that of nonpowder liquid matrices. These data demonstrate the increased heat tolerance of the bacterium specifically in amorphous powders and indicate that this characteristic is not dependent on fat and other components commonly found in infant formula. The information is important in designing strategies to deal with contamination of powders with Enterobacteriacae, including pathogenic Cronobacter spp.

Morfologia e quantificação da microbiota intestinal do curimbatá (Prochilodus lineatus) e do cascudo cinza (Pterygoplichthys anisitsi) cultivados em cativeiro / Morphology and quantification of intestinal microbiota of curimbatá (Prochilodus lineatus) and the gray armored catfish (Pterygoplichthys anisitsi) reared in captivity

Todos os animais vivem em íntima associação com micro-organismos que desempenham importantes funções em seu desenvolvimento normal. Nos vertebrados, a mais populosa e complexa comunidade de micro-organismos reside no trato intestinal. O intuito do estudo foi quantificar, classificar e verificar morfologicamente a população microbiana intestinal de duas importantes espécies de peixes de água doce, o curimbatá (Prochilodus lineatus) e o cascudo cinza (Pterygoplichthys anisitsi). As amostras foram coletadas por meio de raspagens da mucosa intestinal, diluídas seriadamente até 10-4, semeadas em placas contendo ágar soja tripticaseína (TSA) e ágar chocolate (AC) para contagem de bactérias totais e identificação morfológica por Gram, em aerobiose e em anaerobiose facultativa, respectivamente. As contagens de bactérias totais mostraram resultados que variaram entre 10³ e 10(4)ufc.mL-1. Os tipos morfológicos encontrados foram cocos, leveduras e bastonetes Gram negativos e positivos. Estudos adicionais sobre os padrões de colonização microbiana e a morfologia dos micro-organismos aderidos à mucosa intestinal foram possíveis com o uso da microscopia eletrônica de varredura (MEV), sendo encontradas formas variadas de micro-organismos, tais como leveduras, formas cocoides e bacilares flageladas e não flageladas. A microbiota intestinal do curimbatá e a do cascudo cinza provaram ser bastante diversas e populosas, com o predomínio de micro-organismos Gram negativos.

All animals exist in intimate associations with microorganisms that play important roles in the hosts' normal development. In vertebrates, the most populous and complex community of microbes resides in the digestive tract.The aim of this research was to morphologically quantify, classify and verify the composition of intestinal microbiota of two species of freshwater fish, the Prochilodus lineatus and the Pterygoplichthys anisitsi. The samples were collected by scrapings of intestinal mucosa, serially diluted to 10-4, plated on tryptic soy agar (TSA) and chocolate agar (CA) for total bacterial counting and morphological identification by Gram, in aerobiosis and facultative anaerobiosis conditions, respectively. In the total bacterial counting results ranged between 10³ to 10(4) cfu.mL-1. The morphological types found were cocci, yeasts and Gram negative and positive rods. Additional studies about patterns of microbial colonization and the morphology of the adhered microorganisms to the intestinal mucosa were possible using the scanning electron microscopy (SEM) and several forms of microorganisms, such as yeasts, cocci and bacillary shapes flagellated and non-flagellated were found. The intestinal microbiota of P. lineatus and P. anisitsi was diverse and populous, with a predominance of Gram negative microorganisms.

In vitro and in vivo toxicological evaluation of extract and fractions from Baccharis trimera with anti-inflammatory activity.

ETHNOPHARMACOLOGICAL RELEVANCE: Baccharis trimera (Less) DC. (Asteraceae), popularly known in Brazil as "carqueja", have been used in folk medicine to treat gastrointestinal, hepatic and renal diseases, and inflammatory processes as rheumatism. AIM OF THE STUDY: To evaluate the in vitro and in vivo toxicological effects of anti-inflammatory Baccharis trimera aqueous extract and fractions. MATERIALS AND METHODS: Aqueous extract of Baccharis trimera (AEBt) was produced by infusion in boiling water. After lyophylization AEBt was extracted with 80% ethanol, originating the ethanolic supernatant fraction (EFBt) and the aqueous sediment fraction (AFBt). Anti-inflammatory properties of AEBt, EFBt or AFBt (3, 30 or 300 µg/kg b.w.) were evaluated by the carrageenan-induced mouse paw edema using indomethacin (10mg/kg) as positive control. The growth of rat hepatoma cells (HTC) and human embryo kidney epithelial cells (HEK) was determined by protein staining assay. Cytotoxicity was assayed by the tetrazolium salt (MTT) reduction. Cyclosporin was used as reference cytotoxic drug for spleen cells and doxorubicin for HTC and HEK cells. For in vivo toxicological evaluation SW male mice were daily and oral (gavage) treated with extract/fractions at 4.2mg/kg or 42 mg/kg during 15 days. After treatment liver or kidney cells were submitted to comet assay to determine the DNA damage index, and the glutathione S-transferase activity was assayed towards ETHA (class Pi) and CDNB (several classes). Mutagenicity was evaluated by the Ames test using Salmonella typhimurium strains TA97, TA98, TA100, and TA102. RESULTS: The anti-inflammatory effects of EFBt were higher than those of AEBt or AFBt. Mice treatment (3-300 µg/kg) with AFBt reduced the paw edema (3h) at lower levels (29.2-37.3%; P<0.01), than those observed for AEBt (44.7-54.2%; P<0.001), EFBt (49.3-58.2%; P<0.001) or indomethacin (64.6%, P<0.001, 10mg/kg). The growth of kidney cells (HEK) was inhibited by AEBt (IC(50) 182.6 µg/ml), EFBt (IC(50) 78.1 µg/ml) and AFBt (IC(50) 86.2 µg/ml), with lower effects on HTC hepatic cell (IC(50) 308.8 µg/ml, 396.5 µg/ml and 167.9 µg/ml, respectively). As evaluated by MTT test, AFBt exhibited cytotoxicity for HEK cells (IC(50) 372.5 µg/ml), but none for HTC ones; by the way, AFBt stimulated spleen cells (EC(50) 2.2 µg/ml) while cyclosporine, a cytotoxic reference drug inhibited them with IC(50) of 0.42 µg/ml; the IC(50) for doxorubicin for HEK and HTC cells was 0.28 µg/ml and 14.4 µg/ml, respectively, at 96h. No mutagenic potential was observed. Mice treatment with AEBt or AFBt at 42 mg/kg for 15 days altered the kidney relative weight, but not at 4.2mg/kg. Baccharis trimera did not change liver, spleen or popliteal lymph node relative weight. DNA damage index of kidney cells was observed on mice treated with AEBt/AFBt, but not on animals treated with EFBt, while DNA lesions were detected on liver cells only after AFBt treatment. The general activities of hepatic GST and Pi GST were reduced by EFBt and AFBt treatment, respectively. CONCLUSIONS: Baccharis trimera did not show mutagenicity, inhibited the GST activity, a hepatic detoxification enzyme, and induced in vivo (genotoxicity) and in vitro toxicological effects to kidney cells.

Moroccan national study on prevalence of mental disorders: a community-based epidemiological study.

OBJECTIVE: The main objective of this study was to assess the prevalence of common mental disorders in the Moroccan general population. METHOD: On a systematic representative randomized sample, the Moroccan Arabic version of the Mini International Neuropsychiatric Interview (MINI) was used to assess the prevalence of mood, anxiety, substance, and alcohol abuse disorders. RESULTS: Among 5498 subjects interviewed, 40.1% had at least one current mental disorder. Current major depressive disorder was the most common (26.5%), and at least one anxiety disorder was found in 37% of the sample. Mental disorders were more frequent among female, urban, divorced, and unemployed subjects. CONCLUSION: Mental disorders are common in the Moroccan general population, particularly mood and anxiety disorders.

Heme-induced Trypanosoma cruzi proliferation is mediated by CaM kinase II.

Trypanosoma cruzi, the etiologic agent of Chagas disease, is transmitted through triatomine vectors during their blood-meal on vertebrate hosts. These hematophagous insects usually ingest approximately 10mM of heme bound to hemoglobin in a single meal. Blood forms of the parasite are transformed into epimastigotes in the crop which initiates a few hours after parasite ingestion. In a previous work, we investigated the role of heme in parasite cell proliferation and showed that the addition of heme significantly increased parasite proliferation in a dose-dependent manner [1]. To investigate whether the heme effect is mediated by protein kinase signalling pathways, parasite proliferation was evaluated in the presence of several protein kinase (PK) inhibitors. We found that only KN-93, a classical inhibitor of calcium-calmodulin-dependent kinases (CaMKs), blocked heme-induced cell proliferation. KN-92, an inactive analogue of KN-93, was not able to block this effect. A T. cruzi CaMKII homologue is most likely the main enzyme involved in this process since parasite proliferation was also blocked when Myr-AIP, an inhibitory peptide for mammalian CaMKII, was included in the cell proliferation assay. Moreover, CaMK activity increased in parasite cells with the addition of heme as shown by immunological and biochemical assays. In conclusion, the present results are the first strong indications that CaMKII is involved in the heme-induced cell signalling pathway that mediates parasite proliferation.

Sexagem histológica e desempenho de Oreochromis niloticus testando diâmetros de ração de acordo com o aparato bucal / Histological gender diagnosis and performance of Oreochromis niloticus testing diameters of crumble diet according to oral apparatus

Estudou-se o tamanho da boca de larvas de tilápia-do-nilo e testou-se o efeito de diferentes granulometrias da ração sobre o ganho de peso, comprimento e sobrevivência das larvas aos 30 e 60 dias de arraçoamento. Avaliou-se também o método de sexagem por meio de microscopia de luz aos 35 dias de idade. A medida da boca das larvas apresentou valores médios de 918,2±152,9μm aos cinco dias de idade. De acordo com esse dado, testaram-se três granulometrias: 0,25, 0,35 e 0,50mm. Aos 30 e aos 60 dias de arraçoamento, 10 por cento das larvas foram medidas, pesadas e contadas para cálculo da taxa de sobrevivência. O tamanho dos grânulos testados não afetou o desempenho das larvas de tilápia nilótica com alimentação iniciada aos cinco dias pós-eclosão. Quanto à sexagem histológica aos 35 dias de idade, as gônadas apresentaram-se, em sua maioria, indiferenciadas. Recomenda-se que essa análise deva ser realizada de acordo com o tamanho dos animais e não com a idade.

The mouth size of Nile tilapia larvae and the effect of different diameters of crumble fish food over weight gain, total length, and survival of larvae after 30 and 60 days of feeding were studied. The method of gender diagnosis based on light microscopy at 35 days of age was also evaluated. The larval mouth measurement presented average values of 918.2±152.9μm at five days of age. Based on this information, three granule sizes were tested: 0.25, 0.35, and 0.50mm. At 30 and 60 days of feeding, 10 percent of larvae were measured, weighed, and counted to calculate the survival rate. It was verified that the crumble size did not affect the performance of Nile tilapia larvae when feeding was initiated five days after hatching. In relation to the histological gender diagnosis at 35 days of age, most of gonads were undifferentiated. Therefore, it is recommended that this analysis should be carried out according to the size of animals instead of their age.

Anti-Trypanosoma cruzi activity of Pterodon pubescens seed oil: geranylgeraniol as the major bioactive component.

In the search for new therapeutic agents for Chagas' disease, we screened extracts obtained from the Brazilian plant Pterodon pubescens found commercially in the medicinal flora market. We investigated the potential trypanocidal effect of the oleaginous ethanolic extract of P. pubescens seeds and its fractions (PF1, PF1.1, PF1.2, and PF1.3) and of geranylgeraniol (GG-OH), the sole component of the hexane fraction (PF1.2). In experiments with bloodstream trypomastigotes of Trypanosoma cruzi, performed at 37 degrees C in culture medium, PF1.2 and GG-OH showed similar potency, while the oleaginous extract from P. pubescens seeds and the other fractions were about three times less active. GG-OH inhibited the proliferation of intracellular amastigotes, at concentrations which do not affect the mammalian host cell. Transmission electron microscopy and flow cytometry analysis indicate the mitochondrion, an organelle that plays a central role in apoptosis, of both epimastigotes and of trypomastigotes as the major target of GG-OH. On the other hand, the ultrastructural images of the endoplasmic reticulum profiles, myelin-like figures, and concentric membranous arrangements inside damaged mitochondrion are suggestive of an autophagic pathway leading to parasite death. Because the different forms of cell death share some morphological features such as mitochondrial collapse, further studies are needed to disclose the trypanocidal action of GG-OH.

Lupine sensitisation in a population of 1,160 subjects.

Lupin is part of the Mediterranean diet and is also used as a thickener of food products. It has been recognised as a cause of serious allergic reactions. This study aims at determining the prevalence of lupin sensitisation in 1,160 subjects consulting allergologists. This population performed skin prick tests (SPT) to lupin. In case of positivity, a clinical questionnaire was applied and the subjects were tested for other legumes, common inhalants and latex. We find a 4.1% sensitisation rate to lupin, a 75% co-sensitisation between lupin and legumes, a 82.1% co-sensitisation between lupin and pollen and a 28.5% co-sensitisation between lupin and latex. To conclude, we documented a high lupin sensitisation in a selected population, thus stressing the importance of lupin as a food allergen.

Heme requirement and intracellular trafficking in Trypanosoma cruzi epimastigotes.

Epimastigotes multiplies in the insect midgut by taking up nutrients present in the blood meal including heme bound to hemoglobin of red blood cell. During blood meal digestion by vector proteases in the posterior midgut, hemoglobin is clipped off into amino acids, peptides, and free heme. In this paper, we compared the heme and hemoglobin uptake kinetics and followed their intracellular trafficking. Addition of heme to culture medium increased epimastigote proliferation in a dose-dependent manner, while medium supplemented with hemoglobin enhanced growth after 3-day lag phase. Medium supplemented with globin-derived peptides stimulated cell proliferation in a dose-independent way. Using Palladium mesoporphyrin IX (Pd-mP) as a fluorescent heme-analog, we observed that heme internalization proceeded much faster than that observed by hemoglobin-rhodamine. Binding experiments showed that parasites accumulated the Pd-mP into the posterior region of the cell whereas hemoglobin-rhodamine stained the anterior region. Finally, using different specific inhibitors of ABC transporters we conclude that a P-glycoprotein homologue transporter is probably involved in heme transport through the plasma membrane.

DNA vaccines against dengue virus based on the ns1 gene: the influence of different signal sequences on the protein expression and its correlation to the immune response elicited in mice.

We analyzed four DNA vaccines based on DENV-2 NS1: pcENS1, encoding the C-terminal from E protein plus the NS1 region; pcENS1ANC, similar to pcENS1 plus the N-terminal sequence from NS2a (ANC); pcTPANS1, coding the t-PA signal sequence fused to NS1; and pcTPANS1ANC, similar to pcTPANS1 plus the ANC sequence. The NS1 was detected in lysates and culture supernatants from pcTPANS1-, pcENS1- and pcENS1ANC-transfected cells and not in cells with pcTPANS1ANC. Only the pcENS1ANC leads the expression of NS1 in plasma membrane, confirming the importance of ANC sequence for targeting NS1 to cell surface. High levels of antibodies recognizing conformational epitopes of NS1 were induced in mice immunized with pcTPANS1 and pcENS1, while only few pcENS1ANC-inoculated animals presented detectable anti-NS1 IgG. Protection against DENV-2 was verified in pcTPANS1- and pcENS1-immunized mice, although the plasmid pcTPANS1 induced slight higher protective immunity. These plasmids seem to activate distinct patterns of the immune system.

Pathologic study of a fatal case of dengue-3 virus infection in Rio de Janeiro, Brazil.

Dengue hemorrhagic fever (DHF) is a severe febrile disease, characterized by abnormalities in hemostasis and increased vascular permeability, which in some cases results in hypovolemic shock syndrome and in dengue shock syndrome. The clinical features of DHF include plasma leakage, bleeding tendency and liver involvement. We studied the histopathological features of a fatal case of dengue-3 virus infection. The patient, a 63-year old male, presented with an acute onset of severe headache, myalgia and maculopapular rash. Tissue fragments (liver, spleen, lung, heart, kidney and lymph nodes) were collected for light microscopy studies and stained by standard methods. Histopathology revealed severe tissue damage, caused by intense hemorrhage, interstitial edema and inflammation. Some tissue sections were also processed with the immunoperoxidase reaction, which revealed the dengue viral antigen. Dengue-3 virus was isolated and identified with electron microscopy in a C6/36 cell culture inoculated with the patient's serum. Viral particles were detected in the infected cell culture.

Pathologic study of a fatal case of dengue-3 virus infection in Rio de Janeiro, Brazil

Dengue hemorrhagic fever (DHF) is a severe febrile disease, characterized by abnormalities in hemostasis and increased vascular permeability, which in some cases results in hypovolemic shock syndrome and in dengue shock syndrome. The clinical features of DHF include plasma leakage, bleeding tendency and liver involvement. We studied the histopathological features of a fatal case of dengue-3 virus infection. The patient, a 63-year old male, presented with an acute onset of severe headache, myalgia and maculopapular rash. Tissue fragments (liver, spleen, lung, heart, kidney and lymph nodes) were collected for light microscopy studies and stained by standard methods. Histopathology revealed severe tissue damage, caused by intense hemorrhage, interstitial edema and inflammation. Some tissue sections were also processed with the immunoperoxidase reaction, which revealed the dengue viral antigen. Dengue-3 virus was isolated and identified with electron microscopy in a C6/36 cell culture inoculated with the patient's serum. Viral particles were detected in the infected cell culture.