Histopathological and immunological characteristics of placentas infected with chikungunya virus.

Although vertical transmission of CHIKV has been reported, little is known about the role of placenta in the transmission of this virus and the effects of infection on the maternal-fetal interface. In this work we investigated five placentas from pregnant women who became infected during the gestational period. Four formalin-fixed paraffin-embedded samples of placenta (cases 1-4) were positive for CHIKV by RT-PCR. One (case 5) had no positive test of placenta, but had positive RT-PCR for CHIKV in the serum of the mother and the baby, confirming vertical transmission. The placentas were analyzed regarding histopathological and immunological aspects. The main histopathological changes were: deciduitis, villous edema, deposits, villous necrosis, dystrophic calcification, thrombosis and stem vessel obliteration. In infected placentas we noted increase of cells (CD8+ and CD163+) and pro- (IFN-γ and TNF-α) and anti-inflammatory (TGF-ß and IL-10) cytokines compared to control placentas. Moreover, CHIKV antigen was detected in decidual cell, trophoblastic cells, stroma villi, Hofbauer cells, and endothelial cells. In conclusion, CHIKV infection seems to disrupt placental homeostasis leading to histopathological alterations in addition to increase in cellularity and cytokines overproduction, evidencing an altered and harmful environment to the pregnant woman and fetus.

A Chikungunya Virus Multiepitope Recombinant Protein Expressed from the Binary System Insect Cell/Recombinant Baculovirus Is Useful for Laboratorial Diagnosis of Chikungunya.

Chikungunya virus (CHIKV) is an arbovirus currently distributed worldwide, causing a disease that shares clinical signs and symptoms with other illnesses, such as dengue and Zika and leading to a challenging clinical differential diagnosis. In Brazil, CHIKV emerged in 2014 with the simultaneous introduction of both Asian and East/Central/South African (ECSA) genotypes. Laboratorial diagnosis of CHIKV is mainly performed by molecular and serological assays, with the latter more widely used. Although many commercial kits are available, their costs are still high for many underdeveloped and developing countries where the virus circulates. Here we described the development and evaluation of a multi-epitope recombinant protein-based IgG-ELISA (MULTREC IgG-ELISA) test for the specific detection of anti-CHIKV antibodies in clinical samples, as an alternative approach for laboratorial diagnosis. The MULTREC IgG-ELISA showed 86.36% of sensitivity and 100% of specificity, and no cross-reactivity with other exanthematic diseases was observed. The recombinant protein was expressed from the binary system insect cell/baculovirus using the crystal-forming baculoviral protein polyhedrin as a carrier of the target recombinant protein to facilitate recovery. The crystals were at least 10 times smaller in size and had an amorphous shape when compared to the polyhedrin wild-type crystal. The assay uses a multi-epitope antigen, representing two replicates of 18 amino acid sequences from the E2 region and a sequence of 17 amino acids from the nsP3 region of CHIKV. The recombinant protein was highly expressed, easy to purify and has demonstrated its usefulness in confirming chikungunya exposure, indeed showing a good potential tool for epidemiological surveillance.

Subsets of Cytokines and Chemokines from DENV-4-Infected Patients Could Regulate the Endothelial Integrity of Cultured Microvascular Endothelial Cells.

Introduction: It is a consensus that inflammatory mediators produced by immune cells contribute to changes in endothelial permeability in dengue. We propose to relate inflammatory mediators seen in dengue patients with the in vitro alteration of endothelial cells (ECs) cultured with serum from these patients. Methods: Patients with mild (DF) to moderate and severe dengue (DFWS/Sev) were selected. ELISA quantified inflammatory mediators. Expression of adhesion molecules and CD147 were evaluated in the ECs cultured with the patient's serum by flow cytometry. We assessed endothelial permeability by measuring transendothelial electrical resistance in cocultures of ECs with patient serum. Results: Dengue infection led to an increase in inflammatory mediators-the IL-10 distinguished DF from DFWS/Sev. There were no changes in CD31, CD54, and CD106 but decreased CD147 expression in ECs. DFWS/Sev sera induced a greater difference in endothelial permeability than DF sera. Correlation statistical test indicated that low IL-10 and IFN-γ and high CCL5 maintain the integrity of ECs in DF patients. In contrast, increased TNF, IFN-γ, CXCL8, and CCL2 maintain EC integrity in DFWS/Sev patients. Conclusions: Our preliminary data suggest that a subset of inflammatory mediators may be related to the maintenance or loss of endothelial integrity, reflecting the clinical prognosis.

Was It Chikungunya? Laboratorial and Clinical Investigations of Cases Occurred during a Triple Arboviruses' Outbreak in Rio de Janeiro, Brazil.

The co-circulation of chikungunya virus (CHIKV), dengue virus (DENV) and Zika virus (ZIKV) in Rio de Janeiro (RJ), Brazil, caused a challenging triple epidemic, as they share similar clinical signs and symptoms and geographical distribution. Here, we aimed to investigate the clinical and laboratorial aspects of chikungunya suspected cases assisted in RJ during the 2018 outbreak, focusing on the differential diagnosis with dengue and zika. All suspected cases were submitted to molecular and/or serological differential diagnostic approaches to arboviruses. A total of 242 cases suspected of arbovirus infection were investigated and 73.6% (178/242) were molecular and/or serologically confirmed as chikungunya. In RT-qPCR confirmed cases, cycle threshold (Ct) values ranged from 15.46 to 35.13, with acute cases presenting lower values. Chikungunya cases were mainly in females (64%) and the most frequently affected age group was adults between 46 to 59 years old (27%). Polyarthralgia affected 89% of patients, especially in hands and feet. No dengue virus (DENV) and Zika virus (ZIKV) infections were confirmed by molecular diagnosis, but 9.5% (23/242) had serological evidence of DENV exposure by the detection of specific anti-DENV IgM or NS1, and 42.7% (76/178) of chikungunya positive cases also presented recent DENV exposure reflected by a positive anti-DENV IgM or NS1 result. A significantly higher frequency of arthritis (p = 0.023) and limb edema (p < 0.001) was found on patients with CHIKV monoinfection compared to dengue patients and patients exposed to both viruses. Lastly, phylogenetic analysis showed that the chikungunya cases were caused by the ECSA genotype. Despite the triple arboviruses' epidemic in the state of RJ, most patients with fever and arthralgia investigated here were diagnosed as chikungunya cases, and the incidence of CHIKV/DENV co-detection was higher than that reported in other studies.

Increased circulating levels of High Mobility Group Box 1 (HMGB1) in acute-phase Chikungunya virus infection: Potential disease biomarker.

BACKGROUND: Chikungunya virus (CHIKV) causes a febrile syndrome with intense and debilitating arthralgia that can persist for several months or years after complete virus clearance. As there is no specific antiviral treatment or vaccine against CHIKV, identification of serological markers that help clinical management of CHIKV patients is urgent. The High Mobility Group Box 1 (HMGB1) protein is secreted to extracellular milieu and triggers an intense inflammatory process by inducing the overexpression of pro-inflammatory cytokines. HMGB1 plays an important role in several virus diseases as well as in rheumatoid arthritis. OBJECTIVES: This study focus on the investigation of HMGB1 serum levels in a sera panel from CHIKV-infected patients in an attempt to assess its potential as a biomarker for chikungunya clinical management. STUDY DESIGN: Eighty CHIKV-positive samples and 32 samples from healthy donors were subjected to a quantitative HMGB1 ELISA assay to assess the HMGB1 circulating levels. RESULTS: HMGB1 levels were significantly higher in CHIKV-positive samples (516.12 ng/mL, SEM ± 48.83 ng/mL) compared to negative control (31.20 ng/mL, SEM ± 3.24 ng/mL, p < 0.0001). Circulating levels of HMGB1 persisted elevated during the whole acute-phase of disease and correlated with virus titer (p < 0.05). CONCLUSIONS: The present study is the first to describe increased serum levels of HMGB1 in CHIKV infection and its positive correlation with virus titer, suggesting its potential use as a biomarker for diagnosis and treatment of chikungunya fever.

Different Profiles of Cytokines, Chemokines and Coagulation Mediators Associated with Severity in Brazilian Patients Infected with Dengue Virus.

The incidence of dengue in Latin America has increased dramatically during the last decade. Understanding the pathogenic mechanisms in dengue is crucial for the identification of biomarkers for the triage of patients. We aimed to characterize the profile of cytokines (IFN-γ, TNF-α, IL-1ß, IL-6, IL-18 and IL-10), chemokines (CXCL8/IL-8, CCL2/MCP-1 and CXCL10/IP-10) and coagulation mediators (Fibrinogen, D-dimer, Tissue factor-TF, Tissue factor pathway inhibitor-TFPI and Thrombomodulin) during the dengue-4 epidemic in Brazil. Laboratory-confirmed dengue cases had higher levels of TNF-α (p < 0.001), IL-6 (p = 0.005), IL-10 (p < 0.001), IL-18 (p = 0.001), CXCL8/IL-8 (p < 0.001), CCL2/MCP-1 (p < 0.001), CXCL10/IP-10 (p = 0.001), fibrinogen (p = 0.037), D-dimer (p = 0.01) and TFPI (p = 0.042) and lower levels of TF (p = 0.042) compared to healthy controls. A principal component analysis (PCA) distinguished between two profiles of mediators of inflammation and coagulation: protective (TNF-α, IL-1ß and CXCL8/IL-8) and pathological (IL-6, TF and TFPI). Lastly, multivariate logistic regression analysis identified high aspartate aminotransferase-to-platelet ratio index (APRI) as independent risk factors associated with severity (adjusted OR: 1.33; 95% CI 1.03-1.71; p = 0.027), the area under the receiver operating characteristics curve (AUC) was 0.775 (95% CI 0.681-0.869) and an optimal cutoff value was 1.4 (sensitivity: 76%; specificity: 79%), so it could be a useful marker for the triage of patients attending primary care centers.

Zika Induces Human Placental Damage and Inflammation.

In Brazil, an epidemic of Zika virus (ZIKV) infections was declared in 2015 that coincided with alarming reports of microcephaly in newborns associated with mother infection. Although the virus has placental tropism, changes in the tissue morphology and immunity of infected patients have not yet been elucidated. Here, we investigated the histopathological and ultrastructural changes along with the immunological profile and the BDNF expression in rare placental material. Tissues were obtained in the 2015-2016 Brazilian epidemic, of ten ZIKV-infected patients during pregnancy, five resulting in cases of fetal microcephaly and five non-microcephaly, compared to five non-infected control placentae. Viral antigens were only detected in samples from the ZIKV infected patients. Infected placentae presented histopathological severe damage, while the ultrastructural evaluation showed abnormal organelles, such as clusters of virus-like particles consistent with the ZIKV dimensions. Increased infiltration of CD68+ and TCD8+ cells, expression of MMPs, cytokines (IFN-γ and TNF-α) and other immunological mediators (RANTES/CCL5 and VEGFR-2) confirmed excessive inflammation and vascular permeability dysfunction. An evaluation of BDNF showed a decrease that could modulate neuronal damage in the developing fetus. The placental changes caused by ZIKV are not pathognomonic, however, the data provide evidence that this infection leads to severe placental injury.

Zika Virus Infects Human Placental Mast Cells and the HMC-1 Cell Line, and Triggers Degranulation, Cytokine Release and Ultrastructural Changes.

Zika virus (ZIKV) is an emergent arthropod-borne virus whose outbreak in Brazil has brought major public health problems. Infected individuals have different symptoms, including rash and pruritus, which can be relieved by the administration of antiallergics. In the case of pregnant women, ZIKV can cross the placenta and infect the fetus leading to congenital defects. We have identified that mast cells in the placentae of patients who had Zika during pregnancy can be infected. This led to our investigation on the possible role of mast cells during a ZIKV infection, using the HMC-1 cell line. We analyzed their permissiveness to infection, release of mediators and ultrastructural changes. Flow cytometry detection of ZIKV-NS1 expression 24 h post infection in 45.3% of cells showed that HMC-1 cells are permissive to ZIKV infection. Following infection, ß-hexosaminidase was measured in the supernatant of the cells with a notable release at 30 min. In addition, an increase in TNF-α, IL-6, IL-10 and VEGF levels were measured at 6 h and 24 h post infection. Lastly, different intracellular changes were observed in an ultrastructural analysis of infected cells. Our findings suggest that mast cells may represent an important source of mediators that can activate other immune cell types during a ZIKV infection, which has the potential to be a major contributor in the spread of the virus in cases of vertical transmission.

Pneumonia por toxoplasmose em paciente imunocompetente / Toxoplasma pneumonia in an immunocompetent patient: case repor

Com grande distribuição mundial e incidência significativa, a toxoplamose é uma doença comum em mamíferos e pássaros, causada pelo protozoário Toxoplasma gondii. No homem, o parasitismo na fase proliferativa intracelular pode se apresentar sem sintomas, ou causar clínica transitória caracterizada por febre, fadiga e linfadenopatia. Por se tratar de patologia com sintomas inespecíficos e comuns a muitas outras, é fundamental a correta pesquisa de diagnósticos diferenciais, como citomegalovírus e Epstein-Barr. Relatamos o caso de um jovem e hígido, que desenvolveu pneumonia e, após confirmação sorológica para toxoplasmose e o tratamento adequado, apresentou melhora clínica.

With great worldwide distribution and significant incidence, toxoplamosis is a common disease in mammals and birds, caused by the protozoan Toxoplasma gondii. In humans, the parasitism in its intracellular proliferative phase may present no symptoms, or cause a transient condition characterized by fever, fatigue, and lymphadenopathy. Because it is a pathology with nonspecific symptoms that are common to many other conditions, it is fundamental to find the correct research of differential diagnoses, such as for Cytomegalovirus and Epstein Barr. We report a case of a young and healthy man who developed pneumonia and, after serological confirmation for toxoplasmosis and the appropriate treatment, presented clinical improvement

Paciente com Chikungunya evoluindo com Síndrome de Angústia Respiratória do Adulto grave / Chikungunya evolving with SRDA Chikungunya patient progressing to severe Adult Respiratory Distress Syndrome

Objetivo: Demonstrar casos de Chikungunya cujos paciente evoluíram com Síndrome da Angústia Respiratória do Adulto. Métodos: Estudo descritivo e documental cuja a amostra foi composta por pacientes internados em um hospital no município de Campos dos Goytacazes, diagnosticados com sorologia IgM positiva para febre do vírus Chikungunya, que evoluíram para Síndrome da Angústia Respiratória do Adulto. Foram feitas análises de prontuários e de imagens radiológicas, além de revisão de literatura. Resultados: Foram incluídos três pacientes no estudo, sendo que um evoluiu ao óbito e os outros dois obtiveram recuperação de suas funções após o quadro agudo da doença. Conclusão: A Chikungunya é uma doença recente em território nacional, com possível evolução para quadros graves, especialmente em sua fase aguda. Por essa razão, estudos aprofundados são necessários para maior conhecimento e entendimento da patologia e de suas factíveis complicações.

Objective: To report cases of Chikungunya that progressed with Acute Respiratory Distress Syndrome. Methods: This is a descriptive and documental study, the sample of which consisted of patients who were hospitalized, in the city of Campos dos Goytacazes, diagnosed with positive IgM serology for Chikungunya fever, which progressed to Acute Respiratory Distress Syndrome. Medical records and radiological images were analyzed, and literature reviewed. Results: Three patients were included in the study, with one of them progressing to death, and the other two having their functions recovered after acute illness. Conclusion: Chikungunya is a recent disease in the national territory, with possible progression to severe conditions, especially on its acute phase. For this reason, in-depth studies are necessary for a better knowledge and understanding of the pathology and its likely complications

Perfil epidemiológico dos pacientes com evolução subaguda e crônica de infecção por Chikungunya / Epidemiologic profile of patients with subacute and chronic Chikungunya infection evolution

OBJETIVO: Analisar o perfil epidemiológico dos pacientes com evolução subaguda e crônica da infecção por Chikungunya, e avaliar suas principais alterações laboratoriais. MÉTODOS: Estudo observacional, realizado por meio da análise de 31 prontuários de pacientes atendidos em um centro de referência de doenças imunoinfecciosas, no Estado do Rio de Janeiro, no período de janeiro a maio de 2016. Foram selecionados prontuários de pacientes com Chikungunya em fase subaguda ou crônica com diagnóstico confirmado por sorologia IgM, de ambos os sexos e de todas as faixas etárias. As seguintes variáveis foram consideradas: sexo, idade, leucócitos, plaquetas, velocidade de hemossedimentação, e aspartato aminotransferases e alanina transaminase (AST/ALT). RESULTADOS: A faixa etária predominante foi de 50 a 69 anos (64,5%). Pertenciam ao sexo feminino 83,9% dos pacientes. Dentre as alterações laboratoriais, destacaram-se elevação da velocidade de hemossedimentação (46,15%), leucopenia (37%), elevação de transaminases (30,8% AST e 23% ALT) e trombocitopenia (11,1%). CONCLUSÃO: Observou-se a importância epidemiológica na determinação de pacientes potencialmente capazes de desenvolver tais sequelas, representada principalmente pela artralgia incapacitante. A identificação desse grupo, caracterizado como pacientes do sexo feminino e de faixa etária entre 50 e 69 anos, pode ser de grande valia para a prevenção da evolução mórbida dessa arbovirose, sabendo que tais pacientes necessitam de cuidados especiais e acompanhamento clínico mais rigoroso.(AU)

OBJECTIVES: To analyze the epidemiological profile of patients with subacute and chronic Chikungunya infection, and to evaluate their main laboratory abnormalities. METHODS: This is an observational study, carried out through the analysis of 31 medical records of patients attended at a Reference Center for Immunopathological Diseases in the state of Rio de Janeiro, from January to May 2016. Patients with Chikungunya were selected in the subacute or chronic phase with diagnosis confirmed by IgM serology; both genders and all age ranges. The following variables were considered: gender, age, leukocytes, platelets, erythrocyte sedimentation rate, and aspartate aminotransferases and alanine transaminase (ASTALT). RESULTS: The predominant age range was 50 to 69 years (64.5%), with 83.9% of the patients being female. Among the laboratory alterations, high erythrocyte sedimentation rate (46.15%), leukopenia (37%), high transaminases (30.8% AST, and 23% ALT), and thrombocytopenia (11.1%) were observed. CONCLUSIONS: The epidemiological importance in the determination of patients potentially capable of developing such sequels, represented mainly by incapacitating arthralgia, was observed. The identification of this group, characterized as female patients, and between 50 and 69 years of age, may be of great value in the prevention of the morbid progression of this arbovirosis, knowing that such patients require special care, and stricter clinical follow-up.(AU)

Placental Histopathology and Clinical Presentation of Severe Congenital Zika Syndrome in a Human Immunodeficiency Virus-Exposed Uninfected Infant.

In the large Zika virus (ZIKV) epidemic that occurred in Brazil in 2015, the intrauterine fetal exposure to ZIKV was associated with a significant risk of developing microcephaly and neurological disorders in the infected infants. ZIKV-associated disease has since been reported in 24 countries in the Americas. At present, definitive evidence is lacking regarding the intrauterine co-exposure to ZIKV and other viral infections and whether the coinfection impacts the risk of acquiring either infection or disease severity. Here, we provide evidence of intrauterine exposure to both ZIKV and human immunodeficiency virus (HIV) infections, causing congenital Zika syndrome in an HIV-exposed uninfected infant. Clinical, imaging and laboratory examinations of the pregnant woman and the newborn were performed. Histopathology, ZIKV/HIV-specific immunoassays, and ultrastructural evaluation of the placenta were performed. The Zika-asymptomatic, HIV-positive pregnant woman underwent ultrasounds revealing fetal cerebral ventriculomegaly, microcephaly, and brain atrophy. Her baby girl was born small for gestational age and with the neurological sequelae of congenital Zika syndrome. The evaluation of the abnormally large term placenta revealed severe damage to the maternal decidua and chorionic villi, cells positive for ZIKV-specific antigens but not for HIV antigens, and intracellular membranous clusters of virus-like particles approximately 25 nm in diameter. The rapid progression and severity of the congenital Zika syndrome may be related to the uncontrolled HIV disease in the mother. The poor inflammatory response observed in the placenta may have reduced the inherent risk of mother-to-child transmission of HIV.

Acometimento visceral e ocular simultâneo em infecção por toxocara canis acompanhado de farmacodermia / Concurrent visceral and ocular involvementin toxocara canis infection and pharmacodermia

A toxocaríase humana é uma infecção parasitária de distribuição mundial causada pelos nematelmintos das espécies Toxocara canis e Toxocara cati, presentes no intestino do cão e do gato, respectivamente. Clinicamente, na maioria das vezes, é assintomática, porém pode apresentar-se de duas formas: visceral ou ocular. Visceralmente, gera uma síndrome hipereosinofílica crônica, acompanhada por leucocitose e hepatomegalia, podendo ocorrer algum grau de infiltrado pulmonar e febre. Na toxocaríase ocular, ocorre uveite intermediária ou posterior, podendo haver formação de granuloma, geralmente unilateral. O acometimento misto é raro, o que motivou este relato. Trata-se de paciente de 19 anos, sexo masculino, que apresentou como sintoma inicial perda da acuidade visual em olho esquerdo. Recebeu tratamento, sem melhora, com sulfametoxazol + trimetoprima e corticoide, fazendo farmacodermia. Evoluiu com diarreia, febre, dor abdominal e hepatoesplenomegalia. Descartadas infecções agudas por toxoplasmose, sífilis, vírus da imunodeficiência humana (HIV), citomegalovirose e dengue; apresentou leucocitose com hipereosinofilia. Foi solicitada sorologia para toxocaríase, confirmando esta infecção. Após o tratamento, apresentou completa remissão dos sintomas. O objetivo aqui foi debater os fatores confundidores, diagnósticos diferenciais, necessidade de exames complementares específicos e conduta terapêutica, de acordo com o quadro clínico.(AU)

Human toxocariasis is a worldwide parasitic infection caused by ascarid nematodes species: Toxocara canis and Toxocara cati, that are present in the intestines of dogs and cats, respectively. Although clinically, most human infections are asymptomatic, two syndromes of human toxocariasis are recognized: visceral and ocular. The visceral form is a hypereosinophilic syndrome accompanied by leukocytosis, hepatomegaly, some degree of pulmonary infiltrate and fever. In ocular toxacariasis there is intermediate or posterior uveitis, and there may be granuloma formation, usually unilateral. The simultaneous involvement of the two forms is rare, which is what, motivated this report. It is a 19-year-old male patient who initially presented loss of visual acuity in the left eye. He received treatment, without improvement, with sulfamethoxazole-trimethoprim and corticoid, causing a pharmacodermia. He developed diarrhea, fever, abdominal pain and hepatosplenomegaly. It was discarded acute infections by toxoplasmosis, syphilis, human immunodeficiency virus (HIV), cytomegalovirus and dengue. The patient also manifested leukocytosis with hypereosinophilia. Serological testing for toxacariasis was requested, diagnosing the infection. After treatment, he progressed with full symptoms remission. The aim of this study was to discuss confounding factors, differential diagnoses, the need for specific complementary exams and therapeutic management, according to the clinical aspects.(AU)

Dengue Virus Induces NK Cell Activation through TRAIL Expression during Infection.

Dengue is an acute febrile illness with a wide spectrum of signs and symptoms ranging from mild to severe forms characterized by plasma leakage that can be fatal. NK cells are one of the main effectors in early infection and may play an important role in dengue pathogenesis. We investigated NK cell involvement during dengue infection. A higher frequency of NK cell subsets and TRAIL+NK cells was found in mild DF cases when compared to that in severe cases or healthy donors. NK activation markers such as CD107a and TLR3 were upregulated in patients' cells compared to those in healthy donors. In addition, IL12 related to NK cell activation were upregulated in mild DF cases. In vitro PBMC culture models show that DENV-stimulated and IFNα-stimulated NK cells were able to express TRAIL, suggesting an indirect activation of cells, regarding TRAIL expression. Type I IFN receptor blockage on DENV-stimulated PBMCs showed TRAIL expression on NK cells is partially IFNα dependent. In addition, during PBMC stimulation, TRAIL expression on NK cells was inversely correlated with DENV-positive monocytes. Therefore, we observed DENV-induced activation of NK cell populations. A higher activation of NK cells would promote limited viral spread, resulting in decreased inflammatory response, contributing to protection against dengue severity.

Association of rs1285933 single nucleotide polymorphism in CLEC5A gene with dengue severity and its functional effects.

Outbreaks of the Zika, dengue, and chikungunya viruses, especially in the Americas, pose a global threat due to their rapid spread and difficulty controlling the vector. Extreme phenotypes are often observed, from asymptomatic to severe clinical manifestations, which are well-studied in dengue. Host variations are also important contributors to disease outcomes, and many case-control studies have associated single nucleotide polymorphisms (SNPs) with severe dengue. Here, we found that the TC genotype and T-carriers for SNP rs1285933 in the C-type lectin superfamily member 5 (CLEC5A) gene was associated with severe dengue in a Northern Brazilian population (OR=2.75 and p-value=0.01, OR=2.11 and p-value=0.04, respectively). We also tested the functional effect of the CLEC5A protein and found that it is upregulated on the surface of human monocytes after in vitro dengue infection. CLEC5A was correlated with viral load inside the monocytes (Spearman r=0.55, p=0.008) and TNF production in culture supernatants (Spearman r=0.72, p=0.03). Analysis of mRNA in blood samples from DENV4-infected patients exhibiting mild symptoms showed that CLEC5A mRNA expression is correlated with TNF (r=0.67, p=0.0001) and other immune mediators. Monocytes from rs1285933 TT/TC individuals showed lower CLEC5A expression compared to CC genotypes. However, in these cells, CLEC5A was not correlated with TNF production. In summary, we confirmed that CLEC5A is genetically associated with dengue severity outcome, playing a central role during the immune response triggered by a dengue viral infection, and rs1285933 is a relevant SNP that is able to regulate signaling pathways after interactions between the dengue virus and CLEC5A receptors.

Evidence of dengue virus replication in a non-traumatic spleen rupture case.

The present report describes a case of splenic rupture due to dengue, a rare complication of dengue that should be considered in any patient with suspected dengue disease who started with left upper quadrant abdominal pain and hypotension. The pathophysiology of this entity is not yet well elucidated, but one of the theories present in the literature is that it is due to a depletion of coagulation factors and platelets leading to intra-splenic hemorrhage and rupture. The RT-PCR technique detected serotype 1 and histopathological studies of the spleen revealed significant atrophy of lymphoid follicles and extensive hemorrhage areas. Besides histopathological observations, virus replication was investigated by detection of dengue antigens, especially the non-structural 3 protein (NS3) in endothelial cells and splenic macrophages. This important complication has serious clinical repercussions and high mortality, due to the diagnostic difficulty and many factors that usually confuse or delay its diagnosis. Therefore, it is of the utmost importance to recognize their manifestations and their management to try to best minimize their consequences and mortality.

Analysis of Clinical and Laboratory Alterations Related to Dengue Case Severity: Comparison between Serotypes 2 and 4 in Brazil.

The increase in severe dengue (SD) cases has caused great impact on public health and has concerned authorities of countries where the disease is endemic and epidemics reach high proportions. The recognition of progression signs of this severe disease during the initial febrile phase can be difficult, since the symptoms are often indistinguishable from other febrile diseases. The aim of this study was to evaluate the clinical manifestations and laboratory findings in patients from two dengue outbreaks and their association with the disease. The study was conducted in patients (n = 153) with signs and symptoms consistent with dengue occurred during two distinct epidemics, 2010 and 2013, in the city of Campos dos Goytacazes, Rio de Janeiro, Brazil. According to the 2009 World Health Organization criteria, patients were classified as dengue without warning signs ([DwoWS] 60.6%, 57/94), dengue with warning signs ([DwWS] 30.9%, 29/94), and SD (4.25%, 4/94). Patients with DwWS/SD presented lower platelet and leukocyte counts and higher transaminase levels when compared with the DwoWS ones. Interestingly, patients from the epidemic of 2010 caused by dengue virus 2 (DENV-2) had lower platelet counts than patients of the 2013 epidemic caused by DENV-4. Furthermore, plasma leakage, gastrointestinal bleeding, and pleural effusion, hallmarks for a more severe disease, were also more frequently observed in those cases. Although previous studies may have extensively reported the wide range of the clinical aspects of dengue, the characterization of DENV-4 is desirable considering the burden of the disease during epidemics, especially for the health units and hospitals performing patient's management.

Increased circulating procoagulant and anticoagulant factors as TF and TFPI according to severity or infecting serotypes in human dengue infection.

Tissue Factor (TF) is the initiator of coagulation and Tissue Factor Inhibitor (TFPI) is the physiological inhibitor of the TF/FVIIa complex. Circulating levels of TF and TFPI were quantified in dengue patients and the relationships with disease severity and infecting serotype analysed. A significant decrease in TF and TPFI plasma levels was observed in mild DF patients compared with severe dengue. Furthermore, both factors were associated with haemorrhagic manifestations. Finally, TF levels were significantly increased in DENV-1/2 infected patients as compared with DENV-4. These findings suggest that activation of TF-pathway is an important component of DENV -related coagulation disorders.

Aminotransferases analysis in patients with dengue serotype IV / Análise das aminotransferases em pacientes com dengue sorotipo IV

OBJECTIVE: To evaluate the aminotransferase levels inpatients with dengue serotype IV. METHODS: We conducteda cross-sectional analysis of 3,596 people with laboratory confirmed dengue. RESULTS: The analysis showed that 49.2%(1,768) had normal aminotransferase levels, 43.4% (1,559)had changes in levels by up to three times the upper limit ofnormal, 7.0% (253) had elevations three times the referencevalue and 0.4% (16) developed acute hepatitis. Patients withthrombocytopenia and hemoconcentration had major changesin the levels of enzymes (p<0.05). Women were more susceptibleto those changes than men (p<0,05). The average AST and ALTwere higher in dengue hemorrhagic type (147.51±137.74U/Land 111.54±81.27U/L) as compared to dengue fever(55.18±52.06U/L and 60.39±57.50U/L) (p<0.05). Leukopenicpatients had aminotransferases average lower compared toindividuals without any drop in overall white blood cellcount (p<0.05). A total of 224 patients were submitted tohospitalization, with the AST average of 88.18±100.47U/Land ALT 77.95±88.38U/L, which is higher than the average inambulatory patients (p<0.05). CONCLUSION: A significantnumber of patients was found with altered levels of enzymes,which requires clinical laboratory monitoring for an extendedperiod.

OBJETIVO: Avaliar os níveis de aminotransferases em pacientes com dengue sorotipo IV. MÉTODOS: Foi realizada uma análise transversal de 3.596 pessoas com dengue confirmada laboratorialmente. RESULTADOS: A análise mostrou que49,2% (1.768) tinham níveis normais de aminotransferases,43,4% (1.559) mostraram alterações nos níveis em até três vezes o limite superior da normalidade, 7,0% (253) apresentaram elevações três vezes o valor de referência e 0,4% (16) desenvolveram hepatite aguda. Os doentes com trombocitopenia e hemoconcentração tiveram grandes alterações nos níveis enzimáticos(p<0,05). As mulheres foram mais suscetíveis a essas alterações do que os homens (p<0,05). As médias de AST e ALT foram maiores na dengue do tipo hemorrágica (147,51±137,74U/Le 111,54±81,27U/L) em comparação com dengue clássica(55,18±52,06U/L e 60,39±57,50U/L) (p<0,05). Pacientes leucopênicos tiveram a média de aminotransferases menor em comparação com os indivíduos sem qualquer queda na contagem global de glóbulos brancos (p<0,05). Um total de 224pacientes foi submetido a hospitalização, com a média de AST88,18±100,47U/L e ALT 77,95±88,38U/L, o que é mais elevado do que a média em doentes ambulatórios (p<0,05). CONCLUSÃO:Um número significativo de pacientes foi identificado com níveis alterados de enzimas, o que exige acompanhamento clínico e laboratorial por um período prolongado.

Polymyositis due to Toxoplasma gondii in an immunocompetentpatient / Polimiosite por Toxoplasma gondii em paciente imunocompetente

Toxoplasmosis is a worldwide zoonosis caused by the protozoan Toxoplasma gondii, which is mostly acquired by food contaminated with sporulated oocysts present in the feces of cats. The majority of patients evolve without symptoms, if immunocompetent. Its diagnosis is mainly clinical, confirmed by serological study. We report a rare occurrence of toxoplasmosis polymyositis in an immunocompetent patient, emphasing confounding factors, differential diagnoses, complementary tests and therapeutic management, according to the clinical picture.

A toxoplasmose é uma zoonose cosmopolita causada pelo protozoário Toxoplasma gondii, adquirida por alimentos contaminados com oocistos esporulados, presentes nas fezes de felinos. A maioria dos pacientes evolui de forma assintomática, principalmente os imunocompetentes. Possui diagnóstico principalmente clínico, confirmado pela sorologia. Descrevemos uma rara apresentação de polimiosite por toxoplasmose em paciente imunocompetente, enfatizando os fatores confundidores, diagnósticos diferenciais, exames complementares e conduta terapêutica, de acordo com o quadro clínico.