Marialva Tereza Araujo

In situ immune response and mechanisms of cell damage in central nervous system of fatal cases microcephaly by Zika virus

Zika virus (ZIKV) has recently caused a pandemic disease, and many cases of ZIKV infection in pregnant women resulted in abortion, stillbirth, deaths and congenital defects including microcephaly, which now has been proposed as ZIKV congenital syndrome. This study aimed to investigate the in situ immune response profile and mechanisms of neuronal cell damage in fatal Zika microcephaly cases. Brain tissue samples were collected from 15 cases, including 10 microcephalic ZIKV-positive neonates with fatal outcome and five neonatal control flavivirus-negative neonates that died due to other causes, but with preserved central nervous system (CNS) architecture. In microcephaly cases, the histopathological features of the tissue samples were characterized in three CNS areas (meninges, perivascular space, and parenchyma). The changes found were mainly calcification, necrosis, neuronophagy, gliosis, microglial nodules, and inflammatory infiltration of mononuclear cells. The in situ immune response against ZIKV in the CNS of newborns is complex. Despite the predominant expression of Th2 cytokines, other cytokines such as Th1, Th17, Treg, Th9, and Th22 are involved to a lesser extent, but are still likely to participate in the immunopathogenic mechanisms of neural disease in fatal cases of microcephaly caused by ZIKV.

Clinical and pathological importance of vacA allele heterogeneity and cagA status in peptic ulcer disease in patients from North Brazil.

We have examined the prevalence of gene cagA and vacA alleles in 129 patients, 69 with gastritis and 60 with peptic ulcer diseases from North Brazil and their relation with histopathological data. vacA and cagA genotype were determined by polymerase chain reaction. Hematoxylin-eosin staining was used for histological diagnosis. 96.6% of the patients were colonized by Helicobacter pylori strains harboring single vacA genotype (nont-mixed infection). Among them, 11.8% had subtype s1a, 67.8% had subtype s1b, and 17% subtype s2. In regard to the middle region analysis, m1 alleles were found in 75.4% and m2 in 21.2% of patients. The cagA gene was detected in 78% patients infected with H. pylori and was associated with the s1-m1 vacA genotype. The H. pylori strains, vacA s1b m1/cagA-positive, were associated with increased risk of peptic ulcer disease and higher amounts of lymphocytic and neutrophilic infiltrates and the presence of intestinal metaplasia. These findings show that cagA and vacA genotyping may have clinical relevance in Brazil.

Zika virus epidemic in Brazil. I. Fatal disease in adults: Clinical and laboratorial aspects

BACKGROUNDnZika virus (ZIKV) was first detected in Brazil in May 2015 and the country experienced an explosive epidemic. However, recent studies indicate that the introduction of ZIKV occurred in late 2013. Cases of microcephaly and deaths associated with ZIKV infection were identified in Brazil in November, 2015.nnnOBJECTIVESnTo determine the etiology of three fatal adult cases.nnnSTUDY DESIGNnHere we report three fatal adult cases of ZIKV disease. ZIKV infection in these patients was confirmed by cells culture and/or real-time reverse transcriptase polymerase chain reaction (RT-qPCR) and by antigen detection using immunohistochemical assay. Samples of brain and other selected organs taken at autopsy from three patients were also analyzed by histopathological and immunohistological examination.nnnRESULTSnThe first patient, a 36-year-old man with lupus and receiving prednisone therapy, developed a fulminant ZIKV infection. At autopsy, RT-qPCR of blood and tissues was positive for ZIKV RNA, and the virus was cultured from an organ homogenate. The second patient, a previously healthy female, 16 years of age, presented classic symptoms of Zika fever, but later developed severe thrombocytopenia, anemia and hemorrhagic manifestations and died. A blood sample taken on the seventh day of her illness was positive RT-PCR for ZIKV RNA and research in the serum was positive for antinuclear factor fine speckled (1/640), suggesting Evans syndrome (hemolytic anemia an autoimmune disorder with immune thrombocytopenic purpura) secondary to ZIKV infection. The third patient was a 20-year-old woman hospitalized with fever, pneumonia and hemorrhages, who died on 13days after admission. Histopathological changes were observed in all viscera examined. ZIKV antigens were detected by immunohistochemistry in viscera specimens of patients 1 and 3. These three cases demonstrate other potential complications of ZIKV infection, in addition to microcephaly and Guillain-Barre syndrome (GBS), and they suggest that individuals with immune suppression and/or autoimmune disorders may be at higher risk of developing severe disease, if infected with ZIKV.

MYC in gastric carcinoma and intestinal metaplasia of young adults

MYC has a key role in gastric carcinogenesis. We evaluated MYC copy number and protein expression in nonneoplasic, intestinal metaplasia, and gastric cancer samples from five young adults. We observed a significant increase of MYC amplification with the evolution of carcinogenesis process. MYC overexpression was observed in intestinal metaplasia and neoplastic tissue from all patients with intestinal-type gastric cancer and from no patients with diffuse type. MYC copy number and expression can be biomarkers of gastric malignance. Gastric cancer (GC) is the fourth most frequent cancer type and the second highest cause of cancer mortality worldwide [1]. In Par a state, northern Brazil, elevated incidence of GC has been verified [2]. Gastric adenocarcinoma is divided mainly into intestinal and diffuse types according to Laur en classification [3]. The intestinal type progresses through a number of sequential steps, beginning with atrophic gastritis, followed by intestinal metaplasia (IM), intraepitelial neoplasia, and carcinoma [4]. On the other hand, diffuse-type generally does not evolve from precancerous lesions [5], even though some patients presented diffuse-type together with IM, which raised doubts about the association between IM and GC development [6]. Thus, it remains unclear whether IM is a premalignant condition or a marker for an increased risk of malignancy. GC affects mainly older patients. Early-onset GC ( 40 years) is rare, and genetic factors may be more important in these patients. It is suggested that young patients with GC have a poorer prognosis than elderly patients [7]. MYC is a proto-oncogene commonly deregulated in gastric cancer, which is involved mainly in cell cycle regulation and growth arrest [8]. To investigate whether MYC alterations can be used as a cancer biomarker, we evaluated MYC copy number and protein expression in formalinfixed, paraffin-embedded samples microdissected into non-neoplasic mucosa, IM, and neoplastic tissue from five patients with early-onset GC. These samples were obtained at Par a State Jo~ao de Barros Barreto University Hospital, with the approval of the hospital Ethics Committee. Table 1 shows the cases along with their clinicopathologic characteristics. Fluorescence in situ hybridization (FISH) was performed according to the method of Raiol et al. [9], using the MYC region probe (ONPON0824; Bioagency Biotechnology, S~ao Paulo, Brazil) to evaluate this gene copy

Association of cytokine gene polymorphisms and serum concentrations with the outcome of chronic hepatitis B

OBJECTIVEnThe present paper investigated possible correlations between the clinical presentation of hepatitis B and the TNF-α -308G/A, IFN-γ +874A/T, TGF-beta1 -509C/T, and IL-10 -1081A/G polymorphisms and associated serum levels of these cytokines.nnnMETHODSnFifty-three hepatitis patients were selected and divided into two groups: A - inactive (n=30) and B - chronic hepatitis/cirrhosis (n=23). The control group consisted of 100 subjects who were positive for anti-HBc and anti-HBs. The serum concentrations of the cytokines were determined by immunoenzymatic assays. The polymorphisms of the cytokines genes were assessed by PCR and PCR-SSP.nnnRESULTSnThe mean serum levels of IFN-γ of the control group were significantly higher than those of groups A and B, whereas the mean levels TGF-beta1 were significantly higher in groups A and B in comparison with the control. In the case of IL-10, the mean serum level recorded in the control group was significantly higher than that of group B. The TNF-α -308AG genotype was considerably more frequent in group B (43.3%) than the control (14.4%).nnnCONCLUSIONnHigher serum levels of IFN-γ and TGF-beta1 were associated with chronic hepatitis B, and lower serum levels of IL-10 were found in patients with the active disease. Furthermore the presence of allele A of the TNF-α -308 polymorphism suggest a risk of the progressive disease.

In situ inflammasome activation results in severe damage to the central nervous system in fatal Zika virus microcephaly cases

HighlightsZIKV induce activation inflammasome in situ in fatal cases of microcephaly.Recognition of the ZIKV PAMPs by NLRP1, NLRP3, and AIM2 provoke release of caspase 1.Caspase 1 activation converts cytokines IL‐1&bgr;, IL‐18, and IL‐33 to bioactive form.Response induced by Caspase 1, iNOS, and cytokines enhance neuroinflammatory process. Abstract Zika virus (ZIKV) has caused substantial concern worldwide owing to its association with severe birth defects, such as microcephaly and other congenital malformations. Inflammasomes, i.e., multi‐protein complexes that induce inflammation and pyroptosis, are predicted to contribute to the immune response to this flavivirus. Accordingly, in this study, the in situ inflammasome response was evaluated in fatal cases of ZIKV‐linked microcephaly. Brain tissue samples were collected from eight babies, including four ZIKV‐positive microcephalic neonates who died after birth and four flavivirus‐negative neonatal controls who died of other causes and whose central nervous system (CNS) architecture was preserved. In the ZIKV‐positive newborn/stillbirth babies, the major histopathological alterations included atrophy of the cortical layer, a predominance of mononuclear cell infiltration in the Virchow–Robin space, neuronal necrosis, vacuolization and neuronal degeneration, neuronophagy, and gliosis. An immunohistochemical analysis of tissues in the neural parenchyma showed significantly higher expression of the receptors NLRP1, NLRP3, and AIM2, cytokines IL‐1&bgr;, IL‐18, and IL‐33, and enzymes caspase 1, iNOS, and arginase 1 in ZIKV‐positive microcephaly cases than in flavivirus‐negative controls. These results suggest that inflammasome activation can aggravate the neuroinflammatory response and consequently increase CNS damage in neonates with fetal neural ZIKV infection and microcephaly.

Correlation between Apoptosis and in Situ Immune Response in Fatal Cases of Microcephaly Caused by Zika Virus

Zika virus (ZIKV) is a single-stranded positive-sense RNA flavivirus that possesses a genome approximately 10.7 Kb in length. Although pro-inflammatory and anti-inflammatory cytokines and apoptotic markers belonging to the extrinsic and intrinsic pathways are suggested to be involved in fatal cases of ZIKV-induced microcephaly, their exact roles and associations are unclear. To address this, brain tissue samples were collected from 10 individuals, five of whom were diagnosed as ZIKV positive with microcephaly and a further five were flavivirus-negative controls that died because of other causes. Examination of material from the fatal cases of microcephaly revealed lesions in the cerebral cortex, edema, vascular proliferation, neuronal necrosis, gliosis, neuronophagy, calcifications, apoptosis, and neuron loss. The expression of various apoptosis markers in the neural parenchyma, including FasL, FAS, BAX, BCL2, and caspase 3 differed between ZIKV-positive cases and controls. Further investigation of type 1 and 2 helper T-cell cytokines confirmed a greater anti-inflammatory response in fatal ZIKV-associated microcephaly cases. Finally, an analysis of the linear correlation between tumor necrosis factor-α, IL-1β, IL-4, IL-10, transforming growth factor-β, and IL-33 expression and various apoptotic markers suggested that the immune response may be associated with the apoptotic phenomenon observed in ZIKV-induced microcephaly.

Case report: Niemann-Pick disease with liver failure manifestations

OBJECTIVE: To report the case of a patient with Niemann-Pick disease who developed chronic liver failure, and to thus draw attention to this nosological entity. CASE REPORT: A female patient, 34 years of age, presented with splenomegaly and was referred to the Fundação Santa Casa de Misericórdia do Pará hospital, in the City of Belém, Pará State, Brazil, with a diagnosis of Niemann-Pick disease confirmed by enzymatic dosages for investigation of her liver disease. The patient developed cirrhosis after five years with decompensated chronic liver failure and was referred for liver transplantation. FINAL CONSIDERATIONS: It is noted that this disease, although rare, can be a cause of chronic liver disease and should be diagnosed early, as the patients life expectancy depends on the magnitude and impact of the symptoms.

Informe de caso: enfermedad de Niemann-Pick con manifestaciones de insuficiencia hepática

OBJECTIVE: To report the case of a patient with Niemann-Pick disease who developed chronic liver failure, and to thus draw attention to this nosological entity. CASE REPORT: A female patient, 34 years of age, presented with splenomegaly and was referred to the Fundação Santa Casa de Misericórdia do Pará hospital, in the City of Belém, Pará State, Brazil, with a diagnosis of Niemann-Pick disease confirmed by enzymatic dosages for investigation of her liver disease. The patient developed cirrhosis after five years with decompensated chronic liver failure and was referred for liver transplantation. FINAL CONSIDERATIONS: It is noted that this disease, although rare, can be a cause of chronic liver disease and should be diagnosed early, as the patients life expectancy depends on the magnitude and impact of the symptoms.