Adverse birth outcomes associated with Zika virus exposure during pregnancy in São José do Rio Preto, Brazil.

OBJECTIVES: We aimed to report the first 54 cases of pregnant women infected by Zika virus (ZIKV) and their virologic and clinical outcomes, as well as their newborns' outcomes, in 2016, after the emergence of ZIKV in dengue-endemic areas of São Paulo, Brazil. METHODS: This descriptive study was performed from February to October 2016 on 54 quantitative real-time PCR ZIKV-positive pregnant women identified by the public health authority of São José do Rio Preto, São Paulo, Brazil. The women were followed and had clinical and epidemiologic data collected before and after birth. Adverse outcomes in newborns were analysed and reported. Urine or blood samples from newborns were collected to identify ZIKV infection by reverse transcription PCR (RT-PCR). RESULTS: A total of 216 acute Zika-suspected pregnant women were identified, and 54 had the diagnosis confirmed by RT-PCR. None of the 54 women miscarried. Among the 54 newborns, 15 exhibited adverse outcomes at birth. The highest number of ZIKV infections occurred during the second and third trimesters. No cases of microcephaly were reported, though a broad clinical spectrum of outcomes, including lenticulostriate vasculopathy, subependymal cysts, and auditory and ophthalmologic disorders, were identified. ZIKV RNA was detected in 18 of 51 newborns tested and in eight of 15 newborns with adverse outcomes. CONCLUSIONS: Although other studies have associated many newborn outcomes to ZIKV infection during pregnancy, these same adverse outcomes were rare or nonexistent in this study. The clinical presentation the newborns we studied was mild compared to other reports, suggesting that there is significant heterogeneity in congenital Zika infection.

Zika Virus Infection and Solid Organ Transplantation: A New Challenge.

Public health concerns exist surrounding the epidemic of the Zika virus (ZIKV) and the rapid growth of transplantation in developing countries, including endemic zones of active arbovirus transmission, as well as travel to such regions by potential organ donors and recipients. Few data exist regarding the clinical characteristics of ZIKV infection in immunocompromised hosts. Laboratory screening protocols for transplantation to differentiate ZIKV infections from other endemic viral diseases and for the detection of possible donor-derived infection have not been stated. The diagnosis of ZIKV infection remains a challenge, fueled by the lack of standardized commercially available diagnostic tests and validated reference diagnostic laboratories, as well as the limited duration of ZIKV viremia. In this small series, ZIKV infection in renal and liver recipients presented without rash, conjunctivitis, or neurological symptoms, and with abnormal graft function, thrombocytopenia, and bacterial superinfection. We report the first case series of ZIKV infection in solid organ recipients, with a description of clinical and laboratory features and therapeutic management.

Isolation and Characterization of Madariaga Virus from a Horse in Paraíba State, Brazil.

Madariaga virus (MADV), the new species designation for the South American isolates of eastern equine encephalitis virus (EEEV), is genetically divergent and substantially different in ecology and pathogenesis from North American EEEV strains. We isolated and characterized a MADV isolate obtained from a horse in Brazil. Our results support previous phylogenetic studies showing there are three genetically distinct MADV lineages. The MADV isolate from Paraíba State belongs to the South American lineage III and is closely related to Peruvian, Colombian and Venezuelan isolates.

A phylogenetic analysis using full-length viral genomes of South American dengue serotype 3 in consecutive Venezuelan outbreaks reveals a novel NS5 mutation.

Dengue virus currently causes 50-100 million infections annually. Comprehensive knowledge about the evolution of Dengue in response to selection pressure is currently unavailable, but would greatly enhance vaccine design efforts. In the current study, we sequenced 187 new dengue virus serotype 3 (DENV-3) genotype III whole genomes isolated from Asia and the Americas. We analyzed them together with previously-sequenced isolates to gain a more detailed understanding of the evolutionary adaptations existing in this prevalent American serotype. In order to analyze the phylogenetic dynamics of DENV-3 during outbreak periods; we incorporated datasets of 48 and 11 sequences spanning two major outbreaks in Venezuela during 2001 and 2007-2008, respectively. Our phylogenetic analysis of newly sequenced viruses shows that subsets of genomes cluster primarily by geographic location, and secondarily by time of virus isolation. DENV-3 genotype III sequences from Asia are significantly divergent from those from the Americas due to their geographical separation and subsequent speciation. We measured amino acid variation for the E protein by calculating the Shannon entropy at each position between Asian and American genomes. We found a cluster of seven amino acid substitutions having high variability within E protein domain III, which has previously been implicated in serotype-specific neutralization escape mutants. No novel mutations were found in the E protein of sequences isolated during either Venezuelan outbreak. Shannon entropy analysis of the NS5 polymerase mature protein revealed that a G374E mutation, in a region that contributes to interferon resistance in other flaviviruses by interfering with JAK-STAT signaling was present in both the Asian and American sequences from the 2007-2008 Venezuelan outbreak, but was absent in the sequences from the 2001 Venezuelan outbreak. In addition to E, several NS5 amino acid changes were unique to the 2007-2008 epidemic in Venezuela and may give additional insight into the adaptive response of DENV-3 at the population level.

Infantile reflexes and their effects on dental caries and oral hygiene in cerebral palsy individuals.

The aim of this study was to evaluate the presence of pathological primitive reflexes and their effects on dental caries and oral hygiene in cerebral-palsied individuals. A group of 124 non-institutionalized patients (57 males) attending Lar Escola Sao Francisco Rehabilitation Center, who had a medical diagnosis of spastic CP (age range 3-17 years, mean+/-s.d.=8.6+/-4.3) were selected for dental clinical evaluation according to WHO (Oral health surveys: basic methods, WHO, Geneva, 1997) and oral hygiene using the oral hygiene index proposed by Greene and Vermillion (J Am Dent Assoc, 1964, 68, 7). The presence/absence of the pathological primitive oral reflexes of rooting, suckle-swallow, biting and gagging was evaluated by observing patients' reaction after a stimulus. Results were statistically analysed by Fisher's exact, Chi-square and Kruskal-Wallis tests. A significantly higher percentage of the biting reflex in patients with quadriplegia was observed, and also the absence of this reflex in patients with hemiplegia (P=0.006). Patients with hemiplegia presented significantly lower DMF index values. Kruskal-Wallis test showed no statistical difference (P=0.335; 0.677; 0.202) for OHI-S among the quadriplegia, diplegia and hemiplegia for all dentitions. Our data suggest that the more severe the neurological damage is, the more frequent is the presence of the biting reflex and consequently, the higher is the risk of oral diseases in this population due to the difficulty to perform an adequate oral hygiene.

Detection of herpesvirus DNA by the polymerase chain reaction (PCR) in vitreous samples from patients with necrotising retinitis.

AIMS: Viral uveitis and retinitis, usually caused by herpesviruses, are common in immunosuppressed patients. The diagnosis of viral anterior uveitis and retinitis is usually clinical. The polymerase chain reaction (PCR) has been used for the diagnosis of some viral infections, especially those caused by herpesviruses. This paper reports the use of PCR in the diagnosis of viral retinitis in vitreous samples from Brazilian patients. METHODS: PCR was used for the diagnosis of necrotising retinitis in vitreous samples from patients from the Hospital São Geraldo, Universidade Federal de Minas Gerais, Brazil. The vitreous samples were collected by paracentesis and stored until analysis. Samples were analysed by PCR using specific primers designed to amplify herpes simplex virus 1 (HSV-1), varicella zoster virus (VZV), or human cytomegalovirus (HCMV). In a case of anterior uveitis, PCR was performed with a sample from the anterior chamber. RESULTS: Herpesvirus DNA was amplified in 11 of 17 samples. HCVM DNA was detected in nine samples but DNA from HSV-1 and VZV were detected only once each. CONCLUSION: These results strongly suggest that PCR could be used for a rapid complementary diagnosis of viral uveitis and retinitis. A prospective study to evaluate the PCR results, clinical evolution, and treatment is imperative to corroborate the real value of PCR in diagnosis and how it could help the clinicians' approach.

Comparison of virus isolation and various polymerase chain reaction methods in the diagnosis of mucocutaneous herpesvirus infection.

We compared two polymerase chain reaction (PCR) assays (simple and multiplex) and viral isolation to detect herpes simplex virus 1 (HSV-1) and varicella-zoster virus (VZV) in 15 clinical specimens from 13 patients with mucocutaneous herpetic infections. HSV-1 or VZV DNA was detected in 13 specimens by simple PCRs (HSV-1 or VZV PCR) and in 12 specimens by multiplex PCR. On the other hand, viral isolation was positive for 9 specimens only. The PCR protocols used in this study are not only more sensitive and faster than the traditional viral isolation and conventional PCR protocols but also can distinguish rapidly HSV-1 from VZV. We propose the PCRs described here for rapid and precise identification of etiological agents of mucocutaneous herpetic infections.

PCR-based diagnosis of a case of herpetic whitlow in an AIDS patient.

Herpetic infections are common complications in AIDS patients. The clinical features could be uncommon and antiviral chemotherapy is imperative. A rapid diagnosis could prevent incorrect approaches and treatment. The polymerase chain reaction is a rapid, specific and sensible method for DNA amplification and diagnosis of infectious diseases, especially viral diseases. This approach has some advantages compared with conventional diagnostic procedures. Recently we have reported a new PCR protocol to rapid diagnosis of herpetic infections with suppression of the DNA extraction step. In this paper we present a case of herpetic whitlow with rapid diagnosis by HSV-1 specific polymerase chain reaction using the referred protocol.

[Epidemiological, social, and sanitary aspects in an area of the Rio Negro, State of Amazonas, with special reference to intestinal parasites and Chagas' disease]. / Aspectos epidemiológicos, sociais e sanitários de uma área no Rio Negro, estado do Amazonas, com especial referência às parasitoses intestinais e à infecção chagásica.

A cross-sectional study was carried out on the residents of one in every four dwellings in the town of Barcelos (in the northern part of the State of Amazonas, on the right bank of the Rio Negro, 490 kilometers from Manaus by river), in order to evaluate social and sanitary conditions and specific indicators for intestinal parasites and Chagas' infection. During the survey, two questionnaires were applied, a household one to evaluate social and sanitary aspects, and an individual one, for social and epidemiological evaluation of the population conditions. A conglomerate family sample of 171 dwellings was studied. From each of the 658 habitants, a sample was requested for stool examination by Lutz sedimentation and Baermann-Moraes-Coutinho techniques modified by Willcox & Coura (1989), and blood was collected in filter paper for immunofluorescence test by Camargo (1966) and Souza & Camargo (1966) methods modified by Petana & Willcox (1975). The stool examination showed 69.4% of samples with one or more parasites. Ascaris lumbricoides was predominant with 51% of positivity and Entamoeba histolytica, although surveyed by a non-specific method, was present in 19.7%. Surprisingly, 20.1% of the 658 sera samples were reactive for T. cruzi antibodies at a dilution of 1:20 and 13.7% at 1:40. There was a strong correlation between this result and the level of human contact with wild triatomines, known locally as "piasava lice", and we succeeded in isolating by xenodiagnosis one strain of T. cruzi from one patient, a sixty-one-year old man (n. 209 -1), a native of the area, with positive serology for Chagas' disease and who worked in agriculture and transporting piasava and was very familiar with "piasava lice".

Membrane fusion during secretion. A hypothesis based on electron microscope observation of Phytophthora Palmivora zoospores during encystment.

Interpretation of freeze-fracture and thin-section results shows that fusion of the peripheral vesicle with the plasmalemma of a Phytophthora palmivora zoospore occurs at several discrete sites and results in the formation and expansion of a particle-free bilayer membrane diaphragm and in the appearance of a polymorphic network of membrane-bounded tunnels, the lumina of which are continuous with the cytoplasm. The outer half of the bilayer membrane diaphragm appears continuous with the outer half of the plasma membrane; the inner half of the bilayer membrane diaphragm with the inner half of the peripheral vesicle membrane; and the inner half of the plasmalemma with the outer half of the peripheral vesicle membrane. Interpretation of our results leads us to formulate a hypothesis for a sequence of several intermediate stages involved in membrane fusion. The initial fusion event is viewed as a local catastrophe (Thom, R. 1972. Stabilité Structurelle et Morphogenèse. W. A. Benjamin Inc., Reading, Mass.) involving the sudden reorganization of apposed elements of the inner half of the plasmalemma and the outer half of the peripheral vesicle membrane. Fusion of apposed components at the rim of the perimeter of fusion results in the formation of a toroid hemi-micelle which provides continuity between the inner half of the plasmalemma and the outer half of the peripheral vesicle membrane. Simultaneously, apposed components at the site of fusion may reorganize into an inverted membrane micelle. A bilayer membrane diaphragm is then formed by apposition and flowing of components form the outer half of the plasmalemma and the inner (exoplasmic) half of the peripheral vesicle membrane. The existence of large areas of membrane contact before fusion may lead to several fusion events and the formation of a polymorphic network of membrane-bound tunnels.