Establishment and cryptic transmission of Zika virus in Brazil and the Americas.

Transmission of Zika virus (ZIKV) in the Americas was first confirmed in May 2015 in northeast Brazil. Brazil has had the highest number of reported ZIKV cases worldwide (more than 200,000 by 24 December 2016) and the most cases associated with microcephaly and other birth defects (2,366 confirmed by 31 December 2016). Since the initial detection of ZIKV in Brazil, more than 45 countries in the Americas have reported local ZIKV transmission, with 24 of these reporting severe ZIKV-associated disease. However, the origin and epidemic history of ZIKV in Brazil and the Americas remain poorly understood, despite the value of this information for interpreting observed trends in reported microcephaly. Here we address this issue by generating 54 complete or partial ZIKV genomes, mostly from Brazil, and reporting data generated by a mobile genomics laboratory that travelled across northeast Brazil in 2016. One sequence represents the earliest confirmed ZIKV infection in Brazil. Analyses of viral genomes with ecological and epidemiological data yield an estimate that ZIKV was present in northeast Brazil by February 2014 and is likely to have disseminated from there, nationally and internationally, before the first detection of ZIKV in the Americas. Estimated dates for the international spread of ZIKV from Brazil indicate the duration of pre-detection cryptic transmission in recipient regions. The role of northeast Brazil in the establishment of ZIKV in the Americas is further supported by geographic analysis of ZIKV transmission potential and by estimates of the basic reproduction number of the virus.

Role of inosine triphosphate pyrophosphatase gene variant on fever incidence during zidovudine antiretroviral therapy.

Zidovudine, the antiretroviral drug used to treat HIV infection, commonly causes adverse effects, such as systemic fever and gastrointestinal alterations. In the present study, the potential role of inosine triphosphate pyrophosphatase (ITPA) gene variant on the incidence of adverse events during antiretroviral therapy (ART) of HIV with zidovudine was discussed. Individuals from Northeastern Brazil (N = 204) receiving treatment for HIV-1 infection were recruited. Zidovudine-related adverse effects developed during the treatment were registered. The rs1127354 polymorphism in the ITPA gene was genotyped using real-time PCR to assess whether this single nucleotide polymorphism was associated with the occurrence of zidovudine-related adverse effects. We observed a significant association between the ITPA variant genotype and the reported systemic fever (odds ratio = 7.17, 95% confidence interval = 1.19-43.15; P = 0.032). Zidovudine use could indirectly lead to an increase in the levels of inosine monophosphate in an antimetabolite-like manner, which is converted to inosine triphosphate (ITP). The rs1127354 variant caused a decrease in ITPA activity, thereby leading to ITP accumulation. This in turn resulted in cytotoxicity, which was manifested by neutropenia and fever. Therefore, we hypothesized a pharmacogenetic model involving the ITPA variant genotype in multifactorial components that act together to determine the onset of zidovudine-related adverse effects.

Full-length genomic and molecular characterization of Canine parvovirus in dogs from North of Brazil.

With the objective of characterizing Canine parvovirus (CPV) from some suspected fecal samples of dogs collected from the Veterinarian Hospital in Belém city, five positive samples were found by PCR assay and an update molecular characterization was provided of the CPV-2 circulation in Belém. Through sequencing of the complete DNA sequences (NS1, NS2, VP1, and VP2 genes), the CPV-2 strain was identified as CPV-2b (Asn426Asp) circulating in Belém. The CPV-2b strain with a different change at the position Tyr324Leu was detected in all samples assessed and thus reported for the first time for the scientific community. Phylogenetic analysis indicated that Belém CPV-2b and CPV-2a strains would be related to a cluster with samples after the 1990s, suggesting that CPV-2b in Belém originated from CPV-2a circulating in Brazil after the 1990s. Potential recombination events were analyzed using RDP4 and SplitsTree4; therefore, results suggest that CPV-2 sequences here described were not potentially recombination events. Continuous monitoring and molecular characterization of CPV-2 samples are needed not only to identify possible genetic and antigenic changes that may interfere with the effectiveness of vaccines but also to bring a better understanding of the mechanisms that drive the evolution of CPV-2 in Brazil.

In vitro neurotoxicity evaluation of piperazine designer drugs in differentiated human neuroblastoma SH-SY5Y cells.

Abuse of synthetic drugs is widespread worldwide. Studies indicate that piperazine designer drugs act as substrates at dopaminergic and serotonergic receptors and/or transporters in the brain. This work aimed to investigate the cytotoxicity of N-benzylpiperazine, 1-(3-trifluoromethylphenyl)piperazine, 1-(4-methoxyphenyl)piperazine and 1-(3,4-methylenedioxybenzyl)piperazine in the differentiated human neuroblastoma SH-SY5Y cell line. Cytotoxicity was evaluated after 24 h incubations through the MTT reduction and neutral red uptake assays. Oxidative stress (reactive oxygen and nitrogen species production and glutathione content) and energetic (ATP content) parameters, as well as intracellular Ca(2+), mitochondrial membrane potential, DNA damage (comet assay) and cell death mode were also evaluated. Complete cytotoxicity curves were obtained after 24 h incubations with each drug. A significant decrease in intracellular total glutathione content was noted for all the tested drugs. All drugs caused a significant increase of intracellular free Ca(2+) levels, accompanied by mitochondrial hyperpolarization. However, ATP levels remained unchanged. The investigation of cell death mode revealed a predominance of early apoptotic cells. No genotoxicity was found in the comet assay. Among the tested drugs, 1-(3-trifluoromethylphenyl)piperazine was the most cytotoxic. Overall, piperazine designer drugs are potentially neurotoxic, supporting concerns on risks associated with the abuse of these drugs.

Severe bronchiectasis in a patient with common variable immunodeficiency.

BACKGROUND: Bronchiectasis are common in Common Variable Immunodeficiency. These patients are prone to infection, leading to progressive lung destruction and accelerated FEV1 decline. CLINICAL CASE: 40 year-old man, with recurrent respiratory infections, autoimmunity and diarrhea since age 7. At 17 CVID was diagnosed and IVIgG was started. During the following years, respiratory symptoms progressively worsened and bronchiectasis was found on thoracic computed tomography. Bronchoscopy revealed Pseudomonas aeruginosa in bronchoalveolar lavage and bronchial secretions cultures. Eradication therapy led to clinical improvement. DISCUSSION: This case report stresses the importance of regular microbiological screening and appropriate antibiotherapy. Early/aggressive treatment may significantly impact on patients' evolution.

Monocyte activation is a feature of common variable immunodeficiency irrespective of plasma lipopolysaccharide levels.

Common variable immunodeficiency disorders (CVID), the most frequent cause of symptomatic primary immunodeficiency, are defined by impaired antibody production. Notwithstanding, T cell activation and granulomatous manifestations represent the main causes of CVID morbidity even in patients receiving immunoglobulin (Ig) G replacement therapy. Additionally, gut pathology is a frequent feature of CVID. In this study, we investigated monocyte imbalances and their possible relationship with increased microbial translocation in CVID patients. Monocyte subsets were defined according to CD14 and CD16 expression levels and evaluated in terms of human leucocyte antigen D-related (HLA-DR), CD86 and programmed death-1 molecule ligand 1 (PD-L1) expression by flow cytometry, in parallel with the quantification of plasma lipopolysaccharide (LPS) and serum levels of soluble CD14 (sCD14), LPS-binding protein (LBP) and anti-LPS antibodies. CVID patients (n=31) featured significantly increased levels of serum sCD14 and an expansion of CD14(bright) CD16(+) monocytes in direct correlation with T cell and B cell activation, the latter illustrated by the frequency of the CD21(low) CD38(low) subset. Such alterations were not observed in patients lacking B cells due to congenital agammaglobulinaemia (n=4). Moreover, we found no significant increase in circulating LPS or LBP levels in CVID patients, together with a relative preservation of serum anti-LPS antibodies, in agreement with their presence in commercial IgG preparations. In conclusion, CVID was associated with monocyte imbalances that correlated directly with T cell activation markers and with B cell imbalances, without an association with plasma LPS levels. The heightened monocyte activated state observed in CVID may represent an important target for complementary therapeutic strategies.

Production of low-cholesterol butter with Lacticaseibacillus paracasei immobilized in calcium-alginate beads.

This study focused on the application of three strains of Lacticaseibacillus paracasei to assimilate cholesterol in cream and butter. The strains were enclosed in calcium-alginate beads and incubated in cream at 30 °C for 15 h. Immobilization of lactobacilli cultures in calcium-alginate beads resulted in a 23% reduction in cholesterol (p < 0.05) in cream, whereas a negligible reduction was observed in cream fermented with free cells. Butter with a 44% reduction in cholesterol was produced from fermented cream by L. paracasei L2A21K5 entrapped in alginate beads. No significant (p > 0.05) changes in the fatty acid profile were observed in the low-cholesterol butter, except for a slight but significant increase in n-3 fatty acids (p < 0.05). In addition, the indices of atherogenicity and thrombogenicity were significantly reduced in the low-cholesterol butter (p < 0.05). Panelists rated the low-cholesterol butter as good in appearance, consistency, and flavor.

Review of gastric cancer risk factors in patients with common variable immunodeficiency disorders, resulting in a proposal for a surveillance programme.

Common variable immunodeficiency disorders (CVIDs) are the most frequent symptomatic primary immunodeficiencies in adults. They comprise a heterogeneous group of pathologies, with frequent non-infectious complications in addition to the bacterial infections that usually characterize their presentation. Complications include a high risk of malignancy, especially lymphoma and gastric cancer. Helicobacter pylori infection and pernicious anaemia are risk predictors for gastric cancer in the general population and probably in patients with CVIDs. Screening for gastric cancer in a high-risk population appears to improve survival. Given the increased risk of gastric cancer in patients with CVIDs and prompted by a case of advanced gastric malignancy in a patient with a CVID and concomitant pernicious anaemia, we performed a review of the literature for gastric cancer and conducted a cohort study of gastric pathology in 116 patients with CVIDs under long-term follow-up in Oxford. Regardless of the presence of pernicious anaemia or H. pylori infection, patients with CVIDs have a 10-fold increased risk of gastric cancer and are therefore a high-risk population. Although endoscopic screening of all patients with CVIDs could be considered, a more selective approach is appropriate and we propose a surveillance protocol that should reduce modifiable risk factors such as H. pylori, in order to improve the management of patients with CVIDs at risk of gastric malignancy.

European all-cause excess and influenza-attributable mortality in the 2017/18 season: should the burden of influenza B be reconsidered?

OBJECTIVES: Weekly monitoring of European all-cause excess mortality, the EuroMOMO network, observed high excess mortality during the influenza B/Yamagata dominated 2017/18 winter season, especially among elderly. We describe all-cause excess and influenza-attributable mortality during the season 2017/18 in Europe. METHODS: Based on weekly reporting of mortality from 24 European countries or sub-national regions, representing 60% of the European population excluding the Russian and Turkish parts of Europe, we estimated age stratified all-cause excess morality using the EuroMOMO model. In addition, age stratified all-cause influenza-attributable mortality was estimated using the FluMOMO algorithm, incorporating influenza activity based on clinical and virological surveillance data, and adjusting for extreme temperatures. RESULTS: Excess mortality was mainly attributable to influenza activity from December 2017 to April 2018, but also due to exceptionally low temperatures in February-March 2018. The pattern and extent of mortality excess was similar to the previous A(H3N2) dominated seasons, 2014/15 and 2016/17. The 2017/18 overall all-cause influenza-attributable mortality was estimated to be 25.4 (95%CI 25.0-25.8) per 100,000 population; 118.2 (116.4-119.9) for persons aged 65. Extending to the European population this translates into over-all 152,000 deaths. CONCLUSIONS: The high mortality among elderly was unexpected in an influenza B dominated season, which commonly are considered to cause mild illness, mainly among children. Even though A(H3N2) also circulated in the 2017/18 season and may have contributed to the excess mortality among the elderly, the common perception of influenza B only having a modest impact on excess mortality in the older population may need to be reconsidered.

Morphological characterization of the anuran integument of the Proceratophrys and Odontophrynus genera (Amphibia, Anuran, Leptodactylidae).

The morphological characteristics of the leptodactylid integument of Proceratophrys and Odontophrynus genera were investigated by means of stereoscopic, low vacuum scanning electron and light microscopy. The integument surface of Proceratophrys boiei, Proceratophrys laticeps and Proceratophrys appendiculata exhibited several projections, while the integument of Odontophrynus americanus had rounded elevations with smooth profile. Light microscopic observations showed the basic integument morphology for all anurans, i.e., an epidermis and a dermis, which is subdivided into a spongious layer and a compact layer. The epidermis is formed by basal, intermediary and cornified layers. However, in Proceratophrys genus the cornified layer had an irregular outline, while in O. americanus the external surface was smooth. In the spongious dermis, mucous and venom exocrine glands were observed, but in O. americanus an exclusive glandular type with apocrine secretory pattern was identified. The integument morphology showed peculiar characteristics that may be helpful for genus distinction. Thus, morphological methods may be considered as an efficient means to characterize and to differentiate anuran genera.

Maternal aggression in Wistar rats: effect of 5-HT2A/2C receptor agonist and antagonist microinjected into the dorsal periaqueductal gray matter and medial septum.

The objective of the present study was to assess the role of the 5-HT2A/2C receptor at two specific brain sites, i.e., the dorsal periaqueductal gray matter (DPAG) and the medial septal (MS) area, in maternal aggressive behavior after the microinjection of either a 5-HT2A/2C receptor agonist or antagonist. Female Wistar rats were microinjected on the 7th postpartum day with the selective agonist alpha-methyl-5-hydroxytryptamine maleate (5-HT2A/2C) or the antagonist 5-HT2A/2C, ketanserin. The agonist was injected into the DPAG at 0.2 (N = 9), 0.5 (N = 10), and 1.0 microg/0.2 microl (N = 9), and the antagonist was injected at 1.0 microg/0.2 microl (N = 9). The agonist was injected into the medial septal area (MS) at 0.2 (N = 9), 0.5 (N = 7), and 1.0 microg/0.2 microl (N = 6) and the antagonist was injected at 1.0 microg/0.2 microl (N = 5). For the control, saline was injected into the DPAG (N = 7) and the MS (N = 12). Both areas are related to aggressive behavior and contain a high density of 5-HT receptors. Non-aggressive behaviors such as horizontal locomotion (walking) and social investigation and aggressive behaviors such as lateral threat (aggressive posture), attacks (frontal and lateral), and biting the intruder were analyzed when a male intruder was placed into the female resident's cage. For each brain area studied, the frequency of the behaviors was compared among the various treatments by analysis of variance. The results showed a decrease in maternal aggressive behavior (number of bites directed at the intruder) after microinjection of the agonist at 0.2 and 1.0 microg/0.2 microl (1.6 +/- 0.7 and 0.9 +/- 0.3) into the DPAG compared to the saline group (5.5 +/- 1.1). There was no dose-response relationship with the agonist. The present findings suggest that the 5-HT2A/2C receptor agonist has an inhibitory effect on maternal aggressive behavior when microinjected into the DPAG and no effect when microinjected into the MS. Ketanserin (1.0 microg/0.2 microl) decreased locomotion when microinjected into the DPAG and MS, but did not affect aggressive behavior. We interpret these findings as evidence for a specific role of 5-HT2A/2C receptors in the DPAG in the inhibition of female aggressive behavior, dissociated from those on motor activity.

Energy requirements for growth in male and female Saanen goats.

The aim of this study was to investigate the energy requirements of female and intact and castrated male Saanen goats. Animals were randomly assigned to 1 of 2 experiments designed to investigate the energy requirements for maintenance and gain. To determine the maintenance requirements, 85 goats were used (26 intact males, 30 castrated males, and 29 females) with an initial BW of 30.3 ± 0.87 kg. Thirty goats (8 intact males, 9 castrated males, and 13 females) were slaughtered to be used as the baseline group. The remaining goats were assigned in a split-plot design using a 3 × 3 factorial arrangement (3 sexes-intact males, castrated males, and females-and 3 DMI levels-ad libitum and restricted fed to 75 or 50% of the ad libitum intake). The NE was obtained using 65 goats (20 intact males, 22 castrated males, and 23 females) fed ad libitum in a completely randomized design. Eight intact males, 9 castrated males, and 13 females were slaughtered at 30.5 ± 1.53 kg BW. Seventeen goats (6 intact males, 6 castrated males, and 5 females) were slaughtered at 38.1 ± 0.49 kg BW. The remaining goats were slaughtered at 44.0 ± 0.50 kg BW. The NE did not differ between the sexes ( = 0.59; 258.5 kJ/kg BW), resulting in a ME for maintenance of 412.4 kJ/kg BW. The estimated energy use efficiency for maintenance was 0.627. During the growth phase, NE differed between the sexes ( < 0.001); intact males, castrated males, and females showed an average NE equal to 15.2, 18.6, and 22.7 MJ/kg of empty weight gain, respectively. The energy requirements for growth differed between the sexes. The difference was found to be due to distinct NE and partial efficiency of ME utilization for growth in intact and castrated males and females during the late growth phase. This study may contribute to adjustments in feeding system energy recommendations regarding the NE and NE found for goats during the late growth phase.

Ischaemic colitis (necrotizing colitis, pseudomembranous colitis) in acute schistosomiasis mansoni: report of two cases.

Two cases of ischaemic necrosis of the sigmoid colon (necrotizing colitis) are reported in 2 brothers aged 7 and 4 years, diagnosed within a 10 d interval. The children had bathed in streams suspected to be contaminated by Schistosoma mansoni about 50-60 d before the onset of acute disease. Both patients had been previously exposed to schistosome-infected streams without showing signs or symptoms of infection. Before admission, S. mansoni eggs had not been found in the stool. Both patients presented with an apparently identical, relatively symptomatic clinical course with rapid evolution to an acute abdomen. Laparotomy disclosed, in both patients, extensive necrosis (ischaemic necrotizing colitis of schistosomal aetiology) of about 20 cm in the first child and 8 cm in the second, extending from part of the descending colon to the sigmoid. The patients were successfully operated upon (hemicolectomy plus colostomy). The histopathological findings were similar in both patients. Ischaemic necrosis with complete destruction of the mucosa and part of the submucosa was detected in the first case; in the necrotic areas a few eggs of S. mansoni were seen, with no granulomatous reaction, but surrounded by cell shadows, pycnotic nuclei and amorphous material. Necrosis extended to the muscular layer and serosa, in which schistosome granulomas in the necrotic-exudative phase were seen, as well as diffuse granulocytic exudate and fibrin. Sections of tissue from both patients contained numerous eggs and granulomas all in the same exudative phase in regional lymph nodes and near the thrombotic vessels. After surgery, the 2 patients progressed similarly. About 10 d after hospital discharge, the patients received anti-schistosomal treatment with oxamniquine. No further sign of infection was detected at subsequent recall visits.

Bidder's organ of Bufo ictericus: a light and electron microscopy analysis.

Male toads of the Bufonidae Family have rudimentary ovaries designated Bidder's organs, and if the testes are removed this organ develops into a functional ovary, representing a morphological strategy for the reproduction of the species. The Bidder's organ of Bufo ictericus was examined using routine and histochemical techniques by light microscopy and transmission electron microscopy. Each Bidder's organ presented a typical ovarian morphology, being composed of a cortex and a medulla. Bidderian follicles in different stages of development were visualized in the cortex, where they are better developed. The germ cells exhibit a large oocyte with a round-shaped nucleus. The Bidderian follicles are supported by a loose net of reticular fibers. In the medullar region, collagen fibers were immersed in the matrix rich in blood vessels that also contained a small quantity of neutral glycoproteins rich in hexose and/or sialic acid and carboxylated polymers with a characteristic distribution of glycosaminoglycans. The oocyte and the follicular cells were separated by a narrow space containing microvilli. The oocyte exhibit a well developed smooth endoplasmic reticulum, a poorly developed Golgi apparatus, and occasional lysosomes. Concentric cisternal complexes are often visualized; however, their morphological significance remains unclear. The peroxisomes display a fine granular matrix without a crystalline core, with a weak 3,3'-diaminobenzidine-reaction. Intimate association between peroxisomes, peroxisomes and lipid inclusions was observed in the oocyte, suggesting its participation in yolk metabolism.

The cholinergic and dopaminergic systems of the prelimbic sector of frontal cortex and memory in the rat.

Rats were trained to perform delayed non-matching to sample (a working/representational memory task) and visual discrimination (a reference/dispositional memory task) in a T-maze, and implanted bilaterally with cannulae in the prelimbic cortex. The rats were tested postoperatively after bilateral 1-microliter injections of vehicle (Krebs-Ringer), sulpiride (10 micrograms/microliter) or scopolamine (18 micrograms/microliter). Sulpiride had no effect on the performance of either task, whereas scopolamine interfered only with the performance of delayed non-matching to sample. We conclude that dopaminergic mechanisms in the prelimbic cortex are not involved in either type of memory and that cholinergic, but not dopaminergic, mechanisms are important for working/representational memory processes.

Praziquantel inhibits Schistosoma mansoni attachment in vitro.

Male adult Schistosoma mansoni worms were placed in a glass dish containing Tyrode solution and observed for 15 min after addition of praziquantel (0.01 to 1 microM). Praziquantel promoted a concentration- and time-dependent inhibition of sucker-mediated attachment of the worm. Attachment inhibition was correlated with shortening of the parasite. We propose that the rapid and total inhibition of worm attachment observed in vitro with 1 microM praziquantel indicates that therapeutic concentrations of this drug should promote a rapid hepatic shift, in vivo, which may facilitate host tissue reaction.

Clinical and diagnostic aspects of intestinal microsporidiosis in HIV-infected patients with chronic diarrhea in Rio de Janeiro, Brazil.

The objectives of this study were to determine both the prevalence of microsporidial intestinal infection and the clinical outcome of the disease in a cohort of 40 HIV-infected patients presenting with chronic diarrhea in Rio de Janeiro, Brazil. Each patient, after clinical evaluation, had stools and intestinal fragments examined for viral, bacterial and parasitic pathogens. Microsporidia were found in 11 patients (27.5%) either in stools or in duodenal or ileal biopsies. Microsporidial spores were found more frequently in stools than in biopsy fragments. Samples examined using transmission electron microscopy (n=3) or polymerase chain reaction (n=6) confirmed Enterocytozoon bieneusi as the causative agent. Microsporidia were the only potential enteric pathogens found in 5 of the 11 patients. Other pathogens were also detected in the intestinal tract of 21 patients, but diarrhea remained unexplained in 8. We concluded that microsporidial infection is frequently found in HIV infected persons in Rio de Janeiro, and it seems to be a marker of advanced stage of AIDS.

[IQ in hydrocephalus and myelomeningocele. Implications of surgical treatment]. / QI em pacientes com hidrocefalia e mielomeningocele. Implicações do tratamento cirúrgico.

Myelomeningocele occurs in 0.4 for 1000 neonates and is associated with hydrocephalus in 85-90%, and reports on cognition are sparsely found in literature. Forty five children with treated hydrocephalus and myelomeningocele were studied in regard of IQ, and statistically correlated to functional motor level, age of the first shunt, number of revisions of shunt, infection of the shunt and circumference of the head. The medium age was of 7.5 years (3-15 years), 16 males and 29 females. Three (6.6%) had a IQ score > 110, 11 (24.4%) had a score between 100-110, 8 between 85-100 (17.7%), 16 (35.5%) between 85-100 (17.7%) and 7 (15.5%) between 50-70. IQ directly correlated with motor level, having better cognitive results the children with minor functional motor disabilities. Cognition was best in children operated until the seven day of life (t 0.0099), with progressive worse results in children operated after the first month of life, no significance was observed in children operated in the period 7 to 31 days (t 0.1013). Worse results were observed in the group of patients with infection of shunts (t 0.0146). Results were progressively worse with reoperations. The best results in relation of the circumference of the head were seen with children in the medium range (t 0.0115); intermediate results were seen in patients between the medium range and-1SD (t 0.00130) and medium range and +1SD. The worse results were seen in patients at the extremes of > 1SD (t 0.0269) and < ISD (t 0.0042). According to cognitive results the surgical treatment of hydrocephalus have to be done until the first month of life, avoiding reoperations and infections that have unfavorable impact in IQ.