Comparison of two eukaryotic systems for the expression of VP6 protein of rotavirus specie A: transient gene expression in HEK293-T cells and insect cell-baculovirus system

The VP6 protein of rotavirus A (RVA) is a target antigen used for diagnostic assays and also for the development of new RVA vaccines. We have compared the expression of VP6 protein in human embryonic kidney (HEK293-T) cells with results obtained using a well-established insect cell-baculovirus system. The recombinant VP6 (rVP6) expressed in HEK293-T cells did not present degradation and also retained the ability to form trimers. In the insect cell-baculovirus system, rVP6 was expressed at higher levels and with protein degradation as well as partial loss of ability to form trimers was observed. Therefore, HEK293-T cells represent a less laborious alternative system than insect cells for expression of rVP6 from human RVA.     

Musculature of Seison nebaliae Grube, 1861 and Paraseison annulatus (Claus, 1876) revealed with CLSM: a comparative study of the gnathiferan key taxon Seisonacea (Rotifera)

The somatic muscular systems of two species of Seisonacea (Rotifera), Seison nebaliae and Paraseison annulatus, are described using fluorescently labelled phalloidin in combination with confocal laser scanning microscopy. Their overall muscular arrangement is similar and consists of segmentally organised longitudinal fibres that extend the length of the body and are surrounded by semi-circular (= incomplete) bands. However, differences in the musculature between the two species are present and possibly reflect specific adaptations in feeding strategy and locomotion related to the occupation of individual niches on their host, the leptostracan crustacean N. bipes. For example, S. nebaliae has semi-circular muscles in the head region only, while P. annulatus possesses incomplete circular muscles also in the trunk region; furthermore, there are also differences in the arrangements and number of longitudinal muscles. The muscular systems of all rotifer species examined so far are compared in order to establish the ground pattern of the last putative ancestor as well as to seek for traits of systematic importance. Results from both species corroborate earlier hypotheses on the arrangement of muscles in the putative common ancestor of Rotifera, which suggested an orthogonal arrangement consisting of a series of probably continuous (not segmental) inner longitudinal muscles, surrounded by semi-circular fibres, ventrally opened. However, significant morphological and ecological variations among taxa investigated so far show that a consistent correlation between muscular traits and specific ecological features and/or phylogeny is still far from being clear. Hence, musculature of additional taxa, representing the systematic width and occupying a diverse range of habitats, should be investigated.     

Association between number of teeth, edentulism and use of dentures with percentage body fat in south Brazilian community-dwelling older people

doi: 10.1111/j.1741‐2358.2010.00411.x Association between number of teeth, edentulism and use of dentures with percentage body fat in south Brazilian community‐dwelling older people Objective: To evaluate if poor oral status was associated with percentage body fat in a representative sample of south Brazilian community‐dwelling elderly. Background: Evidence suggests that elders with extensive tooth loss and edentulism present with compromised chewing function, which results in dietary changes. However, few studies have assessed whether poor oral status is associated with percentage body fat in community‐dwelling elderly. Materials and methods: A random sample of 471 south Brazilians =>60 years of age was evaluated. Measurements included a questionnaire to assess socio‐demographic, behavioural, general, and oral health data. Percentage body fat was measured with a Tanita TBF 612 digital scale, which measures foot‐to‐foot bioelectrical impedance; and oral status assessment, by means of oral examinations assessing the number of teeth and use of dental prostheses. Correlates of risk of inadequate percentage body fat were assessed by means of multinomial logistic regression. Results: Edentulous participants were more likely to be underfat [OR: 3.11 (1.27–7.61)] or overfat/obese [OR: 1.82 (1.05–3.16)]. Conclusion: The present study shows that edentulous older people had higher odds for inadequate percentage body fat. The maintenance of teeth had a crucial role in increasing the chance of having an adequate body fat percentage in the study population.     

Candidemia in a public hospital in Northeastern Brazil: Epidemiological features and risk factors in critically ill patients

BackgroundCandidemia is a life-threatening fungal infection characterized by the presence of Candida in the blood.AimsTo describe the clinical–epidemiological features and main risk factors among patients with candidemia admitted to Intensive Care Unit.MethodsA cross-sectional, retrospective and observational study was performed between January 2015 and July 2016. Laboratory reports and medical records from ICU patients admitted to a public hospital in northeastern Brazil were analyzed.ResultsThere were 1573 admissions and 67 of them were positive for candidemia. The majority of patients were male (53.3%) and remained at the hospital for more than seven days (86.6%). Non-C. albicans Candida infections (60%) were predominant. Broad-spectrum antibiotic therapy was prescribed in 98.4% of the cases. The most frequent underlying diseases were sepsis (73.3%), presence of solid tumors (15%), respiratory condition (60%), urinary tract disease (56.6%) and gastrointestinal tract diseases (23.3%). Surgeries were carried out on 43% of the patients, consisting of 23.3% abdominal surgeries, with a mortality rate of 92.8%. Risk factors were venous central access (93.3%), mechanical ventilation (81.6%), nasoenteral tube (83.3%), nasogastric tube (25%), indwelling bladder catheter (88.3%), diabetes mellitus (55%) and tracheostomy (36.6%). Statistical analysis correlated the use of indwelling bladder catheter with a higher mortality rate (r = 0.07412, p = 0.0353).ConclusionsThe current study reveals the high case fatality rates among critically ill patients suffering from candidemia admitted to ICU. Herein, we highlight the importance of identifying non-C. albicans Candida species and reinforce the idea of carrying out epidemiological surveillances and antifungal susceptibility tests.     

Use of leukocytes as treatment for endometritis in mares experimentally infected with Streptococcus equi subsp. zooepidemicus

This study compared four treatments for bacterial endometritis in mares experimentally infected with Streptococcus zooepidemicus. Twenty-five mares were used, 20 resistant and five susceptible to endometritis. Mares would be in estrus when infected. Twenty-four hours after inoculation, clinical, bacteriological and cytological examinations were performed and repeated until the first occurrence: negative cytology and no Streptococcus growth or the seventh day post-infection. All mares showed clinical signs of endometritis and were assigned to one of the following treatments: (1) intrauterine infusion of fresh leukocytes; (2) intrauterine infusion of frozen-thawed leukocytes; (3) intrauterine infusion of lysed leukocytes; (4) intrauterine infusion of recombinant human interleukin-8 (rhIL-8); (5) control. Mares were submitted to all treatments, with at least a 14-day interval between treatments in a Latin square design. Treatment did not affect (P=0.121) time needed for resistant mares to eliminate bacteria. Time needed for elimination of bacteria was similar in susceptible mares treated with fresh and frozen leukocytes (P=0.333). Susceptible mares treated with frozen leukocytes also did not differ from those treated with lysed leukocytes (P=0.227) for time to eliminate bacteria, but were significantly different (P>0.02) from those treated with rhIL-8 and control. In resistant mares, physical clearance ability was probably the responsible for bacterial elimination. Intrauterine infusions in susceptible mares with viable or lysed leukocytes associated or not to opsonizing factors, reduced the time to elimination of bacteria. Infusions with bactericidal effect (functional neutrophils and granules) was likely effective and responsible for the more rapid elimination of bacteria in susceptible mares.     

Seasonality and interspecific competition shape individual niche variation in co‐occurring tetra fish in Neotropical streams

The drivers of intraspecific niche variation and its effects on species interactions are still unclear, especially in species‐rich Neotropical environments. Here, we investigated how ecological opportunity and interspecific competition affect the degree of individual trophic specialization and the population niche breadth in tetra fish. We studied the four ecologically similar species (Psalidodon aff. gymnodontus, P. aff. paranae, P. bifasciatus, and Bryconamericus ikaa) in subtropical headwater streams (three sites with two co‐occurring species and three sites with only one species). We sampled fish in two contrasting seasons (winter/dry and summer/wet) and quantified their trophic niches using gut content analysis. Psalidodon bifasciatus was the only species distributed over all the sampled streams. We observed seasonal differences in population trophic niche breadth of P. bifasciatus just when this species co‐occurred with P. aff. gymnodontus. These findings confirm the complex nature of the effects of interspecific competition, depending, for instance, on the identity of the competitor. The degree of individual specialization of P. bifasciatus was higher in the winter, and it was not influenced by the presence of another species. Conversely, the other two Psalidodon species studied presented greater individual specialization in the summer, when fish consumed a higher proportion of allochthonous items (terrestrial insects and seeds), and there were no effects only for B. ikaa. Herein, our results suggest that seasonality in food‐resource availability is a major driver of niche variation and it has the potential to play an important role in how these similar tetra species interact and coexist. Abstract in Portuguese is available with online material.     

Hyper-IgD and periodic fever syndrome: a new MVK mutation (p.R277G) associated with a severe phenotype

Hyperimmunoglobulinemia D and periodic fever syndrome (HIDS; MIM# 260920) is a rare recessively-inherited autoinflammatory condition caused by mutations in the MVK gene, which encodes for mevalonate kinase, an essential enzyme in the isoprenoid pathway. HIDS is clinically characterized by recurrent episodes of fever and inflammation. Here we report on the case of a 2 year-old Portuguese boy with recurrent episodes of fever, malaise, massive cervical lymphadenopathy and hepatosplenomegaly since the age of 12 months. Rash, arthralgia, abdominal pain and diarrhea were also seen occasionally. During attacks a vigorous acute-phase response was detected, including elevated erythrocyte sedimentation rate, C-reactive protein, serum amyloid A and leukocytosis. Clinical and laboratory improvement was seen between attacks. Despite normal serum IgD level, HIDS was clinically suspected. Mutational MVK analysis revealed the homozygous genotype with the novel p.Arg277Gly (p.R277G) mutation, while the healthy non-consanguineous parents were heterozygous. Short nonsteroidal anti-inflammatory drugs and corticosteroid courses were given during attacks with poor benefits, whereas anakinra showed positive responses only at high doses. The p.R277G mutation here described is a novel missense MVK mutation, and it has been detected in this case with a severe HIDS phenotype. Further studies are needed to evaluate a co-relation genotype, enzyme activity and phenotype, and to define the best therapeutic strategies.     

Proteomic characterization of seeds from yellow lupin (Lupinus luteus L.)

Yellow lupin (Lupinus luteus L.) is a legume crop containing a large amount of protein in its seeds. In this study, we constructed a seed‐protein catalog to provide a foundation for further study of the seeds. A total of 736 proteins were identified in 341 2DE spots by nano‐LC‐MS/MS. Eight storage proteins were found as multiple spots in the 2DE gels. The 736 proteins correspond to 152 unique proteins as shown by UniRef50 clustering. Sixty‐seven of the 152 proteins were associated with KEGG‐defined pathways. Of the remaining proteins, 57 were classified according to a GO term. The functions of the remaining 28 proteins have yet to be determined. This is the first yellow lupin seed–protein catalog, and it contains considerably more data than previously reported for white lupin (L. albus L.).