Evaluation of the persistence of infectious human noroviruses on food surfaces by using real-time nucleic acid sequence-based amplification

Noroviruses (NoV) are the major cause of nonbacterial gastroenteritis. However, there is no published study to ascertain their survival on foodstuffs which are directly related to human health risk. In the present study, we developed a rapid, simple, and sensitive real-time nucleic acid sequence-based amplification (NASBA) combined with an enzymatic treatment for distinguishing infectious from noninfectious human NoV. The developed method was validated using spiked ready-to-eat food samples. When feline calicivirus (FCV) was used as a NoV surrogate in the preliminary assays, it appeared more sensitive to heat inactivation and enzymatic pretreatment than the human NoV. This suggests that FCV may not be an ideal model for studying NoV. Our results reveal clearly that the developed enzymatic pretreatment/real-time NASBA combination successfully distinguished the infectious from heat-inactivated NoV. Moreover, we demonstrate that NoV survived for at least 10 days on refrigerated ready-to-eat foods, such as lettuce and turkey. However, the survival rate was higher on turkey than on lettuce, probably because of their different surface natures. The approach developed in this study may be suitable for more in-depth studies of the persistence and inactivation of human NoV and may be applied to other nonculturable RNA viruses. Moreover, the evaluation of infectious NoV survival provided valuable information concerning its persistence on ready-to-eat food.     

Purification and characterization of aprotinin from porcine lungs

Aprotinin, the most studied serine proteinase inhibitor, was isolated from porcine lung for the first time. The purified porcine aprotinin had an Mr value of ∼7 kDa. It cross-reacted with polyclonal serum anti-commercial aprotinin. About 1 μg porcine aprotinin inhibited 6 μg trypsin whereas 1 μg commercial soybean inhibitor inhibited only 1 μg trypsin. The aprotinin gene was also isolated from porcine lung: the deduced amino acid sequence showed 74% identity to bovine aprotinin.     

Systematic revision of the genus Petromica Topsent (Demospongiae: Halichondrida), with a new species from the southwestern Atlantic

The status, scope and classification of the sublithistid demosponge genus Petromica Topsent are revised through morphological analysis of museum specimens of all seven species (including proposed synonyms and varieties), two of which were collected and observed in situ along the Brazilian coast (P. ciocalyptoides (Van Soest & Zea) and P. citrina sp. n.). The synonymy of Petromica and Monanthus Kirkpatrick with priority to the former is justified due to the consistent presence of monocrepid rhizoclone desmas and oxeas in an halichondroid arrangement, and to the lack of co-variance in other morphological characters among the species studied (presence and shape of papillae, surface texture, ectosomal skeleton and desma shape). The proposed synonymy of P. grimaldii Topsent and P. massalis Dendy is refuted due to differences in habit and spicule shape between the two species. Three forms described as varieties of Monanthus plumosus Kirkpatrick are raised to species level: P. plumosa (Kirkpatrick), P. tubulata (Kirkpatrick) and P. digitata (Burton). Phylogenetic analysis indicates that two possibly monophyletic clades may be recognized within Petromica, although with low bootstrap support (35–59%): (P. ciocalyptoides, P. citrina) and (P. grimaldii, P. massalis) (P. plumosa) (P. tubulata) (P. digitata). The classification of Petromica within the Halichondriidae (order Halichondrida) is supported by the confused reticulation of long oxeote spicules with ascending spicule tracts, present in all species of the genus.     

Laminin induces formation of neurite-like processes and potentiates prolactin secretion by GH3 rat pituitary cells

Tumor-derived GH3 rat pituitary cell lines are widely utilized to study mechanisms of prolactin secretion and responsiveness to secretagogues. These cells served here as a model with which to study relationships between shape and function. When GH3 cells were routinely grown in serum-supplemented medium, they exhibited the polygonal phenotype of epithelial cells, with scarce secretory granules. In contrast, when seeded in a serum-free medium, they attached loosely and contained more secretory granules. In both cases, they released prolactin in a nonpolarized manner. We show in the present work that laminin extracted from Englebreth-Holm-Swarm (EHS) tumors was a potent attachment and spreading factor for GH3/B6 cells seeded in serum-free medium. Moreover, it induced the formation of neurite-like processes, which were increased in number and length by chronic treatment with a specific secretagogue, thyroliberin (TRH). These changes in cell shape were correlated with a potentiation of prolactin secretion, both basal and TRH-stimulated. Furthermore, using immunocytochemistry and electron microscopy, we revealed--at the dilated tip of processes--an accumulation not only of prolactin, but also of synaptophysin, a vesicle membrane marker, and of several organelles, such as secretory granules, smooth vesicles, dense bodies and mitochondria. The cytoplasmic processes contained long parallel bundles of microtubules and showed a strong immunoreactivity for beta 2-tubulin. In addition, we found immunocyto-chemical evidence for the presence of 200-k Da neurofilament protein in GH3/B6 cell processes as well as in neurites of cultured hypothalamic neurons. We conclude that, in GH3/B6 cells, laminin induced the differentiation of neurite-like processes, which were the site of polarized organelle transport and exhibited some neuronal markers.     

Plasticity of gonad development in hermaphroditic sparids: ovotestis ontogeny in a protandric species,Lithognathus mormyrus

Synopsis Heterosexual gonad development in a sparid species, Lithognathus mormyrus, was studied by histological and cytological examination, during the first three years of life. Gonad bisexuality is achieved after two months of development, according to the cytological dynamics known in sparids. In one-year-old fishes, a variability in the gonad morphology of the juvenile is shown: three different types of ovotestis have been identified within the same cohort: ovotestes with testicular prevalence (25%), testicular and ovarian equivalence (20%), and ovarian prevalence (55%). This morphological variability of the juvenille ovotestes was consistent with the histological analysis of the sexual structure of the adult stock at the first sexual maturity, which constituted 55.5% of functional males (stemming from the first types of ovotestis) and 44.5% of primary females (from the third type). The plasticity of sexual expression in sparids is emphasized, revealing the potentialities of the ovotestis.     

Effect of Thermal Processing and Maceration on the Antioxidant Activity of White Beans

Phenolic compounds, which naturally occur in beans, are known to have antioxidant activity, which may be partially lost during the processing of this legume. This study evaluated the effect of thermal processing and maceration on the phenolic acid and flavonoids profile and content and on the antioxidant activity of white beans. According to the results obtained from the 2,2-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) method, there were no significant differences among treatment groups analysed. When was using 1,1-diphenyl-2-pycrylhydrazyl method (DPPH), beans cooked without maceration present the higher antioxidant activity, and raw beans the lower. The phenolic acids found in greater amounts were gallic acid and chlorogenic acid. Kaempferol was only detected in the soaked and cooked samples; catechin and kaempferol-3-rutinoside were found in the highest concentrations. Quercetin and kaempferol-3-glucoside were not affected by the cooking process, either with or without maceration. In general, the heat treatment increased the antioxidant activity.     

Heterogeneity of Multimedia Exposures to Neurotoxic Elements (Al,As, Cd,Pb, Mn,and Hg) in Breastfed Infants from Porto Velho,Brazil

Infant exposure to neurotoxic elements is a public health issue that needs monitoring with regard to breast milk composition. We studied six neurotoxic elements in breast milk samples at different stages of lactation in mothers from Porto Velho, Brazil. We used a flow-injection mercury system (FIMS) to determine total Hg concentrations and an inductively coupled plasma optical emission spectrometer (ICP-OES) to determine the concentrations of Al, As, Cd, Pb, and Mn in 106 donors of a human milk bank. Association rules analyses were applied to determine the pattern of binary and ternary mixtures of the measured exposants. The metal concentration was mostly below the limit of detection (LOD) for Cd (99%), Pb (84%), and Hg (72%), and it was above the LOD for As (53%), Mn (60%), and Al (82%), respectively. Median concentrations (dry weight) of Al, As, Hg, Mn, and Pb were 1.81 μg/g, 13.8 ng/g, 7.1 ng/g, 51.1 ng/g, and 0.43 μg/g, respectively. Al is singly the most frequent element to which infants are exposed. Occurring binary combination (> LOD) was 56% for Al-Mn, 41% for Al-As, 22% for Al-Hg, and 13% for Al-Pb. In 100% of neonates, exposure to Al-ethylmercury (EtHg) occurred through immunization with thimerosal-containing vaccines (TCV). Association rules analysis revealed that Al was present in all of the multilevel combinations and hierarchical levels and that it showed a strong link with other neurotoxic elements (especially with Mn, As, and Hg). (a) Nursing infants are exposed to combinations of neurotoxicants by different routes, dosages, and at different stages of development; (b) In breastfed infants, the binary exposures to Al and total Hg can occur through breast milk and additionally through TCV (EtHg and Al); (c) The measured neurotoxic elements were found at low frequencies in breast milk and at concentrations that pose no public health concerns for milk banking.