Vitellin-binding proteins in the nematode Oscheius tipulae (Nematoda, Rhabditida)

We describe the first application of a non-radioactive ligand-blotting technique to the characterization of proteins interacting with nematode vitellins. Chromatographically purified vitellins from the free-living nematode Oscheius tipulae were labeled with fluorescein in vitro. Ligand-blotting assays with horseradish peroxidase-conjugated anti-fluorescein antibodies showed that labeled vitellins reacted specifically with a polypeptide of approximately 100 kDa, which we named P100. This polypeptide is a specific worm`s vitellin-binding protein that is present only in adult worms. Blots containing purified O. tipulae vitellin preparations showed no detectable signal in the 100 kDa region, ruling out any possibility of yolk polypeptides self-assembling under the conditions used in our assay. Experiments done in the presence of alpha-methyl mannoside ruled out the possibility of vitellins binding to P100 through mannose residues. Triton X-114 fractionation of whole worm extracts showed that P100 is either a membrane protein or has highly hydrophobic regions. (C) 2008 Elsevier Inc. All rights reserved.

BYC, an atypical aspartic endopeptidase from Rhipicephalus (Boophilus) microplus eggs

An aspartic endopeptidase was purified in our laboratory from Rhipicephalus (Boophilus) microplus eggs [Logullo, C., Vaz, I.S., Sorgine, M.H., Paiva-Silva, G.O., Faria, F.S., Zingali, R.B., De Lima, M.F., Abreu, L., Oliveira, E.F., Alves, E.W, Masuda, H., Gonzales, J.C., Masuda, A., and Oliveira, P.L., 1998. Isolation of an aspartic proteinase precursor from the egg of a hard tick, Rhipicephalus (Boophilus) microplus. Parasitology 116, 525-532]. Boophilus yolk cathepsin (BYC) was tested as component of a protective vaccine against the tick, inducing a significant immune response in cattle [da Silva, VI., Jr., Logullo, C., Sorgine, M., Velloso, F.F., Rosa de Lima, M.F., Gonzales, J.C., Masuda, H., Oliveira, P.L., and Masuda, A., 1998. Immunization of bovines with an aspartic proteinase precursor isolated from Rhipicephalus (Boophilus) microplus eggs. Vet. Immunol. Immunopathol. 66,331-341]. In this work, BYC was cloned and its primary sequence showed high similarity with other aspartic endopeptidases. In spite of this similarity, BYC sequence shows many important differences in relation to other aspartic peptidases, the most important being the lack of the second catalytic Asp residue, considered to be essential for the catalysis of this class of endopeptidases. When we determined BYC cleavage specificity by LC-MS, we found out that it presents a preference for hydrophobic residues in P1 and P1` in accordance to most aspartic endopeptidases. Also, when analyzed by circular dicroism, BYC presented high beta sheet content, also a characteristic of aspartic endopeptidases. On the other hand, although both native and recombinant BYC are catalytically active, they present a very low specific activity, what seems to indicate that this peptidase will digest its natural substrate, vitellin, very slowly. We speculate that such a slow Vn degradative process might constitute an important strategy to preserve egg protein content to the hatching larvae. (c) 2007 Elsevier Inc. All rights reserved.

The Estrous Cycle Modulates Small Leucine-Rich Proteoglycans Expression in Mouse Uterine Tissues

In the pregnant mouse uterus, small leucine-rich proteoglycans (SLRPs) are drastically remodeled within a few hours after fertilization, suggesting that ovarian hormone levels modulate their synthesis and degradation. In this study, we followed by immunoperoxidase approach, the presence of four members of the SLRP family (decorin, lumican, biglycan, and fibromodulin) in the uterine tissues along the estrous cycle of the mouse. All molecules except fibromodulin, which predominates in the myometrium, showed a striking modulation in their distribution in the endometrial stroma, following the rise in the level of estrogen. Moreover, notable differences in the distribution of SLRPs were observed between superficial and deep stroma, as well as between the internal and external layers of the myometrium. Only biglycan and fibromodulin were expressed in the luminal and glandular epithelia. All four SLRPs were found in cytoplasmic granules of mononucleated cells. The pattern of distribution of the immunoreaction for these molecules in the uterine tissues was found to be estrous cycle-stage dependent, suggesting that these molecules undergo ovarian hormonal control and probably participate in the preparation of the uterus for decidualization and embryo implantation. In addition, this and previous results from our laboratory suggest the existence of two subpopulations of endometrial fibroblasts that may be related to the centrifugal development of the decidua. Anat Rec, 292:138-153, 2009. (c) 2008 Wiley-Liss, Inc.

BIOCHEMICAL VARIABILITY BETWEEN VENOMS FROM DIFFERENT HONEYBEE (APIS-MELLIFERA) RACES

1. The comparison of molecular exclusion cromatography profiles of venoms from sting apparatuses of Apis mellifera ligustica, Apis mellifera adansonii and Africanized honey-bees in Sephadex G-100 revealed both qualitative and quantitative differences.2. The venoms from A.m. ligustica and A.m. adansonii presented, respectively, three and two peaks characteristic of each sub-species, while Africanized honey-bee was characterized by the absence of eight peaks common to the former.3. The polypeptides with M(r) in the range from 100,000 to 7500 da correspond respectively to 62.0%, 66.6% and 68.7% of total proteins from the venon of A.m. ligustica, A.m. adansonii and Africanized honey-bees, while the peptidic fraction with M(r) range from 4100 to 2000 da corresponds to 11.4%, 32.4% and 10.2% of venom protein, respectively.

Morfo-anatomia de plântulas e número cromossômico de Cybistax antisyphilitica (Mart.) Mart. (Bignoniaceae)

Plântulas de Cybistax antisyphilitica (ipê-verde), espécie arbórea que ocorre no Estado de São Paulo, Brasil, foram estudadas morfoanatomicamente e citogeneticamente. As plântulas são eudicotiledôneas, fanerocotiledonares e epigéias. Suas sementes apresentam alas hialinas, assimétricas e de textura papirácea. A raiz é axial com tecido epidérmico irregular. O hipocótilo é verde, glabro e apresenta os elementos do xilema em diferenciação. Os cotilédones são verdes, foliáceos, reniformes, com mesofilo heterogêneo, epiderme pilosa e feixes vasculares colaterais. Os eófilos são glabros, peciolados, de filotaxia oposta, dorsiventrais, hipoestomáticos e possuem mesofilo heterogêneo e assimétrico. Não há diferenças anatômicas significativas entre os eófilos e os metáfilos. O sistema vascular do pecíolo dos metáfilos dispõe-se em forma de ferradura. A espécie apresenta número cromossômico mitótico 2n = 40 com comprimento cromossômico geral médio de 1,042 µm ± 0,140 e amplitude variando de 0,58 µm até 1,60 µm.

Responsiveness of the interrenal tissue of Jundia (Rhamdia quelen) to an in vivo ACTH test following acute exposure to sublethal concentrations of agrichemicals

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Role of brain nitric oxide in the thermoregulation of broiler chicks

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Variação periódica da triiodotironina (T3) plasmática e sua ação na reprodução induzida do matrinxã, Brycon cephalus (Günther, 1869) em cativeiro

O matrinxã, Brycon cephalus, espécie nativa oriunda da Bacia Amazônica, apresenta características adequadas para a piscicultura. Entretanto, trata-se de peixe reofílico, sendo necessário manejo adequado para induzir à reprodução. O objetivo deste trabalho foi caracterizar o perfil da triiodotironina (T3) plasmática no matrinxã, durante 16 meses (outubro/97 a janeiro/99), relacionando-o com a maturação sexual, além de testar a ação do T3 associado ao extrato pituitário de carpa na reprodução induzida da espécie. O experimento foi conduzido no Centro Nacional de Pesquisa de Peixes Tropicais - CEPTA, Pirassununga, SP, e no Departamento de Morfologia e Fisiologia Animal, da Faculdade de Ciências Agrárias e Veterinárias de Jaboticabal - UNESP. Foram amostrados, mensalmente, 8 a 12 peixes, de ambos os sexos, totalizando 173 animais, de onde retirou-se sangue para dosagem do T3 plasmático. As gônadas foram removidas para análise histológica com identificação do sexo e determinação do estádio de maturação. Analisaram-se testículos e ovários de 161 peixes em corte transversal, tendo predominado machos (63,35%) e o estádio sexual imaturo entre ambos. A concentração plasmática de T3 foi maior de dezembro a janeiro, para machos e fêmeas, coincidindo com o período de maior atividade reprodutiva da espécie e maiores temperaturas da água. em janeiro/99, reprodutores de matrinxã foram induzidos com extrato de pituitária de carpa (EPC) associado à administração de T3 (20 mg/kg em 0,1 mL de suspensão oleosa). Os resultados sugeriram que o T3 atuou sinergicamente à gonadotropina do extrato hipofisário e que o tratamento agudo de triiodotironina com o EPC pode estimular o eixo hipotálamo-hipófise-ovário. O tempo de eclosão das larvas provenientes das fêmeas tratadas com o T3 foi menor e o crescimento inicial e a sobrevivência dessas larvas, maiores.

The Developing Left Superior Cervical Ganglion of Pacas (Agouti Paca)

In this study the main question investigated was the number and size of both binucleate and mononucleate superior cervical ganglion (SCG) neurons and, whether post-natal development would affect these parameters. Twenty left SCGs from 20 male pacas were used. Four different ages were investigated, that is newborn (4 days), young (45 days), adult (2 years), and aged animals (7 years). By using design-based stereo-logical methods, that is the Cavalieri principle and a physical disector combined with serial sectioning, the total volume of ganglion and total number of mononucleate and binucleate neurons were estimated. Furthermore, the mean perikaryal (somal) volume of mononucleate and binucleate neurons was estimated using the vertical nucleator. The main findings of this study were a 154% increase in the SCG volume, a 95% increase in the total number of mononucleate SCG neurons and a 50% increase in the total volume of SCG neurons. In conclusion, apart from neuron number, different adaptive mechanisms may coexist in the autonomic nervous system to guarantee a functional homeostasis during ageing, which is not always associated with neuron losses. Anat Rec, 292:966-975, 2009. (C) 2009 Wiley-Liss, Inc.

The adrenergic regulation of the cardiovascular system in the South American rattlesnake, Crotalus durissus

The present study investigates adrenergic regulation of the systemic and pulmonary circulations of the anaesthetised South American rattlesnake, Crotalus durissus. Haemodynamic measurements were made following bolus injections of adrenaline and adrenergic antagonists administered through a systemic arterial catheter. Adrenaline caused a marked systemic vasoconstriction that was abolished by phentolamine, indicating this response was mediated through alpha-adrenergic receptors. Injection of phentolamine gave rise to a pronounced vasodilatation (systemic conductance (G(sys)) more than doubled), while injection of propranolol caused a systemic vasoconstriction, pointing to a potent alpha-adrenergic, and a weaker beta-adrenergic tone in the systemic vasculature of Crotalus. Overall, the pulmonary vasculature was far less responsive to adrenergic stimulation than the systemic circulation. Adrenaline caused a small but non-significant pulmonary vasodilatation and there was tendency of reducing this dilatation after either phentolamine or propranolol. Injection of phentolamine increased pulmonary conductance (G(pul)), while injection of propranolol produced a small pulmonary constriction, indicating that alpha-adrenergic and beta-adrenergic receptors contribute to a basal regulation of the pulmonary vasculature. Our results suggest adrenergic regulation of the systemic vasculature, rather than the pultrionary, may be an important factor in the development of intracardiac shunts. (c) 2007 Elsevier B.V. All rights reserved.

Efeitos do glyphosate e sulfometuron-methyl no crescimento e na qualidade tecnológica da cana-de-açúcar

Os herbicidas, mesmo quando usados em doses reduzidas ou utilizados como maturadores, podem alterar a morfofisiologia da planta, o que pode levar a modificações qualitativas e quantitativas na produção. O presente estudo objetivou avaliar a eficiência agronômica e os efeitos, durante o crescimento da cana-soca, da aplicação de glyphosate e sulfometuron-methyl em baixas doses. O delineamento experimental utilizado foi o de blocos casualizados, com quatro repetições. Os tratamentos foram constituídos pelos herbicidas sulfometuron-methyl e glyphosate em diferentes doses e misturas e por uma testemunha (sem aplicação dos produtos). Uma linha de plantas de cana-de-açúcar foi destinada à aferição da qualidade tecnológica, sendo estabelecido 1 m aleatório a cada época de amostragem. Os colmos coletados foram submetidos ao desponte na altura da gema apical e à desfolha; em seguida, foram encaminhados para processamento segundo a metodologia do Sistema de Pagamento de Cana pelo Teor de Sacarose (SPCTS), sendo considerados os parâmetros tecnológicos: pol cana (PCC), pureza do caldo (PUI), açúcar total recuperável (ATR) e Brix. Nas soqueiras de cana-de-açúcar, realizaram-se análises de crescimento (altura e perfilhos). As avaliações foram realizadas na pré-colheita (30 dias após aplicação dos maturadores) e 30, 60, 90, 120, 150 e 180 dias após a colheita. Os herbicidas glyphosate e sulfometuron-methyl propiciaram melhoria da qualidade tecnológica da matéria-prima,com incrementos significativos na pureza do caldo e no Brix. A aplicação dos produtos não interferiu na produtividade e no teor de açúcar. Houve efeito estimulante no perfilhamento quando se usou glyphosate na dose de 400 mL ha-1 e redução em crescimento (altura) no início do desenvolvimento da cana, porém, com o tempo, o efeito não se manteve.

Estratégia de vida do peixe anual Hypsolebias antenori (Cyprinodontiformes: Rivulidae) do semiárido brasileiro

This work deals with the life strategy of an endangered annual fish, Hypsolebias antenori (Cyprinodontiformes: Rivulidae), of the Brazilian semiarid region. The eggs of these fish hatch at the onset of the rainy season, grows rapidly and they reproduce during the rainy season. When the water puddles dry out, the entire population dies. The resistant eggs which are buried in the bottom of the dried pools go through diapause stages, during which time the embryonic development becomes temporarily arrested. With the onset of the next rainy season, the eggs hatch and a new generation is formed. Specimens of H. antenori were captured during 2011 and 2013, in temporary water pools located in the hydrographic basin of river Jaguaribe in Ceará, Brazil. Sex ratio, the length-weight relationship, the growth type, first sexual maturity, anatomy and histology of the digestive tract, development of gonads, reproductive strategy, karyotypic pattern of the species, and the conservation status of H. antenori were investigated. The results of this study are presented in the form of eight articles. The first article is about the fish faunal composition of the hydrographic basins of Rio Grande do Norte, Brazil, wherein the record of H. antenori is included. The second article deals with the sex ratio, secondary sexual characteristics of males, the length-weight relationship and the type of growth. Males show a pattern of intense coloration with well developed fins. The sex ratio showed a significant predominance of females (1M:1.7 F). Males were larger in length and weight. The equation of weight and total length relationship was Wt=0.0271Lt3,8937, showing a positively allometric growth, indicating greater increase in weight than in length. The third article discusses the anatomy and histology of the digestive tract of H. antenori. It is considered as a generalist feeder with characteristics of omnivore, which utilizes different food sources. The fourth article discusses the stages and phases of gonad development and type of spawning of H. antenori. The fifth article is about the r reproductive strategy adopted by H. antenori which helps in successful reproduction over a short period of life. The sixth article deals karyotypic pattern of the species, constituting the first cytogenetic contribution to the genus. The seventh article discusses about the risk of extinction of this species which suffers a series of threats, such as, habitat loss through land use, deforestation, construction of reservoirs, pollution due to domestic and industrial sewage, besides pesticides and agrochemicals. Furthermore, decreasing rainfall and intensification of aridity due to global climate changes, interferes with the reproductive cycle. The eighth article deals with aggressive behavior adopted between males and among females during reproduction. All temporary water pools sampled during this study were in high degree of degradation, mainly due to human action. There is a great need for conservation measures to protect the populations of annual fish, including the creation of protected areas in the semiarid ephemeral aquatic environments of Brazil

Fibular Nerve Injury After Small Saphenous Vein Surgery

Superficial nerve injuries are very common during varicose vein surgery. In contrast, deep nerve injuries are rare and reported especially when surgery involves the small saphenous vein (SSV). The deep motor nerves most commonly injured are the tibial nerve and the peroneal nerve, which are directly or indirectly affected by extrinsic compression, stretching, or healing process involvement. In this report, two cases of common fibular nerve injury after SSV stripping are described, including treatment used and patient outcomes. Nerve damage mechanisms, anatomy, and prevention strategies are also discussed. In conclusion, fibular nerve damage may occur during SSV stripping. Preventive measures include careful preoperative ultrasonographic investigation of the anatomy of the vein, determining location of the saphenopopliteal joint, and careful dissection far from fibular nerve and restricted to the popliteal fossa.