Effect of Inflammatory Stimuli on the Silver Staining Pattern of the Rat Carotid Endothelium

Silver nitrate stains the intercellular junctions of the endothelium and other cytoplasmic or membrane components. Two protocols are described for the silver staining of rat carotid endothelium that exclude the use of pressurized fixatives and simplify the technique previously described for rat aorta. The entire surface of the carotid endothelium was examined and several parameters (stigmata, granularity, clustering of anionic sites, transversal lines, weakening of silver lines and leukocyte adhesion) were evaluated. We studied the pattern of silver staining in two situations: (1) endothelial activation and (2) neurogenic inflammation. Endothelial activation was produced by the intravenous administration of a proinflammatory albumin or polyinosinic acid. Both products cause a marked increase in leukocyte adhesion concomitant with a decrease in argyrophilia and a weakness or loss of silver lines. Neurogenic inflammation, which is mediated by substances released from sensory nerves, was induced by the intravenous administration of substance P or capsaicin. Both stimuli produced an increase in argyrophilia and weakness or loss of silver lines. Substance P caused a clustering of anionic sites, whereas this phenomenon was more discrete with capsaicin. Nearly 80% of all examined rats (controls and inflammatory stimuli treated) showed endothelial membrane disruptions formed by clusters of cells often in the shape of streaks aligned with the long axis of the vessel. The detection of these discontinuities is important, as loss of endothelial integrity is central in the initiation of pathological events.     

Horseradish peroxidase: a reliable or a misleading tool for the investigations on the origin of the proteins of the aqueous humor?

The concept of the blood-aqueous barrier is largely based on the use of horseradish peroxidase (HRP). The present investigation was designed to check its reliability as a macromolecular tracer, especially with regard to the transport of plasma proteins. Rabbits were killed 5 min to 24 h after being intravenously injected with HRP. The tracer diffused rapidly, reaching the aqueous humor of the eye in 3 min or less and was detected at high concentration in the narrow space between the outer epithelial layer of the ciliary epithelium and the wall of the pervious capillaries in the stroma of the processes. HRP appeared to migrate from the blood to the posterior chamber, permeating the tight junctions, viz., the anatomical basis of the blood-aqueous barrier. It was detected at higher concentration at the anterior surface of the iris, at short time intervals; this was interpreted as penetration of the tracer from the aqueous humor of the anterior chamber. The choroid was also labeled in continuation with the reaction in the stroma of the pars plana of the ciliary body which, in turn, sometimes reached the iris root. Therefore, the pervious blood vessels of the choroid could be a source of macromolecules for the iris root. HRP also induced the formation of lysosomes in the ciliary epithelium. This can hardly be accepted as the way in which plasma proteins are physiologically transported to the aqueous humor. However, the pathway of HRP migration over short time intervals seems to be in agreement with previous research indicating that the entrance of serum albumin into the posterior chamber is the first step of its incorporation into the aqueous humor. Received: 7 June 1996 / Accepted: 15 January 1997     

Developmental Exposure to Fluoxetine Modulates the Serotonin System in Hypothalamus

The selective serotonin reuptake inhibitor (SSRI) fluoxetine (FLU, Prozac®) is commonly prescribed for depression in pregnant women. This results in SSRI exposure of the developing fetus. However, there are knowledge gaps regarding the impact of SSRI exposure during development. Given the role of serotonin in brain development and its cross-talk with sex hormone function, we investigated effects of developmental exposure to pharmacologically relevant concentrations of FLU (3 and 30 nM (measured)) on brain neurotransmitter levels, gonadal differentiation, aromatase activity in brain and gonads, and the thyroid system, using the Xenopus tropicalis model. Tadpoles were chronically exposed (8 weeks) until metamorphosis. At metamorphosis brains were cryosectioned and levels of serotonin, dopamine, norepinephrine, and their metabolites 5-hydroxyindoleacetic acid, 3,4-dihydroxyphenylacetic acid, and homovanillic acid were measured in discrete regions (telencephalon, hypothalamus and the reticular formation) of the cryosections using high-performance liquid chromatography. Exposure to 30 nM FLU increased the concentration of 5-hydroxyindoleacetic acid in hypothalamus compared with controls. FLU exposure did not affect survival, time to metamorphosis, thyroid histology, gonadal sex differentiation, or aromatase activity implying that the effect on the serotonergic neurotransmitter system in the hypothalamus region was specific. The FLU concentration that impacted the serotonin system is lower than the concentration measured in umbilical cord serum, suggesting that the serotonin system of the developing brain is highly sensitive to in utero exposure to FLU. To our knowledge this is the first study showing effects of developmental FLU exposure on brain neurochemistry. Given that SSRIs are present in the aquatic environment the current results warrant further investigation into the neurobehavioral effects of SSRIs in aquatic wildlife.     

Ecoepidemiology of Haemonchus contortus bahiensis, ecotype present in sheep of arid zones of Venezuela

The frequency distribution of female Haemonchus contortus bahiensis Grisi, 1974 in sheep from Venezuelan arid zones is 15.32% for the type with vulvar flap, 51.6% for the vulvar-knob and 33.07% for smooth type. A Shannon-Weaver diversity index corresponding to 1.44 bits was calculated for these forms with similarities in the general size, egg-size and in the number of the longitudinal cuticular ridges. An aggregated kind of distribution in the host population according to the K parameter of a negative binomial distribution was recorded for male and female worms. A complicated interaction was observed between the abundance, aggregation and prevalence of this ecotype and the importance of these findings is discussed with regard to host-parasite relationships.     

l-Alanine uptake in brush border membrane vesicles from the gill of a marine bivalve

Summary Brush border membrane vesicles were prepared from mussel gills using differential and sucrose density gradient centrifugation. These vesicles contained both the maximal Na⁺-dependent alanine transport activity found in the gradient and the maximal activities of -glutamyl transpeptidase and alkaline phosphatase. Electron micrographs showed closed vesicles of approximately 0.1–0.5 m diameter. Transport experiments using these vesicles demonstrated a transient 18-fold overshoot in intravesicular alanine concentration in the presence of an inwardly directed Na⁺ gradient, but not under Na⁺ equilibrium conditions. A reduced overshoot (10-fold) was seen with an inwardly directed K⁺ gradient. Further studies revealed a broad cation selectivity, with preference for Na⁺, which was characteristic of alanine transport but not glucose transport in these membranes. The apparent amino acid specificity of the uptake pathway(s) was similar to that of intact gills and supported the idea of at least four separate pathways for amino acid transport in mussel gill brush border membranes. The apparent Michaelis constant for alanine uptake was approximately 7m, consistent with values forK _t determined with intact tissue.     

Opportunistic zygomycotic infections

This is a literature review of 361 opportunistic fungal infections caused by the Zygomycetes. The clinical and laboratory diagnosis, pathogenesis, management, treatment, and outcome of infection are discussed. The Zygomycetes are a group of opportunistic fungi (orders Mucorales and Entomophthorales) which cause severe infections which may be fatal. Early clinical recognition, prompt diagnostic procedures, control of underlying disease and treatment with high doses of amphotericin B and aggressive surgery increases survival in an otherwise lethal infection.     

Subcellular localization of calcium in sporangiophores of Phycomyces blackesleeanus

The in situ localization of Ca²⁺ in stage I sporangiophores of the fungus Phycomyces blakesleeanus was achieved with the potassium pyroantimonate technique. Precipitates of calcium-antimonate were present in mitochondria, vacuoles, endoplasmic reticulum and adjacent cytoplasm, Golgi-like bodies, and nuclei but not cell walls. Material treated with the calcium chelator EGTA lacked these precipitates. The preferential localization of Ca²⁺ in mitochondria, endoplasmic reticulum and vacuoles suggests that these organelles modulate the level of this cation in sporangiophores of P. blakesleeanus.Abbreviations EGTA ethyleneglycol-bis-(-aminoethyl ether) N,N, tetraacetic acid     

The physiology of sperm recovered from the human cervix: acrosomal status and response to inducers of the acrosome reaction

Cervical mucus was collected from 35 women after artificial insemination. Mucus collections were performed at 1 h, 1 day, 2 days, or 3 days following insemination. Sperm viability was greater than 80% at all recovery times as assessed by exclusion of the supravital dye Hoechst 33258. Virtually 100% of the viable sperm were acrosome-intact at all times as assessed with a fluorescein isothiocyanate-conjugated pea lectin. Sperm were recovered from the mucus after migration into the Biggers, Whittin, and Whittingham medium in vitro. Sperm did not undergo the acrosome reaction in response to human follicular fluid immediately after migration from the mucus but did respond to this agonist after 6 h of incubation in vitro. Sperm recovered at all times after insemination had the same pattern of response to follicular fluid. Sperm that penetrated a column of cervical mucus in vitro also responded to follicular fluid with an increase in acrosome reactions after migration from the mucus and incubation for 6 h in vitro. Unlike the sperm that migrated from cervical mucus, sperm that were separated from semen by Percoll density centrifugation did not undergo the acrosome reaction when challenged with follicular fluid after 6 h but did respond after 24 h incubation. Sperm that migrated from cervical mucus had a similar increase in acrosome reactions after 6 h incubation, regardless of whether the acrosome reaction agonist was follicular fluid or disaggregated human zona pellucida.(ABSTRACT TRUNCATED AT 250 WORDS)     

Performance of nymphal and adult Nezara viridula on selected alternate host plants

Laboratory studies with the southern green stink bug, Nezara viridula (L.) (Hemiptera: Pentatomidae), feeding on seeds or fruits of some alternate host plants, demonstrated differences in nymphal survival and development time, and body weight at adult emergence. Lowest mortalities (25%) were observed for nymphs on fruits of dakota mustard [Brassica kaber (DC.)] and siberian motherwort (Leonurus sibiricus L.). On fruits of castor bean (Ricinus communis L.) nymph mortality was ca. 60%, and on mature seeds of L. sibiricus ca. 75%. Total development time from second instar to adult ranged from ca. 26 days on B. kaber to ca. 42 days on R. communis. Fresh body weight at adult emergence was greatest for nymphs on L. sibiricus and lowest on R. communis. Adult survival after 30 days and mean longevity was greater on L. sibiricus than on the remaining foods, being drastically reduced on R. communis. In general, reproductive performance on all foods was similar, except R. communis, on which no female reproduced. Female weight gain of N. viridula was generally greater on L. sibiricus On R. communis, adults lost weight during their first week and showed the lowest weight gain at the end of day 29. Field observations coupled with these laboratory studies suggest that in the Warta area (northern Paraná state, Brazil, 23° 11 S), L. sibiricus and B. kaber are important alternate food plants in the seasonal phenology of N. viridula. R. communis, however, is of poor nutritional quality, serving primarily as a refuge.

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Résumé L'alimentation au laboratoire de N. viridula avec les graines et les fruits de diverses plantes a entraîné différentes mortalités larvaires, différentes vitesses de développement et des différences dans les poids des adultes à l'émergence. Les mortalités les plus faibles (25%) on été observées avec les fruits de Brassica kaber DC. et de Leonurus sibiricus L.Avec les fruits de Ricinus communis L., la mortalité larvaire était de 60%; sur graines mûres de L. sibiricus, elle était de 75%. La durée du développement du second stade à l'adulte a varié de 26 jours sur B. kaber à 42 jours sur R. communis. Le poids frais des adultes à l'émergence était le plus élevé avec L. sibiricus et le plus faible avec de R. communis. La longévité des adultes au-delà de 30 jours était la plus élevée avec L. sibiricus, et était fortement réduite avec R. communis. Dans l'ensemble, la reproduction était de même importance avec les différents aliments, à l'exception de R. communis avec lequel les femelles ne se reproduisaient pas. Le gain de poids des femelles était généralement supérieur avec L. sibiricus; avec R. communis les adultes ont perdu du poids pendant la première semaine et ont présenté le plus faible gain à la fin du 29ème jour.Des observations dans la nature, associées à ces expériences de laboratoire, laissent supposer que dans la zone de Warta (au Nord de l'état de Paraná, 23° 11 S, au Bresil), L. sibiricus et B. kaber sont des plantes hôtes alternatives importantes dans la phénologie saisonnière de N. viridula. Cependant, R. communis, de faible valeur alimentaire, sert avant tout de refuge.