Oligomeric State and Thermal Stability of Apo- and Holo- Human Ornithine δ-Aminotransferase

Human ornithine δ-aminotransferase (hOAT) (EC 2.6.1.13) is a mitochondrial pyridoxal 5′-phosphate (PLP)-dependent aminotransferase whose deficit is associated with gyrate atrophy, a rare autosomal recessive disorder causing progressive blindness and chorioretinal degeneration. Here, both the apo- and holo-form of recombinant hOAT were characterized by means of spectroscopic, kinetic, chromatographic and computational techniques. The results indicate that apo and holo-hOAT (a) show a similar tertiary structure, even if apo displays a more pronounced exposure of hydrophobic patches, (b) exhibit a tetrameric structure with a tetramer-dimer equilibrium dissociation constant about fivefold higher for the apoform with respect to the holoform, and (c) have apparent T_m values of 46 and 67?°C, respectively. Moreover, unlike holo-hOAT, apo-hOAT is prone to unfolding and aggregation under physiological conditions. We also identified Arg217 as an important hot-spot at the dimer–dimer interface of hOAT and demonstrated that the artificial dimeric variant R217A exhibits spectroscopic properties, T_m values and catalytic features similar to those of the tetrameric species. This finding indicates that the catalytic unit of hOAT is the dimer. However, under physiological conditions the apo-tetramer is slightly less prone to unfolding and aggregation than the apo-dimer. The possible implications of the data for the intracellular stability and regulation of hOAT are discussed.     

Long‐term population dynamics of a coral reef gastropod and responses to disturbance

The age‐specific density of the red‐lipped stromb Strombus luhuanus (Mollusca: Gastropoda) was monitored over 13 years (1981–1993) at four locations on the intertidal reef flat at Heron Island, Great Barrier Reef. Densities were highly variable, but there were persistent, location‐specific differences in population density, age structure and adult body size, the latter indicating that the populations were not extensively linked by adult movement. There was relatively high recruitment at most locations in 1984, 1989 and 1993, each occurring approximately 2 years after El Niño/Southern Oscillation events, although recruit density during these years varied in both space and time. The studied strombs experienced three disturbance events: (i) experimental harvesting at two locations (1984–1985); (ii) siltation from a harbour dredging operation (1987–1988); and (iii) a severe cyclone (1992). Resilience to harvesting at a local scale (0.5–2 ha) was high: density had recovered within a year, due to immigration of adults and older juveniles. Strombus luhuanus responded much more strongly to broad‐scale changes to its environment than to localized harvesting. After dredging, there was a progressive density decline coupled with low recruitment at two locations, and a later decline at a third location, followed by a recruitment‐driven rebound after the cyclone. Generalized environmental effects of siltation and the cyclone were also reflected in substantial changes in algal cover. Long‐term variations in environmental conditions probably cause high temporal variation over large spatial scales through effects on the survival of larvae or recruits. Localized short‐term field monitoring of such species would give a misleading picture of key factors affecting population dynamics.     

Morphological and histological study of castration-induced degeneration and androgen-induced regeneration in the mouse prostate

Degenerative and regenerative changes in the ductal architecture of the ventral and dorsolateral prostates (VP and DLP) of the adult mouse were investigated in microdissected specimens over a time-course of 14 days following castration and subsequently during 14 days of administration of testosterone propionate. After castration, about 35% of the ductal tips and branch-points were lost in distal regions (usually near the capsule) in both prostatic lobes. By contrast, in more proximal regions of the prostate (closer to the urethra), the ducts survived in an atrophic condition. The ductal morphology that had been lost in the distal regions completely regenerated after testosterone propionate was administered to the castrated males. In the VP, androgen replacement simply returned the gland to its former size with moderate ductal distension; in the DLP, excessive epithelial infoldings and ductal distension were elicited in the distal regions of the ducts after 14 days of treatment with testosterone propionate. These results suggest that androgenic responsiveness and dependency are different in distal versus proximal ducts. Distal ducts are exquisitely androgen-dependent and androgen-sensitive; in proximal regions, androgen-dependency is not as strict.     

Trypsin and α-amylase inhibitors are differentially induced in leaves of amaranth (Amaranthus hypochondriacus) in response to biotic and abiotic stress

Seeds of Amaranthus hypochondriacus L. are known to accumulate a trypsin-inhibitor (ATI) member of the potato-I inhibitor family and an α -amylase inhibitor (AAI), possessing a knottin-like fold. They are believed to have a defensive role due to their inhibition of trypsin-like enzymes and α -amylases of insect pests. In this work, both inhibitory activities were found in leaves of young A. hypochondriacus plants. High constitutive levels of foliar inhibitory activity against bovine trypsin and insect α -amylases were detected in in vitro assays. Trypsin inhibitory activity was further increased by exposure to diverse treatments, particularly water stress. Salt stress, insect herbivory and treatment with exogenous methyl jasmonate (MeJA) or abscisic acid (ABA) also induced trypsin inhibitor activity accumulation, although to a lesser degree. In gel and immunoblot analyses showed that foliar trypsin inhibitor activity was constituted by at least three different inhibitors of approximately 29, 8 (including ATI) and 3 kDa, respectively. These inhibitors showed differing patterns of accumulation in response to diverse treatments. On the other hand, significant increases in α -amylase inhibitor activity and AAI levels were detected in leaves of insect-damaged, MeJA- and ABA-treated A. hypochodriacus plantlets, but not in those subjected to water- or salt-stress. A differential induction of trypsin inhibitor activity and α -amylase inhibitor accumulation in response to insect herbivory by two related species of lepidopterous larvae was observed, whereas mechanical wounding failed to induce either inhibitor. The overall results suggest that trypsin and α -amylase inhibitors could protect A. hypochondriacus against multiple types of stress.     

Salmonella serotypes isolated from human enteric processes in Recife, Pernambuco, during 1978-1980

From 13,196 faecal cultures made in Recife-Pernambuco during the period from 1978 to 1980, 1,720 strains of Salmonella were isolated. Serological typing on 1,387 of the isolates recognized 63 serotypes, 73.18% of which belonged to group B. The prevalent serotypes adding up to 1,231 strains (88.75% of the total of the isolates) were: S. typhimurium, S. saint-paul, S. poona, S. derby, S. agona, S. newport, S. oranienburg, S. infantis, S. tshiongwe and S. ndolo.     

Tumor promoter chloroform is a potent protein kinase C activator

The major interaction site for tumor-promoting phorbol esters is the calcium-activated, phospholipid-dependent protein kinase (protein kinase C), a key-element in signal transduction. Binding of phorbol esters results in enzyme activation which mediates, at least in part, the action of these agents. We have investigated the effects of tumor promoter chloroform on protein kinase C activity. Like thrombin and 12-O-tetradecanoylphorbol-13-acetate (TPA), chloroform was able to activate protein kinase C in intact rabbit platelets. In addition, chloroform stimulated enzyme activity as well as TPA binding capacity in cell-free system. Scatchard analysis of the data has shown that chloroform increased the number of phorbol ester binding sites. Structurally related compounds, carbon tetrachloride and methylene chloride, activated the enzyme similarly.     

A transient voltage-dependent outward current in cultured cerebellar granules

Granule cells were dissociated from rat cerebella with a procedure that yields a 98% pure cell population. Potassium currents in these cells were studied using the patch-clamp technique. Depolarizing pulses of 10 mV step and 100 ms duration from a holding potential of –80 mV elicited two different potassium outward currents: a transient, low-voltage activated component and a long lasting, high-voltage activated component. At +30 mV, the total current reached an amplitude of 2 nA (mean value of 15 experiments). The reversal potential of the transient current, estimated by measuring tail currents, was –77 mV, close to that predicted by the Nernst equation. The transient current was half inactivated with a holding potential of –78 mV and completely inactivated with –50 mV or more positive holding potentials. Finally, the current decay could be fitted by the sum of two exponentials with time constants of about 20 and 250 ms.     

Structure and function of hemoglobin in antarctic fishes and evolutionary implications

Summary The hematological features of cold-adapted, red-blooded Antarctic teleosts has prompted this study on the relationship between hemoglobin molecular structure and oxygen-binding properties. The hemolysates from 21 species of 5 families contained one component (Hb 1), often accompanied by an additional, minor one (Hb 2, 5%–10% of total). On the other hand, 3 species of Zoarcidae, a non-endemic family, had 4–5 components. All purified hemoglobins from the former group, but only 1–2 of the 4–5 hemoglobins of Zoarcidae, showed a strong Root effect (pH regulation of oxygen binding). Globins from each hemoglobin have been purified and characterised with respect to molecular structure in several species. The similarity between the complete amino acid sequence of one -chain and those of non-Antarctic -chains is lower than that among the latter sequences, suggesting independent pathways of evolution.Presented at the 5th SCAR Symposium on Antarctic Biology, Hobart, Australia (August 29th-September 3rd, 1988)     

Prevalence of nits and lice in samples of cut hair from floors of barbershops and beauty parlors in Belo Horizonte, Minas Gerais State, Brazil

A louse survey based on samples of cut hair collected from floors of barbershops and beauty parlors was conducted in Belo Horizonte, Minas Gerais State, Brazil, from October 1984 to April 1985, as an alternative way to determine the prevalence of pediculosis capitis in the population. Of 475 samples examined for nits, nymphs, or adults of Pediculus capitis, 140 were infested (29.5%). A total of 58 lice and 3,553 nits were found in 33,632.9 g of hair collected, giving a ratio of 0.10 nit/g. Almost 29% of the nits were viable and capable of being transmitted after hatching. There was significant difference among the infestation rates by socioeconomic levels, and samples from barbershops with male customers were the most infested. Based upon the number of haircuts in each sample, we estimated that 5 or 6% of the population might be infested by this species.     

Biology of Triatoma vitticeps (Stal, 1859) under laboratory conditions (Hemiptera: Reduviidae: Triatominae). I. Evolutive cycle

Observations were made on the evolutive cycle of Triatoma vitticeps, held under laboratory conditions and fed weekly in mice. Of the 435 eggs obtained, from 4 virgin couples, 149 were purposed for the biological cycle study and 286 to evaluate their resistance to starvation, which shall be a second part of this work. Only 50 specimens reached the adult stage in a period of means (S) = 270 +/- 45 days. At the incubation time, the first and second instars were of less than a month for each, while the third, fourth and fifth instars requires approximately one, two and three months, respectively. The search for the first meal occurred clearly on the 3rd, 6th and 10th day. During all the stages, more than 50% of the specimens had only one blood-meal, except the fifth one, when two blood-meals were required. In relation to the time-lapse between the presenting of the blood-meal and the beginning of feeding, as well as the length of the blood-meal, it was observed that these increased gradually according to the stage. From the 423 blood-meals performed, 390 were not followed by defecation in the settled period of 10 min. Under this point of view, T. vitticeps seems to be a poor transmissor of T. cruzi. The experiment was carried out for 13 months and by this time the averages of minimum and maximum temperatures and the humidity were 25 +/- 2 degrees C - 28 +/- 2 degrees C and 80 +/- 2%, respectively. The material belongs to the triatomine colony held at the Oswaldo Cruz Institute, Department of Entomology.