José Roberto Botelho de Souza

Chronic treatment with sildenafil stimulates Leydig cell and testosterone secretion

The phosphodiesterase type 5 (PDE5) inhibitor, Sildenafil, is a novel, oral treatment approach for pulmonary hypertension. As Leydig cells present PDE5, this study was conducted to investigate the effects of the chronic treatment with Sildenafil (25 mg/kg) on male Swiss Webster mice steroidogenesis. After a 4‐week long experimental design, Leydig cells were analysed by morphological and immunocytochemical procedures. Serum testosterone was assayed by radioimmunoassay. Leydig cells presented noteworthy ultrastructural alterations, such as a vesicular smooth endoplasmic reticulum, large vacuoles scattered through the cytoplasm, enlarged mitochondria with discontinue cristaes and whorle membranes with vesicles at the periphery, which are typical characteristics of an activated steroid‐secreting cell. Important immunocytochemical labelling for steroidogenic acute regulatory protein, cytochrome P450 side‐chain cleavage enzyme and testosterone were detected in isolated Leydig cells. In addition, Sildenafil‐treated mice showed significant increased levels of total testosterone. The results obtained in the present study are consistent with the hypothesis that the accumulation of cyclic guanosine monophosphate by PDE5 inhibition could be involved in the androgen biosynthesis stimulation. Important clinical implications of hormonal disorders should be taken into account for patients with pulmonary hypertension.

Phytal Marine Nematode Assemblages and their Relation with the Macrophytes Structural Complexity in a Brazilian Tropical Rocky Beach

The nematofauna from the seaweeds Sargassum polyceratium Montagne, Hypnea musciformis Küetzing, Padina gymnospora Küetzing and the seagrass Halodule wrightii Ascherson was studied in March 2001, at Pedra do Xaréu, Pernambuco, Brazil, in order to investigate the associations of organisms, as well as the relation between the plant architecture and the associated fauna. Soft sediments adjacent to phytal environments were also investigated. Thirty-eight genera were found, including two new records (Odontanticoma sp. and Wieseria sp.) for the Brazilian coast. Multivariate analyses were carried out in order to verify the relation between nematode assemblages on plants and in sediments. The results showed that these assemblages were different in structure and composition between macrophytes and sediments. They were different in structure but not in composition among macrophytes, so the nematode biodiversity was related to the structural features of macrophytes’ habitats.

Eclosion rate, development and survivorship of Aedes albopictus (Skuse)(Diptera: Culicidae) under different water temperatures

In tropical areas, where vector insects populations are particularly numerous, temperature usually range between 25 degrees C and 35 degrees C. Considering the importance of such temperature variation in determining mosquitoes population dynamics, in this work the developmental, eclosion and survival rates of the immature stages of Aedes albopictus (Skuse) were compared under constant 25, 30 and 35 degrees C (using acclimatized chambers) and environmental (25 degrees C to 29 degrees C) temperatures. The hatching rate was considered as total number of larvae recovered after 24h. The development period as well as larval and pupal survival rate were evaluated daily. Eclosion rate was significantly higher under environmental temperature than under the studied constant temperatures, suggesting that temperature variation may be an eclosion-stimulating factor. The mean eclosion time increased with the temperature, ranging from 2.8h (25 degrees C) to 5.2h (35 degrees C). The larval period was greatly variable inside each group, although it did not differ significantly amongst groups (11.0 +/- 4.19 days), with individuals showing longer larval stages in water at 35 degrees C (12.0 +/- 4.95 days) and environmental temperature (13.6 +/- 5.98 days). Oppositely, survival was strongly affected by the higher temperature, where only one individual lived through to adult phase. The results suggest that population of Ae. albopictus from Recife may be adapting to increasing of environmental temperatures and that the limiting temperature to larval development is around 35 degrees C.

Reproductive biology of Echinometra lucunter (Echinodermata: Echinoidea) in a northeast Brazilian sandstone reef

The edible sea urchin Echinometra lucunter (Linnaeus, 1758) is a very common species on the sublittoral-midlittoral in Brazilian rocky shores. The aim of this work was to describe the gametogenesis and reproductive strategy of the E. lucunter population at Muro Alto beach in the Northeast coast of Brazil from August 2004 to August 2005. A total of 240 specimens were collected on the sandstone reef flat from a tidepool during spring low tides. The overall sex ratio was1.12:1,withoutsignificanttemporalvariationexceptinOctober2004. Firsts sexual maturity ocurred in individuals from a diameter of 20.8 mm. There was not a significant difference in gonad index between females and males during the sampling period. The females gonad index variation was associated with a well-defined spawning, corroborated by the histological analysis of the gonads, which demonstrates sex differences of the gamete production. By contrast, the males showed no clear pattern. It is suggested that continuous reproduction with seasonal peaks in the E. lucunter population occurs at Muro Alto beach.

Spatial distribution of the estuarine ichthyofauna of the Rio Formoso (Pernambuco, Brazil), with emphasis on reef fish

The objectives of the present study were to identify species of reef fish that use the Rio Formoso estuary (northeastern Brazil) as a refuge and natural nursery and to describe the spatial distribution of the estuary fish fauna in rainy and dry seasons. A total of 5475 specimens, across 78 species and 39 families, were analyzed; 51.3% of the species were of reef origin. Among these, Eucinostomus melanopterus (Bleeker, 1863), E. gula (Cuvier, 1830), and Sphoeroides testudineus (Linnaeus, 1758), in this order, were the most abundant in the upper estuary; S. greeleyi Gilbert, 1900, E. melanopterus, and Lutjanus synagris (Linnaeus, 1758) were the most abundant in the mid estuary; and E. gula and Albula vulpes (Linnaeus, 1758) in the lower estuary. The percentages of reef species were 39.2%, 54.2% and 66.7% for the upper, mid and lower regions, respectively. The greatest diversity was found in the upper estuary and greatest abundance occurred in the mid region. The type of sediment was a strong determinant of the spatial distribution of fish fauna. The most abundant species were found in a mesohaline (5-18) to euhaline (30-40) salinity regimen, suggesting some capacity for osmotic regulation. The fish fauna of the Rio Formoso estuary receives a direct influence from the reefs and coastal region between Sirinhaem and Tamandare, thereby providing a greater richness of reef fish.

Population dynamics and secondary production of Euzonus furciferus Ehlers (Polychaeta, Opheliidae) in an exposed sandy beach of Southern Brazil

The distribution, population dynamics and secondary production of the polychaete Euzonus furciferus was studied in Atami Beach (Southern Brazil), from February 1992 to March 1993. Euzonus furciferus Ehlers, 1897 is the only deposit feeder species of the upper intertidal region. The organisms were present in the sandy beach all over the year, concentrated in a narrow band, and reaching maximal densities of 3,029 individuals m-2. Peaks of abundance occurred in September 1992, with a mean of 681.8 individuals m-2. Recruitment occurred in winter, with a peak in July. Mean annual biomass was estimated in 0.218 gAFDW m-2, with a total annual production of 0.466 gAFDW m-2 y-1, giving a P/B ratio of 2.13 y-1. Similar values were found for a filter-feeder polychaete inhabiting the same beach, suggesting that general food disposability of each particular environment is more important for production than the trophic strategic employed by those organisms exploring this environment.

Effect of maternal diet rich in omega-6 and omega-9 fatty acids on the liver of LDL receptor-deficient mouse offspring.

BACKGROUND Omega-6 fatty acids are important to fetal development. However, during gestation/lactation, these fatty acids may contribute toward the development of fat tissue. Omega-9 fatty acids are associated with a reduction in serum lipids and protection from liver disease. OBJECTIVES The present study investigated the effect of the maternal intake of omega-6 and omega-9 in hypercholesterolemic mothers on the liver of the offspring. METHODS LDL receptor-deficient mice were fed a diet rich in either omega-6 (E6D) or omega-9 (E9D) for 45 days prior to mating and until the birth of the offspring, evaluating the effect on the offspring liver in comparison to a standard diet (STD). RESULTS Mothers fed with the E6D experienced an increase in total cholesterol (TC) and the offspring exhibited an increase in TC, hepatic triglycerides (TG), and CC-chemokine ligand (CCL)2/monocyte chemoattractant protein (MCP)-1 as well as a reduction in HDL. Histological analysis on this group revealed steatosis, leukocyte infiltrate, and increased CCL2/MCP-1 expression. The ultrastructural analysis revealed hepatocytes with lipid droplets and myofibroblasts. The offspring of mothers fed the standard diet exhibited low serum TC, but microvesicular steatosis was observed. The offspring of mothers fed the E9D exhibited lower serum and hepatic TG as well as higher LDL in comparison to the other diets. The histological analyses revealed lower steatosis and leukocyte infiltrate. CONCLUSIONS The findings suggest that hypercholesterolemic mothers with a diet rich in omega-6 fatty acids predispose their offspring to steatohepatitis, whereas a diet rich in omega-9 has a protective effect.

Hydrocarbons in sediments along a tropical estuary-shelf transition area: Sources and spatial distribution

Estuaries generally act as sediment traps and may retain a range of contaminants associated to this matrix. Aliphatic hydrocarbons (AHs) were investigated in Capibaribe Estuarine System and adjacent shelf, Northeast of Brazil, to evaluate the contamination and to better understand its functionality related to the coast. Fourteen sediment samples were analyzed, using gas chromatography with flame ionization detection. Total AHs concentrations ranged from 7.5 to 190.3μgg-1 and n-alkanes ranged from below detection limit (<DL) to 9.47μgg-1. The highest concentrations were in the upper portion of the estuary, which receives domestic and industrial effluents. The observed UCM and other geochemical markers, indicated contamination by petroleum hydrocarbons. The concentration decrease (about 90%) towards the adjacent shelf suggested an estuarine high retention capacity but dilution and degradation processes cannot be neglected. Similar AHs characteristics reported in sediments from the adjacent shelf suggested that this system may also export contaminants.

Effect of new thiazolidine derivatives LPSF/GQ-02 and LPSF/GQ-16 on atherosclerotic lesions in LDL receptor-deficient mice (LDLR(-/-)).

BACKGROUND Atherosclerotic cardiovascular disease is a chronic inflammatory condition. Thiazolidinediones (TZDs) are used to enhance sensitivity to insulin and have demonstrated a protective effect over a variety of cardiovascular markers and risk factors. Controversially, the TZDs are associated with the development of heart failure. Thus, lines of research have invested in the search for new molecules in order to obtain more selective and less harmful treatment alternatives for the pathogenesis of atherosclerosis and its risk factors. METHODS Animals were fed a diet rich in fat for 10 weeks. In the last 2 weeks, animals received either pioglitazone, LPSF/GQ-02, or LPSF/GQ-16 daily through gavage. At the end of the treatment, blood was collected for biochemical analysis and the aortas were dissected for subsequent analyses. RESULTS No changes in the blood lipid profile were found following the use of the drugs in comparison to the control. However, the new thiazolidine derivatives were more efficient in improving insulin resistance in comparison to pioglitazone and the control group. Morphometric analyses revealed that neither pioglitazone nor LPSF/GQ16 led to satisfactory effects over atherosclerosis. However, LPSF/GQ-02 led to a reduction in area of the atherosclerotic lesions. Ultrastructural analyses revealed extensive degeneration of the endothelium and an increase in apoptotic cells in the subendothelial space following the use of pioglitazone and LPSF/GQ-16. However, LPSF/GQ-02 caused minimal cell alterations in the aortic endothelium. Regarding markers, endothelial nitric oxide synthase (eNOS) and matrix metalloproteinase 9 (MMP-9), LPSF/GQ-16, and pioglitazone exerted similar effects, increasing the expression of MMP-9, and had no effect on the expression of eNOS compared with the control group. On the other hand, LPSF/GQ-02 was effective in reducing the expression of MMP-9 and increased eNOS significantly. CONCLUSIONS The results suggest that the new thiazolidine derivative LPSF/GQ-02 is a promising candidate for the treatment of atherosclerosis.

Relationship between litter characteristics and female size in Tityus stigmurus (Scorpiones, Buthidae)

Abstract Tityus stigmurus (Thorell 1876) is one of the most medically important scorpion species in Brazil, but many basic aspects of its life history are unknown. Here the pattern of female reproductive investment was examined, along with development of the 1st and 2nd instars and the relationship between 2nd instar mass and molting to the 3rd instar. Relative to other buthid scorpions, T. stigmurus has a smaller litter (average 10 young) and a shorter 1st-instar period (average 4 days) and 2nd-instar period (average 68 days). Neither litter size nor offspring mass showed a relationship to female size. A significant positive correlation was observed between total litter mass and litter size. The minimum mass required for successful molting to the 3rd instar was 34.0 mg. Overall, female reproductive resources in T. stigmurus appear to be applied to the production of more but not heavier offspring.