Chronic-arginine supplementation improves endothelial cell vasoactive functions in hypercholesterolemic and atherosclerotic monkeys

Chronic exposure to l-arginine results in regression of atherosclerotic lesions and reversal of endothelial dysfunction. We investigated whether chronic l-arginine supplementation induces regression of atherosclerotic lesions and reversal of endothelial dysfunction in atherogenic rhesus monkeys and the mechanism which leads to these effects. About 12 male rhesus monkeys were fed 1% cholesterol and 18 g butter for 6 months to create an experimental model of hypercholesterolaemia and atherosclerosis (Group I) and 12 monkeys were fed standard stock diet for 6 months (Group II). After, 6 months these two groups were further divided into 2 sub-groups which in addition to their respective diets were fed 2.5% l-arginine in drinking water for additional 6 months (Group III and Group IV). Systemic nitric oxide (NO) formation was assessed as plasma nitrite and cGMP formation every 3 months. Oxygen free radical (OFR) generation and malondialdehyde production as an index of lipid peroxidation were determined. Changes in isometric tension were compared in isolated ring segments of thoracic aorta from normal and hypercholesterolemic animals.Cholesterol feeding progressively reduced plasma nitrite and cGMP generation (p<0.05). Dietary l-arginine partly restored the levels of plasma nitrite and cGMP (p<0.05) but did not change plasma cholesterol levels. l-arginine significantly reduced aortic intimal thickening, blocked the production of carotid and coronary intimal plaques and completely preserved endothelium-dependent vasodilator function. Further, l-arginine significantly inhibited generation of the reactive oxygen species (ROS) generation and lipid peroxidation.Chronic oral supplementation with l-arginine blocks the progression of plaques via restoration of nitric oxide synthase substrate availability and reduction of vascular oxidative stress. (Mol Cell Biochem 269: 1–11, 2005)     

Cut-off values for normal sperm morphology and toxicology for automated analysis of rat sperm morphology and morphometry

We used automated sperm morphology analysis to investigate rat sperm morphometry and morphology in Sprague-Dawley and Wistar rats in three research centers to develop normal baseline values for sperm morphometry and to quantify the percentage of morphologically normal sperm in healthy rats. The participating centers were IRSN in Paris, France (Sprague-Dawley rats), University of the Western Cape, South Africa (Wistar rats) and Stellenbosch University (Wistar rats), South Africa. All three centers used identical sperm isolation techniques from the cauda epididymis, the same staining protocols, identical computer-aided sperm morphometry analysis (CASMA) software and microscopes with similar optics. With CASMA, fully automated analysis of the different parts of stained sperm, e.g., head, acrosome, mid-piece, can be performed, many sperm morphometric features can be measured accurately and eventually normal sperm morphology can be defined. We found that it is possible to distinguish sperm morphometric characteristics of Sprague-Dawley and Wistar rats. We also developed cut-off values for evaluating the percentage normal sperm in these two rat strains using the automatic analysis mode. Normal sperm morphology varied between 67 and 74% by contrast with previous findings of > 90%.     

Preparation of soybean seed samples for FT-IR microspectroscopy

Typical preparation of seed samples for infrared (IR) microspectroscopy involves imbibition of the seed for varying time periods followed by cryosectioning. Imbibition, however, may initiate germination even at 4° C with associated changes in the chemistry of the sample. We have found that it is possible to section seeds that are sufficiently hard, such as soybeans, on a standard laboratory microtome without imbibition. The use of dry sectioning of unimbibed seeds is reported here, as well as a comparison of different mounting media and modes of analysis. Glycerol, Tissue-Tek, and ethanol were used as mounting media, and the quality of the resulting spectra was assessed. Ethanol was the preferred mountant, because it dried quickly with no residue and thus did not interfere with the spectrum of interest. Analysis in transmission mode using barium fluoride windows to hold the samples was compared with transmission-reflection analysis with sections mounted on special infrared-reflecting slides. The two modes of analysis performed well in different regions of the spectrum. The mode of analysis (transmission vs. transmission-reflection) should be based on the components of greatest interest in the sample.     

The seminiferous epithelial cycle and spermat ogenesis in rams (ovis aries )

The efficiency of spermatogenesis and degenerations of different spermatogenic cells under normal conditions of the environment have been investigated in rams. The meiotic divisions and the position of first-generation spermatids in haematoxylin-eosin stained testicular preparations were used to identify eight stages of the seminiferous epithelial cycle (SEC). The stages of relatively long duration (i.e., 1,2,3,4,8) were sub-divided. The percent-ages of frequency for the 14 stages reported were also studied. Three generations of type A (A(1), A(2), A(3)), one generation of type intermediate (In) and two generations of type B (B(1), B(2)) spermatogonia were recognized. A(2) and B(2) spermatogonia as well as primary and secondary spermatocytes did not degenerate. Contrarily, A(1), A(2), A(3), In and B(1) spermatogonia showed 25, 13.7, 27.3 and 21.2% degenerations respectively. We concluded that compared with the previously used eight-stage classification, subdividing stages with long durations as done in this study facilitates investigating the degenerations of spermatogenic cells. The efficiency of spermatogenesis in rams was 47.58% since one A(3) spermatogonium produces 30.45 spermatids/spermatozoa against the expected number of 64.     

Correlation between steroids and plasmin in rat ovaries during ovulatory process.

Progesterone (0.5 mg/rat) and estradiol-17 beta (10 micrograms/rat) injected(im) to adult female albino rats on the morning of proesterous significantly enhanced the ovarian plasmin activity as measured by the fibrin plate method at the time of ovulation which was confirmed to be at 2.30 a.m. by estimation of ovarian plasmin activity at definite intervals before and after ovulation. The ovarian plasmin activity showed a gradual increase towards ovulation and reached a maximum level at 2.30 a.m. and again decreased after ovulation. However, the nonsteroidal estrogen antagonist, tamoxifen induced inhibitory effects on the ovarian plasmin activity as compared to control and estradiol-17 beta treatment. Thus these studies reveal a positive relationship between steroids and the ovarian plasmin activity during ovulation.     

Comparison of glucose-6-phosphate dehydrogenase activity in healthy and atretic follicles in rat ovary

Changes in the glucose-6-phosphate dehydrogenase activity have been determined in relation to atresia of Graafian follicles in the rat ovary. Induction of atresia in follicles either due to absence of hCG in the hormonally stimulated immature ovaries or by repeated injections of pentobarbitone sodium to proestrous rats caused significant rise in the enzyme activity. Measurement of enzyme activity in isolated follicular compartments of healthy and atretic follicles revealed that it is significantly higher in the thecal tissue than the granulosa. Increase in enzyme activity in the atretic follicles than the healthy ones occurs due to its rise both in theca and granulosa cells. The significance of these changes in the enzyme activity in healthy and atretic follicles are discussed in relation to the precocious luteinization of cells in the follicular envelope with the onset of atresia.     

Morphology of the post-ovulatory follicle of house sparrow.

The morphology of the post-ovulatory follicle (or corpus luteum) in the sparrow (Passer domesticus) ovary has been investigated with special reference to the origin of luteal cells which finally fill the fillicular activity. The development and degeneration of luteal cell mass has been described in three phases. The luteal cell mass consists of hypertrophied granulosa luteal cells during the first phase and of both granulosa and thecal luteal cells during the second phase. During the second phase owing to their different staining reactions, both types of luteal cells can be differentiated. In the advanced stages of regression, i.e. during the third phase, the whole luteal cell mass consists of thecal luteal cells and connective tissue elements as the granulosa luteal cells had degenerated and disappeared by this stage.     

Fate of aluminium and nickel in soil. Evaluation through lysimeters under laboratory conditions

ABSTRACT

Nanomaterials (Nms) applications and environmental deposition are continuously increasing. Aluminum (Al) and nickel (Ni) fate in soil, both from gamma alumina-based Nms and as chloride salts were evaluted through lysimeters. After 85 days of treatment, which included irrigation and collection of eluates, the soil of each lysimeter was divided into four sections. The metal concentration was analyzed in eluates, soil samples, and extracts. Al and iron (Fe) present in soil eluted from Control lysimeter. Al from Nms suspension treatment was quantified in the eluates since 30 days on. Ni eluted upon solid salt deposition on top of one device. These results indicate that Al and Ni applied under certain conditions on soil, could leach and reach groundwater. The total concentration and bioavailability (extractable metals) of Al and Fe in soils showed similar patterns. Ni was retained only in the soil of devices treated with chloride salts. Bioavailability % results were of concern for Ni under certain conditions of treatment: 15.57% and 11.08% in two chloride salt-treated lysimeters versus 0.55% and 0.47% of those in control and treated with Nms lysimeters. Conducting studies with different kinds of soil and longer treatment periods should be useful to understand Nms-metals fate in the environment. The results presented here constitute important evidences both for significant metal release from Nms and elution and for considerable Ni bioavailability, after deposition on soil in the form of Nms or as a chloride salt, respectively. Then, possible toxic effects could occur through exposure of aquatic and terrestrial organisms.