Phagocytosis of bacteria by polymorphonuclear leukocytes: a freeze-fracture, scanning electron microscope, and thin-section investigation of membrane structure

The changes in membrane structure of rabbit polymorphonuclear (PMN) leukocytes during bacterial phagocytosis was investigated with scanning electron microscope (SEM), thin-section, and freeze-fracture techniques. SEM observations of bacterial attachment sites showed the involvement of limited areas of PMN membrane surface (0.01-0.25μm(2)). Frequently, these areas of attachment were located on membrane extensions. The membrane extensions were present before, during, and after the engulfment of bacteria, but were diminished in size after bacterial engulfment. In general, the results obtained with SEM and thin-section techniques aided in the interpretation of the three-dimensional freeze-fracture replicas. Freeze-fracture results revealed the PMN leukocytes had two fracture faces as determined by the relative density of intramembranous particles (IMP). Membranous extensions of the plasma membrane, lysosomes, and phagocytic vacuoles contained IMP's with a distribution and density similar to those of the plasma membrane. During phagocytosis, IMPs within the plasma membrane did not undergo a massive aggregation. In fact, structural changes within the membranes were infrequent and localized to regions such as the attachment sites of bacteria, the fusion sites on the plasma membrane, and small scale changes in the phagocytic vacuole membrane during membrane fusion. During the formation of the phagocytic vacuole, the IMPs of the plasma membrane appeared to move in with the lipid bilayer while maintaining a distribution and density of IMPs similar to those of the plasma membranes. Occasionally, IMPs were aligned to linear arrays within phagocytic vacuole membranes. This alignment might be due to an interaction with linearly arranged motile structures on the side of the phagocytic vacuole membranes. IMP-free regions were observed after fusion of lysosomes with the phagocytic vacuoles or plasma membrane. These IMP-free areas probably represent sites where membrane fusion occurred between lysosomal membrane and phagocytic vacuole membrane or plasma membrane. Highly symmetrical patterns of IMPs were not observed during lysosomal membrane fusion.     

Cytochemical Localization of Plasma Membrane Enzyme Markers During Interiorization of Tachyzoites of Toxoplasma gondii by Macrophages

The enzyme activity of Mg-ATPase, Na⁺-K⁺-ATPase, 5'-nucleotidase and NAD(P)H-oxidase was cytochemically detected at the ultrastructural level in mouse peritoneal macrophages infected with untreated and with specific antibody-coated Toxoplasma gondii tachyzoites. The Mg⁺⁺-ATPase and 5'-nucleotidase were distributed throughout the macrophages'plasma membrane but were not observed in the membrane lining endocytic vacuoles containing ingested parasites; however, Na⁺-K⁺-ATPase activity was detected in the macrophages'plasma membrane as well as in the parasitophorous vacuoles that contained untreated or specific antibody-coated parasites. Reaction product, indicative of NAD(P)H-oxidase, was detected in the parasitophorous vacuoles that contained only-specific antibody-coated parasites.     

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)     

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.