The possible alleviating effect of garlic supplement on the neural retina in a rat model of hypercholesterolemia: A histological and immunohistochemical study

The purpose of this work was to prove that oxidative stress is the main mechanism responsible for retinal neurodegenerative changes, subsequent apoptosis, and inflammatory cytokine release in rats fed with a high cholesterol diet (HCD) and determine the role of garlic in alleviating these changes. Forty rats were equally divided into four groups: control, garlic-treated (positive control), HCD, and HCD + garlic-treated (HCD + G). By the end of the experiment (24 weeks) blood samples were collected for assessment of serum lipid profile, oxidative stress parameters, and plasma levels of IL-6 and TNF-α. Both eyes of the rats were enucleated; one was used for light microscopic examination and the other for electron microscopic examination. There was a significant increase in the levels of serum lipids, oxidative stress parameters, IL-6 and TNF-α, and area of expression of caspase-3 in the HCD group compared to both the control and HCD + G groups. Histological examination revealed degenerative changes in all layers of the neural retina in the HCD group. Garlic administration resulted in a significant improvement in the biochemical, immunohistochemical, and histological characteristics of hypercholesterolemic rats. These findings support the hypotheses that garlic has strong antioxidant, anti-apoptotic, and anti-inflammatory properties. Garlic ameliorates the neurodegenerative changes in the neural retina of hypercholesteremic rats.Key words: Hypercholesterolemia, garlic, neural retina     

Lipid accumulation in the rat liver: a histological and biochemical study.

The sequential pattern of lipid accumulation and associated biochemical changes were studied in two commonly used experimental models of nutritional fatty liver in rats. Female rats were maintained for 8 weeks on high fat, low protein diets containing adequate methionine and choline, and drinking water ad libitum (Diet 1), or deficient in methionine and choline and containing 20% ethanol as a substitute for drinking water (Diet 2). Histologically, there was a progressive increase in liver lipids, mainly in the periportal areas. Occasional foci of liver cell necrosis with lipogranuloma formation occurred in areas of severe fatty change. These changes appeared earlier and were more marked in rats maintained on Diet 2. Electron micrographs revealed large lipid droplets in the liver cells, which sometimes contained myelin figures. The mitochondria were enlarged, distorted and appeared as amorphous structures with disorientated cristae in rats on Diet 1, whereas they had a condensed conformation in rats maintained on Diet 2. Rough endoplasmic reticulum was fragmented and degranulated particularly in rats on Diet 1, and smooth endoplasmic reticulum showed hyperplasia and vesiculation in rats on Diet 2. There was a progressive increase in the total liver lipids and triglycerides in both the groups of rats. This fatty change was accompanied by a significant increase in hepatic 3-hydroxybutyrate, acetoacetate, malate, 2-oxoglutarate, citrate, lactate, ammonia, glutamate, alanine and aspartate, and a significant decrease in oxaloacetate, urea and glucose concentrations. The mass action ratios for alanine aminotransferase, aspartate amino transferase, and glutamate dehydrogenase, generally moved in a parallel direction. Hepatic ATP content was considerably reduced accompanied by a decrease in [ATP]/[ADP] ratios and a significant increased in [lactate]/[pyruvate] and [3-hydroxybutyrate]/[acetoacetate] ratios. There was a corresponding decrease in the [NAD+]/[NADH] ratios both in the cytoplasmic and mitochondrial compartments. These biochemical changes were particularly severe in rats maintained on Diet 1 and Diet 2 for 8 weeks. There was a very good relationship between impaired mitochondrial and endoplasmic reticulum functions, redox and phosphorylation states, and the relevance of their changes to the fate of fatty liver cells.     

Metabolic effects of repeated short-term exposures to heat in the rat

Repeated daily exposures of rats to a hot ambient temperature (40-41 degrees C) for 20 days induced activation of the oxidative enzymes in the thigh muscle and of lactate dehydrogenase in the myocardium. Heat-stressed rats also showed increased tolerance to cold. These changes, it is suggested, are caused by the increased sympathetic activity bouts associated with the heat stresses. The interscapular brown adipose tissue, however, showed adaptive changes similar to those induced by warm acclimation. The activity of oxidative enzymes was decreased and the fat droplets became large and unilocular.     

A study of intercellular bridges during spermatogenesis in the rat

A morphological evaluation of intercellular bridges was undertaken during rat spermatogenesis. The dimensions and relationships of the bridges were shown to vary during different phases of spermatogenesis. Cellular divisions of spermatogonia and spermatocytes resulted in the partitioning of pre-existing bridges by complex structures termed bridge partitioning complexes, which are described in detail, as is the process whereby new bridges are formed. The structure of premeiotic bridges was generally consistent; however, during spermiogenesis, the structure of bridges and bridge contents were modified at specific phases of their development. The plasma membrane density associated with the cytoplasmic aspect of early step 1 spermatids separated into multiple dense bands that encircled the peripheral aspect of late step 1 spermatid bridges. By step 2 of spermiogenesis, these dense bands became associated with several cisternae of endoplasmic reticulum, which later coalesced into a single saccule that completely encircled the bridge structure by step 4. At steps 10-13 of spermiogenesis, the single saccule of endoplasmic reticulum vesiculated into many smaller cisternae. Also, filament-bounded densities (measuring 10-12 nm in diameter) appeared within the bridge channel. At step 17 of spermiogenesis, the filament-bounded densities were no longer apparent, but an anastomosing network of endoplasmic reticulum, often in the configuration of a sphere, occupied the entire central region of the bridge. In step 19 spermatids, the smooth endoplasmic reticulum within the bridge channel and the multiple cisternae lining the bridge density were gradually displaced. The subsurface density of bridges gradually lost its prominence. Some cytoplasmic lobes were connected by extremely narrow (approximately 22 nm) cytoplasmic channels. Similar-appearing channels were seen on the surface zone of cytoplasmic lobes or residual bodies, this observation suggesting that channels were sites of severence of bridges. Just prior to the separation or disengagement of the spermatid from the cytoplasmic lobe, selected bridges appeared to open to form large masses. After spermiation, residual bodies were not found joined by bridges; but from the size of some of the residual bodies, it was suspected that they were formed by coalescence of more than one cytoplasmic lobe. Freeze-fracture demonstrated few intramembranous particles on either the P or E face of the plasma membrane forming the bridge; this finding suggested bridge structures restricted free lateral movement of membrane constituents across the bridge.(ABSTRACT TRUNCATED AT 400 WORDS)     

A quantitative study of spermatogenesis in the developing rat testis

Quantitative (stereological) studies were performed to determine the number of germ cells in the developing rat testis. Sprague-Dawley rats aged 1-70 days were fixed by immersion or perfusion and embedded in Epon Araldite. Blocks of tissue were sectioned at 1.5 microns and stained with toluidine blue dye. Sections were systematically scanned and the areal density of nuclear profiles counted using an unbiased counting frame. Numerical density and absolute number of germ cells in the processed block were then estimated. Corrections for processing shrinkage were determined by comparing the volume of processed and unprocessed samples. The results demonstrate the necessity of determining absolute number rather than volume density (or areal density) in comparing germ cell numbers. In these experiments, spermatogonial numbers stabilized in the range 18.4-23.6 million per testis on Day 30. The number of primary spermatocytes that were first apparent on Day 15 increased rapidly to 54.6 million per testis on Day 30 and then slowly to 73.6 million on Day 70. Round spermatids were first apparent on Day 25 and increased rapidly to 85.7 million per testis on Day 40, then continued to increase to 151.9 million on Day 70. The study provides both methods and baseline data for future experiments involving manipulation of the spermatogenic potential of the testis.     

Effect of cannabinoids on spermatogenesis in vivo: a cytological study

The cytogenetic effects of delta 9-tetrahydrocannabinol (THC) and cannabinol (CBN) (10 mg/kg) were investigated in hybrid mice of genotype (C57BL x C3H)F1. Mice were treated for 5 consecutive days with the specific cannabinoid; 16 days after the last treatment the meiotic cells were evaluated. Analysis of the spermatocyte bivalents at the first meiotic metaphase failed to reveal any numerical or structural abnormality. Contrary to previous reports we failed to find any major meiotic abnormalities associated with THC and CBN treatments. There was no evidence of ring or chain figures. The centromeric banding procedure (C banding) was used to support observations of Giemsa stained cells. It has previously been reported that cannabinoids suppress the incorporation of 3H-uridine into pachytene spermatocytes and round spermatids; the absence of chromosome aberrations in the present study suggests that an adverse effect appears to be present, but may be at a level other than cytogenetic.     

The effect of Methallibure on spermatogenesis in Poecilia reticulata

Methallibure treatment in Poecilia reticulata resulted in an interruption of spermatogenesis. This inhibition occurred at the stage of transformation of secondary spermatogonia to primary spermatocytes. The possible endocrine nature of this inhibition is discussed and it is suggested that treatment with Methallibure might provide a suitable alternative to surgical hypophysectomy in the study of pituitary-gonad relationships in teleosts.     

The effect of bull seminal ribonuclease on spermatogenesis in the rabbit

Bull seminal ribonuclease is a very strong antigen in rabbits. In most animals the antibodies were formed after the first course of immunizations. Antibodies were of IgG immunoglobulin character. When bull seminal ribonuclease itself, and in a complex with IgG immunoglobulin fraction, was injected into rabbit testes, lower weight testes and inhibition of seminiferous epithelium development were observed for a few weeks. Similar changes were evoked by subcutaneous injections of the enzyme. During the inhibition of spermatogenesis by the enzyme no histological changes in the intertubular tissue of testes or changes in androgen concentration in the blood serum were detected.     

Response of rat skin to high-dose unidirectional x-ray microbeams: a histological study

There is growing interest in evaluating microbeam radiation therapy as a potential clinical modality. Microbeam radiation therapy uses arrays of parallel, microscopically thin (<100 microm) planes of synchrotron-generated X rays (microplanar beams, or microbeams). Due to the relatively low beam energies involved in microbeam radiation therapy (a median beam energy of 120 keV was used in the present study), the dose penetration of microbeams in tissue is lower than that used in conventional radiotherapy. This lower energy necessitates using a significantly elevated dose to the skin's surface during clinical microbeam therapy to ensure an adequate dose distribution in the target tumor. The findings of the present study, using a rat skin model, indicated that the skin had an extremely high tolerance to microbeam radiation at doses considerably in excess of those that were therapeutically effective in preclinical studies. A histological study was undertaken to evaluate the biological mechanisms underlying this high tolerance. The irradiation configuration employed single-exposure, unidirectional microbeams 90 microm wide, with 300 microm beam spacing on-center. The in-beam skin-surface absorbed doses were in the range 835-1335 Gy. Monte Carlo simulations of the dose distribution indicated that the "valley" dose, i.e. the radiation leakage between adjacent microbeams, was about 2.5% of the in-beam dose. The high tolerance of the rats' skin to microbeams and the rapid regeneration of the damaged segments of skin were attributed to the surviving clonogenic cells situated between the adjacent microplanar beams. In the epidermis, clonogenic cells in the hair follicular epithelium appeared to play a key role in the regeneration process.