Dietary supplementation of antioxidants improves semen quality of IVF patients in terms of motility, sperm count, and nuclear vacuolization.

BACKGROUND: This study aimed to investigate the influence of an oral antioxidative supplementation on sperm quality of in vitro fertilization (IVF) patients, as analyzed by sperm motility according to the WHO criteria and motile sperm organelle morphology examination (MSOME). METHODS: Semen samples were collected from 147 patients before undergoing an IVF/intracytoplasmic morphologically-selected sperm injection (IMSI) cycle and 2 - 12 months after an antioxidative supplementation. Semen analysis was evaluated according to WHO and MSOME criteria. Spermatozoa were grouped according to the size of nuclear vacuoles within the sperm's heads. Patients were divided into oligoasthenoteratozoospermic (OAT) and non-OAT men. Between first and second semen analysis, patients were supplemented orally with an antioxidative preparation. RESULTS: After the antioxidative therapy we observed a significant reduction in the percentage of immotile sperm cells in the patients. Additionally, the percentage of class I spermatozoa according to MSOME criteria was significantly higher after antioxidative supplementation. In OAT patients the percentage of class I sperm was found to be increased, although not significantly. However, we observed a drastic improvement in sperm motility as well as in total sperm count in this group. CONCLUSION: The results demonstrated a considerable improvement in semen quality, notably in OAT patients. Considering the putative relationship between semen quality on the one hand and reactive oxygen species on the other, the observed changes in the sperm parameters indicate that a decline in semen quality, and even subtle morphological changes, might be associated with oxidative stress. Our findings suggest that an antioxidative and micronutrient supplementation has a remarkable benefit for IVF patients having restricted sperm parameters, in particular.

Autophagy after experimental intracerebral hemorrhage.

Autophagy contributes to ischemic brain injury, but it is not clear if autophagy occurs after intracerebral hemorrhage (ICH). This study examined whether ICH-induced cell death is partly autophagic. It then examined the role of iron in inducing this form of cell death after ICH. Male, adult Sprague-Dawley rats received an infusion of autologous whole blood or ferrous iron into the right basal ganglia. Control rats (sham) had a needle insertion. The rats were killed at 1, 3, 7, or 28 days later. Some rats were treated with either deferoxamine or vehicle after ICH. Microtubule-associated protein light chain-3 (LC3), a biomarker of autophagosome, and cathepsin D, a lysosomal biomarker, were measured by Western blot analysis and immunohistochemistry. Immunofluorescent double-labeling was used to identify the cell types expressing cathepsin D. Electron microscopy was performed to examine the cellular ultrastructure changes after ICH. We found that conversion of LC3-I to LC3-II, cathepsin D expression, and vacuole formation are increased in the ipsilateral basal ganglia after ICH. Intracerebral infusion of iron also resulted in enhanced conversion of LC3-I to LC3-II and increased cathepsin D levels. Deferoxamine (an iron chelator) treatment significantly reduced the conversion of LC3-I to LC3-II and cathepsin D levels after ICH. Our results demonstrated that autophagy occurs after ICH, and iron has a key role in ICH-induced autophagy. This also suggests that iron-induced autophagy may play a role in brain injury in other diseases associated with iron overload.

Cerebral vacuolation induced in rats by the administration of LUP-3FDC, an anti-tuberculosis cocktail.

A study was conducted to determine the subacute oral toxicity of LUP-3FDC (a cocktail composed of rifampicin, isoniazid and pyrazinamide) and LUP-Q1 (gatifloxicin sesquihydrate) as well as the potential effects of their combination when administered as repeated sublethal oral (gavage) doses for a period of 90 days in seven (7) groups of Wistar rats. Three (3) additional groups were allowed to live for 28 days after the end of treatment to evaluate the potential reversibility of any toxic effects observed. Mortality was observed at all dose levels. General body weakness and hind limb paralysis (attributable to peripheral neuropathy) were observed in animals administered 1400mg/kg/day LUP-3FDC, 800mg/kg/day LUP 3-FDC+300mg/kg/day LUP Q1 and 1400mg/kg/day LUP-3FDC+300mg/kg/day LUP-Q1. The administration of LUP-3FDC at doses of 1100 or 1400mg/kg/day or a combination of 1400mg/kg/day LUP-3FDC and 300mg/kg/day LUP-Q1 induced an increased incidence of vacuolation in the brain compared to control animals. This effect, which was observed predominantly in the cerebellar roof nuclei, was attributed to the isoniazid component of the compound. Vacuoles were located primarily in the myelinated areas close to cerebellar roof nuclei, but were also seen in the olfactory tubercle, thalamus, cerebral cortex, septum and basal ganglia. Females were more susceptible to this change; vacuoles were still evident in males and females 28 days after the cessation of compound administration. The rat cerebellum is prone to develop vacuolation with age; isoniazid may "accelerate" or "enhance" this tendency in young rats.

Ioxaglate-induced light and electron microscopic alterations in the renal proximal tubular epithelium of rats.

Vacuolization of the proximal tubular epithelial cells was produced in rats by the intravenous administration of the radiographic contrast medium ioxaglate at high multiples of the human diagnostic dose. Samples of the renal cortex and outer zone of the medulla were examined by light and electron microscopy. We observed enlargement, confluence, and migration of vacuoles containing pleomorphic dense material and distinct inclusion bodies. With time, vacuolization disappeared, though single vacuoles partly engaged in extruding their contents into the tubular lumen were still visible. We concluded that radiographic contrast medium at high dose levels can produce a reversible disturbance in the transport vesicular system of the proximal tubular epithelial cells without affecting the specific cell organelles.

Intraretinal silicone oil vacuoles after macular hole surgery with internal limiting membrane peeling.

PURPOSE: To report the detection of intraretinal silicone oil vacuoles after the use of a silicone oil tamponade for macular hole surgery with internal limiting membrane (ILM) peeling. DESIGN: Observational case report. METHODS: A 57-year-old woman with a recurrent macular hole in the left eye underwent macular hole surgery with ILM peeling and silicone oil tamponade. After early silicone oil emulsification was detected, the silicone oil was removed. RESULTS: Follow-up ophthalmoscopic examination and optical coherence tomography imaging revealed intraretinal silicone oil vacuoles in the area of ILM peeling. CONCLUSIONS: Internal limiting membrane defects may facilitate the entry of silicone oil into the retina, leading to accumulation of oil vacuoles. The use of silicone oil in macular hole surgery with ILM peeling may complicate the postoperative outcome.

Identification of hibernating myocardium by acoustic microscopy.

Hibernating myocardium is viable myocardium that recovers after revascularization. The observation of loss of contractile proteins (myofibrils) and accumulation of glycogen in hibernating cardiomyocytes provide the basis for diagnosing hibernating myocardium. In this pilot study, acoustic microscopy was used to identify the cellular structure of normal vs. hibernating myocardium. Sections cut at 5-microm of archival paraffin blocks on glass slides were used for this study. Acoustic microscopy of normal cardiomyocytes showed intracellular linear echoes suggestive of myofibrils, and cardiomyocytes of hibernating myocardium revealed absence of myofibrils and dense intracellular echoes that corresponded to glycogen accumulation on optical microscopy. This modality of visualization allows a definitive diagnosis of hibernating myocardium.

Establishment and characterization of a new canine B-cell leukemia cell line.

A new cell line derived from a spontaneous canine leukemia was established and designated GL-1. The cells have been cultured in a floating fashion and passaged for over two years. They were round with rich cytoplasm containing many rough endoplasmic reticula and mitochondria. Peroxidase staining was negative. The nuclei of many cells were round, but segmented nuclei were seen frequently. The doubling time of the cells was 27.3 hr and they had 78 chromosomes. Surface marker analysis using monoclonal antibodies (MABs) and flowcytometry revealed that GL-1 possessed CD45 and surface IgG. However, the cells did not react with MABs detecting T-cell markers. These results indicate that GL-1 has a lymphocytic lineage and is derived from a B-cell leukemia.

The arachiform vacuolar body: an overlooked shared character in the ascospores of a large monophyletic group within Parmeliaceae (Xanthoparmelia clade, Lecanorales).

Sections of apothecia were used to study the internal morphology of ascospores in the largest monophyletic clade within Parmeliaceae composed of Xanthoparmelia and related genera. The results were compared with fertile representative species of most other parmelioid clades. All the Xanthoparmelia species had spores with a single smooth vacuole, which was peanut-shaped, with different degrees of constriction in the equatorial plane. This differs from the ellipsoid vacuole of other parmelioids. In the Xanthoparmelia clade, sexual reproduction seems much more common than in other parmelioids. Thus, we suggest that the presence of this unique spore morphology might contribute to the evolutionary success of this monophyletic group. Further, the discovery of this useful ascospore character demonstrates that detailed ascospore morphological studies significantly enhance molecular phylogenetic analyses. Ascospore features may be more taxonomically significant in Parmeliaceae than hitherto considered.

X-linked vacuolated myopathy: membrane attack complex deposition on muscle fiber membranes with calcium accumulation on sarcolemma.

Cytochemical localization of calcium on muscle fibers from patients with X-linked vacuolated myopathy demonstrated strong sarcolemmal or vacuolar calcium deposits in histologically abnormal muscle fibers. All membrane attack complex-positive fibers demonstrated such calcium accumulation. Total content of calcium was significantly elevated in muscles from patients with X-linked vacuolated myopathy compared to control muscles. Taken together, these results suggest that calcium accumulation on sarcolemma could be probably secondary to membrane attack complex deposition on the cell surface.