Seventeen sperm samples were evaluated by transmission electron microscopy (TEM) before and after swim-up separation. DNA-fragmentation was tested by terminal d-UTP nick end labeling (TUNEL) in unselected and selected semen samples, and the results were analyzed in relation to sperm ultrastructural characteristics detected by TEM. A significant improvement in mean numbers and percentages of structurally normal sperm was observed after swim-up selection, corresponding to a significant decrease in the percentage of necrotic and apoptotic sperm, while the percentage of sperm with immature nuclei did not change significantly. TUNEL indicated a significant decrease in chromatin-fragmented sperm after swim-up. Swim up selection based on sperm motility excludes many sperm with ultrastructural evidence of necrosis (absent or reacted acrosome, disrupted chromatin, broken plasma membrane) and apoptosis (misshapen nuclei with marginated chromatin), as confirmed by TUNEL analysis. Nevertheless, immature sperm with elliptical or roundish nuclei, misshapen acrosomes and uncondensed chromatin remain part of fertilizing pool.

Adult↗

PubMedDate not supplied

Notulae seminologicae. 1. New combinations of Kartagener's syndrome.

In this note new consequences of the Kartagener's syndrome are described. In males the syndrome involved a diffused sperm immaturity; in one female severe skeletal defects were present probably resulting from bad organization of the renal apparatus.

Adult↗

PubMedDate not supplied

Notulae seminologicae. 4. Mathematical evaluation of submicroscopical alterations in spermatozoa of sterile men with varicocele.

In this paper the mathematical formula recently proposed by Baccetti & Mirolli in this journal is used for the quantitative electron microscopical evaluation of six submicroscopical defects frequently present in the spermatozoa ejaculated by patients affected by various degrees of varicocele. All these characteristics are independent from each other and are in some way related to imperfect sperm maturation. They concern the shape and position of the acrosome, the status of the chromatin, of the mitochondria, of the axoneme, the individuality of the cell. The incidence in the sperm population of these submicroscopical characters is clearly related to the degree of the affection as estimated by usual clinical parameters. This demonstrates that the electron microscopical sperm analysis evaluated by the present formula is a very sensitive tool in revealing fine prognostical parameters.

Acrosome↗

PubMedDate not supplied

Apoptosis in the OC-k3 immortalized cell line treated with different agents.

The aim of this study is to outline the mechanisms leading cochlear cells to die. We utilized an immortalized cell line (OC-k3 cells) derived from the organ of Corti of transgenic mice in order to perform in-depth biochemical studies with no limitations on sample size and number. We probed these cells with cisplatin and gentamicin, two drugs which display in vivo undesired ototoxic side-effects. We investigated cell viability, reactive oxygen species (ROS) production and glutathione (GSH) levels and tested the effects of different concentrations of cisplatin and gentamicin from 0 to 48 h. Results show that cells undergo a dose- and treatment-time-dependent apoptosis characterized by nuclear fragmentation, integrity of the cell membrane and mitochondria, and absence of DNA endonuclease activity. During the early part of treatment, ROS production increases and intracellular GSH decreases, probably due to the activation of protein kinase C alpha. Use of antioxidants such as N-acetylcysteine, GSH and vitamin C rescues cells from apoptosis almost completely. Overall, these data indicate that ROS generation might play a central role in inducing inner ear cell apoptosis and may have an additive role in the ageing process.

Animals↗

PubMedDate not supplied

Fhit protein expression in human gastric cancer and related precancerous lesions.

The FHIT gene is altered in several types of tumors and abnormal expression of Fhit protein have also been reported in some preneoplastic lesions. We have determined the Fhit expression on histological samples of 26 patients affected by preneoplastic lesions who developed a gastric cancer within 2 years. The expression of the Fhit protein was always present in all preneoplastic lesions, while the Fhit protein immunostaining was distributed unevenly in 10 cases and completely lost in 6. The complete loss of Fhit expression only in areas of neoplastic low differentiation suggests that FHIT gene takes part in late gastric carcinogenesis.

Acid Anhydride Hydrolases↗

PubMedDate not supplied

Cisplatin ototoxicity in the Sprague Dawley rat evaluated by distortion product otoacoustic emissions.

The present study has evaluated the use of distortion product otoacoustic emission (DPOAE) responses in the detection of cisplatin-induced ototoxicity in a Sprague Dawley rat animal model. The cisplatin was administered as a 16 mg/kg, dose introduced by a slow 30-min intraperitoneal infusion. Data from three DP-gram protocols, DPOAE input-output responses at 8 kHz, and auditory brainstem responses (ABRs) at 8, 12 and 16 kHz were collected before and 72 h after treatment. The post-treatment ABRs at 16 kHz showed the greatest mean threshold shift of 33.6 dB. The post-treatment DP-gram data showed significant reduction of the signal to noise ratios in the majority of the frequencies tested, across all tested protocols. The data suggest that the most sensitive DPOAE procedure for the early detection of the cisplatin-induced ototoxic damage is the DPOAE I/O protocol. Morphological analyses indicated that the inner hair cells remained intact, while several types of alterations were observed in the arrangement of the stereocilia in the outer hair cells.

Animals↗

PubMedDate not supplied

Inositide-modifying enzymes: a cooperative role in regulating nuclear morphology during differentiation of myeloid cells.

Differentiation and functional response of mature myeloid cells require cytoskeleton remodelling in a dynamic system that involves subcellular organization and regional signalling. Within the myeloid lineage, neutrophils constitute a cell type in which different cell compartments, and predominantly the nucleus, undergo distinctive large changes involving actin reorganization. In the context of the progressive elucidation of the nuclear structure and composition that has been achieved in the last two decades, it is now clear that the nucleus possesses an ordered and dynamic skeletal structure which shares many properties with the cytoskeleton, and the full set of substrates and enzymes that participate in the inositol lipid metabolism. Consolidated evidence indicate that the changes in cytoskeleton assembly are regulated also by phosphoinositides in a way dependent on their local concentration and availability. Indeed, enzymes able to affect the amount and phosphorylation of inositol lipids can play fundamental roles in determining the architectural transitions of the cell. The expression pattern and the changes of activity of PLC and PI 3-K in the nucleus during differentiation of tumoral myeloid precursors suggest that these enzymes play a crucial role in modifying the intranuclear pool of phosphoinositides, which in turn induce the changes in nucleoskeleton associated to granulocytic maturation. It can be speculated that defective control of nucleoskeleton assembly is one of the causes of dysregulated cell maturation or differentiative block in the course of myeloid leukemias. Inositide modifying enzymes can thus be regarded as potential targets for molecularly designed therapeutic intervention on hematological malignancies.

Actins↗

PubMedDate not supplied

Effect of phospholipids on the activity of DNA polymerase I from E. coli.

Sphingomyelin, phosphatidylethanolamine and lecithin at concentrations which destabilize the DNA helix enhance the DNA polymerase activity, but sphingosine and phosphatidylserine have only a moderate effect. MgCl2 was shown to modify the action of sphingomyelin on the DNA polymerase activity. The role of phospholipids in the DNA replicating process was analyzed.

DNA Nucleotidyltransferases↗

PubMedDate not supplied

Notulae seminologicae. 2. The 'short tail' and 'stump' defect in human spermatozoa.

In this note several cases of stunted tails involving the total sperm population in sterile humans are described. Half of the cases are classified as 'short tailed' spermatozoa, the other half as 'stump defect' previously described in bulls. Both defects are referred in details at electron microscopical level.

Acrosome↗