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Biomedical subjects

M Barnet

Publications and source records attributed to M Barnet.

34 records · Page 2Linked to original sources
PubMed1981

Antigen-specific detection of soluble immune complexes in conglutinin-binding assays.

A modification of the conglutinin-binding assay has been used for antigen identification in circulating immune complexes (IC). The complexed antigen was identified, in conglutinin-bound complexes, by antibodies or antibody fragments, specifically directed against the antigen. These antibody preparations were either radiolabelled or detected with 125I-protein A. This system was found efficient using in vitro-formed bovine serum albumin--anti-bovine serum albumin and tetanus toxoid--anti-tetanus toxoid soluble immune complexes at equivalence or at slight antigen or antibody excess. In addition, with 125I-IgG--anti-HBs and with 125I-F(ab')2--anti-HBs, we examined 44 sera from 41 patients with viral type B hepatitis and the presence of HBs was observed in 18 (40.9%) IC containing tested samples. Therefore, this method appears applicable to clinical investigation.

Animals↗
PubMed1980

Some properties of acid and alkaline phosphatase in seminal fluid and isolated sperm.

Activities of acid and alkaline phosphatases were examined in spermatozoa isolated from 177 semen samples differing in sperm counts. Alkaline phosphatase was also determined in seminal fluid. The enzymes were assayed using disodium p-nitrophenyl phosphate as substrate and were studied with respect to susceptibility to various concentrations of tartrate (acid) and to heat (alkaline). Electrophoretic separation of alkaline phosphatase from seminal fluid was performed using an Helena apparatus. The results showed that acid phosphatase activity in spermatozoa decreased with increase in sperm densities and that elevation of tartrate from 0.028 to 0.17 M usually correlated an inhibition of the enzyme from 72% to 78% (mean values). Alkaline phosphatase was very low in sperm and generally below the sensitivity of the method used. Activity of alkaline phosphatase in seminal fluid showed a tendency to increase with the increase in sperm counts, but the significance of differences between groups was not statistically valid. Exposure of seminal fluid to 55 degrees C for 16 min resulted in enzyme inactivation of about 90% and in this respect the alkaline phosphatase resembles the enzyme of bone origin. The electrophoretic pattern, however, did not confirm this view and the type of alkaline phosphatase in seminal fluid is not clear.

Acid Phosphatase↗
PubMed1980

Properties of spermatozoa from normospermic and oligospermic human semen fractionated on columns of discontinuous gradients of albumin.

One hundred and twenty-eight samples of human semen, divided into four groups according to sperm counts, were fractionated on two types of columns of discontinuous gradients of bovine serum albumin (3%, 15%, or 7.5%, 17.5%). In addition to morphology and motility, protein and DNA contents of spermatozoa as well as microscopical pictures of their sediments were examined in order to assess sperm sensitivity to damage in the course of common isolation procedures. The proportions of morphological normalcy and motility grades within the groups were higher in the bottom column fractions than in the middle ones; both showing prevalence over parent semen. Motility percentages were also higher in the bottom than in middle fractions, but no prevalence over parent semen was observed. Protein and DNA contents of spermatozoa were similar in the two fractions within the same group. Comparison between groups revealed an increase in percentages of normal morphology and motility and a decrease in protein and DNA with elevation of sperm density. Microscopic examinations revealed similar degrees of contamination in both fractions which were more abundant in oligospermia than in normospermia. It is suggested that sperms with better morphology and motility are as sensitive to common laboratory procedures as cells of lower quality originating from the same specimen.

Cell Fractionation↗
PubMed1979

High sperm densities and the quality of semen.

Thirty semens with sperm densities above 200 million per milliliter from patients with fertility disorders were examined for volume, motility, vitality, and morphology of spermatozoa, and for fructose content as well as for percentile decreases with time of activity and viability. In addition, blood levels of follicle stimulating hormone, luteinizing hormone, testosterone, and prolactin were estimated. Percentages of motility, vitality, and morphological normality, motility grades, and percentile decreases in both activity and viability were found to be comparable with those of oligospermic specimens. On the average, volume of semens were lower by about 30% than in normo- and oligospermia. 42% of the investigated semens had low fructose values (below 1 mg/ml). In about 60% of patients the prolactin levels were low (2-5 ng/ml). Follicle stimulating hormone, luteinizing hormone, and testosterone were either within normal range or slightly decreased. Despite the above abnormalities, the overall numbers of "normal" spermatozoa per ejaculate were not inferior to those usually present in normospermic specimens, suggesting that factors of presently unknown nature, involved in regulation of both proliferation and spermatogenesis, might be responsible for fertility disorders in these conditions.

Follicle Stimulating Hormone↗
PubMed1979

Sensitivity of human spermatozoa to various isolation procedures: differences in relation to sperm counts.

The aim of the present work was to examine the sensitivity of human spermatozoa from oligospermic and normospermic semen samples to manipulations in the course of isolation by generally used procedures. To this end we performed protein and DNA estimations in spermatozoa isolated by procedures differing in rigorousness (washings with Ringer's buffer and a medium containing ficoll) as well as microscopic examinations of washed sediments. The results showed that the washed sediments contained, in addition to spermatozoa, materials containing protein and DNA, possibly deriving from the breakage of sperm cells. These materials increased in amount with the decrease of original sperm counts. We suggest that abnormal spermatozoa, whose percentage usually increases with the severity of oligospermia, are more sensitive to damage than the normal ones.

Buffers↗
PubMedDate not supplied

Motility, vitality and percentages of morphologically abnormal forms of human spermatozoa in relation to sperm counts.

The present work was designed to study the proportions of motile and vital sperms as well as of various types of morphologically defective spermatozoa in both normozoospermic and oligozoospermic human semens, aiming at examining whether with the decrease in sperm counts there is a predilection of certain pathologies of these cells. It was found that, generally, the percentages of non-motile and non-vital spermatozoa and of all abnormal forms examined increase with the degree of oligozoospermia. However, no predilection of particular forms could be observed. It is suggested that morphological abnormalities of spermatozoa could be only a partial expression of pathological processes involved in fertility disorders and that the number of biologically defective spermatozoa may be higher than that assessed by routine laboratory techniques.

Adult↗
PubMedDate not supplied

Low energy narrow band non-coherent infrared illumination of human semen and isolated sperm.

Sixty-two samples of human normal and abnormal semen and 23 samples of washed sperm were illuminated for 4 minutes with a narrow band non-coherent infrared device (BioBeam). Immediately after illumination the following parameters were examined in comparison with appropriate controls: motility, viability and morphology, fructose content of semen, acrosome reaction of sperm, viscosity of seminal plasma and analysis of the protein pattern. The mean value of motility grade calculated for the total number of 62 semen samples was 2.58 +/- 0.79 versus 2.17 +/- 0.75 (non illuminated controls), P less than 0.005. The mean value of motility grade of 23 samples of washed sperm was 2.89 +/- 0.77 versus 2.43 +/- 0.62, respectively (P less than 0.01). In two illuminated seminal plasma specimens, the viscosity was decreased by 8.8% and 14.8%, the protein content and pattern remained, however, unchanged. Neither the percentages of motile, viable and morphologically normal sperm, nor the fructose content of semen were found to be affected by the illumination.

Acrosome↗
PubMedDate not supplied

Semen volume and fructose content of human semen. Survey of the years 1980-1989.

The communication presents percentages of semen with low (less than 1 ml) and high (greater than or equal to 6 ml) volume and of semen with decreased fructose content (less than 1 ml) in the total number of 8,128 human specimens analyzed during the years 1980-1989. Percentages of low volume specimens and of specimens with decreased fructose content were found to increase with time. High volume semen lacked such trend.

Adolescent↗
PubMedDate not supplied

Total and free testosterone and estradiol in human semen.

Total and free testosterone (T) and estradiol (E2) were determined by a radioimmunoassay technique in 30 normozoospermic and oligozoospermic human semen samples. The mean amounts of total T ranged from 25.1 +/- 7.4 to 32.5 +/- 12.2 ng/100 mL, and those of the free hormone from 7.0 +/- 2.8 to 7.9 +/- 2.3 ng/100 mL. The values for E2 were 10.5 +/- 4.4 to 11.2 +/- 4.5 pg/mL and 2.3 +/- 1.1 to 3.2 +/- 2.6 pg/mL, respectively. There was no correlation between sperm density of semen and the levels of T and E2. It is suggested that the nonspecific binding of T and E2 in semen provides a physiological reservoir for active, free T and E2 and that an appropriate ratio of free T to free E2 in semen may be crucial in the processes that lead to fertilization.

Adult↗
PubMedDate not supplied

Sialic acid content of human spermatozoa in relation to age and quality of sperm.

Sialic acid (SA) has been estimated in human spermatozoa separated according to age by the use of a discontinuous gradient of human serum albumin (HSA), and according to quality (viability and motility) by the use of a discontinuous gradient of bovine serum albumin (BSA). The amount of SA in the "mature" sperm was 3.5 +/- 0.7 (SD) micrograms/mg protein, whereas in the immature sperm subpopulation, enriched with morphologically abnormal sperm, it was 6.9 +/- 3.4 micrograms/mg protein (P = .05). SA estimated in spermatozoa separated according to quality had values of 3.6 +/- 1.2 micrograms/mg protein in sperm of higher quality and 4.1 +/- 1.3 micrograms/mg protein in that of lower quality; this difference was not of statistical significance. It is suggested that changes in SA levels might reflect differences in the degree of sialylation of protein in the subpopulation of sperm, and thus be related to various stages in the fertilization process.

Cell Separation↗
PubMedDate not supplied

Separation of human semen on Percoll gradients: effect on percentage of motile and morphologically normal sperm and proportion of acrosome reacted sperm.

OBJECTIVES: To examine whether sperm motility and morphology correlate with ability to undergo the acrosome reaction in vitro. METHODS: Sixty-one seminal specimens were separated on a discontinuous Percoll gradient (70%, 40%). Percentages of motile and morphologically normal sperm, their motility grade, and percentages of acrosome-reacted sperm before and following acrosome reaction inducing treatment were estimated. RESULTS: Percentage of motile sperm from normozoospermic specimens increased from whole semen (47.6 +/- 5.6) to sediments (59.6 +/- 13.0). Motility grade exhibited a similar trend (2.1 +/- 0.5 and 3.6 +/- 0.5-scale of 1-4). Percentage of morphologically normal sperm increased from semen (27.3 +/- 10.9) to sediment (53.8 +/- 14.8). All differences were statistically significant, (P < 0.00). In abnormal semen the motility grade and percentage of morphologically normal sperm also increased from semen to sediment (1.9 +/- 0.4 to 3.2 +/- 0.8 and 17.7 +/- 7.8 to 37.1 +/- 15.4 respectively, (P < 0.001). Percentages of motile sperm were similar. In normozoospermic sediments, both non-treated or treated to induce the acrosome reaction, proportions of acrosome-reacted sperm were significantly higher than in semen (18.3 +/- 6.9% and 20.0 +/- 6.5% vs. 6.8 +/- 4.2% respectively, P < 0.001). A similar trend was obtained using abnormal aliquots; however, the acrosome-reacted sperm accounted for only 9.0 +/- 5.1% and 10.8 +/- 4.3% versus 5.4 +/- 2.8% (P < 0.025). CONCLUSIONS: Morphology and motility of sperm were correlated with the proportion of acrosome-reacted sperm.

Acrosome↗
PubMedDate not supplied

Percentages of acrosome-reacted sperm in human semen close to liquefaction and following acrosome-reaction inducing treatment.

OBJECTIVES: To compare spermatozoa from normal and subnormal seminal specimens in their ability to undergo the acrosome reaction, as defined by the triple-staining method. METHODS: Normal (39) and abnormal (23) specimens were examined by staining promptly after liquefaction. In a total of 32 specimens, the reaction was examined before and after acrosome-reaction-inducing treatment, which was essentially incubation of isolated sperm in BWW containing 0.3% BSA and 4 mM calcium chloride. RESULTS: Percentage of vital acrosome-reacted sperm (true acrosome reaction) of untreated specimens: normal, 10.0 +/- 4.9%, abnormal, 5.3 +/- 3.6% (P < .001). After treatment: increase in reacted sperm from normal specimens, 9.0 +/- 7.1% to 22.7 +/- 14.0% (P = .005); abnormal specimens, 5.4 +/- 2.9% to 7.3 +/- 4.2% (NS). CONCLUSIONS: Low capability of sperm to undergo the acrosome reaction might be a decisive factor in the low fertility potential characterizing the majority of abnormal semen. Effort in infertility treatment should be directed to increasing the proportion of acrosome-reacted sperm.

Acrosome↗