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.

Inhibition of sialyl transferase activity by gossypol acetic acid in human seminal plasma.

Seventeen aliquots of 15 microliters seminal plasma of human origin were incubated with 25 micrograms, 15 micrograms, 5 micrograms and 2.5 micrograms gossypol acetic acid (GAA), dissolved in BWW containing 0.9% sodium chloride and 0.9% benzyl alcohol, at 37 degrees C for 60 minutes. Following incubation, activity of sialyl transferase (S.T.) was determined by a procedure involving incorporation of radioactive sialic acid into asialofetuin. The activity of S.T., expressed as cpm per hour per 15 microliters seminal plasma was compared to controls consisting in incubation of sperm in BWW containing 0.9% benzyl alcohol and 0.9% sodium chloride. The activity of S.T. in controls represented 61.3 +/- 8.0%-67.7 +/- 8.9% of the activity obtained by incubation with BWW only. GAA was found to exert a dose-dependent inhibition of S.T. activity, ranging from 38.3 +/- 20.6% to 53.4 +/- 19.4% (with 25 micrograms) and from 11.3 +/- 14.8% to 21.9 +/- 14.8% (with 2.5 micrograms).

Sialyltransferase activity in AF10 myeloma cell line.

A STA was observed in a human-derived myeloma cell line, AF10, that produces IgE paraprotein. The STA in the culture medium of the AF10 myeloma cell line was associated in the 24 to 48 hr period of incubation with IgE biosynthesis, and increased thereafter up to 72 hr, while IgE remained stable. These data support our previous observations that human myeloma cells are associated with a relatively high production of sialyltransferase, which is released to the medium, presumably by a mechanism of shedding.

Separation of sub-populations of sperm with higher fertility potential from normal and pathological semen by peanut agglutinin.

Incubation of human sperm from semen with counts below 20 x 10(6)/ml with peanut agglutinin (PNA) resulted in agglutination of about 70% of sperm. An inverse correlation was found between non-agglutinating (PNA) sperm and sperm density (r = 0.48, p less than 0.01) and a direct correlation with acrosome-damaged sperm (r = 0.83, p less than 0.001). Binding of 125I-PNA to sperm revealed high, and possibly also low affinity binding sites on sperm from both normozoospermic and oligozoospermic origins. Quantitative transmission electron microscopy analysis revealed that agglutinated, PNA+, sperm had lower frequency of acrosomeless spermatozoa than PNA(-)-sperm (44% versus 77%) and higher fertility score (+0.18 +/- 0.1 and -1.5 +/- 0.87 respectively, p less than 0.01). Removal of PNA- sperm from oligozoospermic semen may increase the fertility score of the remaining sperm.

Andrological parameters in men with high sperm counts and possible correlation with age.

Two hundred eighty six human semen samples with sperm counts above 200 million/ml, were divided into 3 groups according to semen volume: (1) less than 1 ml (low), (2) 1 ml-5 ml (normal), and (3) greater than or equal to 6 ml (high). Each group was examined for routine andrological parameters. Also examined was a possible correlation between sperm counts, semen volumes, and ages of patients. The quality of motility (percent and grade) and viability of sperm were generally lowest in specimens with a volume below 1 ml and highest in those of normal volume. Proportions of morphological normality exhibited a similar trend, lacking, however, statistical significance. The mean fructose content of low-volume semen was significantly decreased in comparison with those of normal volume. Patients with low-volume semen and sperm counts above 200 million/ml were older than those with similar sperm counts and normal volume. The mean ages of all patients with high sperm counts of semen were higher than of those with lower sperm counts. It was assumed that in contradiction to specimens with high sperm counts and low volume, those whose volumes range from 1 ml-5 ml should be considered normal with respect to motility, viability, and morphological normality.

Sialyltransferase activity in plasma cells of multiple myeloma.

A marked elevation of sialyltransferase activity (STA) was observed in a solid tumor of plasma cells, which had been removed from a patient with multiple myeloma (MM), as compared to normal lymphatic tissues. STA was also determined in mononuclear bone marrow cells of 10 patients with MM and found to be 12 times higher than that of bone marrow mononuclear cells from 5 patients with non-malignant disorders (with less than 1% plasma cells in the bone marrow aspirate). A significant correlation was found between STA and the number of plasma cells in the bone marrow aspirate.

Andrological parameters in human semen of high (greater than or equal to 6 ml) and low (less than or equal to 1 ml) volume.

A study was carried out to evaluate 792 semen of high (greater than or equal to 6 ml) and low (less than or equal to 1 ml) volume and sperm counts ranging from 0.1 to 200 x 10(6) per ml. Particularly emphasized were: motility percentage and grade, percentage viability, morphologically normal sperm and immature cells, and the concentration of fructose in semen (mg/ml). The Duncan multiple range test and the Kruskal-Wallis test with multiple comparison of ranks were used in the statistical analyses. The results show that abnormal semen volume influence neither the basic tendency of andrological parameters to change in accordance with sperm counts of semen nor to correlate with each other. The quality of motility was significantly higher in specimens of high semen volume as compared to those of low semen volume. The concentration of fructose in semen was generally higher in specimens of high volumes than in those of low volumes, including semen devoid of sperm. It was assumed that in general increased semen volume does not affect the quality of other andrological parameters, thus being probably unrelated with fertility potential. The considerably lower values of both motility and viability characterizing the semen of low volume would suggest a lower fertility potential of specimens affected by this type of abnormality.

The influence of seminal plasma and polyaminic substances on the motility of isolated human sperm.

The purpose of the study was to examine the influence on the motility patterns of isolated human sperm of seminal plasma originating from semen of high and low motility, as well as of seminal plasma of azoospermic origin. Also assessed was the effect of incubation of sperm in the presence of solutions of the polyamines spermine, spermidine, and putrescine. The mean values of the obtained motility percentage and grade were related to the quality of sperm motility in semen and the quality of seminal plasma in that respect. The best results were obtained with specimens containing greater than or equal to 50% motile sperm and having a motility grade greater than or equal to 2. No relationship was found between the concentrations of polyamines in the seminal plasma and sperm motility of respective semen, as well as between the levels of polyamines and their effect on isolated sperm motility. It appears that the participation of polyaminic substances in the process of motility could be contributive and dependent on additional factors present in the semen.

Sialyltransferase activity and sialic acid levels in multiple myeloma and monoclonal gammopathy.

A significant elevation of peripheral blood mononuclear cell sialyltransferase activity (STA) was observed in 14 patients with multiple myeloma (MM), and compared to 7 patients with monoclonal gammopathy of undetermined significance (MGUS) and to 10 controls. Serum sialyltransferase was significant higher in MM patients as compared to controls. It was also higher than in MGUS patients, but the difference here was not statistically significant. STA was also determined in mononuclear bone marrow cells of 5 patients with MM (with 50 to 100% plasma cells in the bone marrow aspirate) and found to be 19 times higher than that of bone marrow mononuclear cells from 6 patients with non-malignant disorders (with less than 1% plasma cells in the bone marrow aspirate). No significant differences were observed in peripheral blood mononuclear cell sialic acid levels between MM, MGUS and controls.

Beta-endorphin and calcitonin in human semen.

The levels of beta-endorphin (beta-E) and calcitonin were estimated in 36 samples of seminal plasma from semen of normospermic, oligozoospermic, and azoospermic origins and in pools of isolated sperm. The mean levels in plasma calculated for all samples examined were 192 +/- 224 pg/ml for beta-E and 754 +/- 397 pg/ml for calcitonin. The amounts in sperm were as follows: for beta-E in pools with sperm counts of 0.1-10 x 10(6)/ml, 157.2 +/- 99.7 pg/10(8) and 27.9 +/- 23.6 pg/ml protein; in pools of greater than 10-30 x 10(6)/ml, 71.2 +/- 41.5 pg/10(8) and 6.5 +/- 1.2 pg/mg protein; in pools of greater than 30-200 x 10(6)/ml, 24.9 +/- 9.7 pg/10(8) and 61 +/- 1.9 pg/mg protein. For calcitonin the amounts were: 501.2 +/- 170.8 pg/10(8) and 27.4 +/- 21.5 pg/mg protein, correspondingly. It was suggested that beta-E and calcitonin present in seminal plasma are synthesized mostly in a compartment of the male reproductive system. The high cellular beta-E and calcitonin levels would be involved in the process of motility through their effect on calcium transport.

Sialic acid content in mouse myeloma cells and derived B-cell hybridomas with different metastatic potentials.

In a previous work we have reported on the metastatic capacities of mouse myeloma cells (NSO) and of two hybridomas derived from them. The present study was designed to examine the sialic acid content of these cell types, their attachment to glass and their capping in the presence of wheat germ agglutinin (WGA). The results showed no significant differences in sialic acid content between NSO and Hybridoma A (Hy A) cells which are non and low metastatic, respectively, whereas Hybridoma B (Hy B), which is highly metastatic, contained 1.4-1.6-fold higher sialic acid compared to the other two cell types. NSO cells exhibited a high, Hy A a moderate and Hy B a negligible attachment to glass. Capping experiments showed a low response of NSO cells, a moderate response of Hy A and a high response of Hy B in the presence of WGA. The results are discussed with regard to the metastatic potential of these cell types taking into consideration the properties cell surface sialic acid is known to endow to cells, determining various aspects of their behaviour.

Sialic acid content and sialyltransferase activity in human lymphocytes with advancing age.

Sialic acid and sialyltransferase activity were determined in lymphocytes obtained from the blood of 78 healthy male volunteers aged 20-80 years. When grouping was made in double decades, statistical evaluation using the Duncan procedure indicates that sialic acid did not show significant differences between groups, whereas the sialyltransferase activity was significantly higher in the group aged 41-60 years as compared to the group aged 20-40 years and the group aged 61-80 years, both at the 0.05 level.

Sialic acid and neuraminidase activity in rat kidneys 6 months after uninephrectomy.

Sialic acid and neuraminidase activity were determined in the cortex of the remnant kidneys of six uninephrectomized rats. As controls served either the kidneys removed at operation or age-matched kidneys from eight sham operated rats. Six months after uninephrectomy the kidneys became hypertrophied and their mean weight was about 40% higher than age-matched kidneys. Blood urea and creatinine and protein levels in 24-hr urine collections were significantly higher in the experimental animals as compared to those of the same animals before nephrectomy and to sham operated rats, indicating a marked impairment of kidney function. The mean concentration of sialic acid in the cortex of hypertrophied kidneys was not statistically different from either that of the removed or that of the age-matched kidneys. Neuraminidase activity expressed as either per gram fresh tissue or per milligram protein, was not different in the removed and in the hypertrophied kidney. The activity, however, in the latter was significantly lower than in the age-matched kidneys. Whether this finding can be associated with the impairment of kidney function in rats 6 months after uninephrectomy remains to be studied.

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.

Sialic acid in platelets of schizophrenic patients.

1. Nerve cell membrane sialic acid is involved in the activity of nervous tissue by its capacity to bind Ca ions and positively charged biogenic amines. 2. Blood platelets may serve as a model for amine-storing neurons. 3. The purpose of the present study was to determine the sialic acid content of platelets of schizophrenic patients in view of reports showing a reduced serotonin uptake by their platelets. 4. To this end platelets were isolated from the blood of 26 schizophrenic patients (13 males and 13 females) of various diagnostic subtypes and from 21 healthy subjects, and sialic acid was determined after hydrolysis at 80 degrees for 1 h in 0.1 NHCl. 5. The results showed significantly lower contents of sialic acid in the patients as compared to controls calculated both per 10(8) cells and per mg protein (18% and 25% lower, respectively) and appear to be in line with the reduced serotonin uptake in their cells in schizophrenia. 6. There were no appreciable differences between sexes and between the various subtypes of this disease.

Sialic acid content and sialyltransferase activity in lymphocytes from neonates.

Venous blood from 24 healthy volunteers, 20-40 years old, and umbilical cord blood from 28 healthy full-term neonates, immediately after delivery, was used for the determination of lymphocyte sialic acid content and sialyltransferase activity. A significantly increased content of lymphocyte sialic acid and an increased sialyltransferase activity were observed in neonates as compared to adults. It is suggested that the increased sialic acid content of fetal lymphocytes may contribute to a possible masking effect of antigenic sites on fetal immunocompetent cells.

Iron deficiency caused by 7 weeks of intensive physical exercise.

The present study was designed to evaluate the effect of an intensive physical training program involving both isometric and isotonic activities on the body iron status of 8 females and 11 males (age 20 +/- 1 year). The training was carried out over a 7 week period and included 8 h of varying physical activities each day. Venous blood samples were obtained from the subjects prior to the beginning of the training, on day 2 and in weeks 2, 4, 6 and 7 of the program. Blood samples were analyzed for iron, ferritin and hemoglobin (Hb) concentrations, total iron binding capacity (TIBC) and red blood cell count (RBC). Iron levels of males and females decreased 65% after 2 weeks of training (p less than 0.001). At the end of the training program 5 males and 6 females had lower than normal iron values (less than 13.4 mumol.l-1). TIBC increased 25% in women and 18% in men following 2 and 4 weeks of training (p less than 0.001) and remained at this elevated level throughout the training period. Ferritin levels decreased 50% in both sexes after 4 weeks of exercise (p less than 0.05) and remained at this level until the end of the training. Hb and RBC decreased 8-10% in both sexes during the training period. In two of the women anemia occurred after 4 weeks of training. The development of latent iron deficiency in a substantial number of participants after a relatively short period of training is uncommon and may reflect the high intensity of exercise required in this program.(ABSTRACT TRUNCATED AT 250 WORDS)

Sialyl transferase in human semen.

The activity of the enzyme sialyl transferase, which is responsible for the transfer of sialic acid, has been examined in the following human seminal plasma: (1) originating from semen with sperm counts above 30 X 10(6)/ml and exhibiting normal values of other andrological parameters; (2) from semen with sperm counts below 25 X 10(6)/ml and with other andrological parameters of poor quality; (3) from semen devoid of sperm. Additionally, enzyme activity was examined in fractionated (split) semen. The activity in plasma based on measurement of radioactivity incorporated into asialofetuin, following incubation in the presence of 14C sialic acid, was expressed as cpm both per volume and per mg protein. There was a tendency for decreased activity with the decrease in sperm density of semen. The difference, however, lacked statistical significance. Enzyme activity in split semen was significantly lower in the first than in the second fraction.

Modulation of carcinoembryonic antigen release by HT-29 colon carcinoma line in the presence of different agents.

In this study we followed the effects of various differentiating agents on the expression of carcinoembryonic antigen (CEA) released into the medium by a colon carcinoma cell line HT-29. Butyric acid 1 mM markedly increased the level of CEA (12-fold in comparison to control levels). 12-O-tetradecanoyl-phorbol-13-acetate (TPA) 50 ng/ml and 5-azacytidine 4 x 10(-6) M increased the amount of CEA, 2- and 1.5-fold respectively. On the other hand retinoic acid 10(-5) M, N methyl-formamide 1% and N,N hexamethylene bisacetamide 2.5 mM decreased CEA 2-, 4- and 3-fold respectively. Our results emphasize that various differentiating agents affect CEA levels differently. Thus changes in CEA levels appear not to be reliable as a marker of a more differentiated phenotype.

Lactate dehydrogenase and alkaline phosphatase isoenzymes and protein-bound sialic acid in regenerating rat liver.

Lactate dehydrogenase (LDH) and alkaline phosphatase (AP) isoenzyme patterns and protein-bound sialic acid content were compared between normal, regenerating rat liver 10 days after partial hepatectomy and fetal rat liver. For this purpose, liver from ten adult rats and two pools of ten fetal livers each were examined. Isoenzymes were separated by electrophoresis on cellulose acetate and their percent distribution calculated after quantitation by densitometry of the bands. LDH-5 and LDH-4 combined represented in all the tissues examined 90%-94% of the total activity. LDH-5/LDH-4 ratios were nearly equivalent in the normal and regenerated liver (7.14, 6.41), but substantially lower in fetal liver (2.50). Two bands of AP were visualized in electropherograms. AP-1/AP-2 ratio was lower in regenerated liver (1.57) as compared to normal liver (2.27) and still lower in fetal liver (1.06). Protein-bound sialic acid was, on protein basis, slightly but not significantly higher in regenerated liver (1.71 microgram/mg protein) than in normal liver (1.43), and significantly higher in fetal liver (1.87). The relatively small differences in isoenzyme patterns and in protein-bound sialic acid between regenerated and normal liver as compared to those between fetal and normal tissue add support to the view that the cells in regenerated liver are not of embryonic origin.