PubMed Health⌕ Search

Biomedical subjects

R Golan

Publications and source records attributed to R Golan.

At least 37 records · Page 2Linked to original sources

Phase imaging of moving DNA molecules and DNA molecules replicated in the atomic force microscope.

Phase imaging with a tapping mode atomic force microscope (AFM) has many advantages for imaging moving DNA and DNA-enzyme complexes in aqueous buffers at molecular resolution. In phase images molecules can be resolved at higher scan rates and lower forces than in height images from the AFM. Higher scan rates make it possible to image faster processes. At lower forces the molecules are imaged more gently. Moving DNA molecules are also resolved more clearly in phase images than in height images. Phase images in tapping mode AFM show the phase difference between oscillation of the piezoelectric crystal that drives the cantilever and oscillation of the cantilever as it interacts with the sample surface. Phase images presented here show moving DNA molecules that have been replicated with Sequenase in the AFM and DNA molecules tethered in complexes with Escherichia coli RNA polymerase.

Aluminum Silicates↗

Effects of pivalic acid and sodium pivalate on L-carnitine concentrations in the cauda epididymidis and on male fertility in the hamster.

In this study, administration of pivalic acid or its sodium salt was found to decrease the L-carnitine concentration in the epididymal lumen of the hamster; it also tested whether this decrease affected sperm cell motility, chromatin structure, or fertilizing capacity. Provision of pivalic acid or its sodium salt (20 mM or 40 mM) in the drinking water of mature male golden hamsters for 30 days reduced (by 72%, 75%, and 83% in three experiments) the L-carnitine concentration of the cauda epididymidis but did not inhibit sperm chromatin condensation, as assessed by flow cytometry. The treatments did not alter the location of motile sperm in the epididymidis nor did they appreciably affect the motility of sperm obtained from the distal cauda epididymidis. The numbers and percentage of ova that reached the 2-cell stage 36-40 h after uterine insemination with spermatozoa from control and treated hamsters served as a measure of sperm fertility. Treatment with pivalic acid or sodium pivalate did not render male hamsters infertile although it appeared to reduce the fertilizing ability of their spermatozoa. These results suggest that the high concentration of L-carnitine present in the lumen of the cauda epididymidis is not required for maturation of sperm chromatin or development of sperm motility.

Animals↗

Evaluation of chromatin condensation in human spermatozoa: a flow cytometric assay using acridine orange staining.

The quality of sperm chromatin is an important factor in fertilization and is especially critical where one spermatozoon is artificially selected for fertilizing an egg (as in intracytoplasmic sperm injection). In this study, flow cytometry after staining of human spermatozoa with Acridine Orange was used to study chromatin structure. A method is described for estimating the percentage of cells in a human sperm sample that have completed epididymal maturation in regard to chromatin condensation. Of the 121 samples of the semen that were examined, nine contained a higher percentage of hypocondensed spermatozoa and six samples contained elevated amounts of hypercondensed spermatozoa. In addition to aberrancies in chromatin condensation other defects showed up as satellite populations of spermatozoa with higher than normal ratios of red/green fluorescence after Acridine Orange staining. Such defects were found in 15 semen samples. The use of swim-up and Percoll gradient centrifugation methods was shown to improve the percentage of spermatozoa with normal chromatin structure in some samples with poor initial quality.

Acridine Orange↗

Epididymal maturation of chromatin in spermatozoa from control monkeys (Macaca fascicularis) and those treated with Cetrorelix, a gonadotropin-releasing hormone antagonist.

In order to examine changes in sperm chromatin upon epididymal maturation in the macaque epididymis (Macaca fascicularis), spermatozoa were obtained from six regions of the duct and examined for the state of their chromatin condensation by flow cytometry after staining with acridine orange. To see whether changes were affected by androgens, tissue was obtained from five monkeys treated with the gonadotropin releasing hormone (GnRH) antagonist Cetrorelix. Spermatozoa were recovered from treated and control animals after 16 days (at hemicastration) and another 9 days of treatment. Chromatin condensation of epididymal spermatozoa from controls displayed an increase upon maturation. After 16 days of GnRH-antagonist treatment, spermatozoa in the caput epididymidis displayed greater fluorescence than those from controls, but this was reduced during epididymal transit to values found in the distal epididymal regions of the controls. It is concluded that epididymal chromatin condensation 1) is normal in GnRH-antagonist-treated monkeys as long as sperm are being produced and 2) can compensate for poor testis function so that spermatozoa with normal states of chromatin condensation are found in the distal cauda epididymidis and probably the ejaculate.

Animals↗

Inhibition of Escherichia coli beta-galactosidase by 2-nitro-1-(4,5-dimethoxy-2-nitrophenyl) ethyl, a photoreversible thiol label.

1-Nitro-2-phenylethene (beta-nitrostyrene, 1) which is a thiol-protecting reagent (Jung, G., Fouad, H. and Heusel, G. (1975) Angew. Chem. Int. Ed. Engl. 14, 817-818), was demonstrated in this work to be an irreversible inhibitor of beta-galactosidase (EC 3.2.1.23), an enzyme known to be inhibited by some thiol reagents or through modifying a methionine residue at the active site. No reversal of the inhibition was observed upon subsequent incubation with mercaptoethanol or irradiation (350 nm). 1-(4,5-dimethoxy-2-nitrophenyl)-2-Nitroethene 2) was also shown to be an irreversible inhibitor (94% inhibition, pH 8.3) of the enzyme. Kcat values of beta-galactosidase at pH 8.3 with o-nitrophenyl beta-D-galactopyranoside (ONPG) as the substrate and at the highest inhibitor concentrations employed for compound 1 (4.06 x 10(-4) M) ranged from 1.67 x 10(4) S-1 after 30 min of preincubation to <0.07 x 10(4) S-1 after 180 min preincubation. For compound 2 (9.5 x 10(-5) M) Kcat values ranged from 2.70 x 10(4) S-1 following 30 min preincubation to 1.15 x 10(4) S-1 after 180 min of preincubation; the changes in Km(app), however, were small. The activity was not recovered following incubation with mercaptoethanol. Since compound 2 and the inhibited enzyme are 2-nitrobenzyl derivatives, they are expected to be photosensitive and indeed, irradiation of the inhibited enzyme in the presence of mercaptoethanol resulted in recovery (89%, pH 8.3) of the enzyme activity.

Enzyme Inhibitors↗

Changes in chromatin condensation of human spermatozoa during epididymal transit as determined by flow cytometry.

Inasmuch as caput epididymal and even testicular spermatozoa are now being used to generate pregnancies by direct injection into the oocyte, differences in the chromatin of spermatozoa from proximal and distal locations in the epididymis were studied. Acridine Orange staining was used to investigate chromatin structure in human spermatozoa which had left the testis and were undergoing maturation in the epididymis. Measurement of green and red fluorescence intensities of human spermatozoa by flow cytometry demonstrated a decrease in binding of Acridine Orange to DNA as the spermatozoa traversed the epididymis. Using spermatozoa from the cauda epididymis as the standard, the percentages of spermatozoa from the efferent duct, proximal corpus epididymis, midcorpus epididymis, distal corpus epididymis, proximal cauda epididymis and distal cauda epididymis that had matured with regard to chromatin condensation were 28 +/- 5, 39 +/- 3, 49 +/- 5, 64 +/- 5, 69 +/- 6 and 74 +/- 4% respectively. It may be concluded that eggs fertilized by ejaculated spermatozoa receive a more highly condensed form of chromatin than that received by eggs inseminated with proximal epididymal or testicular spermatozoa.

Acridine Orange↗

Cardiovascular responses to beta-blockade and 5 degrees C cold air stress.

To determine the cardiovascular responses to beta-blockade and cold air stress, six males were randomly exposed at rest to three drug conditions (placebo, nonselective beta-blockade (propranolol), and selective beta-blockade (atenolol)) in each of two environments (5 and 25 degrees C) for 1 h. Cardiac output was lower on beta-blockade than on the placebo in both the 25 and 5 degrees C environments. Cardiac output on propranolol (4.2 +/- 0.3 L.min-1) at 5 degrees C was lower than on atenolol (4.7 +/- 0.4 L.min-1, p < 0.05). Mean arterial pressure was greater (p < 0.05) at 5 than 25 degrees C for each drug condition. There was no drug effect on total peripheral resistance at 25 degrees C. At 5 degrees C, total peripheral resistance on both beta-blockers (propranolol 1942.7 +/- 169.9 dyn.s.cm-5 (1 dyn = 10 microN); atenolol 1706.7 +/- 160.0 dyn.s.cm-5) was higher (p < 0.05) than on the placebo (1485.3 +/- 111.8 dyn.s.cm-5). Total peripheral resistance was also higher on propranolol than atenolol (p < 0.05). In conclusion, cold air stress interacts with beta-blockade to elevate total peripheral resistance by decreasing cardiac output while having little effect on mean arterial pressure. These effects are greater on nonselective than on selective blockade.

Adult↗

Energy expenditure variations in soldiers performing military activities under cold and hot climate conditions.

This study assessed the energetic status of soldiers exposed to intense physical activities in cold and warm weather. Thirty subjects participated in a two-phase study group A (n = 18) in the winter phase and group B (n = 12) in the summer phase. Energy expenditure (EE) was measured by the doubly labeled water technique; after a single, oral dosing of 2H(2)18O, daily urine samples were collected for 12 successive days. Energy intake (EI) was assessed from detailed food records analyzed by computerized food charts. Energy balance was calculated as the difference between EI and EE for each subject. Mean (+/- SE) daily EE was 4,281 +/- 170 and 3,937 +/- 159 kcal/day for the winter and summer groups, respectively. Daily EI was 2,792 +/- 124 kcal/day in group A and almost identical in group B. A negative energy balance of 1,422 +/- 163 kcal/day and 924 +/- 232 kcal/day (not significant) was calculated for groups A and B, respectively. Energy expenditure is primarily determined by the level of activity rather than by climate conditions; EI is insufficient to offset the high energy requirements under these conditions.

Adult↗

Orthodox Jews have a lower incidence of malignant melanoma. A note on the potentially protective role of traditional clothing.

The incidence of malignant melanoma (MM) in orthodox and non-orthodox Jews in Israel during the years 1970-1982 was compared through an assessment of incidence rates in the orthodox and non-orthodox neighborhoods in the city of Jerusalem and by a comparison of the rates in 2 cities on the outskirts of Tel Aviv, one of which has a strongly orthodox population. Within the city of Jerusalem the incidence of MM for the European-American born in the orthodox neighborhoods was significantly lower than in the rest of the city in males, and of borderline significance in females. A similar pattern was seen upon comparison of the orthodox city of Bnei Brak to its neighbor city, Givatayim. The findings are consistent with the role of sun exposure in the etiology of malignant melanoma.

Clothing↗

Alkaline phosphatase in human semen: an investigation using enzyme inhibitors and gel electrophoresis.

Human seminal alkaline phosphatase was investigated with respect to its electrophoretic mobility, heat lability, and susceptibility to inhibition by phenylalanine, tartrate, and homoarginine. Total alkaline phosphatase activity in 30 samples of human semen was measured colorimetrically, using p-nitrophenylphosphate as substrate. Using linear regression analysis, no significant correlation was found between the enzyme activity and the sperm count, sperm motility, semen volume, and the concentrations of seminal inositol and fructose. The alkaline phosphatase activity was higher in the earlier portion of split ejaculate samples. Sodium DL-tartrate (42 mmol/l), which inhibits acid phosphatase, did not inhibit seminal alkaline phosphatase significantly. L-Homoarginine (10 mmol/l), an inhibitor of the liver and bone isoenzymes, inhibited the seminal enzyme (53%), whereas L-phenylalanine (12 mmol/l), a strong inhibitor of placental alkaline phosphate, decreased activity by about 10%. Electrophoresis of semen samples on agarose revealed a broad band which was not sharpened after treatment with neuraminidase. Semen total alkaline phosphatase was essentially totally inactivated by heating at 56 degrees C for 15 min or 10 min at 65 degrees C; similar behaviour has been reported for the liver and bone isoenzymes. Electrophoresis after heating did not reveal a residual band of heat-stable placental-like alkaline phosphatase. Semen alkaline phosphatase appears to contain more than one isoenzyme, but placental-like alkaline phosphatase cannot be more than a minor component.

Alkaline Phosphatase↗

Prevalence of HIV antibodies in transsexual and female prostitutes.

Human immunodeficiency virus (HIV) prevalence was studied in an unselected group of 216 female and transsexual prostitutes. Subjects were asked about age, biological sex, marital status, children, length of occupation, sexual practices, and drug abuse history. Blood was drawn on site. All 128 females who did not admit to drug abuse were seronegative; 2 of the 52 females (3.8%) who admitted to intravenous drug abuse were seropositive. In contrast, 11.1% of the 36 male transsexuals (including 3 out of 32 non-drug abusers) were seropositive. The results support the notion that vaginal transmission of HIV is less effective than anal transmission.

Adolescent↗

Career plans for male-dominated occupations among female seniors in religious and secular high schools.

This study investigated the hypothesis that the religious beliefs of young women significantly affect their career planning. All female seniors (N = 315) in one public religious and two public secular high schools in Israel responded to inventories examining their (1) orientation to homemaking or career, (2) interest in male-dominated occupations, and (3) preference for male-dominated occupations. Results from regression analyses indicated that young women from the secular schools, as compared to those from the religious school, expressed greater interest in and preference for male-dominated occupations. In addition, religious orientation, more than other background variables, had predictive power for interest in and preference for male-dominated occupations.

Adolescent↗

Influence of various substrates on the acetylcarnitine:carnitine ratio in motile and immotile human spermatozoa.

Human spermatozoa were incubated in albumin-containing Hepes-buffered modified Ringer's solution, in the presence or absence of externally supplied substrates. The acylated forms of carnitine were identified by bioautography. Incubation of the cells with propionate or n-valerate resulted in increased content of propionylcarnitine, but n-butyrate, isobutyrate, n-valerate, isovalerate, hexanoate or heptanoate did not result in the appearance of acylcarnitine of chain length C4-C7. The addition of methionine, valine or isoleucine (whose catabolic pathways should produce propionyl-CoA) to the incubation medium did not increase propionylcarnitine. In all cases acetylcarnitine was the major acylcarnitine in human spermatozoa. The ratio of acetylcarnitine:carnitine remained relatively constant in spermatozoa incubated without external substrate for up to 4 h. No significant change in the ratio was observed when glucose, fructose or citrate were present in the incubation medium. Sorbitol decreased the ratio slightly and aspartic acid slightly increased it. A more pronounced increase in the ratio was caused by lactate or pyruvate. This increase was observed in motile spermatozoa but not in samples from asthenospermic men, indicating that metabolic utilization of pyruvate and lactate may differ in motile and immotile cells.

Acetylcarnitine↗

Carnitine and acetylcarnitine in motile and immotile human spermatozoa.

Human sperm were found to contain acetylcarnitine, carnitine, and only traces of propionylcarnitine and four-carbon acylcarnitines. Carnitine and acetylcarnitine were present in higher concentrations in sperm than in the corresponding seminal fluid samples, and the degree of acylation of carnitine was greater in sperm than in seminal plasma. The ratio of acetylcarnitine/carnitine was 1.77 +/- 0.69 in extracts of sperm from samples with a low degree of motility (0-10% motile), whereas it was 4.70 +/- 1.58 in samples which were 40-80% motile. The possible significance of this difference with regard to the degree of acylation of coenzyme A is discussed.

Acetylcarnitine↗

Carnitine and short-chain acylcarnitines in the lumen of the human male reproductive tract.

Bioautography of human semen demonstrated the presence of L-carnitine, acetylcarnitine, propionylcarnitine and C4-acylcarnitines (butyrylcarnitines). In studies designed to ascertain the organs secreting these compounds into semen it was found that: Quantitative analyses of semen obtained pre- and post-vasectomy showed markedly decreased L-carnitine concentrations, slightly decreased acetylcarnitine levels, and unchanged propionyl- and butyrylcarnitine concentrations as a result of vasectomy. The earlier portion of human split ejaculate samples characteristically contained higher concentrations of carnitine and its propionyl- and butyryl esters and a lower concentration of acetylcarnitine than did the later portion. Vas deferens luminal fluid contained carnitine and each of the short-chain acyl derivatives, as shown by bioautography. Prostatic fluid contained only small amounts of carnitine and its derivatives. Spermatozoa contained much more acetylcarnitine than free carnitine, and only trace amounts of the other acylcarnitines. It is concluded that carnitine in semen is secreted primarily by the epididymis, that a proportion of the acetylcarnitine derives from the seminal vesicles and epididymis, and that propionylcarnitine and butytylcarnitine enter semen, at least in part, together with the vas deferens contents.

Adult↗

A comparative study of carnitine and acylcarnitine concentration in semen and male reproductive tract fluids.

1. L-carnitine and its short-chain acyl derivatives were measured in semen of boar, man, rooster, bull, squirrel monkey and ram. 2. Carnitine was present in concentrations of 220, 476, 540, 1880, 3330 and 3820 muM respectively, and acetylcarnitine concentrations were 2, 191, 354, 709, 259 and 2750 muM. Propionyl and C4-acyl carnitines were found in small quantities. 3. Analysis of sequential "Split-ejaculate" samples of boar semen showed that carnitine concentration correlated with sperm count, suggesting an epididymal origin for this substance in this species. 4. Bioautographic analysis of samples obtained from ram and rat indicated that carnitine and acetylcarnitine were present in epididymal fluid and in fluids from accessory glands of the male reproductive tract.

Animals↗

Glutathione peroxidase activity in various types of blood cells in multiple sclerosis.

Previous studies demonstrated a significantly lower mean activity of glutathione peroxidase (GSH-Px) in erythrocytes of patients with multiple sclerosis than in control groups of normal subjects or patients with various neurological disorders. The present investigation has demonstrated that, in contradistinction to erythrocytes, a normal activity of GSH-Px is found in lymphocytes, granulocytes and platelets of multiple sclerosis patients. These results were obtained both with hydrogen peroxide, which serves as a specific substrate for selenium dependent GSH-Px, and t-butyl hydroperoxide which reacts both with selenium dependent and independent GSH-Px.

Blood Cells↗