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Biomedical subjects

A Makler

Publications and source records attributed to A Makler.

At least 37 records · Page 2Linked to original sources

Endogenous digoxin-like immunoreactivity measured in seminal fluid from a normal male population.

Endogenous digoxin-like immunoreactivity (EDLI) has been detected in different biological fluids and in several pathophysiological conditions. In this study, using radioimmunoassay we reported for the first time the existence of bound and unbound EDLI in normal seminal fluid. The unusual finding was the detection of unbound EDLI in the seminal fluid, while this reactivity was undetected in plasma. Two main hypotheses are presented: (1) local secretion of unbound EDLI and/or (2) passive diffusion from plasma to the seminal fluid of unbound EDLI and subsequent local concentration.

Adult↗

Sealed mini-chamber of variable depth for direct observation and extended evaluation of sperm motility under the influence of various gases.

A new chamber for microscopical observation of living cells, e.g. spermatozoa, under hermetically sealed conditions and over an extended period is described. Motile spermatozoa were serially observed for several hours and the effect of various gases on sperm motility has been studied. The chamber could also be used to study the effect of various toxic gases in the fields of microbiology and toxicology.

Atmosphere Exposure Chambers↗

Induction of spermatogenesis and fertility in hypogonadotropic azoospermic men by intravenous pulsatile gonadotropin-releasing hormone (GnRH).

Gonadotropin-releasing hormone (GnRH) has only recently become a helpful tool in the medication of hypogonadotropic hypogonadism (HH). Two azoospermic patients with HH who had previously been treated with hCG/hMG because of delayed puberty and each of whom had fathered a child after previous gonadotropin therapy were referred due to secondary failure of hCG/hMG treatment to induce spermatogenesis and fertility. A pulse study where blood was drawn every 15 minutes for LH, FSH and PRL RIAs was performed in each patient, and afterwards a bolus of i.v. GnRH was injected to assess gonadotropin responsiveness. A portable GnRH pump was connected to each patient so that it administered 5-20 micrograms of GnRH i.v. every 89 minutes. Spermatogenesis was first detected after 42 and 78 days respectively in the 2 treated HH men and 4 1/2 months from the start of treatment their wives became pregnant. No thrombophlebitis or other complications of the i.v. therapy occurred. In the case of the first patient, the semen was washed and concentrated and intra-uterine inseminations were carried out in an attempt to shorten the time needed to achieve fertility. The first pregnancy was successfully terminated at 38 weeks with the delivery of 2 heterozygotic normal male babies. The second pregnancy ended in spontaneous delivery of a healthy female. We conclude that i.v. pulsatile, intermittent GnRH administration is a safe, efficient and highly successful means of treating azoospermic men with HH.

Adolescent↗

Semen evaluation following preparation for in vitro fertilization of human oocytes.

Semen preparation is an important step of in vitro fertilization (IVF) and can affect the success of this procedure. Prior to oocyte insemination, spermatozoa are washed to remove seminal plasma which is believed to contain decapacitation factors. This study was undertaken to evaluate the effect of preparation on semen quality and subsequent successful IVF. Oocytes were recovered from 12 hMG/hCG-stimulated women by laparoscopy, and 6 h later semen specimens were obtained from the male partners. After liquefaction, 1 ml of semen was centrifuged twice in Ham's F10 medium supplemented with 10% of homologous serum, and the final suspension was used to inseminate the preovulatory eggs. In the initial and washed-sperm suspensions, motility was evaluated by the MEP method, and the occurrence of acrosome reaction and sperm viability were evaluated by the triple-stain technique. Fertilization was documented by the formation of two pronuclei. Washing caused a significant decrease in the percentage of motile sperm from 68% to 59% but significantly increased mean sperm velocity from 26 to 29 micron/sec (p less than 0.01). The mean fertilization rate was 65%, and no correlation was found with any of the parameters of semen quality before or after washing. Semen preparation for IVF is associated with a decrease in the percentage of motile sperm that does not seem to affect the fertilizing ability of normal spermatozoa but may be of importance in patients with abnormal semen.

Cell Survival↗

An improved sperm immobilization test based on multiple exposure photography.

An improved method for objective evaluation of results in the complement dependent sperm immobilization test, with the aid of the multiple exposure photography (MEP) method is described. With this method it is possible to determine the percentage of residual motility of spermatozoa quantitatively after incubation with sera suspected of having immobilizing activity in the presence of complement. Long term storage of information, reexamination of data and comparison of results after various treatment are routinely possible with this method.

Antibodies↗

Clinical, endocrine and ultrastructural study of XY gonadal dysgenesis. A case report.

About 120 cases of XY gonadal dysgenesis have been reported on. We treated such a patient with bilateral gonadectomy. The gonadal tissue's capacity to respond to hormonal trophic stimulation was assessed. When the gonads were examined ultrastructurally, structures with the morphologic characteristics of stromal ovarian cells, Sertoli's cells and Leydig's cells were found. Because of the potential malignancy of the XY gonads, bilateral gonadectomy and hormonal substitution therapy are recommended for these patients. We prefer to use combined hormone replacement with sequential estrogen and progesterone rather than sequential unopposed estrogen because of the small but increased risk of endometrial hyperplasia and carcinoma after long-standing sequential therapy.

Adolescent↗

A new method for rapid determination of sperm concentration in bull and ram semen.

A simple method for rapid evaluation of bull and ram sperm concentration is described. In this technique, a sample from an undiluted specimen was placed in a special 10-mum counting chamber and examined either by an ordinary or a phase-contrast microscope. The pattern distribution of the observed spermatozoa was matched as closely as possible with one of five pictures of a standard scale. This scale was prepared from serial photomicrographs that ranged from 100 to 1500 million per ml. The number that appeared at the corresponding photograph immediately indicated the sperm concentration of the tested specimen. In this way values up to 1500 million per ml could be determined rapidly with no further procedures. More concentrated specimens needed slight dilution to make them suitable for direct evaluation. Accuracy and reliability were statistically evaluated; the mean deviation from the conventional method was less than 15.2% with confidence level of 95%.

Journal Article↗

Factors affecting sperm motility. IX. survival of spermatozoa in various biological media and under different gaseous compositions.

The role of various environmental conditions on sperm motility and their survival was investigated by incubating washed human spermatozoa in various biologic or artificial media and under different atmospheric compositions. When sperm were analyzed by the multiple exposure photography technique for objective motility determination, it was found that none of the investigated biologic media induced any immediate stimulatory effect on the tested spermatozoa. Exchange of seminal fluid between normal and oligoasthenospermic specimens did not reveal any beneficial or harmful effect on sperm motility. Sperm did not survive for more than 24 hours when kept in their original seminal fluid under any circumstances. The most favorable media tested for prolonged sperm survival were pure human umbilical cord serum or Ham's solution containing 0.5% human albumin. However, even in these media sperm did not survive much beyond 24 hours unless incubated in an atmosphere containing 5% CO2 or a triple mixture of 90% N2, 5% O2, and 5% CO2. Only under these circumstances could sperm stay alive, in some cases up to 5 days.

Carbon Dioxide↗

Improved techniques for collecting motile spermatozoa from human semen. I. A self-migratory method.

A simple apparatus to collect moving sperm by non-traumatic means which can be used for artificial insemination is described. The technique is based on enhancing the process of migration from the seminal fluid into a top-layered artificial medium in an ordinary test tube. This has been achieved by controlling 3 main variables: 1) The dilution of migrated sperm was minimize by using only 0.5 ml of the medium layered onto 1 ml semen; 2) increasing the surface area between these media by turning the test tube from a vertical to almost a horizontal position; 3) stimulating sperm activity by incubation at 37 degrees C under air: 5% CO2 for 30 min. When restored to a vertical position approximately 0.3 ml medium, sufficient for most AIH or IVF procedures, was gently aspirated. The effects of these variables on the rate of sperm migration was tested one at a time, and increments that ranged from 20% to as much as 10-fold were detected. When these 3 variables were optimized and 58 semen specimens analyzed, it was found that motility increased from 42 to 87%, velocity from 24.5 to 27.3 micron per sec, whilst abnormal forms dropped from 37 to 15%. The final concentration of motile sperm was 23 X 10(6)/ml compared to an original mean concentration of 34 X 10(6)/ml, indicating a relative recovery of 68%. Oligoasthenospermic specimens revealed similar changes in sperm motility, velocity, morphology and recovery. However, due to the low initial content of moving sperm (4.8 X 10(6)/ml), their final concentration was also low (2.7 X 10(6)/ml). Such specimens required additional preliminary preparation to increase the sperm concentration prior to the migratory procedure.

Cell Separation↗

Improved techniques for separating motile spermatozoa from human semen. II. An atraumatic centrifugation method.

A simple and atraumatic method for concentrating washed, motile spermatozoa from normal and subnormal semen specimens is described. It incorporates a modified technique of centrifugation in which sperm are spun onto a soft, fluid cushion, thereby minimizing mechanical damage. Following initial semen dilution to 5-10 ml in an artificial medium, the mixture is transferred to a test tube and layered onto 1 ml of oily contrast medium (Lipiodol). After centrifugation at 300 g for 10 min all but 0.5 ml of the supernatant is discarded, and the unshaken test tube is incubated at 37 degrees C for 15-20 min, during which time the motile sperm migrate into the upper 0.5 ml. After this incubation, 0.3 ml of the upper layer is removed which is sufficient for most IVF and AIH purposes. It contains concentrated, washed, motile spermatozoa that are free of debris and most abnormal forms. No change in pH or osmolarity and no diffusion of any iodine from the oil base into the top layer were detected.

Cell Separation↗

Rapid microcomputer-based analysis of semen characteristics from photographs taken by the MEP method.

Use of a microcomputer with the multiple exposure photography (MEP) method for rapid analysis of semen characteristics is described. The computer is programmed to calculate information fed from a digitizing tablet while scanning projected images of photographed sperm. In this way sperm concentration, percentage of motility, average velocity and frequency distribution of sperm velocity, as well as percentage of abnormal forms are immediately obtained. The method was found to be as accurate, less prone to error, and significantly less time consuming than the previous manual measuring method. It can greatly facilitate routine semen analysis in laboratories using the MEP method.

Computers↗

Factors affecting sperm motility. V. Washing and resuspension of human spermatozoa in various artificial media.

Fresh semen specimens from fertile donors were subjected to one-step and two-step washings in six various commonly used artificial media. It was found that washing procedures per se, in most cases, had an immediate and extended harmful effect on sperm motility which was much more prominent after the second washing. However, human albumin, when added, could usually protect spermatozoa from this deleterious effect. None of the tested media showed any stimulatory or "revitalization" effect, and the increase in sperm velocity after one washing in some of these media was attributed to a simple decrease in viscosity of the original seminal fluid. The possible mechanism of the deleterious effect of sperm washings as related to the property of the medium and some implications for practical clinical and research studies are discussed.

Humans↗

Factors affecting sperm motility. VI. Sperm viability under the influence of bacterial growth in human ejaculates.

The influence of bacterial growth on human sperm motility and viability was evaluated objectively with the multiple-exposure photography method. Experimental semen specimens, obtained from normal donors bh nonaseptic means of masturbation, were incubated with antibiotics at room temperature or body temperature for 24 hours. Although bacteria, grew in control specimens, were totally eradicated in all antibiotic-treated specimens, no significant difference was found between these groups with regard to sperm motility throughout the time of incubation. Sperm survival was not inhibited, nor was it extended as a result of suppression of bacterial growth. In both groups, survival time was much shorter in specimens incubated at body temperature than in those kept at room temperature. Sperm motility was not affected after 2 hours of incubation of fresh specimens with concentrations of various pathogenic bacteria similar to those found in severe prostatitis. The question of whether the use of antibiotics in the treatment of asthenospermia per se has a prognostic value is discussed.

Adult↗

Factors affecting sperm motility. VII. Sperm viability as affected by change of pH and osmolarity of semen and urine specimens.

The effects of pH and osmolarity of semen and urine specimens on motility and velocity of human spermatozoa were studied objectively with the aid of the multiple exposure photography (MEP) method. The pH of fresh ejaculates ranged from 7.2 to 8.2 and specimens were slightly hyperosmotic ranging between 300 to 380 mOsm/kg. Gradually changing the pH and osmolarity to either side of normal values led to progressive loss of sperm motility. However, sperm velocity was slightly increased by mild alkalinization and hyperosmolarity. Spermatozoa that became immobilized by acidification regained their motility shortly after pH was restored to normal values. In the majority of instances spermatozoa lost their motility when mixed with fresh urine specimens. Neutralization of urinary pH could not protect them from this effect unless urine osmolarity was also isotonically adjusted. It is suggested that patients with retrograde ejaculation should adequately increase their fluid intake before recovery of sperm from their bladder for artificial insemination.

Humans↗

Factors affecting sperm motility. VIII. Velocity and survival of human spermatozoa as related to temperatures above zero.

The effect of temperature on motility and survival of ejaculated human spermatozoa was investigated with the aid of the multiple exposure photography (MEP) method for objective sperm motility determination. Fresh specimens from healthy donors were analyzed while being heated or cooled gradually, or during their storage at various temperatures from 0 to 48 degrees C. Sperm velocity increased steadily from zero to 50.4 nm/sec between freezing point and body temperature. Thereafter, their activity dropped dramatically and total immobilization occurred at 45 degrees C. The induced immobilization was reversible providing exposure to those extreme temperatures was short enough to prevent permanent damage. Sperm survived up to 24-48 h when stored at 23 degrees C, while at body temperature, their survival in vitro was much shorter and rarely extended beyond 12 h. Their longevity was still shorter at higher or lower temperatures, especially when approaching 48 degrees C. With the aid of the combined supravital staining and MEP methods it was found that temperatures of extreme levels induced mainly immobilization rather than a spermicidic effect. The possible mechanism of thermal effect on sperm motility and some of its practical implications are discussed.

Cell Survival↗