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Hypo-osmotic Swelling Test (hypo-osmotic + swelling_test)
Selected AbstractsCell Subpopulation-related Volumetric Parameters: a Complementary Tool of the Modified Hypo-osmotic Swelling Test on Model of Boar SpermatozoaREPRODUCTION IN DOMESTIC ANIMALS, Issue 5 2000A. Petrounkina Content It is a general property of the intact animal cell to swell rapidly in response to hypo-osmotic conditions. The modified hypo-osmotic swelling test (HOS-test) is an indicative test to evaluate the integrity of the plasma membrane by means of an electronic cell counter, based on the relative increase of the cell volume in response to hypo-osmotic conditions. In this study the relationships between the osmotically induced changes of the cell volume of boar spermatozoa as determined by cell counter and the integrity of the membrane as determined by propidium iodide staining (PI) were studied. Boar sperm cell volume distributions were measured under iso-osmotic (300 mosmolar) conditions and after a hypo-osmotic stress (150 mosmolar). The relative volume shift of mean and modal volume were calculated as a proportion coefficient of modal and mean values of the cell volume distributions by transition from iso-osmotic to hypo-osmotic conditions. The volumetric parameters related to the different cell subpopulations were derived from the different peaks of cell volume distributions. PI-staining techniques were used for comparison. The values of the volume shift and of derived percentages of the osmotically inactive cells were correlated negatively and positively, respectively (p < 0.05) with the percentage of the PI-stained cells. This correlation indicates that a relationship exists between membrane functions of the different cell compartments (sperm head and tail) due to the circumstance that the increase of the cell volume in the HOS-test is associated with the morphological changes in the tail and the PI-staining is associated with the membrane integrity and permeability of the head region. The advantage of computer-assisted volume measurement is that a large number of cells (5000,50 000 spermatozoa) can be measured and evaluated during one procedure and in a very short time. The relative volume shift is a quantitative continuous parameter characterizing the osmotic reactivity and membrane functional competence of a cell population and of subpopulations within one ejaculate. This parameter could be useful to evaluate membrane functional competence rapidly and sensitively. Inhalt Es ist eine generelle Eigenschaft membranintakter tierischer Zellen, mit einer Volumenzunahme auf eine hypoosmotische Belastung zu reagieren. Der auf der relativen Vergrößierungdes Zellvolumens basierende modifizierte hypoosmotischeSchwelltest ist ein indikativer Test zur Beurteilung der Membranintegrität mittels eines elektronischen Partikelzählers. In dieser Studie wurden die Zusammenhänge zwischen der mittels der Propidiumjodid-Färbung bestimmten Zellmembranintegrität und den osmotisch induzierten Veränderungen des Zellvolumens von Eberspermien untersucht. Volumenverteilungen von Eberspermien wurden unter isoosmotischen (300 mosmolar) und hypoosmotischen (150 mosmolar) Bedingungen gemessen. Die relative Volumenverschiebung der modalen und mittleren Werte der Volumenverteilung wurde als Quotient aus Modalwerten der Zellvolumenverteilungen und des mittleren Zellvolumens beim Übergang von isotonen zu hypotonen Bedingungen berechnet. Die auf verschiedene Subpopulationen bezogenen volumetrischen Parameter werden aus den originalen Volumenverteilungen berechnet. Der Betrag der Zellvolumenzunahme und die aus den Volumenverteilungen bestimmten Anteile an Zellen mit beschädigter Geißielmembran korrelierten signifikant negativ bzw. positiv (p < 0,05) mit dem Anteil an den Zellen mit beschädigter Kopfmembran, der sich aus der Propidiumjodid-Färbung ergab. Es wird geschlossen, daßi im Verhalten zwischen den Membranen der verschiedenen Zellkompartimente (Spermienkopf und-Geißiel) ein Zusammenhang besteht. Die beschriebene Methode ermöglicht die Analyse großier Zellpopulationen (5.000,50.000 Zellen). Die relative Volumenverschiebung stellt einen quantitativen kontinuierlichen Parameter dar, der den Membranzustand der Eberspermien einer Spermatozoenpopulation und Subpopulationen innerhalb eines Ejakulates charakterisiert. Diese Parameter können zur schnellen und sensitiven Beurteilung der Membranzustandes eingesetzt werden. [source] Hypo-osmotic swelling test and unexplained repeat early pregnancy lossJOURNAL OF OBSTETRICS AND GYNAECOLOGY RESEARCH (ELECTRONIC), Issue 1 2010Sudhindra M. Bhattacharya Abstract Aim:, To study the relationship of various sperm characteristics and hypo-osmotic swelling test (HOS test) with repeat unexplained early pregnancy loss. Methods:, Semen samples from husbands of 74 couples with a history of repeat early pregnancy loss (group A) were analyzed according to World Health Organization criteria, and a HOS test was performed in each case. Semen samples from 65 husbands with proven fertility (group B) were also studied for comparison. Results:, No statistically significant differences were noted in the age of the husbands, sperm concentration, sperm morphology and percent motile sperm between groups A and B. The mean HOS test scores of the two groups were significantly different (group A: 60.4%; group B: 76.9%; P = 0.01 [normal value: >60%]). In group A, 33.8% of cases (25/74) and in group B, 12.3% of cases (8/65) showed low HOS test scores. Conclusion:, The sperm HOS test may be helpful to screen for any paternal factor associated with repeat embryonic or early fetal loss and in a resource-poor setting, and may be utilized in any clinical laboratory. [source] Effect of N-acetyl-L-cysteine Supplementation in Semen Extenders on Semen Quality and Reactive Oxygen Species of Chilled Canine SpermatozoaREPRODUCTION IN DOMESTIC ANIMALS, Issue 2 2010AJ Michael Contents The objective of this study was to evaluate the quality of chilled dog semen processed with extenders containing various concentrations of N-acetyl-L-cysteine (NAC). Ejaculates from five dogs were collected, pooled and evaluated for concentration, motility, rapid steady forward movement (RSF-movement), viability, acrosomal integrity and by the hypo-osmotic swelling test (HOST). In addition, superoxide anion (O2 -,) production, hydroxyl radicals (OH,) and total reactive oxygen species (tROS) were determined. The pool was divided into five aliquots, which were diluted to a final concentration of 66.66 × 106 spermatozoa/ml with Tris-glucose-egg yolk extender containing one of the following concentrations of NAC (0, 0.5, 1, 2.5 or 5 mm). The semen aliquots were chilled and preserved at 4°C. Semen quality was evaluated after rewarming at 72 h. Sperm motility was significantly higher with the 0.5 mm concentration compared with the control group (p = 0.001). Rapid steady forward movement was higher with the 0.5 and 1 mm concentrations compared with the control and 5 mm group (p < 0.001). Viability and HOST percentages were not significantly altered. Compared with the control, the 5 mm concentration showed significantly reduced percentages of spermatozoa with normal acrosomes (p = 0.049). None of the ROS values at 72 h were significantly affected by the presence of NAC in semen extenders, although all NAC concentrations showed lower O2 -, and OH, values compared with the control. Only the concentrations of 1 and 5 mm inhibited the significant increase of tROS values after 72 h, compared with the fresh semen value. In conclusion, NAC supplementation of semen extenders is beneficial to semen motility of canine spermatozoa during chilling with the 0.5 mm concentration being the most effective, although no significant ROS inhibition was observed at 72 h. [source] Influence of sexual stimulation on sperm parameters in semen samples collected via masturbation from normozoospermic men or cryptozoospermic men participating in an assisted reproduction programmeANDROLOGIA, Issue 3 2000Y. Yamamoto Summary. To evaluate the influence of sexual stimulation via sexually stimulating videotaped visual images (VIM) on sperm function, two semen samples were collected from each of 19 normozoospermic men via masturbation with VIM. Two additional samples were collected from each man via masturbation without VIM. The volume of seminal plasma, total sperm count, sperm motility, percentage of morphologically normal spermatozoa, outcome of hypo-osmotic swelling test and zona-free hamster oocyte sperm penetration assay, and markers of the secretory function of prostate were significantly larger in semen samples collected via masturbation with VIM than masturbation without VIM. The improved sperm parameters in the samples collected via masturbation with VIM may reflect an enhanced prostatic secretory function and increased loading of the vas deferens at that time. In a similar protocol, two semen samples were collected via masturbation with VIM from each of 22 non-obstructed azoospermic men. Semen samples from these men had been occasionally positive in the past for a very small number of spermatozoa (cryptozoospermic men). Two additional samples were collected from each cryptozoospermic man via masturbation without VIM. The volume of seminal plasma, total sperm count, sperm motility, and a marker of the secretory function of prostate were significantly larger in semen samples collected via masturbation with VIM. Fourteen out of the 22 men were negative for spermatozoa in both samples collected via masturbation without VIM. These men demonstrated spermatozoa in both samples collected via masturbation with VIM. Six men with immotile spermatozoa in both samples collected via masturbation without VIM exposed motile spermatozoa in both samples collected via masturbation with VIM. High sexual stimulation during masturbation with VIM results in recovery of spermatozoa of greater fertilizing potential both in normozoospermic and cryptozoospermic men. The appearance of spermatozoa after masturbation with VIM in the vast majority of cryptozoospermic men is of clinical significance in programmes applying intracytoplasmic sperm injections for the management of severe male infertility and obviates the need for testicular biopsy. [source] | ||||||||||