Rosiglitazone plus metformin is effective and well tolerated in clinical practice: results from large observational studies in people with type 2 diabetes.

This study investigated the efficacy during daily practice of rosiglitazone (RSG) added to metformin (MET) in poorly controlled type 2 diabetes mellitus. Two post-marketing observational studies were conducted in Germany over 6 months. RSG (4 mg/day titrated to 8 mg/day as required) was added to existing MET in 11,014 subjects. Subjects were maintained on diet and exercise. Addition of RSG to MET significantly reduced median HbA 1c by 1.3% (8.1 vs. 6.8%; p < 0.0001) and median fasting blood glucose (FBG) by 47.0 mg/dl (171.0 vs. 124.0 mg/dl; p < 0.0001) after 6 months. The proportion of subjects achieving HbA(1c) targets of < or = 6.5 and < or = 7.0% increased from 3.5 to 38.8% and from 13.5 to 63.7%, respectively. Mean systolic and diastolic blood pressure decreased by 7 and 3 mmHg, respectively (p < 0.0001). Mean weight decreased by 1.7 kg and was constant or reduced in most (74.1%) subjects. Addition of RSG to MET significantly reduces median HbA 1c and FBG in clinical practice and is generally well tolerated.

Scintigraphic monitoring of catheter-port systems in type I diabetics with continuous insulin therapy.

OBJECTIVE: We developed a non-invasive scintigraphic procedure to control the function of percutaneous catheter-port systems for continuous insulin therapy. The aim of the study was to evaluate the scintigraphic results and to compare the absorption kinetics of intraperitoneal and umbilical catheter-ports. MATERIAL AND METHODS: Seven patients with intraperitoneal and nine patients with umbilical catheter-port systems implanted into the partially redilated umbilical vein were investigated. All these patients had normal functioning catheter-ports. Additionally, three patients with intraperitoneal and three patients with umbilical catheter-port dysfunction confirmed either by radiography or laparoscopy were studied. After the injection of (99 m)TcO (4)(-) into the port a region-of-interest was drawn around the activity at the tip of the catheter and the half-life of tracer absorption (T(1/2)) calculated. RESULTS: The normal intraperitoneal catheter-port group showed a T(1/2) value of 6.7 +/- 3.2 min and the normal umbilical catheter-port group a T(1/2) of 6.6 +/- 2.0 min. There was no significant difference in T(1/2) between intraperitoneal and umbilicial systems. The dysfunctional catheter-port group (T(1/2) 19.3 +/- 6.7 min) differed ( p = 0.0005) from the normal catheter-port group (T(1/2) 6.7 +/- 3.2 min). On the basis of the normal group an upper threshold value of T(1/2) was calculated to be 11.8 min. CONCLUSIONS: Imaging with (99 m)TcO (4)(-) is an accurate, non-invasive, and quick method to assess the function of insulin catheter-ports. A T(1/2) value > 11.8 min is indicative of a catheter dysfunction. Umbilical and intraperitoneal catheter-ports show similar absorption rates of (99 m)TcO (4)(-).

Application of techniques for sperm selection in fresh and frozen-thawed stallion semen.

The objective of this research was to improve the techniques in processing chilled and frozen-thawed horse semen. In a preliminary experiment (Exp. I), different techniques for sperm selection and preparation [Swim-up, Glass wool (GW) filtration, Glass wool Sephadex (GWS) filtration; Percoll] were tested for their suitability for equine spermatozoa and results were compared with the routine procedure by dilution (Exp. I). In the main experiment (Exp. II), two sperm preparation techniques (GWS, Leucosorb) refering to the results of Exp. I and a previous study of our group (Pferdcheilkunde 1996 12, 773) were selected for processing complete ejaculates either for cooled-storage or cryopreservation. In a third experiment (Exp. III), pregnancy rates from inseminations with semen processed according to the techniques tested in Exp. II were compared with those obtained with semen processed according to routine procedures. In Exp. I (six stallions, six ejaculates/stallion), between 48 and 92% of spermatozoa were lost following the different sperm selection procedures (p < 0.05). Preparation of sperm increased percentage of progressively motile spermatozoa (pms) [Swim-up, GW, GWS vs dilution, Percoll (p < 0.05)] and decreased percentage of sperm head abnormalities [Swim-up, GW, GWS vs dilution, Percoll (p < 0.05)] probably by not improving the quality of individual cells, but by elimination of spermatozoa of inferior quality. In Exp. II (eight stallions, three ejaculates/stallion) Leucosorb and GWS procedures allowed the filtration of large volumes (extended ejaculates) for routine laboratory practice. GWS and Leucosorb filtration resulted in increased motility, membrane integrity and sperm viability after storage of spermatozoa until 48 h at +5 degrees C when compared with control (diluted) and centrifuged semen (p < 0.05). Significantly more spermatozoa were recovered after centrifugation (87.8 +/- 15.4%) compared with GWS (63.5 +/- 18.6%) and Leucosorb filtration (53.6 +/- 22.3%). GWS or Leucosorb procedure resulted in successful cryopreservation of stallion semen without centrifugation for removal of seminal plasma. The per cycle conception rate of inseminated mares using 200 x 106 pms transferred within 8 h after collection of semen was not affected by GWS filtration or Leucosorb separation when compared with centrifugation (n.s.; Exp. III). In conclusion, GWS and Leucosorb filtration results in the improvement of semen quality and should be considered as a method for stallion semen processing. Additional studies are needed for the evaluation of potentially higher fertilizing ability of stallion spermatozoa separated by techniques for sperm selection.

Sperm-cell volumetric measurements as parameters in bull semen function evaluation: correlation with nonreturn rate.

Sperm-cell volume, measured electronically by cell counter, is a parameter providing information about the state and integrity of the plasma membrane by determining cell osmotic reactivity (swelling level). Electronic volume measurement is a modification of the hypo-osmotic swelling test, based on the increase in sperm volume in response to hypo-osmotic stress. In this study the volumetric method was applied to bull ejaculates, and the relationships of volumetric parameters, osmolality of seminal plasma, and concentration of sodium and potassium ions in seminal plasma, with the nonreturn rate (NRR) were examined. Significant correlations were found between volumetric parameters, conventional spermatological parameters, and NRR. The relative volume shift of the mean volume correlated significantly with motility before and after thawing (P < 0.05). NRR correlated significantly with iso-osmotic cell volume (- 0.49; P < 0.05) and with the relative volume shift (0.51, P < 0.05). The prediction level of regression models was improved when volumetric parameters (iso-osmotic cell volume) were included in the multiple regression model. Therefore, using electronic volume measurement as a component for fast, correct and valid (up to 50,000 cells), recording sperm-cell population may help to evaluate ejaculate quality more precisely.

Chlamydia trachomatis in andrologic patients--direct and indirect detection.

BACKGROUND: Chlamydia trachomatis is considered to be the most common sexually transmitted disease in Germany. It is currently unclear whether chlamydial infection causes pathological conditions of the male accessory glands with consequences for male infertility. PATIENTS AND METHODS: Within the framework of several prospective studies the association between sperm quality, male accessory gland function and infection with C. trachomatis was investigated in men of couples with unexplained infertility. Chlamydial infection was determined by serologic methods and by proof of Chlamydia-specific DNA. As a marker of infection the direct determination of granulocytes in the ejaculate or the measurement of the polymorphonuclear (PMN) elastase concentration was used. The male accessory gland function was evaluated using the markers fructose, citric acid and alpha-glucosidase in the seminal plasma. RESULTS: Chlamydia-specific DNA in the ejaculate was present in between 3-5% of the subjects, which corresponds to its prevalence in the normal population. Chlamydia IgA antibodies were demonstrated with a frequency of 38% in seminal plasma (n = 834) using a genus-specific test (rELISA). Using other species-specific tests (MIF, SeroCT, IgA pELISA and ImmunoComb), Chlamydia IgA antibodies were found at frequencies of between 8 and 22%. CONCLUSION: Only in a few individual cases was it possible to show a connection between reduced sperm quality, disturbed male accessory gland function and indication of infection with Chlamydia, bacteria or Ureaplasma.

An improved method of sperm selection by glass wool filtration.

An improved method of sperm selection by glass wool filtration is introduced. After incubation of glass wool filtrates for 30 min at 37 degrees C in a conical-shaped 1.5-ml tube, an enrichment of highly motile spermatozoa was found in the bottom layer of the tube. The effect turned out to be dependent on the conical shape of the tube, as it was not observed in flat-bottomed tubes. Native ejaculates (obtained from 30 men) and their glass wool filtrates were analysed by cell counter, computer-assisted sperm-motility analysis, morphological differentiation and supravital staining of spermatozoa. When 400 microl of ejaculate, diluted with 800 microl of medium, was applied to the top of a column consisting of a 1-ml disposable syringe barrel gently packed with 15 mg of glass wool to a depth of 6 mm, an enrichment of viable spermatozoa was found in the first three 100-microl fractions taken from the bottom of the tube. It is the simplicity of this technique that makes it so easily applicable.

Comparative analysis of organ-specific autoantibodies and celiac disease--associated antibodies in type 1 diabetic patients, their first-degree relatives, and healthy control subjects.

OBJECTIVE: In type 1 diabetes the coexistence with other endocrine diseases and organ-specific autoantibodies has been frequently reported leading to the concept of autoimmune polyendocrine syndrome (APS). In addition, an association of type 1 diabetes with celiac disease has been described. These disorders share a similar genetic background, and first-degree relatives of type 1 diabetic patients may also be affected significantly. Screening for specific antibodies allows early diagnosis of these disorders. RESEARCH DESIGN AND METHODS: In the present cross-sectional study, we analyzed sera from 197 recent-onset type 1 diabetic patients at the time of diagnosis, 882 first-degree relatives, and sera of 150 healthy control subjects for prevalence and co-occurence of the following antibodies (method): insulin autoantibodies (radioimmunoassay); GAD and IA-2 antibodies (radioligand assay); islet cell antibody, anti-adrenal cortex antibodies, and anti-gastric parietal cell antibodies (indirect immunofluorescence); anti-thyroglobulin and anti-thyroid peroxidase antibodies; and gliadin IgG/A and tissue-transglutaminase IgA (enzyme-linked immunosorbent assay). RESULTS: The overall frequency of gastric patietal cell antibodies and adrenal antibodies did not differ significantly among groups. In contrast, type 1 diabetes-associated antibodies and thyroid antibodies were significantly more frequent both in recent-onset type 1 diabetic patients and in the group of first-degree relatives (P < 0.05). The prevalence of gliadin IgG/IgA and transglutaminase IgA was significantly higher in the group of recent-onset type 1 diabetic patients (P < 0.05), but the difference between first-degree relatives and control subjects did not reach statistical significance. Focusing on the coexistence of antibodies, the group of recentonset type 1 diabetic patients presented with 27.4% of the subjects testing antibody-positive-specific for two or more of the envisaged disorders (i.e., type 1 diabetes, autoimmune thyroiditis, and celiac disease) compared with 3.1% in the group of first-degree relatives and 0 of 150 in the control population (P < 0.05). CONCLUSIONS: We conclude that, in an active case-finding strategy, recent-onset type 1 diabetic patients should be routinely screened at least for concomitant autoimmune thyroid disease and additionally for celiac disease. Screening in their first-degree relatives should include at a minimum the search for thyroid autoimmunity in addition to screening for pre-type 1 diabetes.

Cardiac transplantation in patients with insulin-treated diabetes mellitus.

BACKGROUND AND METHODS: As documented earlier the incidence of cardiac mortality in diabetic patients due to coronary artery disease is high. Cardiac transplantation for congestive heart failure due to coronary artery disease, cardiomyopathy, and valvular diseases is obviously a therapeutic option in patients suffering from insulin-treated diabetes mellitus. To shed more light on this problem we performed a retrospective analysis of 40 patients with insulin-treated diabetes mellitus (three type-1; 37 type-2: insulin-treated for at least three months before cardiac transplantation) referred to our transplant unit for cardiac transplantation between March 1989 and December 1996. RESULTS: Orthotopic cardiac transplantation was performed in 40 patients (4 women, 36 men) aged 32-73 years (mean 56 years) with an insulin-treated diabetes mellitus preexisting for 3-348 months (mean 65.1 months). Donor age ranged from 15 to 72 years (mean 35.5 years) matched for body weight and blood group. Overall mortality in this group was 40.0% with an early mortality of 12.5%. CONCLUSIONS: Our results show that type-1/2 insulin-treated diabetes mellitus preoperative to heart transplantation is not a contraindication in patients suffering from end-stage heart failure. Adequate therapy of diabetes mellitus as well as individual immunosuppressive therapy are important in order to minimize additional organ damage caused by the drugs themselves or resulting infectious complications.

Cyclical changes in sperm volume during in vitro incubation under capacitating conditions: a novel boar semen characteristic.

The osmotic reactivity of boar spermatozoa during incubation in vitro was studied using a hypo-osmotic swelling test in conjunction with electronic measurement of cell volume. Sperm populations showed fluctuations in both iso-osmotic cell volume and hypo-osmotic volume response that fitted mathematical models for periodicity. Significant differences of frequency and amplitude were observed during sperm incubation under capacitating conditions as compared with those under non-capacitating conditions. In addition, different boars showed specific differences in their fluctuation characteristics under capacitating conditions. During incubation under capacitating conditions, a decrease in osmotic reactivity was observed that correlated with a decrease in motility, while the absolute value of the earliest maximum of the osmotic-induced response correlated with an increase in the proportion of discharged acrosomes. The time course of the cyclical behaviour of osmotic reactivity may be a useful parameter for assessing boar sperm response to capacitating conditions.

Assessing equine sperm-membrane integrity.

The swelling of cells in a hypo-osmotic medium has been described as an important criterion for assessing the functional integrity of the sperm plasma membrane. The resistance of equine spermatozoa to osmolarity changes was studied by extending 98 semen samples collected from nine stallions in media at five osmolarities (300, 200, 150, 100, and 50 mOsmol l(-1)). The response of the cells was measured by the spermatocrit technique and eosin staining. Spermatocrit determines the increase on spermatozoal volume under hypo-osmotic conditions, a sign of functional integrity of sperm plasma membrane, whereas the eosin staining evaluates the viability of spermatozoa. A significant positive correlation (P<0.01) was observed between spermatocrit values and percentage of eosin-unstained cells. Spermatocrit measurements and eosin staining proved to be useful methods to evaluate the integrity of sperm plasma membrane under hypo-osmotic conditions and could be used as an additional criterion to predict semen preservation ability.

The combination of antibodies to GAD-65 and IA-2ic can replace the islet-cell antibody assay to identify subjects at risk of type 1 diabetes mellitus.

First-degree relatives of type 1 diabetic patients are at increased risk of developing diabetes and, until recently, islet cell antibodies (ICA) have represented the major risk marker used for identification of individuals at increased risk for subsequent progression to diabetes. In order to determine the value of antibodies to GAD-65 and IA-2ic to identify individuals at high risk for type 1 diabetes mellitus, we measured both autoantibodies and ICA in 1436 first-degree relatives of patients with type 1 diabetes. In addition, the sera were analyzed for thyroid, adrenal and gastric-parietal cell autoantibodies as markers for possible polyendocrine involvement. GAD-65 Abs were found in 135 out of 1436 (9.4%) first-degree relatives and in 57 of 98 (58.2%) ICA-positive subjects. IA-2ic were detected in 52 of 1436 (3.6%) first-degree relatives and in 44 of 98 (44.8%) ICA-positive relatives. IA-2ic and/or GAD-65 were detected in 73 of 98 (74.5%) ICA-positive relatives. Interestingly, antibodies to GAD-65 and/or IA-2ic were present in 91.2% of individuals with more than 20JDF-units. Anti-IA-2ic and GAD-65 were positively correlated with high levels of ICA. Anti-IA-2ic and GAD-65 were found in 19% and 48.5% of subjects with ICA levels of 5-20JDF-u but in 68.8% and 76.5% of individuals with ICA of 40JDF-u or more, respectively (p < 0.001), compared to subjects with ICA levels less than 5 JDF-u. When autoantibody frequencies among the relatives were analyzed according to relationship to the proband, the offspring and siblings had a higher frequency of ICA and IA-2ic (p<0.05) than the subgroup of parents. A significant association was observed between IA-2ic and thyroid antibodies. In addition, higher levels of IA-2ic were found in relatives with positive TPO antibodies (p < 0.001); this correlation was particularly strong in offspring and siblings (p < 0.01). Determination of GAD-65 and IA-2ic antibodies may be considered as an alternative to primary ICA-screening, enabling the screening of large populations.

The Giessen-Bad Oeynhausen family study: improved prediction of type I diabetes in a low incidence population of relatives using combinations of islet autoantibodies in a dual step model.

To determine the value of a combined antibody screening for prediction of type I diabetes in a low incidence cohort, we prospectively studied 882 first-degree relatives (485 parents, 382 siblings and 15 offsprings) for up to 11 years who were not preselected for islet cell antibody (ICA) status. During the observation period, 16 individuals developed diabetes. The first serum sample obtained at study entry was analyzed for ICA and antibodies to insulin (IAA), glutamic acid decarboxylase (GADA) and anti-IA-2ic. A multivariate analysis, according to the Cox proportional hazard model considering the joint effects of all baseline variables, selected the four antibodies and the specific family history as significant risk confounding factors (p < 0.05). Further analysis by Kaplan-Meier Life-table methods confirmed a significantly increasing risk of diabetes with the number of autoantibodies present (p < 0.001). In accordance with the Cox model, relatives with more than one affected family member (a multiplex pedigree) and siblings and offsprings vs. parents were at increased risk of IDDM (p < 0.05). In addition to technical problems, a screening strategy based on initial ICA testing has the potential of missing ICA negative subjects among future cases of type I diabetes (19% were ICA negative in the present study) and we therefore set out to evaluate an alternative approach using a dual step strategy with a combination of GADA and anti-IA-2ic for initial screening followed by retesting of positive individuals for ICA and IAA. The combination of GADA and anti-IA-2ic for primary screening (step 1) proved to be more sensitive, identifying 94% of future cases of type I diabetes compared to 81% using ICA as initial test and this antibody combination identified 93% of those individuals with ICA of 20 JDF or more. Retesting of positive individuals for ICA and IAA (step 2) significantly improved the positive predictive value confering a risk of diabetes for siblings and offsprings with more than 2 antibodies within 5 years of 67% (95%CI: 39-90). We conclude that the prognosis of contracting IDDM in relatives is strongly related to the number of autoantibodies present, but the family history should be additionally considered for individual risk assessment. The proposed screening strategy could overcome the inherent problems of the ICA and IAA assays for large-scale screening. In the present study it allows 5-year risk estimates of up to 67% identifying 94% of future cases of type I diabetes.

Human sperm penetration in different media.

Sperm penetration into bovine cervical mucus and hen egg white using capillary tube penetration was investigated to verify the suitability of the capillary tube penetration test with hen egg white as a test of human sperm function. Semen samples from 50 consecutive patients were used for penetration tests and spermatozoa of a further 10 semen samples were penetrated into bovine cervical mucus and hen egg white for special motility assessment by computer-assisted motility analysis. Penetration tests revealed the well-known different ability of spermatozoa to penetrate into cervical mucus and a different penetration of spermatozoa into egg white for two nearly equal groups (n = 24 and n = 26, respectively). One group showed penetration comparable with cervical mucus and one group a very fast penetration up to the limit of the scale of measurement. Motility assessment of spermatozoa that penetrated into cervical mucus and egg white revealed significant differences in straight-line velocity, linearity and lateral head displacement. The number of spermatozoa selected actively during the penetration procedure was significantly higher in cervical mucus than in hen egg white. Spermatozoa selected by bovine cervical mucus and hen egg white exhibited a different motility pattern. There was significantly better linearity and less lateral head displacement in egg white than in cervical mucus. Sperm penetration into hen egg white appeared to be influenced by different sources of egg white.

Lipid peroxidation in human spermatozoa and maintenance of progressive sperm motility.

Washed and deep frozen spermatozoa of 46 patients from an infertility clinic were separated into 3 different groups depending on their progressive motility (expressed as the sperm motile efficiency index according to Ishii et al., 1977), determined 0 and 3 h after liquefaction, and were examined for their lipid peroxidation (LPO) potential by means of the thiobarbituric acid assay. Spontaneous and iron-catalysed generation (after 15, 30 and 60 min incubation) of thiobarbituric acid-reactive substances (TBARS) was measured spectrophotometrically. Spontaneous LPO revealed the highest generation of TBARS in the group of spermatozoa with initially normal progressive motility and decreased maintenance of progressive motility after 3 h of aerobic incubation. Iron-catalysed LPO generally revealed the highest amounts of TBARS after 60 min, especially in the aforementioned group with decreased motility maintenance. The differences between this group and the two other groups were highly significant. Consequently, spermatozoa with initially normal progressive motility but decreased maintenance of motility, generated higher amounts of stable LPO products than others, which suggests that loss of motility under aerobic incubation seems to be the consequence of enhanced LPO processes.

[The significance of A1 and A2 antibodies against beta-casein in type-1 diabetes mellitus]. / Wertigkeit von A1- und A2-Antikörpern gegen beta-Kasein beim Typ-1-Diabetes mellitus.

BACKGROUND AND OBJECTIVE: The hypothesis that cow's milk components may constitute environmental factors in the pathogenesis of insulin-dependent diabetes mellitus (IDDM) is increasingly gaining in importance. This study for the first time determined, in a family study, antibodies against the most common variants of the beta-casein (A1 and A2). PATIENTS AND METHODS: A total of 1,257 sera--from 287 patients with IDDM, 386 siblings, 477 individual parents and 107 healthy controls--were tested by enzyme-linked immunosorbent assay (ELISA) for anti-beta casein A1- and A2-IgG antibodies. RESULTS: Antibodies against casein were present in all four groups, highest titres being found in diabetics. In all four groups there was an inverse correlation between A1 and A2 antibodies and age (p < 0.001). Increased amounts of anti-casein A1 antibodies were found among patients with IDDM and their siblings. On the other hand, the serum samples from parents and control persons contained antibodies against the A2 variant. The preferential binding of sera to the one or other casein variant was in all four groups statistically significant (p < 0.001). CONCLUSION: Because the A1 variant of beta-casein correlates with the onset of IDDM, but can also occur in normal controls, this may confirm the hypothesis of a defective oral immunotolerance to cow's milk antigens in IDDM.

Coping with employment discrimination against diabetics: trends in social medicine and social psychology.

In many European countries, social-medical aspects in the management of diabetes mellitus are not satisfactorily respected. Our contribution reports a study addressing the impact of diabetes on the patient's career and daily work, in order to determine the extent to which diabetics are being discriminated against at work. Type I diabetics were questioned about their experience, and not on the objective burden. A questionnaire was developed to evaluate patients' social and employment problems. Few elements of an education program for Type I diabetes optimizing social skills (social competence) are demonstrated. In a group of 6-8 patients, assertive behavior in the work place is modelled (e.g., for hypoglycaemia, social phobia) by applying psychological methods (behavior modification role-playing). These methods can help diabetic patients to master their discrimination. They learn assertive behavior in social situations with superiors and colleagues and develop self-confidence (self-efficacy). This special education program supports Type I diabetics in coping with employment discrimination.

Extracellular matrix proteins as early markers in diabetic nephropathy.

Basement membrane thickening and mesangial expansion characterize the renal involvement in diabetes mellitus and precede any symptoms of renal dysfunction, e.g., albuminuria and changes in glomerular filtration rate. Since the morphological changes can only be diagnosed by biopsy, this study was designed to investigate whether the urinary excretion of renal extracellular matrix proteins might reflect the morphological alterations. To specify the extent of renal involvement in diabetes, the patients, type I as well as type II diabetics, were classified according to their urinary albumin excretion: normal albumin excretion below 30 micrograms/min, microalbuminuria from 30 to 300 micrograms/min, and overt albuminuria above 200 micrograms/min. Laminin, collagen IV, and fibronectin, all intrinsic components of the renal extracellular matrix, were determined in serum and urine by radioimmunoassay or enzyme-linked-immunosorbent-assay, respectively. The results are given as median values (mean). Additionally, the urinary fragment pattern of fibronectin was analysed qualitatively by immunoblotting. Laminin concentrations in serum and in urine did not change in diabetics. Collagen IV decreased in serum of patients with increased albumin excretion (controls: mean = 255 micrograms/l, normoalbuminuric patients: mean = 56 micrograms/l, microalbuminuric patients: mean = 52 micrograms/l, and patients with overt albuminuria: mean = 70 micrograms/l; alpha < 0.01) and increased in urine (controls, normoalbuminuric and microalbuminuric patients: not detectable, patients with overt albuminuria: mean = 5 ng/12 h; apha < 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)

The spermatozoal volume as indicative of the plasma membrane integrity (modification of the hypoosmotic swelling test). I. Methods.

The aim of the present study was to establish a more differentiating indicator of plasma membrane integrity of spermatozoa than the classic version of the hypoosmotic swelling test according to Jeyendraan. Spermatozoa were prepared by density gradient centrifugation (90% Percoll) to select 'fertilization competent' spermatozoa only. After a second washing procedure sufficiently pure sperm cell suspensions were obtained. The volume distributions of these sperm cells were measured with a Coulter Counter at 25 degrees C after adaptation in 300 mosmolar NaCl solution resp. 150 mosmolar NaCl solution for 5 min. These volume distributions showed significantly different patterns for the isotonic and hypotonic stress situation in the simple salt solution. Moreover, the comparison of the response to hypoosmotic stress showed more than four reproducible characteristic patterns, promising well differentiated results for different sperm populations. The new method for the detection of hypoosmotic swelling effect might be a real and valuable functional parameter.

The spermatozoal volume as indicative of the plasma membrane integrity (modification of the hypoosmotic swelling test). II. Diagnostic approach.

If using a method of statistical sifting based on the Kolmogoroff-Smirnov test for differentiation of volume distribution curves of a modified HOS-test, different curve types are obtained under isotonic as well as under hypotonic conditions. Arrangement of spermatological parameters of the relevant ejaculates according to the individual curve types reveals significant differences for motility. No correlation can be seen with the results of the microscopic HOS-test. Consequences for the use of the modified HOS-test in medical practice are discussed.