Evidence for renal kinins as mediators of amino acid-induced hyperperfusion and hyperfiltration in the rat.

This study examined the role of tissue kallikrein and kinins in renal vasodilation produced by infusion of amino acids (AA). In rats fed a 9% protein diet for 2 wk, intravenous infusion of a 10% AA solution over 60-90 min reduced total renal vascular resistance and increased glomerular filtration rate (GFR) by 25-40% and renal plasma flow (RPF) by 23-30% from baseline. This was associated with a two- to threefold increase in urinary kinin excretion rate. Acute treatment of rats with aprotinin, a kallikrein inhibitor, resulted in deposition of immunoreactive aprotinin in kallikrein-containing connecting tubule cells and inhibited renal kallikrein activity by 90%. A protinin pretreatment abolished the rise in urinary kinins and prevented significant increases in GFR and RPF in response to AA. In a second group of rats pretreated with a B2 kinin receptor antagonist, [DArg Hyp3, Thi5,8 D Phe7]bradykinin, AA infusion raised urinary kinins identically as in untreated controls, but GFR and RPF responses were absent. Aprotinin or the kinin antagonist produced no consistent change in renal function in rats that were not infused with AA.AA-induced increases in kinins were not associated with an increase in renal kallikrein activity. Notably, tissue active kallikrein level fell 50% in AA-infused rats. These studies provide evidence that kinins generated in the kidney participate in mediating renal vasodilation during acute infusion of AA.

Kinin, a mediator of diabetes-induced glomerular hyperfiltration.

Renal kallikrein is increased in diabetic patients and streptozotocin (STZ)-induced diabetic rats with hyperfiltration. Chronic inhibition of renal kallikrein reduces glomerular filtration rate (GFR) and renal plasma flow (RPF) in hyperfiltering STZ-induced diabetic rats. To investigate whether these actions of kallikrein and its inhibition are kinin-mediated, we used a B2-kinin receptor antagonist (BKA). In STZ-induced diabetic rats with hyperfiltration, renal kallikrein excretion rate was significantly increased (P < or = 0.01), and kinin excretion rate was increased 57%, as compared with control rats. Left kidney GFR and RPF were measured before and during a 40-min infusion of BKA (0.5 micrograms.kg-1.min-1) or vehicle. Infusion of the kinin receptor antagonist reduced the GFR and RPF significantly. GFR was reduced by 18%, from an average baseline value of 2.07 +/- 0.11 to 1.70 +/- 0.06 ml/min, P < or = 0.001 (means +/- SE). RPF was reduced by 25%, from 6.74 +/- 0.38 to 5.06 +/- 0.17 ml/min, P < or = 0.001. Total renal vascular resistance was significantly increased during BKA infusion, P < or = 0.001. Vehicle infusion for the same period had no significant effect on GFR, RPF, or renal vascular resistance. These findings further support the hypothesis that increased renal production of kinins contributes to the renal vasodilation of diabetes.

Body mass index and body girths as predictors of mortality in black and white women.

BACKGROUND: The high prevalence of obesity in black women has been hypothesized to contribute to higher rates of coronary heart disease and total mortality. Investigators have recently refined the study of obesity by differentiating anatomic patterns of the physical location of adipose tissue on the body. We examined fat patterning as a predictor of mortality in black women. METHODS: Body mass index (BMI) and body girths were examined as predictors of all-cause and coronary heart disease mortality during 25 to 28 years of follow-up in black and white women in the Charleston Heart Study. RESULTS: The BMI was associated with all-cause and coronary heart disease mortality in white, but not black, women. After controlling for differences in BMI, the risk of all-cause mortality was greater in white women with larger chest and abdominal girths, while midarm girths were inversely associated with mortality. The hazard at the 85th percentile relative to the 15th percentile of abdomen/midarm ratio was 1.44 in models that included BMI, education, and smoking as covariates. In black women, the girths were not predictive of either all-cause or coronary heart disease mortality. CONCLUSIONS: The failure of BMI and fat patterning to predict mortality in black women challenges previously held assumptions regarding the role of overweight in the higher mortality experienced by black women.

Variability in islet cell antibody determination: a quantitative assessment.

The source of variability in the outcome of islet cell antibody (ICA) determinations due to pancreas and serum were independently assessed with a quantitative assay. The effect of choice of pancreatic substrate and serum specimen was assessed by testing for parallelism among twenty-five dilution curves of five sera from diabetic subjects and five pancreata. The five by five format allowed for five tests of pancreatic variability (one serum against five pancreata) and five tests of serum variability (one pancreas against five sera). Analyses of variance and covariance of these dilution curves indicate that test outcome is affected by both the sera (5 tests, p values 10(-6) to 10(-7) and the pancreas (5 tests, p values 10(-3) to 10(-5)). The data support heterogeneity among islet cell antibody isotypes and raise the possibility of diversity of islet cell antigens. Antigenic diversity of pancreatic substrates may interfere with inter- and intra-laboratory comparisons of islet cell antibody determinations. The strong effect of pancreas type on ICA outcome infers possible heterogeneity of the islet cell antigen(s) capable of reacting with human islet cell antibodies.

Levels of transferrin and alpha 2-HS glycoprotein in women with and without endometriosis.

The study objective was to test the hypothesis that elevated levels of transferrin and alpha 2-HS glycoprotein occur in the peritoneal environment of patients with endometriosis that may lead to the observed autoimmunity to these proteins. We set up a double sandwich enzyme-linked immunosorbent assay (ELISA) for measuring levels of transferrin and alpha 2-HS glycoprotein in the serum and peritoneal fluid samples from women with (n = 24-60) and without endometriosis (n = 35-49). Serum and peritoneal fluid levels of alpha 2-HS glycoprotein and peritoneal fluid levels of transferrin were significantly elevated in patients with endometriosis, in contrast to the controls. Serum levels of transferrin in patients, however, were significantly less in the patients than in controls. We conclude that transferrin and alpha 2-HS glycoprotein are present at high concentrations in the peritoneal fluids of patients with endometriosis. This may play a significant role in the autoimmune pathophysiology of endometriosis.

A comparison of the effects of cholesterol on CHD mortality in black and white women: twenty-eight years of follow-up in the Charleston Heart Study.

The relationship between cholesterol and 28-year CHD mortality in women was evaluated in the Charleston Heart Study. Linear, quadratic, and cubic models were investigated using Cox proportional hazards regression analysis. In white women, the linear, quadratic, and cubic terms for cholesterol were significant suggesting an asymmetric J-shaped relationship. In black women, only the linear term in all three models was statistically significant suggesting an increasing CHD mortality rate with increasing cholesterol level. The lack of consistency of results by different statistical analyses in black women make conclusions concerning the nature of the relationship between cholesterol and CHD mortality less strong in black women than in white women. Compared with women having a cholesterol value equal to the mean of the group (241 mg/dl), white women having a cholesterol value one standard deviation above the mean (s = 52.5 mg/dl) had a 60% higher CHD mortality rate (hazard ratio = 1.6, 95% CI: 1.2-2.1). In black women, the estimated hazard ratio for a one standard deviation (s = 47.8 mg/dl) increase in cholesterol is 1.4 (95% CI: 1.03-1.8). The results suggest that the relationship of cholesterol to CHD mortality is different in white and black women. The relationship in white women appears to be curvilinear and represented by an asymmetric curve while the relationship in black women is not curvilinear and the overall pattern of association, while possibly linear, is equivocal.

Black women have smaller abdominal girths than white women of the same relative weight.

Anthropometric measurements were compared in 312 white and 242 black women (mean age 54) who were participants in the Charleston Heart Study. Body mass index (BMI) was greater in black women (27.8 kg/m2) than in white women (24.7 kg/m2) as were body circumferences. However, when the girth measurements were adjusted for BMI, some racial differences were reversed. Age, smoking and BMI-adjusted abdominal girth was smaller in the black women than in the white women (88.9 cm vs 92.2 cm). The ratio of abdomen to midarm circumference was larger in white women than black women (3.24 vs 3.09), and could be interpreted to indicate a less central fat pattern in the black women. This conclusion should be viewed with caution since circumference measurements, though often used in epidemiologic research, do not differentiate between subcutaneous fat and visceral fat.

Predictors of physical disability in elderly blacks and whites of the Charleston Heart Study.

During the 1984/85 recall of the Charleston Heart Study Cohort, physical function data were obtained for 247 white males, 376 white females, 123 black males, 247 black females and 71 high socioeconomic status (SES) black males over 60 years of age. Black females had the highest prevalence of physical disability (55.8%), followed by white females (43.2%), black males (39.0%) and white males (25.8%) and high SES black males (22.3%). Physical disability was 1.5-2.5 times as prevalent among individuals with a history of cardiovascular disease (CVD) than those without such a history. Among individuals without a current history of CVD univariate analyses showed the following as significant (lower 95% CI greater than 1.0) predictors of physical disability: elevated systolic blood pressure in white females, black males, and black females; elevated cholesterol in black females; obesity in black females; and low educational level in white females. Regression analyses indicated that obesity in 1960 accounted for 10.9 and 2.9% respectively of the variability in physical disability scores in 1985 for black females and white females.

Observer variation in the angiocardiographic diagnosis of mitral valve prolapse.

The purpose of this study was to assess the reliability of the angiocardiographic diagnosis of mitral valve prolapse by measuring agreement between observers using defined diagnostic criteria. Sixty high-quality left ventriculograms, selected to include many that showed possible mitral valve prolapse, were reviewed by three angiocardiographers. Disagreement between observers as to positivity occurred in 26 percent and 30 percent of the cases on two reviews, and disagreement as to specific scallop involvement occurred in 68 percent and 78 percent of the cases. A specific observer agreed with himself in interpretation of positive 78 to 82 percent of the time and in the interpretation of specific scallop involvement 55 to 90 percent of the time. These results indicate that in the absence of generally agreed upon quantitative angiocardiographic criteria for the diagnosis of mitral valve prolapse, there is considerable variability among observers in the interpretation of mitral valve prolapse by angiocardiographic studies.

Cytotoxic antibodies to lymphocytes: relationship to sperm immunity.

Sperm and T lymphocytes share antigens, and sera from infertile subjects with sperm immunity have agglutinating antibodies to T lymphocytes. In the present study, cytotoxic antibody titers to sperm (C-Sp) and lymphocytes (C-Ly) were measured in sera, seminal plasma, cervical mucus and vaginal secretions of 52 infertile couples. The samples were divided into C-Sp negative and C-Sp positive groups. C-Ly titers against control T lymphocytes were significantly elevated in C-Sp positive sera of 35 males (P less than 0.005) and 27 females (P less than 0.05) as compared with C-Sp negative sera. C-Ly titers against autologous or partner T lymphocytes were similarly elevated in the sera of both groups. Titers of C-Ly in C-Sp positive secretions (seminal plasma, cervical mucus and vaginal secretions) against autologous and partner, as well as control, T lymphocytes were significantly elevated. In addition, elevated titers of C-Ly to autologous, partner and control B lymphocytes were observed in sera, cervical mucus and vaginal secretions from females with sperm immunity. The infertile males, however, did not have significant B lymphocyte immunity. Immunofluorescence assays revealed a significantly higher incidence of sperm and T lymphocyte antibodies (primarily IgG) in the C-Sp positive than in the C-Sp negative group. Titration of 100 sera from infertile subjects (50 couples) against a panel of lymphocytes of known HLA specificities failed to reveal a pattern of activity against HLA antigens of the A, B and C loci. Absorption of immune sera with sperm or lymphocytes reduced C-Sp and C-Ly titers significantly (P less than 0.001). These results suggest that infertile subjects with sperm immunity produce antibodies to cross-reactive antigens present on sperm and T lymphocytes.

Anti-sperm antibodies, detected by agglutination, immobilization, microcytotoxicity and immunobead-binding assays.

To determine the reliability of tests currently utilized in the detection of sperm-reactive antibodies, sera were provided as unknowns and studied without knowledge of the clinical histories. Four laboratories performed tray agglutination tests (TAT), three complement-dependent immobilization (SIT), and single laboratories sperm cytotoxicity (SCT), passive haemagglutination (PHA) and immunobead binding (IBB). Most investigators demonstrated an excellent correlation between duplicate sample results. Nearly all of the female sera were free of anti-sperm antibodies and positive results did not appear in greater frequency in women with unexplained infertility as compared with other categories. For the male sera, the highest incidence of anti-sperm antibodies in the infertile group (21% positive for sperm-reactive IgGs) was obtained by immunobead binding. The GAT and TAT results gave 7 and 12% positives, except for lower results in one laboratory. Sperm-reactive antibodies were detected most commonly in vasectomized men, with all assays except SCT and PHA. Of the newer techniques studied, IBB results correlated well with TAT, GAT and SIT, while SCT and PHA did not, suggesting that a different group of antibodies, perhaps directed against other sperm-associated antigens, was being detected by the latter procedures. In this light, emphasis was placed on the need to validate whether results of particular methodologies correlated with impaired sperm function and to develop methods that provided evidence for this premise, either on the basis of clinical criteria or altered gamete interaction in vitro.

A computerized sperm motion analysis [published erraturm appears in Fertil Steril 1986 Nov;46(5):980].

Semen samples from 35 fertile men and husbands of 32 infertile couples were evaluated with the use of a rapid computerized sperm motion analysis (CSA). The counts and percentages of motile sperm measured by routine semen analysis (RSA) and CSA were comparable in both groups. In CSA, mean values of swimming speed (microns per second), linearity, and motility index of sperm in the fertile men were significantly higher than those in the infertile group. At 3 hours, the number of sperm moving at a speed of 21 to 30 microns/second were higher in the infertile group than in the fertile group. In contrast, the number of sperm moving at a speed of 31 to 50 microns/second and higher was increased in the fertile group over the infertile group. Good correlations were obtained between RSA and CSA sperm counts and percentages of progressively motile sperm. Sperm counts, percent motility, indices of motility, and mean sperm speed obtained by two observers with the use of CSA correlated well. It seems that the sperm speed and the motility index remain unaltered in repeat semen analyses of fertile men. It is concluded that a computerized analysis of sperm swimming speed is a reliable and rapid mode for evaluating semen samples and offers more discriminatory sperm motion characteristics than the RSA.

Endometrial antigens involved in the autoimmunity of endometriosis.

Serum and peritoneal fluid from five fertile women without endometriosis and serum (n = 23) and peritoneal fluid (n = 12) from infertile women with endometriosis were tested for the presence of antibodies against endometrial tissue antigens by a Western blot analysis. Antigens with molecular weights (MW) of 19, 31, 38, and 42 kd reacted with antibodies in the serum and peritoneal fluid from both fertile and infertile women. Antibodies in 20 of 23 (87%) sera and all 12 (100%) peritoneal fluid samples from endometriosis patients reacted against endometrial antigens with molecular weights (MW) of 26 kd and/or 34 kd. Serum from 10 patients (43%) and peritoneal fluid from 6 patients (50%) also had antibodies to an endometrial antigen with MW of 21.5 kd. Reactivity to other endometrial antigens with MW 16, 24, 48, and 75 kd was also noted in patients with endometriosis. Antibodies in the serum and peritoneal fluid from fertile women failed to react against these antigens. It is concluded that the humoral and local endometrial autoimmunity detected in patients with endometriosis is primarily directed against antigens with MW of 26 and 34 kd.

Time-related changes in the plasma concentrations of prolactin, gonadotropins, sex hormone-binding globulin, and certain steroid hormones in female runners after a long-distance race.

Running is associated with an increase in plasma concentrations of certain anterior pituitary hormones and adrenal steroids. This study reports such increases after a marathon race. Six trained female runners, 26 to 42 years old, participated in a marathon race. Fasting (resting) blood samples were collected a few weeks before the race (baseline) and immediately (0 hour), 1 hour, and 4 hours after the run. The data were analyzed with the use of two-way analyses of variance (F-test), paired t-test, and Page's test. At 0 hour, compared with baseline, significant increases were observed in the plasma concentrations of testosterone (T), dehydroepiandrosterone sulfate (DHEA-S), cortisol (F), free T index (T/SHBG), and prolactin (PRL). At 1 hour, levels of these steroid hormones and PRL declined, some significantly. At 4 hours, levels of all hormones except DHEA-S returned to baseline. No significant changes were observed in concentrations of follicle-stimulating hormone (FSH), luteinizing hormone (LH), and sex hormone-binding globulin (SHBG), as evaluated by F-test. Running-associated changes in plasma hormonal concentrations revert to baseline in four hours, although DHEA-S may take a little longer.

Cytotoxic sperm antibodies inhibit sperm penetration of zona-free hamster eggs.

Zona-free hamster egg sperm penetration assay was used to study the effects of cytotoxic sperm antibodies on egg penetration by the sperm of fertile and infertile men. Twenty-nine fertile and 9 infertile men did not have significant cytotoxic sperm antibodies in their serum and seminal plasma; 7 infertile men were positive for these antibodies in serum and seminal plasma. Two others were positive in sera, and 14 were positive in seminal plasma. Sperm from 18 of 23 (78%) infertile men with sperm antibodies had poor egg penetration (less than or equal to 20%) compared with only 6 of 38 (16%) nonautoimmune men (P less than 0.0001). Sperm from nonautoimmune fertile men were coated with seminal plasma and serum of autoimmune men and serum of isoimmune women, resulting in a significant decrease in hamster egg penetration. Sixteen of 21 (76%) seminal plasma samples with cytotoxic sperm antibodies reduced the control sperm penetration of hamster eggs by greater than or equal to 50%. Coating of sperm from fertile men with serum and seminal plasma samples from non-sperm-immune fertile and infertile subjects did not alter their penetration of hamster eggs. Coating of sperm from autoimmune men with cytotoxic antibody-positive autologous seminal plasma samples resulted in a significant decrease of egg penetration. The inhibitory effect of antibody-positive seminal plasma samples on egg penetration by control sperm was abrogated when the samples were preabsorbed with sperm. It is concluded that cytotoxic sperm antibodies, especially those in seminal plasma, inhibit hamster egg penetration by autologous and control sperm. This may explain in part the incidence of infertility associated with sperm antibodies.

Association of human leukocyte antigens B7 and BW35 with sperm antibodies.

One hundred three infertile couples and 50 fertile couples were tested for the presence of sperm antibodies by passive hemagglutination and cytotoxicity assays and typed for human leukocyte antigens (HLAs) of the A and B loci. Fifty-four percent of the males and 43% of the females with sperm antibodies were found to have the HLA-B7 antigen, in contrast to 19% of a general population (P less than 0.0001). HLA-BW35 (a Workshop antigen of the B locus) was also present at a significantly increased antigenic frequency (P less than 0.00001) in those females with sperm antibodies. Of the 73 infertile couples with sperm antibodies, 79% shared the B7, B8, and/or BW35 alleles. Two of 12 infertile couples and 10 of 50 fertile couples without sperm antibodies shared these alleles. The significant association of HLA-B7 and BW35 alleles with sperm antibodies in infertile subjects suggests a selective disadvantage of these antigens in human reproduction.

Sperm motility on postcoital testing correlates with male autoimmunity to sperm.

To assess whether or not immunologic factors in husbands, wives, or both, influence the motility of sperm in the female reproductive tract, hemagglutination and cytotoxicity sperm antibody (Ab) assays and postcoital tests (PCTs) were performed in 293 infertile couples. More couples without male autoimmunity to sperm (64% of 66; P less than 0.001) had greater than or equal to 10 motile sperm per high power field (adequate PCT), as compared with 26% of 122 couples with untreated male autoimmunity, 19% of 77 couples with corticosteroid-treated male autoimmunity without a pregnancy, and 36% of 28 couples with successfully treated male autoimmunity to sperm. Good correlation was obtained among pregnancy achievement, lack of sperm antibodies, and adequate sperm motility in the PCT (P less than 0.0001). Sperm motility in the PCT correlated positively with sperm motility in the semen and inversely with cytotoxic sperm Ab in the serum and seminal plasma of men and women and hemagglutinating sperm Ab in the cervical mucus samples. Sperm motility in the PCT has a predictive value of 72% for male autoimmunity and 57% for female isoimmunity to sperm in the presence of normal cervical mucus and in the absence of infections.

Antibodies to A19 and Bw35 complexes of human leukocyte antigens are present in infertile subjects with sperm antibodies.

Sera and secretions from 100 couples with unexplained infertility were tested for sperm antibodies by cytotoxicity and passive hemagglutination and also for antibodies to human leukocyte antigen (HLA) by cytotoxicity assays. Lymphocytes of the study subjects were typed for 61 HLA-A and B alleles. Thirteen of 30 (43%) men with sperm autoimmunity also had HLA antibodies in their serum and/or seminal plasma samples, in contrast to 2 of 35 (6%) nonautoimmune males (P = 0.0003). Twenty-five of 35 (71%) sperm antibody-positive infertile women had HLA antibodies in their sera and/or secretions, while only 7 of 31 (23%) women without sperm antibodies were positive (P = 0.00007). Antibodies to HLA-A19 (A26, A29, AW30, AW31, AW32, AW33, and AW34) and Bw35 (B5, B15, B17, and B18) complexes were present in 19 of 22 (86%) infertile men and 44 of 48 (92%) infertile women positive for HLA antibodies (P less than 0.01). The presence of antibodies to HLA-A19 and/or Bw35 in the infertile subjects did not correlate with the presence of HLA-A19 and/or Bw35 in the husbands. The presence of antibodies to HLA-A19 and/or Bw35 in the cervical mucus of the infertile women correlated with their presence in the seminal plasma of their husbands. It is suggested that antibodies to sperm antigens cross-reactive with HLA-A19 and/or Bw35 may be relevant to infertility.

Sex-hormone-binding globulin in clinically hyperandrogenic women: association of plasma concentrations with body weight.

The relationship of sex-hormone-binding globulin (SHBG) with actual body weight (ABW), ideal body weight (IBW), ABW as percentage of the IBW (% IBW), Quetelet index (weight/height2) and plasma concentrations of various androgens and 17 beta-estradiol (E2) were studied in 9 normal and 57 hirsute patients (group 1). In hirsute patients, plasma levels (ng/dl, mean +/- standard error of the mean [SEM]) of testosterone (T; 77 +/- 4), dihydrotestosterone (DHT; 26 +/- 2), androstenedione (delta 4A; 184 +/- 16), and SHBG (0.91 +/- 0.05 micrograms DHT/dl) but not of dehydroepiandrosterone (DHA; 608 +/- 55) and E2 (6.1 +/- 0.1) were significantly different from those in controls. A negative correlation was observed between SHBG and ABW, both in controls (P less than 0.05) and hirsute patients (P less than 0.01). The hirsute patient population was subdivided into two groups: nonobese (group 2; 60 +/- 1 kg; n = 35) and obese (group 3; 96 +/- 2 kg; n = 22). Plasma androgens, T/SHBG (an index of free T) and E2 in groups 2 and 3 (T: 75 +/- 4, 81 +/- 7; DHT: 24 +/- 2, 28 +/- 3; T/SHBG: 85 +/- 7, 105 +/- 11; delta 4A: 203 +/- 13, 155 +/- 16; DHA: 663 +/- 83, 521 +/- 49; E2: 6.1 +/- 1.0, 5.8 +/- 0.9) were similar; yet SHBG in group 3 (0.75 +/- 0.04) was significantly lower than in group 2 (1.0 +/- 0.01). Inverse correlations between SHBG and ABW, % IBW, and ABW/H2 were observed in group 2 but not in group 3. We conclude that a negative relationship exists between SHBG and the body size in nonobese women and that in hirsute patients, obesity leads to a further lowering of SHBG through mechanism(s) probably independent of androgens.

Plasma androgens, 17 beta-estradiol, and sex hormone-binding globulin in patients with hirsutism and/or clitoromegaly.

Plasma androgen, 17 beta-estradiol, and sex hormone-binding globulin (SHBG) levels were measured in 11 normal subjects (group 1), 18 hirsute patients with no clitoromegaly (group 2), 13 hirsute patients with clitoromegaly (group 3), and 8 patients with clitoromegaly but no hirsutism (group 4). Significantly elevated levels of testosterone (T) and dihydrotestosterone (DHT) were found in groups 2 and 3 but not in group 4 when compared with group 1. In contrast, levels of dehydroepiandrosterone (DHEA) in groups 1 and 2 were comparable but significantly lower than those in group 3 and 4; the levels in the latter two groups were not significantly different from each other. In groups 2, 3, and 4, levels of androstenedione (delta 4A) and the factor T/SHBG were significantly elevated whereas SHBG levels were significantly suppressed when compared with those of group 1. The clitoral index correlated (P less than 0.01) with DHEA levels in group 3. It is concluded that clitoromegaly without hirsutism is associated with increased plasma levels of DHEA and delta 4A. In contrast, hirsutism without clitoromegaly is associated with elevated levels of T, DHT, and delta 4A but normal DHEA levels.