Long-term treatment of chronic chagasic cardioneuropathy with a mixture of gangliosides: effect on evolution of circulating antiGM1 antibodies.

Chagas disease involves a cardiac impairment, being the first alterations of autonomic disorders which affect heart rate and blood pressure control. At this stage, diminished heart rate responses to atropine and propranolol are observed. Prior studies have shown that short term ganglioside treatment improves the responses to these agents, but there is no information about the long term effect of gangliosides and the evolution of antiGM1 titers. The effects of long term treatment with gangliosides on autonomic tests in patients with chagasic cardiodisautonomy and the evolution of antiGM1 titers were studied in 90 patients (57 men, 33 women, aged 25-60 years) with positive serology for Chagas disease and electrocardiogram showing sinusal bradycardia and incomplete right branch block, without cardiomegaly, with autonomic alterations by postural and Valsalva's tests. All patients were submitted to a test that consisted of intravenous injection of atropine 0.04 mg/kg followed 3 min later by intravenous injection of propranolol 0.2 mg/kg. During these tests heart rate and blood pressure were recorded continuously. Subsequently, 30 patients were treated with 100 mg/day of a mixture of gangliosides by intramuscular injection during 15 days in a row, followed by 40 mg/day during another 75 days. Another 30 patients received continuous treatment for 12 months. The remaining 30 patients were controls. The antiGM1 antibody circulating titers were determined before the treatment, at the third and 12th month. Seventy-four patients completed the study. Before treatment, the heart rate increased, though slightly, after the injection of atropine. After 3 months of ganglioside treatment a statistically significant increase in the response to atropine was recorded. In the controls at 12 months, the response to atropine remained increased without differences between the patients treated for 3 and 12 months. The control patients did not show any modification of the heart rate response during 12 months. Both ganglioside-treated groups showed an increase in the response to propranolol. The antiGM1 titer distribution was similar in both healthy subjects and chagasic patients. None of the patients had positive antiGM1 titers in basal conditions nor significant modifications after the ganglioside treatment. Chagasic cardioneuropathy was not associated in this study with high antiGM1 antibody titers. Chagasic patients showed a diminished heart rate response to atropine as well as to propranolol. Ganglioside treatment determined an increased heart rate response, particularly after atropine. Increased heart rate response was maintained until 1 year, without differences between the patients treated for 3 and 12 months. No changes in the antiGM1 titers were observed during the ganglioside treatment.

Pregnant women with impaired tolerance to an oral glucose load in the afternoon: evidence suggesting that they behave metabolically as patients with gestational diabetes.

OBJECTIVE: In previous studies we observed the existence of a circadian variation of the blood glucose response to oral glucose in pregnant women with higher values at 4 PM. Some women with increased risk of diabetes with normal oral glucose tolerance tests at 8 AM also had values above maximum normal levels at 4 PM. The aim of this trial was to determine the clinical significance of this impaired tolerance in the afternoon. STUDY DESIGN: Seventy-seven pregnant women with normal risk of diabetes (65 of normal weight and 12 overweight), 75 with increased risk of diabetes (26 overweight), and 12 patients with gestational diabetes were incorporated in the study. All women underwent two oral glucose tolerance tests (1.5 gm/kg) at 31 to 32 weeks' gestation at 8 AM and 4 PM with a 1-week interval. At 33 weeks' gestation a whole-day blood glucose profile was performed with usual food intake; samples were withdrawn before each meal and at 30, 60, and 120 minutes after each meal. The weight of the newborns was recorded. RESULTS: (1) Results of oral glucose tolerance tests confirmed a circadian variation of the response in all groups; (2) 37 women with increased risk of diabetes had higher values after oral glucose tolerance testing than the normal threshold at 4 PM but not at 8 AM; (3) among women with normal risk of diabetes all values were within the normal range despite the circadian variation; (4) blood glucose levels during whole-day profiles were normal in women with normal risk of diabetes and with increased risk of diabetes with normal oral glucose tolerance testing at 4 PM, whereas all women with increased risk of diabetes and impaired tolerance in the afternoon showed hyperglycemic episodes; (5) the percentage of newborns with high weight (>90th percentile) among women with increased risk of diabetes and abnormal oral glucose tolerance tests at 4 PM was similar to the percentage found in women with gestational diabetes and much higher than the one observed in women with normal oral glucose tolerance tests in the afternoon. CONCLUSIONS: The impairment of the response to oral glucose tolerance testing seen in some patients with increased risk of diabetes at 4 PM but not at 8 AM seems of clinical significance in view of the abnormal whole-day blood glucose profile these women had and the weights of the newborns.

A temporal study of somatostatin secretion and its inhibitory effect in genetically diabetic mice (C57BL/KsJ-db/db).

Somatostatin (SRIF) acts as a physiological regulator of insulin and glucagon secretion. This study explored whether alterations in SRIF secretion and activity in perifused pancreas from spontaneously diabetic mice (C57BL/KsJ-db/db) could be correlated with hypersecretion of insulin by the beta-cells. SRIF release upon stimulation with 27.5 mM glucose was biphasic in controls, whereas a first phase peak was absent in 30-to 90-day-old diabetic mice (db/db). Twelve- to 28-day-old db/db mice showed two distinct patterns compared to controls: biphasic hypersecretion in the 12- to 20-day-old group compared to normal secretion in the 22- to 28-day-old group. Basal SRIF secretion showed a tendency to be elevated above controls in some of the db/db age groups, but the difference was not statistically significant. Insulin release from control pancreases was biphasic, whereas in db/db mice, basal hypersecretion, absence of the first peak, and second phase hypersecretion were observed. beta-Cell sensitivity to the inhibitory effect of SRIF was diminished in db/db mice aged 12-90 days. Diazoxide, on the contrary, inhibited insulin secretion from the pancreas of diabetic and normal mice to the same extent. The results suggest that altered secretion of and response to SRIF in C57BL/KsJ-db/db mice might provide an explanation for the anomalies in insulin secretion in the first stages of this type of diabetic syndrome.

D-Tryptophan-6-luteinizing hormone-releasing hormone in the treatment of normogonadotropic oligoasthenozoospermia.

D-Tryptophan-6-LH-RH was assessed in the treatment of patients with normogonadotropic oligoasthenozoospermia in 18 subjects selected on the basis of at least 3 spermiograms, long standing infertility, normal LH, FSH, prolactin and testosterone serum levels and lack of evidence of any other pathologic involvement. Testicular biopsies performed on these patients showed hypospermatogenesis with foci of alteration at the spermatid stage in some of them. D-Trp-6-LH-RH was administered im for 90 days at a dose of 5 micrograms every 2 days, 10 micrograms daily or 10 micrograms every 2 days. There was no significant improvement in the concentration of spermatozoa or in the motility and vitality parameters. Moreover, in 5 patients who received 10 micrograms daily, basal levels of LH and FSH and the response to LH-RH, decreased significantly during treatment. D-Trp-6-LH-RH at the dose used in this study does not seem useful for the treatment of oligoasthenozoospermia normogonadotropic patients.

The effect of a treatment with doxycycline on semen of asthenozoospermic patients with T-mycoplasma genital infection.

The pathogenic role of T-Mycoplasma seminal infection in male sterility is a matter of controversy. The aim of this study was to observe the effect of a specific treatment in asthenozoospermic patients in which the only finding was the presence of T-Mycoplasma in semen. Twenty five patients (age 24 to 49 years) were incorporated. Average time of known infertility was 4.0 +/- 0.5 years. Asthenozoospermia was diagnosed by at least three previous spermatograms. All other studies were negative with the only exception of the tests to study the presence of T-Mycoplasma in semen. Each patient was treated with doxycycline (200 mg/day) during 20 days. Control spermatograms were performed at 30 and 90 days from the initiation of treatment. After treatment the tests to detect the presence of T-Mycoplasma in semen were negative in all patients. A significant increase of the percentages of forwardly progressive spermatozoa and of live and motile spermatozoa was observed together with a significant decrease of the percentage of non motile spermatozoa. Qualitatively eleven patients (44%) achieved a normalization of the spermogram; five patients (20%) obtained significant improvements while nine patients (36%) did not experience any significant change. Although the real significance of T-Mycoplasma in the determination of male sterility is still a matter of controversy, its presence in semen in asthenozoospermic patients should be considered and, consequently, treated.

Pentoxifylline (BL 191) by oral administration in the treatment of asthenozoospermia.

The aim of this trial was to study the effects on semen of pentoxifylline administered chronically per os to patients with normogonadotropic asthenozoospermia. Fifteen infertile men (age 22 to 44 years) were incorporated. Each subject was submitted to a treatment with pentoxifylline per os at a dose of 1,200 mg per day during no less than four months (x plus or minus SE 6.6 plus or minus 0.7 months). Assessment of the results was performed by serial spermatograms performed during and up to three months after treatment. The results showed a significant improvement of the percentages of forwardly progressive spermatozoa and of live and motile spermatozoa. Five of the patients achieved a normalization of semen (frank improvement) and seven a significant improvement in comparison to the initial values. Pregnancy was obtained by two patients. Pentoxifylline could be a useful alternative for the treatment of patients with normogonadotropic asthenozoospermia.

Effect of the phosphodiesterase inhibitor Pentoxyfylline on human sperm motility.

Tyrode fluid and Tyrode fluid plus Pentoxifylline were individually added to aliquots of semen samples obtained from 6 normal men and 6 infertile patients considered to have idiopathic normogonadotropic oligoasthenozoospermia. Pentoxifylline was added to final concentrations of 0.15, 0.30 and 0.60 mM. One aliquot with no addition served as control. Samples were incubated in 37 degrees C and observed by light microscopy at 30 minutes and at 1, 2 and 4 hours after obtaining the material. At observation time, semen quality was evaluated by determining the percentages of forwardly progressive spermatozoa, slowly progressive spermatozoa, "in situ" motile spermatozoa, live and non-motile spermatozoa and dead spermatozoa. Results reported included only the first and last category. Tyrode fluid did not affect significantly the motility and the duration of activity of spermatozoa. Ejaculated human spermatozoa both from normal and asthenozoospermic men added the Pentoxifylline at 0.30 and 0.60 mM showed a longer lasting activity than those of control semen and semen added only with Tyrode fluid.

Effect of D-leucine-6-luteinizing hormone-releasing hormone ethylamide in patients with hypogonadotropic hypogonadism with anosmia.

Four men with hypogonadotropic hypogonadism and anosmia were tested with acute intravenous injections of luteinizing hormone-releasing hormone (LH-RH) and D-leucine-6-LH-RH-ethylamide (D-L eu-6-LH-RH-EA) with a 1-week interval. Each patient was then treated with this drug for 60 days and tested again after this period with an intravenous injection of D-L eu-6-LH-RH-EA. The administration of LH-RH resulted in a significant increase in the LH level in only one patient and in follicle-stimulating hormone (FSH) and testosterone increases in none. The analog D-Leu-6-LH-RH-EA resulted in significant increases in LH levels in two patients, in FSH levels in three, and in testosterone levels in one. Results obtained after treatment were closely similar to those observed before treatment. Clinical improvement in terms of increased libido, erection, pubic hair growth, and testicular size was observed. D-Leu-6-LH-RH-EA could be useful in the treatment of patients with hypogonadotropic hypogonadism, a possibility deserving further studies.

Bromocriptine in the treatment of hyperolactinemic amenorrhea.

Thirty women with secondary amenorrhea and hyperprolactinemia were studied; galactorrhea was present in 25 of them, and 18 were infertile. Serum prolactin (PRL) levels were high in all cases, between 26 and 120 ng/ml. All women were treated with bromocriptine in increasing doses from 2.5 to 5.0 or 7.5 mg daily, according to the response obtained, for 4 months. In 27 patients a PRL determination was performed during treatment; values returned to normal (up to 20 ng/ml) in 23 women and remained high in 4. Galactorrhea disappeared in 21 of 25 women. Ovulatory menses were re-established in 17 patients (56.6%). Seven women became pregnant (38.8%), one of them after bromocriptine and clomiphene were given simultaneously in the same cycle. According to our results and a literature review the following conclusions may be drawn: (1) bromocriptine is a useful therapeutic tool for re-establishing menstruation and inducing ovulation in patients with the hyperprolactinemic-amenorrhea syndrome; (2) the association of bromocriptine and clomiphene could be the next step in the treatment of patients who fail to ovulate with bromocriptine alone.

Release of prolactin during pregnancy: effect of sulpiride.

The aim of this trial was to study the prolactin-releasing capacity of the pituitary during pregnancy by means of an acute stimulation with sulpiride. Thirty women with normal pregnancies were included in the study (first trimester, nine women; second trimester, eleven women thir trimester, ten women). Each woman received an intramusclar injection of 100 mg of sulpiride sulfate between 8 and 9 A.M. Three similar groups received injections of a saline solution. Blood samples were obtained before and 30 and 60 minutes after the injection. In each sample the prolactin concentration was determined by radioimmunoassay. Basal prolactin levels increased during pregnancy. Significant responses to sulpiride were observed during the three stages of pregnancy, and the levels in the second and third trimesters were higher than those in the firsttrimester. The prolactin-releasing capacity of the pituitary, as judged by the response to sulpiride, seems to be maintained during pregnancy.

Effect of two phosphodiesterase inhibitors, cyclic adenosine 3':5'-monophosphate, and a beta-blocking agent on human sperm motility.

Mann-fructose fluid (MF), and MF plus caffeine, MF plus pentoxifylline, MF (dibutyryl cAMP), MF plus propranolol, and MF plus propranolol plus dibutyryl cAMP were individually added to aliquots of semen samples obtained from 18 normal men. These drugs were added to a final concentration of 0.6 mM. One aliquot with no addition served as control. Samples were incubated at 37 degrees C and observed by light microscopy at 30 minutes and at 1,2, and 4 hours after obtained the material. At each observation time, semen quality was evaluated by determinating the percentages of forwardly progresssive spermatozoa, slowly progressive spermatozoa, "in situ" motile spermatozoa, live and nonmotile spermatozoa, and dead spermatozoa. Mann-fructose fluid resulted in a decrease in motility and the duration of activity of spermatozoa, Caffeine seemed to neutralize the deleterus effect on the buffer, whereas pentoxifylline and cAMP seemed to increase the duration of activity of spermatozoa. Propranolol resulted in a dramatic decrease in motility, an effect that could not be neutralized by the simultaneous addition of cAMP.

PIP: Mann-fructose fluid (MF), MF plus caffeine, MF plus pentoxifylline, MF plus dibutyryl cyclic adenosine 3',5'-monophosphate (cAMP), MF plus propranolol, and MF plus propranolol plus dibutyryl cAMP were individually added to aliquots of semen samples obtained from 18 normal men to a final concentration of .6 mM to determine whether these drugs affected percentage, quality, and duration of activity of ejaculated human spermatozoa. At each observation time (30 minutes, 1, 2, and 4 hours), semen quality was evaluated by determining percentages of forwardly progressive, slowly progressive, in situ motile, live and nonmotile, and dead sperm. MF resulted in a decrease in motility and duration of activity compared with controls (P .001). Caffeine seemed to neutralize the deleterious effect, whereas pentoxifylline and cAMP seemed to increase duration of activity. Propranolol resulted in a dramatic decrease in motility, an effect that could not be neutralized by adding cAMP. The effects of adding cAMP seem to confirm that an increase of the intracellular content of this compound could determine a longer-lasting activity of ejaculated human spermatozoa.

D-leucine-6-luteinizing hormone-releasing hormone ethylamide in the treatment of normogonadotropic oligoasthenospermia.

The effectiveness of D-leucine-6-luteinizing hormone-releasing hormone ethylamide (D-Leu-6-LH-RH-EA) in the treatment of idiopathic normogonadotropic oligoasthenospermia was assessed in 17 patients selected on the basis of at least three previous sperm counts; a history of long-standing infertility; normal serum levels of luteinizing hormone, follicle-stimulating hormone, and testosterone; and lack of any evidence of other pathologic involvement. On testicular biopsy all patients showed hypospermatogenesis or arrest at the spermatid stage with the exception of two with more severe disorders. D-Leu-6-LH-RH-EA- was administered intramuscularly for 90 days at a daily dose of 5 microgram (four patients), 10 microgram (four patients), 20 microgram (five patients), and 200 microgram (four patients). The results showed no significant improvement in the parameters considered (number of spermatozoa per milliliter, percentage of live and motile spermatozoa, and percentage of forwardly progressive spermatozoa). D-Leu-6-LH-RH-EA does not seem promising for the treatment of oligoasthenospermic patients.

Pituitary and testicular response to LH-RH and to a long-acting analogue (D-leu-6-LH-RH-ethylamide).

The LH, FSH and testosterone response to 50 microgram i.v. LH-RH and to 2.5, 5.0, 10.0, and 20.0 microgram i.m. D-Leu-6-LH-RH-ethylamide was studied in 6 subjects with normogonadotrophic, idiopathic oligozoospermia. All injections were given between 8:00 and 9:00 a.m. at weekly intervals. Blood samples were collected before and 30 and 45 minutes after injection of LH-RH, and before and 2, 4, 6, 12, and 24 hours after D-Leu-6-LH-RH-ethylamide injections. Serum levels of LH, FSH, and testosterone were measured by triplicate by R.I.A. LH and FSH responses to LH-RH showed peak values at 30 minutes, whereas following injections of D-Leu-6-LH-RH-ethylamide, maximal values were recorded between 4 and 6 hours and were higher than those obtained with LH-RH. A significant rise of serum testosterone levels was observed after all stimuli, with peak values 45 minutes after LH-RH and 4 to 6 hours after D-Leu-6-Lh-RH-ethylamide injections. The testosterone levels obtained following LH-RH and different doses of the analogue did not differ significantly from one another. The same sequence of maximal LH and FSH values was obtained after injections of LH-RH and of D-Leu-6-LH-RH-ethylamide, particularly at doses of 10.0 and 20.0 microgram. This suggests that the gonadotrophin response to these agents would be a true reflection of the individual mode of pituitary response.

Luteinizing hormone (LH), follicle-stimulating hormone, and testosterone responses to consecutive injections of D-leucine-6-LH-releasing hormone ethylamide in normal men.

Luteinizing hormone (LH), follicle-stimulating hormone (FSH), and testosterone (T) responses to three consecutive intravenous injections of D-Leu-6-LH-releasing hormone ethylamide (D-Leu-6-LH-RH-EA) at 3-hour intervals were studied in six healthy, fertile, male volunteers 34.2+/-1.6 years of age. Each man received three injections of 20 microng of D-Leu-6-LH-RH-EA at 6:00 A.M., 9:00 A.M., and 12:00 noon, respectively. Blood samples were obtained before the first injection and at 1, 2, 3 (before the second injection), 4, 5, 6 (before the third injection), 7, 8 and 9 hours after the beginning of the test. Serum levels of LH, FSH, and T were determined by radioimmunoassay with the double-antibody technique. The response to the first injection of D-Leu-6-LH-RH-EA confirmed the longer duration of the stimulation of LH and FSH release caused by this compound as compared with that caused by LH-RH. Serum T levels rose significantly, almost paralleling the variations experienced with gonadotropins. The second injection caused a slight increase in LH and T responses in relation to the first injection. Two and three hours after administration, the third stimulus resulted in hormone levels lower than those obtained with the first two injections. Possible explanations for this finding might be a reduction of pituitary responsiveness as a result of multiple stimulation with D-Leu-6-LH-RH-EA, or spontaneous circadian variation of the pituitary response, or a combination of factors.

Pituitary and ovarian response to acute stimulation with LH-RH in normal and anovulatory women.

The LH FSH estradiol and progesterone responses to acute stimulation with LH-RH were studied in 12 normal women with ovulatory cycles (4 in the initial follicular phase, 4 in the mid-follicular phase and 4 in the late follicular phase) and in two castrated women, two under hormonal contraception, two with ovarian amenorrhea, twelve with central amenorrhea of no detectable origin (6 with normal and 6 with low basal gonadotrophins), eleven anovulatory patients with pseudomenstruation, two with anorexia nervosa, and two with pituitary amenorrhea. Each woman received a rapid i.v. injection of 100 microgram synthetic LH-RH at 9:00 a.m. Serum levels of LH, FSH, estradiol and progesterone were determined by radioimmunoassay in samples collected before and 60, 120, 240 and 480 minutes after injection. The findings were : 1) A significant rise in estradiol and progesterone levels, in addition to LH and FSH elevation, in normal women; 2) A lack of ovarian steroid response in the castrated women and in ovarian amenorrheas, which suggests that the source of steroid response to stimulation is not extragonadal; 3) Significant differences in the responses of the four hormones to LH-RH in the women with central amenorrhea in comparison with the normal group with great variability of results; the steroid response in the presence of a positive LH response might correlate with the severity and/or prognosis of the disorder, a point deserving further study; 4) In anovulatory women with pseudomenstruation, LH responses for the most part normal, and particularly, progesterone responses.

PIP: Simultaneous pituitary and ovarian responses to acute stimulation (100 mcg iv injection) with luteinizing hormone-releasing hormone (LH-RH) in normal women at different times of the menstrual cycle were determined and the results were compared with those obtained in women with anuvulation from different causes. There were 12 normal women, 2 women who had had surgical oophorectomy, 2 who were taking combined hormonal contraceptives, 1 with amenorrhea following pelvic irradiation, 1 with gonadal dysgenesis, and 2 with anorexia nervosa. There were also 12 patients with secondary amenorrhea without detectable pathology. All patients received an iv injection of 100 mcg of synthetic LH-RH. In the normal patients basal LH levels were significantly (p .05) higher on Days 13-14 of the cycle than on Days 4-5. In all 3 phases of the cycle, LH reached peak poststimulation levels within 60 minutes after LH-RH injection. Castrate women showed basal LH levels and LH response profiles similar to normals. There were no estadiol or progesterone responses in this group. Women using hormonal contraception showed low basal levels of both gonadotropins with poststimulation LH response but no follicle stimulating hormone response. Estradiol response in this group was slightly lower and more sustained than in normal women. In patients with ovarian amenorrhea, responses were similar to those in castrated women. In patients with anorexia nervosa findings were normal.