High vegetable and fruit diet intervention in premenopausal women with cervical intraepithelial neoplasia.

OBJECTIVE: To examine whether diet intervention can promote increased vegetable and fruit intake, as reflected in increased plasma carotenoid and decreased plasma total homocysteine concentrations, in premenopausal women with cervical intraepithelial neoplasia, a precancerous condition. DESIGN: Randomized controlled diet intervention study. SUBJECTS: Fifty-three free-living premenopausal women who had been diagnosed with cervical intraepithelial neoplasia, were randomly assigned to an intervention (n = 27) or a control (n = 26) group. INTERVENTION: Individualized dietary counseling to increase vegetable and fruit intake. MAIN OUTCOME MEASURES: Diet was assessed by food frequency questionnaire. Plasma carotenoids and total homocysteine were measured at enrollment and at 6 months follow up. ANALYSIS: Associations between baseline plasma concentrations of carotenoids and homocysteine and influencing factors were examined with multiple regression analysis. Repeated measures analysis of variance was used to test for group by time effects in these plasma concentrations. Plasma carotenoids at baseline and 6 months in the study groups, and differences in homocysteine concentrations from baseline to 6 months, were compared with independent sample t tests. RESULTS: Repeated measures analysis of variance showed significant group by time effects (P<.01) in plasma carotenoid and homocysteine concentrations. In the intervention group, total plasma carotenoids increased by an average of 91%, from 2.04+/-0.13 (mean+/-standard error of the mean) to 3.90+/-0.56 micromol/L and plasma total homocysteine was reduced by 11%, from 9.01+/-0.40 to 8.10+/-0.44 micromol/L (P<.003). Neither changed significantly in the control group. APPLICATIONS: Individualized dietary counseling can effectively promote increased vegetable and fruit intake in premenopausal women. This dietary pattern may reduce risk for cancer and other chronic diseases and also promote an improvement in folate status.

Neuropeptide Y (NPY) levels in alcoholic and food restricted male rats: implications for site selective function.

Neuropeptide Y (NPY) has been implicated in the control of ingestive, cardiovascular, and reproductive function. Blood pressure and sexual function were examined in Long-Evans rats receiving 6% ethanol-containing or calorically matched liquid diets, or rat chow ad lib. After 12 weeks of exposure, rats were sacrificed and plasma hormone levels and NPY content of microdissected brain regions were determined. Neither long-term alcohol ingestion nor caloric restriction were associated with major decrements in copulatory behavior. Long-term alcohol ingestion was associated with decrements in erectile function ex copula. Long-term alcohol ingestion was also associated with: (i) a moderate degree of hypertension; (ii) a failure to gain weight; (iii) decrements in circulating levels of LH, testosterone, and ACTH (but not progesterone); and (iv) increased corticosterone levels. Long-term alcohol ingesting and calorically-restricted rats exhibited alterations in daily feeding patterns. These physiological changes in response to long-term alcohol ingestion or caloric restriction were associated with neural site-selective differences in NPY content. Elevated NPY in the paraventricular nucleus was associated with voluntary (as in alcohol ingestion) or involuntary (as in caloric restriction) reductions in food intake. Differences in NPY in the suprachiasmatic and ventromedial nuclei were associated with the differences in feeding patterns. The decrements in hormone levels were associated with higher levels of NPY in the median eminence and in the arcuate nucleus.

Effects of angiotensin II on sexual function, blood pressure, and fluid intake are differentially affected by AT-1 receptor blockade.

We have previously reported that third ventricular administration of angiotensin II (ANG II) immediately before mating tests suppressed copulatory behavior in male rats. The present studies examine the effects of short- (3 days) and long-term (21 days) intracerebroventricular (i.c.v.) infusion of ANG II (6 microg/h), on parameters of copulatory behavior, fluid intake, and blood pressure in sexually experienced male Long Evans rats. Further, to test the hypothesis that suppression of masculine copulatory behavior by ANG II involves interaction with the angiotensin AT-1 receptor, a highly selective nonpeptide antagonist (L-158,809) was administered in the drinking water (25 mg/liter) to a group of ANG II-infused animals. I.c.v. infusion of ANG II was associated with increases in systolic blood pressure and fluid intake. In copulatory tests after 3, 9 and 15 days of infusion, rats infused with ANG II exhibited increased latencies to the initiation of copulatory behavior and to ejaculation, as well as increased intervals to reinitiate copulatory behavior after the ejaculation. Administration of L-158,809 blocked the effects of i.c.v. infusion of ANG II on systolic blood pressure and fluid intake. Further, L-158,809 attenuated the effects of i.c.v. infusion of ANG II on parameters of copulatory behavior. Data from this study provide support for a modulatory role for ANG II in the regulation of sexual behavior. In addition, this regulation seems to involve the AT-1 receptor.

Central propranolol and pindolol, but not atenolol nor metoprolol, inhibit sexual behavior in male rats.

Are the anti-sexual effects of propranolol and pindolol due to actions within the brain? To answer this, these agents were administered directly into the brain ventricular system (ICV). Additionally, atenolol and metoprolol were evaluated to see whether differential delivery to the brain contributed to the observed lack of effect of systemically administered beta 1-adrenoceptor antagonists. ICV administration of pindolol (45 or 90 micrograms) was followed by a suppression of copulation. At 45 micrograms, inhibition was limited to performance aspects of copulation, whereas at 90 micrograms, decrements in motivational and performance aspects of copulation were evident. ICV administration of propranolol also suppressed copulatory behavior. At 45 micrograms, no significant effects were observed, whereas at 90 micrograms decrements in motivational and performance aspects of copulation were evident. In contrast, ICV administration of the beta 1-adrenoceptor antagonists, atenolol and metoprolol, was not associated with any major modifications in copulatory behavior. We suggest that the inhibitory effects of propranolol and pindolol may involve interactions with 5-HT1A receptors in the CNS. Alternatively, it may be that the adverse effects of pindolol and propranolol are due to the simultaneous blockade of both beta 1- and beta 2-adrenoceptors.

The inhibitory effects of propranolol on genital reflexes in male rats.

We have previously reported that propranolol adversely affects sexual behavior in male rats. To elucidate whether the effects of propranolol might involve decrements in ability, we examined two components of sexual function ex copula--ejaculatory reflex capacity and erectile reflexes. In the first study, we examined the effects of various doses of (+/-)-propranolol (1.25-10 mg/kg) administered subcutaneously. Marked inhibition was observed, evidenced by increases in the latency to ex copula ejaculation and to initial erection and decrements in the number of seminal emissions and in the number of erectile reflexes. Analyses of dose-response relationships indicated that the degree of inhibition increased with increasing dose. In the second study, we evaluated the stereo-selectivity of the responses. Both (+)- and (-)-propranolol (1.25 mg/kg) significantly inhibited ejaculatory reflex potential, and although (+)- and (-)-propranolol significantly inhibited erectile reflexes, (-)-propranolol had a greater effect. The data are interpreted to indicate that a) propranolol-induced sexual dysfunction involves both motivational and ability aspects; and b) propranolol-induced inhibition of genital reflexes may be due, at least in part, to mechanisms other than beta-adrenoceptor blockade.

The inhibition of sexual behavior in male rats by propranolol is stereoselective.

We have previously reported that administration of racemic mixtures of propranolol was associated with a marked inhibition of mating behavior in male rats. To compare the effects of (+)-propranolol, (-)-propranolol, and (+/-)-propranolol in sexually experienced males, rats ejaculating in four or more mating tests were divided into three groups (N = 16 per group) such that no differences in parameters of copulatory behavior were evidence in preexperimental tests. No major effect of propranolol on parameters of behavior associated with initiation of sexual behavior was evident. In contrast, other measures of behavior were profoundly modified. The ejaculatory threshold, indicated by the number of intromissions preceding ejaculation, was increased after (+)- and (+/-)-propranolol, but not (-)-propranolol. The number of mounts without intromission preceding ejaculation was increased only after (+/-)-propranolol. A decrease in copulatory efficacy was evident after (-)- or (+/-)-propranolol, but not after (+)-propranolol. Increases in ejaculation latency, intercopulatory interval, and postejaculatory interval were observed after (-)- and (+/-)-propranolol, but not after (+)-propranolol. In summary, the present data indicate that the (-) isomer of propranolol is the active form necessary for the inhibitory effects of propranolol on male sexual function. We suggest that this inhibition is due to specific receptor-mediated mechanisms, involving beta-adrenoceptors and 5-HT1A receptor interactions.

Chronic ethanol ingestion alters parameters of mid-latency auditory evoked potentials in male rats.

The purpose of this study was to determine the effects of chronic ethanol ingestion on components of mid-latency auditory evoked potentials (MAEPs). Male Sprague-Dawley rats were administered 10% ethanol in drinking water for 10 months. MAEPs were obtained and compared to age-matched controls provided tap water. Data were obtained for varying frequencies (4, 8, 16, 24, and 32 kHz) and intensities (65, 75, and 85 dB sound pressure level). Ethanol treatment was associated with increased latencies, as well as decreased amplitudes of Na and Pa. The effects were most prominent for MAEP component Pa, but also appear for component Na. We suggest that chronic alcohol consumption induces structural and/or neurochemical alterations in substrates for cortical information processing.

Sexual function in altered physiological states: comparison of effects of hypertension, diabetes, hyperprolactinemia, and others to "normal" aging in male rats.

In this review, we examine the changes in sexual function that accompany deviations from "normal" physiological states. We propose that the changes one observes in many altered physiological states should not be viewed in isolation. We describe our paradigms for assessing sexual function, and proceed to evaluate how sexual function changes with hormonal deprivation and aging, in rat models for hypertension, in severe hyperprolactinemia, in streptozotocin-induced diabetes, after chronic alcohol intake, after chronic morphine administration, and after exposure to the heavy metal, cadmium. We will provide evidence for the involvement of adrenergic transmitters and two neuropeptides, neuropeptide Y and somatostatin, in the neuroendocrine regulation of sexual behavior. Finally, we compare and contrast the changes observed relative to the changes seen in "normal" aging in rats. The sequence of age-related changes in sexual function is distinct. The first change observed is a decrement in ex copula erectile reflexes. Next are decreases in ejaculatory threshold, followed shortly by increases in initiation and reinitiation of copulation after ejaculation. This is followed by a decrement in the number of males copulating to ejaculation. Finally, there is a failure to initiate the copulatory process. This sequelae is relatively common, being evident after castration, with hyperprolactinemia, and after exposure to cadmium. The data available for sexual function in hypertension is incomplete and modified by the etiology, but a suggestion for this sequelae is seen in SHR. In contrast, sexual dysfunction associated with chronic morphine administration appears to be due to an initial deficit in motivational aspects. Testosterone reverses sexual dysfunction associated with castration, but not with idiopathic sexual inactivity, nor with sexual dysfunction associated with aging, diabetes, or chronic morphine administration. Comparing sexual function in rat models for hypertension, diabetes and chronic ethanol leads to the conclusion that increases in blood pressure, like decreases in testosterone, cannot be the primary causal factor for sexual dysfunction. Age, hormonal history of the subject, and the age at castration influence changes in sexual function. Age-related sexual dysfunction appears to be contributed to by changes in adrenergic-neuropeptidergic, to include sympathetic, systems. Site-specific administration of NPY induces alterations in parameters of copulatory behavior which mimic those seen in aging and the retention of ejaculatory behavior with aging is associated with site-selective attenuation (or reversal) of age-associated changes in NPY content. Yohimbine enhances copulatory activity in castrated and aging rats, and attenuates or reverses the antisexual effects of clonidine, epinephrine and somatostatin.(ABSTRACT TRUNCATED AT 400 WORDS)

Testosterone-induced copulatory behavior is affected by the postcastration interval.

A well entrenched hypothesis regarding hormonal action is that as the time interval following hormonal deprivation increases there is a corresponding decrease in the sensitivity of the system to the effects of hormone replacement. With this in mind, we examined the effects of a prolonged period of hormonal deprivation (9 mo), and compared these to the effects of a shorter period (1 month), on the restoration of copulatory behavior and seminal vesicle weights. Castration of sexually vigorous male Long-Evans rats at 6 mo of age was followed by the virtual disappearance of ejaculatory behavior within 1 mo. Testosterone (T) was administered (5 mm or 20 mm T-containing Silastic capsules) either 1 or 9 mo after castration, and copulatory tests were conducted 3, 7, 10, 14, and 17 days later. 5 mmT and 20 mmT were equally effective in restoring behavior in the rats treated 1 mo after castration. In contrast, 5 mmT was more effective in inducing copulatory behavior than 20 mmT in the rats treated 9 mo after castration. The time course to maximal effect was longer in the rats given T 9 mo after castration. Rats were sacrificed 21 days after T administration. Expressed seminal vesicle weights and plasma testosterone were increased in a dose-dependent manner independent of the postcastration interval. These data indicate that somatic and behavioral effects of T are differentially modified by the period of preceding hormonal deprivation.

Cadmium-induced sexual dysfunction does not involve increased hepatic metabolism of testosterone nor increased circulating levels of corticosterone.

Sexually experienced male rats were injected IP with 0, 0.3, 1.5, 3.0, or 6.0 mg/kg cadmium chloride. The highest dose was fatal within 48 h of injection. A dose-related deficit in erectile function was observed in ex copula tests 48 h after injection. Copulatory dysfunction was evident in mating tests 72 h after injection. Hepatic mixed function oxidase activity after 0.3 mg/kg injections was not different from controls, but was diminished by 50% in rats treated with 3.0 mg/kg. The higher cadmium doses (1.5 and 3.0 mg/kg) were associated with equivalent and marked reductions in circulating testosterone levels, and lesser decrements in circulating corticosterone levels. The rats treated with 1.5 or 3.0 mg/kg cadmium chloride lost weight (32 +/- 12 and 39 +/- 9 g, respectively). Relative decapsulated adrenal gland weights were markedly increased in rats treated with 3.0 mg/kg. These data indicate that exposure to cadmium is associated with alterations in sexual, hepatic, and adrenal function, with erectile dysfunction occurring at the lowest dose. The effects on copulatory behavior are especially striking because it requires some weeks for copulatory behavior to wane following castration.

Aging-induced decrements in neuropeptide Y: the retention of ejaculatory behavior is associated with site-selective differences.

Neuropeptide Y (NPY) has been implicated in the control of reproductive and cardiovascular function. We observed an age-related decrease in the number of males copulating to ejaculation and a moderate systolic hypertension in middle-aged (16- to 17-month-old) rats. NPY content was examined in microdissected brain nuclei in 5 groups of rats: 2 groups of young rats, 1 heterosexually naive and the other ejaculating in 3 successive mating test; 3 groups of middle-aged rats, 1 heterosexually naive, 1 group that had extensive sexual experience but failed to ejaculate in tests at 16.5 months of age, and the third continuing to ejaculate at 16.5 months of age. NPY levels were found to vary depending on the brain area, the age of the animals, and the maintenance of ejaculatory behavior. In sexually naive middle-aged males, NPY levels were uniformly lower than in younger males. There were no differences in NPY levels of young animals, regardless of sexual experience. In the medial preoptic area, the group that retained ejaculatory behavior through 16.5 months of age, had higher levels of NPY than those observed in young sexually experienced rats. In sexually experienced rats that were no longer ejaculating at 16.5 months of age levels were lower than all other groups except the sexually naive middle-aged group. In the hypothalamic arcuate nucleus, levels were equivalent in the young groups and in the middle-aged rats that retained ejaculatory behavior, being greater than in the middle-aged rats that were no longer ejaculating or were sexually naive.(ABSTRACT TRUNCATED AT 250 WORDS)

Benextramine, a putative neuropeptide Y receptor antagonist, attenuates the termination of receptivity.

Sexual behavior in female rats is dependent on gonadal steroids. In ovariectomized rats, progesterone treatment typically exerts a biphasic effect on copulatory behavior induced by prior treatment with estradiol. Thus, there is an initial augmentation of the facilitative effect of estradiol occurring 4-10 h after progesterone. This is followed by a profound inhibitory effect, essentially terminating receptivity. We hypothesized that the receptivity terminating effect of progesterone could be due to increased synthesis and release of neuropeptide Y in relevant brain regions. Rats were tested for female sexual behavior 4 h after progesterone (52 h postestradiol). Immediately following this test, benextramine was administered (0, 3, or 15 mg/kg, IP). Subsequently, behavioral tests were administered 24, 48, 72, and 96 h postbenextramine. Benextramine treatment attenuated the inhibitory effects of progesterone on receptivity (lordosis quotients and percent of responding animals) without affecting either proceptive or rejection behaviors. These data suggest that blockade of NPY (and alpha-adrenergic) receptors is associated with selective enhancements of specific components of sexual behavior in female rats. Specifically, blockade of NPY receptors by benextramine is associated with continued receptivity.

Hormones and sexual behavior in relationship to aging in male rats.

To determine if the age-related decline in male sex behavior is correlated with hormonal factors, a longitudinal study was conducted. Sexually experienced males were given mating tests every 2 months from 7 through 27 months of age. To study possible relationships between changes in behavior and alterations in hormone levels, blood samples were taken before and after these bimonthly tests. At 23 months, cross-sectional studies were also conducted comparing results to those obtained in 5-month-old males. Significant changes in mating behavior first appeared at 11 months; mount latency, intromission latency, ejaculation latency, postejaculatory interval, and intercopulatory interval were increased. Similarly, detectable decreases in testosterone (T) also occurred at this age. A significant decline in luteinizing hormone (LH) was not seen until 19 months. Correlational analyses revealed small (r less than or equal to -0.29) but significant negative correlations between T and parameters of mating behavior with age. When each age was examined separately, no significant correlations appeared. Plasma T was not predictive of behavioral performance. At 23 months, cross-sectional studies revealed deficits in mounting and penile reflex behavior but ejaculatory reflex capacity was unimpaired. At 28 months, males were decapitated. Only T levels showed a significant effect of age; estradiol, prolactin, and LH were unaffected when compared to 5-month-old males. The data suggest that although there are small and significant negative correlations between circulating testosterone and parameters of mating behavior with advancing age, it is unlikely that the observed decline in testosterone is the primary cause of the age-induced behavioral deficits. It is likely that the major causal factor(s) involves non-hormone-dependent changes within the CNS.

Sexual function and neuropeptide Y levels in selected brain regions in male spontaneously hypertensive rats.

Sexual function was examined in spontaneously hypertensive rats (SHR) from 8 to 20 wk of age and compared with normotensive Wistar-Kyoto (WKY) and Long-Evans rats (LE). Blood pressures (evaluated indirectly) were elevated in SHR (185 +/- 2 and 195 +/- 3 mmHg at 16 and 19 wk of age, respectively) relative to WKY and LE (135-144 mmHg). SHR exhibited good copulatory behavior but displayed fewer erections (less than 20% of the number displayed by WKY or LE) in ex copula tests. At the conclusion of the study (20 wk of age), body weights were lowest in SHR, intermediate in WKY, and greatest in LE. Relative weights of testes were greater in SHR, whereas relative weights of accessory organs, pituitary and adrenal glands, and kidneys were equivalent across strains, as were circulating levels of aldosterone. Circulating levels of testosterone were higher in SHR and WKY than in LE. Neuropeptide Y (NPY) levels in the median preoptic and arcuate nuclei were significantly greater in SHR than in WKY or LE, whereas NPY levels in the medial preoptic area and the suprachiasmatic, hypothalamic dorsomedial, and hypothalamic paraventricular nuclei were equivalent in SHR and WKY, with both greater than LE. No strain differences were evident in the medial nucleus of the amygdala, the bed nucleus of the stria terminalis, the median eminence, the anterior hypothalamic nucleus, or the hypothalamic dorsomedial nucleus.

Suppression of copulatory behavior in male rats following central administration of clonidine.

Adrenergic transmitters have been implicated in the regulation of male sexual behavior. In the present study the contribution of alpha 2-adrenoceptors, located within the central nervous system, was evaluated. Sexually experienced male Long-Evans rats were implanted with a permanent cannula in either the third cerebral ventricle or, unilaterally, in the medial preoptic area. Baseline mating tests were administered 4, 7 and 10 days after surgery and only males ejaculating (at least) in the 7 and 10 day tests were used. Clonidine-evoked dose-dependent decrements in the number of males mounting, intromitting and ejaculating, with administration into the medial preoptic area more effective than into the 3rd ventricle. In those animals mating, administration of 20 nmol into the 3rd ventricle was associated with decreases in the latency to ejaculation and intercopulatory interval, whereas administration of 2 nmol into the medial preoptic area was associated with increases in these parameters and decreases in the ejaculatory threshold. Administration of yohimbine into the medial preoptic area attenuated the effects of systemically-administered clonidine and the systemic administration of yohimbine completely prevented the copulatory suppression induced by administration of clonidine into the medial preoptic area. It is suggested that central alpha 2-adrenoceptors are important in the control of male sexual behavior and that alterations in adrenergic mechanisms in the medial preoptic area may underlie sexual dysfunction of various etiologies.

Structural and antigenic polymorphism of the 35- to 48-kilodalton merozoite surface antigen (MSA-2) of the malaria parasite Plasmodium falciparum.

Merozoite surface antigen MSA-2 of the human parasite Plasmodium falciparum is being considered for the development of a malaria vaccine. The antigen is polymorphic, and specific monoclonal antibodies differentiate five serological variants of MSA-2 among 25 parasite isolates. The variants are grouped into two major serogroups, A and B. Genes encoding two different variants from serogroup A have been sequenced, and their DNA together with deduced amino acid sequences were compared with sequences encoded by other alleles. The comparison shows that the serological classification reflects differences in DNA sequences and deduced primary structure of MSA-2 variants and serogroups. Thus, the overall homologies of DNA and amino acid sequences are over 95% among variants in the same serogroup. In contrast, similarities between the group A variants and a group B variant are only 70 and 64% for DNA and amino acid sequences, respectively. We propose that the MSA-2 protein is encoded by two highly divergent groups of alleles, with limited additional polymorphism displayed within each group.

Steroid sulfatase activity in the skin of the external ear canal.

Steroid sulfatase is an enzyme which has an important role in the complex process of cell desquamation. The specific activity of this enzyme was found to be higher in the epithelium of the deep external auditory canal than in the epithelium of the superficial external canal. We speculate that this steroid sulfatase activity gradient is involved in the outward migration of the keratinocytes of the canal epithelium.

Clonidine suppresses copulatory behavior and erectile reflexes in male rats: lack of effect of naloxone pretreatment.

Adrenergic transmitters and opioid peptides are implicated in the regulation of male sexual behavior. In the present studies we examine a possible interaction between these two neurochemical systems in the regulation of components of male sexual behavior. In mating tests, clonidine (0.25 mg/kg, 6 min pretest) induced a profound deficit in intromissive and ejaculatory behavior whereas naloxone (5 mg/kg, 20 min pretest) evinced a facilitation of ejaculatory behavior, evidenced by decreases in the ejaculation latency in the initial copulatory series and by decreases in ejaculation latency and intercopulatory interval in the second copulatory series. Importantly, prior treatment with naloxone did not prevent or attenuate the copulatory suppression induced by clonidine. In ex copula penile reflex tests, clonidine (0.25 mg/kg, 6 min pretest) decreased the incidence of seminal emission and the number of penile responses (erections, cups and flips). Naloxone (5 mg/kg, 20 min pretest) was without effect on any of the parameters of penile reflex activity and, further, failed to prevent or attenuate the erectile suppression induced by clonidine. A final study evaluated the dose-response relationship of clonidine-induced erectile dysfunction. A similar degree of erectile dysfunction was observed after 0.005, 0.025 or 0.25 mg/kg clonidine, whereas 0.0005 mg/kg was without effect. Previous studies demonstrated a dose-dependent inhibition of mounting, intromissive and ejaculatory behavior, with 0.25 mg/kg selectively eliminating ejaculatory behavior in mating tests. These data demonstrate a dose-dependent inhibition of penile reflexes, with inhibition occurring at doses lower than those required to induce copulatory dysfunction. Further, we suggest that opioid receptors sensitive to naloxone blockade are not involved in the clonidine-induced suppression of copulatory or erectile function.