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Biomedical subjects

B L Lasley

Publications and source records attributed to B L Lasley.

161 records · Page 9Linked to original sources
PubMed1977

The two pools of pituitary gonadotropin: regulation during the menstrual cycle.

Information on the relative activity and on the functional relationships between the acutely releasable (1st) and reserve (2nd) pools of pituitary LH during the course of the normal menstrual cycle was obtained via a 4 h LRF infusion (0.2 mug/min X 4 h). This was immediately followed by 3 pulses of LRF (10 mug at 2 h intervals) to assess further the size of the acutely releasable pool after activation of the reserve pool by the infusion. Our observations indicate that two functional pools of LH are present in all phases of the menstrual cycle and that comparative pool size or activity is influenced profoundly by ovarian steroid feedback as well as by the pattern of input of hypothalamic LRF. From the early to the late follicular phase, in synchrony with the rising levels of E2, the size of the 2nd pool is preferentially augmented. A small increase in the 1st pool activity is not apparent until the late follicular phase when a 5-fold increase in the size of the 2nd pool is also attained. During the mid-luteal phase and in association with relatively high progesterone (P) and E2, th large 2nd pool is maintained as in the late follicular phase but the 1st pool is strikingly smaller. Activation of the 2nd pool of LH by the LRF infusion (priming) increases the acutely releasable LH (1st pool) in all three phases of the cycle, as evidenced by an enhanced response to the 1st but not subsequent pulses of LRF at the end of the infusion as compared with non-infused controls. This priming effect is likely a reflection of activation or "shifting" of LH from the larger 2nd pool to the smaller 1st pool. It is found that this priming effect is greatest during the mid-luteal phase as compared to other phases of the cycle. During the days of mid-cycle LH surge, a dramatic reversal of the relative activity of the two pools in observed and this is manifested by an enormous increase in the activity of the 1st relative to the 2nd pool. In contrast to other phases of the cycle, the release of LH from the 2nd pool is not sustained and this premature decline in LH release despite continuous LRF infusion appears to be due to pituitary depletion of LH as evidenced by the failure of the pituitary response to pulses of LRF immediately following the infusion...

Estradiol↗
PubMed1976

A simple method for the assay of eight steroids in small volumes of plasma.

A simple method is described for the simultaneous radioligand assay of four delta5-3beta-hydroxysteroids adjacent to one another on the biosynthetic pathway (pregnenolone [1], 17alpha-hydroxypregnenolone, dehydroepiandrosterone and 5-androsterone-3beta, 17beta-diol), and their four delta4-3keto products (progesterone, 17alpha-hydroxyprogesterone, 4-androstene-3, 17-dione and testosterone). Two plasma aliquots are extracted and fractionated each for four steroids and individual corrections are made for losses. For fractionation, maximum use is made of the high resolution and reproducibility of celite minicolumns, using propylene glycol as stationary phase, and a discontinuous gradient of ethyl acetate in iso-octane as mobile phase. The fractions are then assayed in the appropriate radioligand end-assay system. Each assay was finally validated by demonstrating coincidence of peaks of immuno- and radioactive steroid in extracts of female plasma. Results in pre-pubertal girls and women in the follicular phase of the menstrual cycle suggest that the major change in adrenal steroid production at puberty may be an increase in 17, 20-desmolase activity. There appears to be little reversal of this change in adrenal function after ovariectomy.

Adult↗
PubMed1976

Transplacental gradients of sex-hormone-binding globulin in human and simian pregnancy.

An attempt has been made to clarify the degree and control of production of the concentration gradient of sex-hormone-binding globulin (SHBG) between human mother and fetus. SHBG was measured by an improved ammonium sulphate precipitation method using 5 alpha-dihydrotestosterone as ligand. Maternal and fetal (cord blood) plasma SHBG was measured at delivery in twenty-three normal, four twin and one anencephalic pregnancies and in non-pregnant subjects. Comparison was also made with two macaque species, the rhesus monkey (M. mulatta) and the bonnet monkey (M. radiata), sampled shortly after delivery. In the human, a highly consistent gradient (mean 21-fold) was found between mother and fetus, which was independent of fetal sex and weight. Fetal but not maternal SHBG was significantly higher in the twin pregnancies. Maternal SHBG was 5-fold higher than in forty non-pregnant women, and fell with a half-life of 7-1 days immediately post-partium. In the two monkey species, SHBG with similar binding-characteristics exists but levels were 2-5-fold higher in the non-pregnant adult females than in women. In pregnancy, maternal SHBG levels were unchanged in the bonnet (nine observations) and fell markedly in the rhesus monkey (twenty-three observations) ; levels in the infants were comparably low in all three species, though more variable in the monkeys. In seven bonnet moneky mother-infant paris sampled again 2 or 2 weeks later, SHBG had risen mardedly (mean 6-1-fold) in the infants as has been shown previously in the human. The species differences in the non-pregnant females and the mothers may in part be related to species differences in oestrogen production. The macaque moneky appears suitable for studying further the mechanism of the neonatal SHBG rise, in which oestrogens cannot be implicated but thyroid hormones may be involved.

Animals↗
PubMed1976

Gonadotropin secretion in response to low and high doses of LRF in normal and hypogonadal women (functional disparity of the gonadotrophs).

A comparison was made of gonadotropin release after an iv bolus of large (150 mug) and small (10 mug) doses of LRF during various phases of the menstrual cycle and in primary hypogonadal women. Gonadotropin release was quantitatively similar to small and large doses of LRF in hypogonadal women with low estradiol levels. In contrast, normal women from the early to late follicular phase of the cycle, the gonadotropin response to a small dose of LRF (10 mug) was not as great as that observed with a large dose of LRF (150 mug) and did not elicit the augmented gonadotropin response of the late follicular phase. During the mid-luteal phase, a significant enhancement of responses to small doses of LRF was observed. The sustained gonadotropin release induced by the large dose of LRF in the late follicular phase was no longer apparent during the mid-luteal phase, although the dose-related discrimination of LH release was maintained. These dose-related disparities in the gonadotropin release to LRF might represent functional expressions of the two-pool dynamics of the gonadotrophs induced by ovarian steroids.

Clinical Trials as Topic↗
PubMed1976

The functional changes of the pituitary gonadotrophs during the menstrual cycle.

Submaximal doses of LRF, administered over a period of several hours, either by repeated pulses (10 mug at 2 h intervals X 5) or by constant infusion (0.2 mug/min X 4 h), have permitted the assessment of changes in the releasable gonadotropin during the menstrual cycle. Quantitations in the acute releasable and ultimately releasable gonadotropins were made which represent, respectively, the sensitivity and reserve of the gonadotrophs. The functional expression of these two components of gonadotropin release exhibited profound changes during the menstrual cycle and were in synchrony with the cyclicity of ovarian steroid levels; during the early follicular phase, both sensitivity and reserve were at a minimum, but with increasing levels of E2, a preferential increase in reserve over sensitivity (P less than 0.005) was found. Although both sensitivity and reserve increased dramatically near the midcycle, the relative change in these two components was reversed from the late follicular phase to the midcycle surge. The presence of this phenomenon may be causally related to the development of a self-priming effect of LRF at this time, as evidenced by an augmentation of gonadotropin release to the 2nd pulse of LRF. Thus, a build-up in pituitary store consequent to the greater increase in reserve than in sensitivity, together with the appearance of the self-priming effect of LRF induced by progressively rising levels of E2, may constitute the essential dynamics required for the development of the midcycle gonadotropin surge. Pituitary sensitivity and reserve continued to be high during the early luteal phase but reduced progressively thereafter. In all studied, FSH responses were less obvious but showed remarkable parallelism to the pattern of LH responses. We have concluded that the functional capacity of the gonadotrophs exhibits a remarkable cyclic change and that the adenohypophysis represents a critical feedback site in the development of pre-ovulatory gonadotropin surge.

Adult↗
PubMed1976

Assessments of the functional capacity of the gonadotrophs in men: effects of estrogen and clomiphene.

Serum LH and FSH responses to serial injections of LRF with small (10 mug x 5), large (150 mug x 5), decremental (300 to 10 mug), and incremental (10 to 300 mug) doses at 2-hour intervals were assessed in eugonadal men. At constant doses, pulses of LRF induced pulsatile LH release which was qualitatively similar but quantitatively greater for the large than for the small dose of LRF. There were no periods of refractoriness or augmentation of subsequent responses from prior exposure to LRF when administered at 2-hour intervals. LH responses to incremental and decremental doses of LRF resulted in corresponding measurable changes in the magnitude of pituitary LH release. The FSH responses to pulses of LRF at all doses tested were uniformly small. These data suggest that analyses of the initial and integrated release (10 h experiment) to pulses of LRF may disclose the functional capacity of the gonadotrophs and that small variations in the endogenous LRF delivered may represent a significant factor in the control of LH release. The small dose (10 mug x 5) of pulses of LRF was utilized in the assessment of estrogen and clomiphene treatments on the functional capacity of the gonadotrophs in normal men. Compared with the pretreatment results, both constant doses of ethinyl estradiol (50 mug/day x 7 days) and incremental doses of estradiol benzoate (100 to 400 mug, twice daily injections x 4 d) induced an attenuation of the initial release as well as of the integrated response 10 h) to pulses of LRF. Clomiphene treatment (100 mg/day x 5 d), likewise, resulted in a reduction of gonadotropin release to all pulses of LRF. These data suggest that circulating estrogen in intact men may have both a negative and a positive feedback effect on the gonadotrophs and that the testicular estrogen secretion as well as the extraglandular sources of estrogen, may play a critical role in the regulations of gonadotropin secretion in man.

Adult↗
PubMed1975

The role of estrogen in the modulation of pituitary sensitivity of LRF (luteinizing hormone-releasing factor) in men.

The role of estradiol in modulating pituitary gonadotropin release in the human male was studied by evaluating the effects of clomiphene on the pituitary gonadotropin response to synthetic LRF (150 mug) in 6 eugonadal men. Administration of clomiphene (100 mg single daily dose time 5 days) induced a significant elevation of the basal levels of LH, FSH, estradiol and testosterone. However, the pituitary release of LH and FSH in response to LRF was markedly diminished by the clomiphene treatment. This finding suggests that in men, as in women, endogenous estradiol provides feedback regulation of gonadotropin output by the pituitary.

Adult↗
PubMed1975

The effects of estrogen and progesterone on the functional capacity of the gonadotrophs.

The functional capacity of the gonadotrophs under the influence of exogenous estrogen and progesterone was assessed by repeated stimulation with submaximal doses of LH-releasing factor (LRF) (10 mug at 2-h intervals) of subjects during the early follicular phase of the cycle and of hypogonadal women. The initial increment of peak serum LH and FSH concentrations after the first administration of LRF, was used to describe the pituitary sensitivity, and the integrated release during the 10 hours of LRF-stimulated pulses was utilized to approximate the pituitary gonadotropin reserve. During the early follicular phase, response to LRF stimulations was relatively stable with corresponding release of LH and FSH. An augmentation of sensitivity, as well as the reserve, for both LH and FSH was elicited by an incremental change in circulating estradiol levels, a change imposed by daily administration of estradiol benzoate for 4 days during the early follicular phase. Under the same conditions, the addition of progesterone (10 mg, im) at the end of the estradiol benzoate treatment induced a marked amplification of the estrogen-augmented pituitary gonadotropin sensitivity and reserve. The pituitary sensitivity, relatively higher than the reserve in hypogonadal subjects, was reversed by the administration of ethinyl estradiol (20-50 mug/day) for 7 days. These data indicate that the functional capacity of the gondotrophs is profoundly modulated by estrogen through relative changes in pituitary sensitivity and reserve, and that progesterone in low doses exhibited an amplifying effect on estrogen-primed gonadotrophs in both the pituitary sensitivity and the reserve.

Estradiol↗
PubMedDate not supplied

Gender differences in the mechanism of dioxin toxicity in rodents and in nonhuman primates.

This study examined the differences in mechanisms of toxicity when adipose cells from males and females were exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Glucose uptake by adipose tissue in vitro was decreased significantly in male guinea pigs within 1 d of intraperitoneal injection of TCDD, but there was no significant effect in females, even at 28 d after treatment. A similar difference between male and female guinea pigs was detected in the effect of TCDD on lipoprotein lipase (LPL) activity, except that a significant decrease in LPL activity was observed 28 d after treatment. Experiments with adipose tissue explants from untreated guinea pigs and macaques revealed similar gender differences in the effect of TCDD in vitro on glucose uptake and LPL activity. Both time-course studies and dose-response studies with TCDD in vitro confirmed the greater sensitivity of male tissues to TCDD toxicity. TCDD induced lipid peroxidation in the adipose tissues of male guinea pigs, while it had no effect in females. 3H-TCDD binding affinity studies in adipose explant tissues showed that tissues from male guinea pigs and monkeys had a higher binding capacity for TCDD than female tissues. TCDD induced a significant reduction in nuclear protein phosphorylation and an increase in cytosolic protein phosphorylation in adipose tissue from male guinea pigs; the effects in female tissues were opposite: nuclear protein phosphorylation increased and cytosolic protein phosphorylation decreased. In a cell-free system in the absence of the nucleus, adipose tissues from male guinea pigs and monkeys responded to TCDD with a rapid stimulation of tyrosine kinase activity but female tissues from both species had a significantly lower and slower response. TCDD induced the DNA binding of AP-1 in adipose tissues of male guinea pigs, but in female tissues TCDD reduced the DNA binding of AP-1. In summary, the results of this study demonstrate gender differences in the response of nonreproductive cells to TCDD. Some of these differences involve different mechanisms of toxicity in both the cytoplasmic and nuclear compartments of the cell.

Adipose Tissue↗
PubMedDate not supplied

Effect of dioxin on ovarian function in the cynomolgus macaque (M. fascicularis).

Ovarian function was evaluated in mature female cynomolgus macaques 443 to 625 days following a single oral exposure (1, 2, or 4 microg/kg BW) to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Urinary estrone conjugates (E1C), pregnanediol-3-glucuronide (PdG), and follicle stimulating hormone (FSH) were measured. Three of four animals in the high dose group had no evidence of menstrual cycles while animals in the low and medium dose groups plus one from the high dose group had cycles that were similar to those of control animals. The noncycling animals had baseline E(1)C concentrations without ovulatory midcycle peaks and monotonic PdG profiles. Mean FSH concentrations during the midfollicular phase of the medium dose group and during the entire cycle of the high dose group were elevated compared to those of the control group and the endometria of the noncycling animals were inactive. These data demonstrate that a single exposure of 4 microg/kg BW TCDD leads to long-term adverse effects on ovarian function in primates.

Administration, Oral↗
PubMedDate not supplied

Effect of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on chorionic gonadotropin secretion by human trophoblasts.

This study tests the idea that the environmental toxicant, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), affects human trophoblast differentiation and alters the secretion of chorionic gonadotropin (CG). Primary cultures of cytotrophoblast cells were incubated under differentiation-inducing and nondifferentiation-inducing conditions in the presence or absence of different concentrations of TCDD. Levels of immunoreactive CG as well as bioactive CG were measured in culture supernatants. TCDD caused a significant increase in the secretion of immunoreactive CG from differentiated trophoblast cultures but had no effect on the secretion of bioactive hormone. The net effect was a TCDD-dependent reduction in the CG bioactive/immunoreactive (B/I) ratio for differentiated trophoblast cultures. TCDD had no effect on immunoreactive or bioactive CG secretion by undifferentiated trophoblasts. Immunocytochemical studies showed that TCDD had no effect on the morphologic differentiation of trophoblast cells as determined by staining nuclei and desmosomal proteins. On the other hand, immunocytochemical staining for CG was increased in cells exposed to TCDD compared to control cells. These in vitro results support earlier in vivo studies in macaques suggesting that trophoblast is a target for TCCD and that TCDD-induced early pregnancy loss is accompanied by a decrease in the CG B/I ratio.

Cell Nucleus↗
PubMedDate not supplied

Mechanism of toxic action of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in cultured human luteinized granulosa cells.

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) caused a significant decrease in estradiol (E2) production when it was administered to human luteinized granulosa cells (hLGCs) in culture. We investigated the involvement of the epidermal growth factor receptor (EGFR) and protein tyrosine kinase (PTK) in this TCDD-induced toxicity. Upregulation in 125I-EGF binding to EGFR was measured after 24 h of TCDD treatment, while downregulation in EGFR binding was measured after 72 h of TCDD treatment. Upregulation of EGFR binding was associated with a significant decrease in postnuclear (7000 x g supernatant) PTK activity, but this activity was stimulated after 72 h of TCDD treatment. TCDD altered the level of tyrosine phosphorylation in proteins with molecular weights 35, 40, 43, 45, 60, and > 205 kDa. TCDD caused a significant increase in postnuclear cAMP-dependent protein kinase (PKA) after 24 h of treatment. The actions of TCDD on protein kinases were partially blocked by the protein synthesis inhibitor, cycloheximide. On the other hand, TCDD increased nuclear PTK and decreased nuclear PKA activity. E2 inhibited the postnuclear and nuclear activity of both PTK and PKA in control samples, but did not affect TCDD actions on either postnuclear or nuclear PTK activity. However, E2 abolished the stimulatory effect of TCDD on PKA activity in postnuclear protein. In the presence of insulin, TCDD did not induce any additional changes in postnuclear or nuclear PTK. Forskolin (FK) alone inhibited postnuclear PTK activity and stimulated its nuclear activity. The addition of TCDD 20 min after FK resulted in an increase in postnuclear PTK, but there was little change in nuclear PTK as compared to the effect of FK alone. The stimulatory effect of TCDD on postnuclear PKA activity was enhanced by insulin and TCDD reversed the negative effect of FK, but there was no effect of either insulin or FK on the inhibition by TCDD of nuclear PKA activity. TCDD decreased the activity of MAP2 kinase and reduced the binding activity of AP-1 DNA when given alone, and also blocked the E2 stimulation of MAP2K. These findings suggest that TCDD may interrupt the endocrine function of hLGCs through the blockage of the mitotic signal directly or indirectly through the interaction of PTK/MAP2K and PKA signaling.

Cell Cycle↗
PubMedDate not supplied

Classification of adverse reproductive effects can be improved by measurements of multiple biomarkers for ovarian toxicity and early fetal loss.

Hormone assays have been developed and applied for monitoring reproductive function using self-collected urine samples in non-clinical populations of women. Early pregnancy loss, menstrual dysfunction, reduced fertility as well as the site of toxicity can now be assessed using daily early morning urine samples. The understanding of the specific limitations of individual assay systems is important, however, to make the best use of these systems. The use of multiple end-points and computer algorithms is suggested to avoid misclassification of adverse reproductive events.

Algorithms↗
PubMedDate not supplied

Introduction: rationale for an update. Reproductive hazards.

More than 104,000 chemical and physical agents have been identified in the workplace, but the effects on reproduction of at least 95% of them have not been assessed. Here, the editors of this volume describe changing demographics in the workplace, which underscore the need for evaluating potential hazards to reproduction. They also review findings regarding occupational reproductive hazards to men and women that have been reported over the past decade.

Adult↗
PubMedDate not supplied

Methods for evaluating reproductive health of women.

The use of urine as a practical sample for evaluating the reproductive health of women in population-based studies is still being developed and applied. The adaptation of enzyme-based assays for the assessment of ovarian steroids as well as simple computer algorithms for interpreting the data both simplify and increase the efficiency of modern reproductive epidemiologic studies. New statistical approaches and new assays have extended physiologic monitoring to include early pregnancy, luteal phase function, and ovulation. While urine is superior to blood sampling, it is important to remember, however, the limitations of urinary hormone measurements when applying them alone to identify specific types of reproductive failure because variations in metabolism and excretion rate still contribute some uncontrollable variation. The rate of progress in developing and refining these methods over the past 5 years indicates that urinary methods of reproductive monitoring will continue to dominate this field and eventually be used in the clinic as well.

Abortion, Spontaneous↗