Neuronal cell bodies in the hypothalamic paraventricular nucleus mediate stress-induced renin and corticosterone secretion

The present studies were undertaken to determine the involvement of neurons in the hypothalamic paraventricular nucleus (PVN) in stress-induced renin secretion. The stressor was a 10-min conditioned emotional response (CER) paradigm. Bilateral electrolytic lesions in the PVN prevented the stress-induced increase in plasma renin activity (PRA), and plasma renin concentration (PRC). Stress-induced corticosterone secretion was also blocked, supporting the histological verification and suggesting that the lesion included corticosterone-releasing factor neurons in the PVN. Stress-induced renin secretion appears to be restricted to the PVN, as electrolytic lesions in the nucleus reuniens, dorsal and caudal to the PVN, did not prevent the stress-induced increase in either PRA or PRC. The next step was to determine whether cell bodies in the PVN or fibers of passage through the PVN mediate the stress-induced increase of these hormones. For this purpose, bilateral stereotaxic injections of the cell-selective neurotoxin ibotenic acid (10 micrograms/microliter; 0.3 microliters per side) were performed 14 days prior to the stress procedure. Histological evaluation of the tissue revealed cell death and lysis in the PVN. Ibotenic acid injection into the PVN prevented the effect of stress on PRA, PRC and corticosterone levels. None of the lesions prevented the stress-induced rise in plasma prolactin concentration. These results suggest that neurons in the PVN play an important role in mediating stress-induced increases in renin and corticosterone but not prolactin secretion.     

Insulin-like growth factor I promotes leiomyoma cell growth in vitro

Objective: Our purpose was to determine whether insulin-like growth factors I and II preferentially stimulate uterine leiomyoma cells versus myometrial cells in monolayer culture.Study design: Leiomyomas and normal myometrium were obtained at hysterectomy from five premenopausal women. Specimens were enzymatically digested for use in primary monolayer cell cultures. By use of serum-free media, insulin-like growth factor I or II was added in 1, 10, and 100 ng/ml concentrations to both cell types with the patient serving as her own control. Cell number, prolactin production, and proliferative index values were measured on day 15 of cell culture.Results: Significant increases in cell number were found in the leiomyoma cultures (p < 0.05) treated with 10 and 100 ng/ml insulin-like growth factors I but not with insulin-like growth factos II. Neither factor exerted a stimulatory effect on myometrial cells.Conclusion: Insulin-like growth factor sI preferentially stimulates leiomyoma cells in monolayer culture. These results suggest an autocrine-paracine role in vivo for this factor in conjuction with gonadal steroids in promoting leiomyoma growth.     

Reproductive hormone levels after pituitary allograft in cyclosporin-treated monkeys

OBJECTIVE: To determine the course of events during the onset of hyperprolactinemic amenorrhea, a nonhuman primate model was sought that did not require suckling or interference with the in situ hypothalamic-pituitary axis. DESIGN: Because removal of the adenohypophysis from hypothalamic influence results in secretion of large quantities of prolactin (PRL) but little of the other adenohypophyseal hormones, we explored the possibility of establishing pituitary allografts in monkeys. Normally cycling female rhesus monkeys were immunosuppressed with a daily regimen of cyclosporin A (CyA; 10 to 15 mg/kg per day) and then subcutaneously grafted with a pituitary from another animal (allograft). Blood samples were obtained daily via saphenous vein puncture during control, only CyA-treatment, and allografted-plus CyA- menstrual cycles. SETTING: Oregon Regional Primate Research Center, Beaverton, Oregon. PARTICIPANTS: Female Macaca mulatta exhibiting regular menstruation. INTERVENTIONS: None. MAIN OUTCOME MEASURES: Prolactin, luteinizing hormone (LH), estradiol (E2), and progesterone (P) levels were determined in harvested serum. RESULTS: Temporary survival of 5 of 11 (45%) allografts was assumed based on elevations in serum PRL. Of the viable grafts, 4 of 5 (80%) resulted in reproductive dysfunction, as first evidenced by delay or loss of the preovulatory rise in E2. When the peak of follicular E2 was delayed, then the LH surge occurred, but it was also delayed. If follicular E2 levels did not peak, then the LH surge was absent as was luteal P production. CONCLUSION: These data suggest that in the etiology of PRL-induced infertility in women, the first event is a suppression of follicular E2 production. In addition, the hypothalamus probably remains responsive to the positive feedback of E2 during early or moderate hyperprolactinemia.     

Hepatic microvascular development in relation to the morphogenesis of hepatocellular plates in neonatal rats

The development of hepatic microvascular heterogeneity after birth, and its temporal relationship to the development of parenchymal cell plates have received little attention. As a result, the morphogenesis of some of the parameters contributing to this heterogeneity in suckling and weaned rats was studied as a function of time between postpartum days 4 and 30 using in vivo light microscopic, electron microscopic, and immunocytochemical methods. During the early suckling period, the sinusoid network is highly anastomotic, with little evidence of zonation, and the parenchymal cell plates contain multiple cells and are irregularly arranged throughout the lobule. Sinusoidal endothelial fenestration is sparse at 4 days, but phagocytic Kupffer cell (KC) function already exists and exhibits zonal heterogeneity, with more cells located in the periportal zone. With increasing age, endothelial fenestrae increase and organize as sieve plates. Widened centrilobular radial sinusoids form through a loss ("drop-out") of intersinusoidal sinusoids (ISS). Concomitantly, the associated cell plates straighten and become one cell thick. Hepatocyte DNA synthesis and mitosis are higher in the periportal zone, which retains thickened cell plates and anastomotic sinusoids. The centrilobular sinusoids may widen to accommodate the increased volume of blood that results from the loss of ISS as well as the increased numbers of periportal sinusoids containing flow that feed these vessels. KC phagocytic activity increases during the suckling period concomitant with an increase of gut-derived endotoxin in the portal blood, which suggests that the KCs may be releasing mediators that affect sinusoid diameter, blood flow, endothelial fenestration, and perhaps parenchymal growth either directly or through the stimulation of growth factors.     

Pharmacy mass casualty disaster plan implemented after the train wreck

The involvement of a hospital pharmacy department in a mass casualty disaster revealed strengths and weaknesses of the department's disaster response plan. Effective communication and notification within the pharmacy department and the need to plan to have pharmacists in the triage area to determine patients' lost medications are important areas that the department had not practiced in disaster drills.     

Protection of breastfeeding in Papua New Guinea

In Papua New Guinea the bottle-feeding of babies has been increasing, predominantly among unemployed women of low educational status. Many women are unaware of their legal right to have breaks at work for the purpose of breastfeeding, and a high proportion of workplaces have no facilities for mothers who wish to breastfeed their children. The laws on the feeding of infants should be updated and implemented, and an effort is needed to explain the benefits of breastfeeding and the rights of working mothers.     

The use of patient mix‐adjusted control charts to compare in‐hospital costs of care

We introduce a technique for patient mixadjusting charts and compared differences between unadjusted and patient mixadjusted results. Our data came from coronary artery bypass graft (CABG) surgery patients at Baptist Medical Center, Oklahoma City, Oklahoma. We first developed an unadjusted control chart to compare monthly changes in CABG surgery costs and then used a published model to patient mixadjust our control chart information. Before adjustment, the average log costs for three of ten months were outside the 90% control limit lines, and there was a trend toward increasing costs. After adjustment, two months had average costs outside the 90% lower control limit lines, and the trend toward increasing costs had been explained by differences in patient acuity.     

Estrogen and progestin receptor immunocytochemistry in lactotropes versus gonadotropes of monkey pituitary cell cultures

The distribution of estrogen receptors (ER), progesterone receptors (PR), PRL, and gonadotropins in different cell types of the monkey pituitary was examined by immunocytochemical (ICC) labeling of pituitary cell cultures. Dispersed monkey pituitary cells were cultured on extracellular matrix and in serum-free medium for 6-14 days. Individual cultures were singly stained for ER, PR, PRL, LH, and FSH or double labeled for PR and one of the protein hormones. ICC reaction product localizes over the nuclei of cells that are positive for the steroid receptors, whereas reaction product localizes over the cytoplasm of cells that are positive for the protein hormones. Sixty-one percent of the parenchymal cells were positive for PRL, while 1-3% were positive for LH or FSH. Sixty-two percent of the parenchymal cells were ER positive. ER staining was localized over the nuclei of two morphologically distinct cell types. One cell type is smaller and more prevalent than the second cell type. Based on single staining for each of the protein hormones, we propose that the smaller cells are lactotropes, and the larger cells are gonadotropes. PR-positive cells averaged 7.7% of the parenchymal cells. Double ICC staining for PR and the protein hormones demonstrated that PR localize in the nuclei of gonadotropes, but not lactotropes, of monkey pituitary cell cultures. The absence of PR in lactotropes is consistent with our observation that progesterone has no direct effect on PRL secretion in monkey pituitary cell cultures. In contrast, the presence of PR in gonadotropes suggests that progesterone may act directly at the pituitary to modulate gonadotropin secretion in the primate. In conclusion, ER are present in both lactotropes and gonadotropes. PR are present in gonadotropes, but not lactotropes, of the primate pituitary.     

Ovarian Regulation of Kisspeptin Neurones in the Arcuate Nucleus of the Rhesus Monkey (Macaca mulatta)

Tonic gonadotrophin secretion throughout the menstrual cycle is regulated by the negative‐feedback actions of ovarian oestradiol (E₂) and progesterone. Although kisspeptin neurones in the arcuate nucleus (ARC) of the hypothalamus appear to play a major role in mediating these feedback actions of the steroids in nonprimate species, this issue has been less well studied in the monkey. In the present study, we used immunohistochemistry and in situ hybridisation to examine kisspeptin and KISS1 expression, respectively, in the mediobasal hypothalamus (MBH) of adult ovariectomised (OVX) rhesus monkeys. We also examined kisspeptin expression in the MBH of ovarian intact females, and the effect of E₂, progesterone and E₂ + progesterone replacement on KISS1 expression in OVX animals. Kisspeptin or KISS1 expressing neurones and pronounced kisspeptin fibres were readily identified throughout the ARC of ovariectomised monkeys but, on the other hand, in intact animals, kisspeptin cell bodies were small in size and number and only fine fibres were observed. Replacement of OVX monkeys with physiological levels of E₂, either alone or with luteal phase levels of progesterone, abolished KISS1 expression in the ARC. Interestingly, progesterone replacement alone for 14 days also resulted in a significant down‐regulation of KISS1 expression. These findings support the view that, in primates, as in rodents and sheep, kisspeptin signalling in ARC neurones appears to play an important role in mediating the negative‐feedback action of E₂ on gonadotrophin secretion, and also indicate the need to study further their regulation by progesterone.