The type 2 corticotrophin-releasing hormone receptor mediates orexin A-induced luteinising hormone suppression in ovariectomised rats.

Orexins are thought to be regulatory factors of the arousal and sleep patterns. They also affect immune, feeding, autonomic and neuroendocrine systems. We have previously shown that intracerebroventricular (i.c.v.) injection of orexin decreases pulsatile luteinising hormone (LH) secretion in ovariectomised (OVX) rats. However, the details of this mechanism have not been fully examined. Intracerebroventricular injection of orexin A also stimulates corticotrophin-releasing hormone (CRH) systems, which have been implicated in the stress-induced suppression of reproductive function. In the present study, we investigated the role of CRH systems in orexin-induced LH suppression. OVX rats were implanted with i.c.v. and intravenous (i.v.) cannulae. After i.c.v. injection of orexin and/or CRH receptor antagonists, blood samples were collected through the i.v. cannula at 6-min intervals for 120 min for LH measurement. Intracerebroventricular injection of orexin A or B (3 nmol/2.5 microl) suppressed pulsatile LH secretion. Coadministration of orexin A and alpha-helical corticotrophic-releasing factor (CRF), a nonselective CRH receptor antagonist (13 nmol/2.5 microl), or astressin(2)B, a selective type2 (CRH-R2) CRH receptor antagonist (28 nmol/2.5 microl), partly restored pulsatile LH secretion. Orexin B-induced LH suppression was not restored by alpha-helical CRF. In addition, i.c.v. injection of orexin A increased CRH and urocortin II (UcnII), but not Ucn mRNA levels, in the hypothalamus. These findings suggest that CRH-R2 mediates orexin A-induced LH suppression and it is possible that CRH and UcnII in the hypothalamus are involved in this pathway.

Serum osteoprotegerin/osteoclastogenesis-inhibitory factor during pregnancy and lactation and the relationship with calcium-regulating hormones and bone turnover markers.

Pregnancy and lactation induce dynamic changes in maternal bone and calcium metabolism. A novel cytokine termed osteoprotegerin (OPG)/osteoclastogenesis-inhibitory factor (OCIF) was recently isolated; this cytokine inhibits osteoclast maturation. To define the effects of pregnancy and lactation on circulating OPG/OCIF in mothers, we studied the changes in the levels of OPG/ OCIF as well as those of calcium-regulating hormones and biochemical markers of bone turnover in the maternal circulation during pregnancy (at 8-11 weeks, at 22-30 weeks, at 35-36 weeks and immediately before delivery) and lactation (at 4 days and at 1 month postpartum). Serum intact parathyroid hormone levels did not change and were almost within the normal range in this period. In contrast, serum 1,25-dihydroxyvitamin D levels increased with gestational age and were above the normal range during pregnancy. After delivery, they fell rapidly and significantly (P<0.01) to the normal range. The levels of serum bone-specific alkaline phosphatase, one of the markers of bone formation, increased with gestational age. After delivery, these levels were further increased at 1 month postpartum. The levels at 1 month postpartum were significantly higher than those at 8-11 and 22-30 weeks of pregnancy (P<0.01 and P<0.05 respectively). The levels of serum C-terminal telopeptides of type I collagen, one of the markers of bone resorption, did not change during pregnancy. After delivery, they rapidly and significantly (P<0.01) rose at 4 days postpartum, and had then fallen by 1 month postpartum. Circulating OPG/OCIF levels gradually increased with gestational age and significantly (P<0.01) increased immediately before delivery to 1.40+/-0.53 ng/ml (means+/-S.D.) compared with those in the non-pregnant, non-lactating controls (0.58+/-0.11 ng/ml). After delivery, they fell rapidly to 0.87+/-0.27 ng/ml at 4 days postpartum and had fallen further by 1 month postpartum. These results suggest that the fall in OPG/OCIF levels may be partially connected with the marked acceleration of bone resorption after delivery.

Effects of leptin on secretion of LH and FSH from primary cultured female rat pituitary cells.

BACKGROUND: Leptin, which is the product of the obese gene, is believed to play important roles in pubertal development and reproductive function in females. In a study using adult male rats, it was found that leptin stimulated secretion of gonadotropin from the pituitary in a dose-related manner. However, there has been no such study in female rats. OBJECTIVE: To investigate the effects of leptin on the production of LH and FSH from the pituitary in female rats, using primary cultured pituitary cells. METHODS: In this study, we determined body weight, serum leptin concentration and serum estradiol (E(2)) concentration in female Wistar rats at 3, 5, 6, 7, 9 and 11 weeks of age, and cultured pituitary cells from 6-week-old female Wistar rats with leptin (0--10(-7) mol/l) and GnRH (0 or 10(-8) mol/l). Then basal and GnRH-stimulated extra- and intracellular LH and FSH were assayed by RIA. RESULTS: Serum leptin concentration increased with increases in body weight and E(2) concentration. The pubertal serum leptin concentration was about 10(-10) mol/l. At a lower or moderate concentration, leptin produced dose-related increases in both basal and GnRH-stimulated extra- and intracellular LH and FSH in pituitary cells. At a concentration of 10 mol/l, leptin significantly (P<0.05) stimulated both basal and GnRH-stimulated extra- and intracellular LH and FSH. However, at greater concentrations, these effects diminished. CONCLUSIONS: These results indicated that leptin induced pituitary cells to produce and secrete both LH and FSH, with or without GnRH. The concentration of leptin that induced the greatest production of gonadotropins by pituitary cells was 10(-10) mol/l, which was the same as the physiological pubertal concentration. Leptin may be involved in the onset of puberty. It is also conceivable that leptin may be a cause of ovulatory failure, not only in weight loss but also in weight gain.

Involvement of chromosomes 4, 11, and 17 in a case of myelodysplastic syndrome.

A case of myelodysplastic syndrome in a 68-year-old male in whose marrow cells two translocations were established, i.e., t(4;11)(q13;q23) and t(11;17)(p?:q11), as well as other karyotypic changes (-6,-18,15p+), is described. The relation and identity of the bands involved in the translocations affecting chromosomes #11 and #17 in leukemias in which these chromosomes are specifically affected, i.e., t(4;11) in acute myelomonocytic leukemia and t(15;17) in acute promyelocytic leukemia, are discussed in relation to the case described.

Pattern visual evoked cortical potentials in patients with toxic optic neuropathy caused by toluene abuse.

PURPOSE: Electrophysiological evaluation of the visual function of patients with toxic neuropathy caused by toluene abuse. METHODS: Fifteen patients (mean age 25.6 years, eight men and seven women) were diagnosed with bilateral optic neuropathy. Pattern visual evoked cortical potentials (PVECPs) and clinical symptoms were investigated. RESULTS: Visual acuities at the initial visit were less than 0.1 in 5 cases and 0.1-1.0 in 10 cases. PVECPs were followed up in the 15 cases. At the first recording, PVECPs were nonrecordable in both eyes of 11 cases, the P100 peak latency was prolonged in both eyes of 3 cases, and only 1 case showed a normal P100 peak latency. After treatment, visual acuities improved more than 2 lines in 6 cases, 3 of whom showed normal P100 peak latency in the PVECPs. Visual prognosis and PVECP changes were identical in both eyes of all patients. CONCLUSIONS: In patients with toluene optic neuropathy, the P100 peak latency of PVECP shortened as visual acuity improved. The VECP abnormalities in these patients suggest that there is a severe effect on the optic nerve after prolonged exposure to toluene.

Treatment of micrognathia with edentulous maxilla by sagittal split mandibular osteotomy and a subperiosteal implant immobilized with transmaxillary screws.

Micrognathia complicated by edentulous maxilla was treated by performing sagittal-split mandibular osteotomy and immobilizing a subperiosteal implant using transmaxillary screws. The patient was a 42-year-old man who had a birdlike facial deformity caused by significant hypoplasia of the mandible. He also demonstrated significant malocclusion attributable to micrognathia and edentulous maxilla caused by resorption of the alveolar bone. These conditions impaired his mastication and articulation, making it impossible for him to eat regular food or carry out normal conversation. A subperiosteal implant was placed on the edentulous maxilla, and was rigidly immobilized to the maxilla using five transmaxillary screws. A prosthesis was then attached to the implant, and by using the implant as the point of reference and the anchor, the mandible was moved forward by sagittal-split mandibular osteotomy. Intermaxillary fixation was subsequently performed. The postoperative course has been favorable, and his facial complexion has improved significantly. One and a half years after his surgery, there has been no sign of complications or malocclusion caused by mandibular retraction. He is now able to eat regular food and speak normally.

Plasma-treated lenowoven polyethylene ribbon for bone fixation at dentoalveolar osteotomy.

Plasma-treated lenowoven polyethylene ribbon was used for splinting between teeth around osteotomy lines in anterior segmental dentoalveolar osteotomy. This fixation was continued for 2 to 3 months, during which there was no damage or fall of the ribbon. In addition, the patients were able to eat their usual food, had much less pain in the teeth and oral mucosa, did not feel uncomfortable in the mouth, had no tooth damage or carious teeth, and had a good aesthetic appearance. In all eight jaws (five patients), accurate and strong bone fixation was achieved based on the planned occlusion. Patient complaints and pain were obviously less with this method than other methods. Polyethylene ribbon is quite useful for bone fixation in anterior segmental dentoalveolar osteotomy.

Anti-atherosclerotic action of elastase--with special reference to its effect on elastic fibres.

In order to determine the anti-atherosclerotic action of elastase, the aortas from four groups of rats--group N fed a normal diet, group D fed an atherogenic diet with vitamin D2, group N + Ela fed a normal diet with elastase, and group D + Ela fed an atherogenic diet and vitamin D2 with elastase--were studied histologically after 3, 6, 9, and 12 weeks of feeding. Group D displayed a remarkable atheromatous change, while the change was insignificant in group D + Ela. The histopathological findings in group D consisted of marked rarefaction, fat deposition, and atheroma in the subendothelial tissue and tunica intima; Van Gieson's staining revealed a conspicuous decrease in elastic fibres, and the remaining elastic fibres had such abnormalities as a tortuous course, bends, and fissures. For group D + Ela, on the other hand, there were few foam cells in the subendothelial tissue and tunica intima and much less marked rarefaction of interstice and fat deposition. Further, the arrangement of elastic fibres was only slightly disordered, and the decreases in elastic fibres and their fissures were moderate. In sharp contrast to group D. which showed marked calcium deposition, group D + Ela was found to have low calcium deposition. No macroscopic and histopathologic abnormalities were found in groups N and N + Ela. Elastase had an inhibitory effect on experimental atherosclerosis in the rat and also regulated the degradation and biosynthesis of elastin, which resulted in the regeneration of elastic fibres.

Orexins, orexigenic hypothalamic neuropeptides, suppress the pulsatile secretion of luteinizing hormone in ovariectomized female rats.

It is well known that feeding disorders are deeply related to reproductive dysfunction, and some feeding regulatory factors may modulate the reproductive function. We examined the effect of orexins, the newly discovered orexigenic hypothalamic neuropeptides, on the pulsatile secretion of LH to clarify their influence on the reproductive function. We administered orexins or saline into the third ventricle of bilaterally ovariectomized (OVX) rats, and measured the serum LH concentration by RIA in blood samples drawn every 6 min for 2 hours to analyze the pulsatile secretion. In the orexin-treated groups, the mean LH concentration and the pulse frequency were significantly reduced (p < 0.01), but the pulse amplitude did not differ significantly. These data indicate that orexins suppress the pulsatile secretion of LH by influencing GnRH neurons at the hypothalamic level.

Genes encoding giant danio and golden shiner ependymin.

Ependymin (EPN) is a brain glycoprotein that functions as a neurotrophic factor in optic nerve regeneration and long-term memory consolidation in goldfish. To date, true epn genes have been characterized in one order of teleost fish, Cypriniformes. In the study presented here, polymerase chain reactions were used to analyze the complete epn genes, gd (1480 bp), and sh (2071 bp), from Cypriniformes giant danio and shiner, respectively. Southern hybridizations demonstrated the existence of one copy of each gene per corresponding haploid genome. Each gene was found to contain six exons and five introns. Gene gd encodes a predicted 218-amino acid (aa) protein GD 93 percent conserved to goldfish EPN, while sh encodes a predicted 214-aa protein SH 91 percent homologous to goldfish. Evidence is presented classifying proteins previously termed "EPNs" into two major categories: true EPNs and non-EPN cerebrospinal fluid glycoproteins. Proteins GD and SH contain all the hallmark, features of true EPNs.

Analysis of predisposing clinical and laboratory findings for the development of endogenous fungal endophthalmitis. A retrospective 12-year study of 79 eyes of 46 patients.

PURPOSE: To analyze the predisposing factors for the development of endogenous fungal endophthalmitis (EFE) for its early diagnosis and treatment. SUBJECTS AND METHODS: Seventy-nine eyes of 46 patients with EFE treated in the 12-year period between 1986 and 1998 were included. A retrospective analysis was conducted in respect to age, sex, underlying disease, visual acuity, findings in the anterior and posterior segments, fungal culture of surgical specimens, fever of unknown origin, neutrophils < or = 500/mL, Cand-tec > or = x 4, beta-D-glucan > or = 20 pg, and final visual acuity. RESULTS: The patients were 34 men (74%) and 12 women (26%) between 18 and 78 years of age (mean 57.2 years). Thirty-three of the 46 patients (72%) also were diagnosed with cancer. Fungal infiltration limited to the retina (Stage I) was noted in 13%, budding in the vitreous cavity (Stage II) in 40%, vitreous opacity (Stage III) in 29%, and retinal detachment with Stage III (Stage IV) in 18% of 79 eyes with EFE. Forty patients (87%) were undergoing intravenous hyperalimentation (IVH). The mean interval between the start of IVH and the onset of disease was 11 days. Vitreous surgery was performed in 26 eyes (33%). Candida albicans was detected from surgical specimens in 38%. Fever of unknown origin was noted in 76%, neutrophils < or = 500/mL in 67%, Cand-tec > or = x 4 in 57%, and beta-D-glucan > or = 20 pg in 90% of subjects. CONCLUSION: In patients susceptible to opportunistic infection, beta-D-glucan > or = 20 pg (90%), IVH (87%), fever of unknown origin (76%), male sex (74%), the presence of cancer (72%), neutrophils < or = 500/mL (67%), and Cand-tec > or = x 4 (57%) were considered to be predisposing factors for the development of EFE.

Efficacy of every-other-day administration of conjugated equine estrogen and medroxyprogesterone acetate on gonadotropin-releasing hormone agonists treatment in women with endometriosis.

We performed a randomized controlled study to determine the efficacy of add-backed therapy by every-other-day administration of 0.625 mg conjugated equine estrogen (CEE) and 2.5 mg medroxyprogesterone acetate (MPA) on GnRH agonists (GnRH-a) treatment in Japanese women with symptomatic endometriosis. At the end of treatment, serum estrone and estradiol levels in the add-back group (n = 11) were significantly higher than those in the control group (n = 10). The assessment of Beecham classification by bimanual examination, serum CA-125 levels, and the frequency of genital bleeding revealed no significant differences between the two groups. The add-back group showed reduced Kupperman indices relative to those of the control group, and could prevent the loss of bone density. These findings led to a conclusion that GnRH-a therapy added back by every-other-day administration of 0.625 mg CEE and 2.5 mg MPA was a safe and effective treatment for Japanese women with endometriosis.

Synthesis and structure-activity relationship of a new series of potent angiotensin II receptor antagonists: pyrazolo[1,5-a]pyrimidine derivatives.

We have already reported 7-oxo-4,7-dihydropyrazolo[1,5-a]pyrimidine-3-carboxylic acid derivatives, which are potent in vitro angiotensin II (AII) antagonists, but have no oral antihypertensive activity. Removal of the carboxylic acid and replacement of the heteroaromatic system afforded potent in vitro antagonists. Removal of the carbonyl oxygen and changing the position of the biphenyltetrazole substituent were critical to the display of oral activity. To improve the in vitro and oral activities, modifications were made of the substituents at the 3- and 5-positions of the pyrazolo[1,5-a]pyrimidine. Structure-activity studies showed the methyl substituent at the 3-position to be essential for potent in vivo activity. We present the design, syntheses, and biological data of a series of pyrazolo[1,5-a]pyrimidine derivatives, which are orally active AII receptor antagonists.