The ability of alpha MSH, a POMC-derived peptide, to antagonize the effects of beta-endorphin (beta EP) on PRL and LH secretion was studied in the primate. Seven ovariectomized rhesus monkeys bearing chronic indwelling third ventricular catheters for peptide infusion were used for these studies. Peripheral blood samples for PRL and LH RIA were obtained every 15 min during a 3-h control period when saline was infused into the ventricle, followed by a 5-h period of peptide infusion at a rate of 25 microliters/h. When beta EP was infused at a dose of 5 micrograms/h, plasma PRL rose from a mean baseline of 3.5 +/- 0.7 ng/ml to a peak of 21.3 +/- 2.2 ng/ml. When the same animals were infused with 20 micrograms alpha MSH together with 5 micrograms beta EP, the peak concentration of PRL was reduced to 8.2 +/- 1.7 ng/ml (P less than 0.001). When a higher dose of beta EP (20 micrograms/h) was infused, PRL rose to a peak of 38.2 +/- 1.8 ng/ml. This response was again markedly blunted, and the peak PRL response was reduced to 7.3 +/- 2.2 ng/ml when 20 micrograms beta EP were infused together with 80 micrograms alpha MSH (P less than 0.001). Analysis of the area under the plasma PRL concentration curves demonstrated a significant reduction in area during the 5-h infusion with beta EP plus alpha MSH compared to that during infusion of beta EP alone. The mean area was reduced from 3480 +/- 570 ng min/ml after 5 micrograms beta EP alone to 1030 +/- 200 after 5 micrograms beta EP plus 20 micrograms alpha MSH and from 6230 +/- 990 after 20 micrograms beta EP to 1020 +/- 320 ng min/ml after 20 micrograms beta EP plus 80 micrograms alpha MSH (P less than 0.01). Des-acetyl alpha MSH (80 micrograms) was also effective in reducing the PRL response to 830 +/- 380 ng min/ml (P less than 0.05). The suppression of pulsatile LH release by beta EP was also attenuated by alpha MSH. During the 5-h infusion of beta EP, total LH secretion was reduced to 65.9 +/- 3.8% of that measured during the 3-h saline infusion compared to 87.2 +/- 2.7% after infusion of beta EP plus alpha MSH (P less than 0.001) or 91.7 +/- 4.1% after beta EP plus des-acetyl alpha MSH (P less than 0.05).(ABSTRACT TRUNCATED AT 400 WORDS)

Inhibition of acetyl-CoA carboxylase (ACC) is one approach used for treating metabolic syndrome. Using partially purified ACC to screen herbs commonly used in Taiwanese folk medicine, we previously showed that an ethanol extract of Polygonum hypoleucum Ohwi (EP) had potent ACC inhibitory activity and partially alleviated metabolic disorders induced by a high fat diet. Since ACC plays a crucial role in de novo lipogenesis, the favorable effects of EP on metabolism were tested under lipogenic conditions in the present study. On incubating high glucose (30 mM)-stimulated HepG2 cells with EP (72.5 or 145 microg/mL), ACC and fatty acid synthase activity, triacylglycerol content, and microsomal triacylglycerol transfer protein mRNA levels were all significantly reduced (P < 0.05, vs vehicle). When EP was given at low, medium, and high dosages (94, 188, and 470 mg/kg) to sucrose water-treated Wistar rats for four weeks, alleviation of symptoms associated with metabolic syndrome, including obesity, insulin resistance, hypertriglyceridemia, and hypertension, accompanied by hepatic ACC inactivation, was seen in the low dosage group. Four compounds (emodin, emodin-8-O-beta-D-glucopyranoside, (+)-catechin, and (-)-epicatechin) isolated from EP were identified as ACC inhibitors. These results confirm that P. hypoleucum Ohwi, acting partly through ACC inhibition, has favorable effects in alleviating metabolic disturbances occurring under lipogenic conditions.

Subjects were patients with schizophrenia or schizoaffective disorder enrolled in extension studies (Study A and Study B) after participating in 12-week studies of long-acting injectable risperidone [Kane, J.M., Eerdekens, M., Lindenmayer, J.-P., Keith, S.J., Lesem, M., Karcher, K., 2003. Long-acting injectable risperidone: efficacy and safety of the first long-acting atypical antipsychotic. Am. J. Psychiatry 160, 1125-1132; Lindenmayer, J.-P., Eerdekens, L., Berry, S., Eerdekens, M., 2004. Safety and efficacy of long-acting risperidone in schizophrenia: a 12-week, multicenter, open-label study in stable patients switched from typical and atypical oral antipsychotics. J. Clin. Psychiatry 65, 1084-1089]. Twelve months of treatment were completed by 55% of Study A patients and 52% of Study B patients. The median modal dose of long-acting injectable risperidone was 50 mg/14 days in both studies. Most frequent adverse events were psychosis, headache, insomnia, agitation, and rhinitis. EPS-related adverse events were reported in 33% of patients in Study A and 22% in Study B. Patients with Clinical Global Impressions ratings of "not ill" and "mild" increased from 14% at baseline to 54% at endpoint in Study A and from 42% to 65% in Study B. It is concluded that treatment with long-acting injectable risperidone for 1 year or longer appeared to be safe and well tolerated in patients with schizophrenia or schizoaffective disorder.

The effects of ethanol and beta-endorphin (beta-EP) on productions of cytolytic factors granzyme B, perforin and IFN-gamma in splenic rat NK cells were determined. Intracranial administration of beta-EP increased protein and mRNA levels of cytolytic factors in NK cells. Chronic ethanol feeding reduced the basal and beta-EP-induced levels of cytolytic factors in NK cells. In vitro treatment of beta-EP on NK cells increased the levels of perforin, granzyme B and IFN-gamma and their mRNA transcripts, whereas ethanol pre-treatment prevented beta-EP effects on cytolytic factors in these cells. These results suggest that beta-EP and ethanol interact to regulate NK cell functions.

OBJECTIVE

The purpose of this study was to assess the quantitative effects of contrast material concentration on hepatic parenchymal and vascular enhancement in multiphasic computed tomography (CT), using multi-detector row helical CT.

METHODS

We designed a prospective randomized study to test two different concentrations of contrast material on five phasic scans of the liver. One hundred patients were randomly assigned to two groups: an iodine concentration of 300 mg/mL in group A and 370 mg/mL in group B. All patients received a fixed volume of 100 mL at a 4 mL/sec injection rate. Enhancement values for the hepatic parenchyma and aorta at three levels (upper, middle, and lower level of the liver), and values for portal and hepatic veins were statistically compared between the two groups.

RESULTS

Hepatic parenchymal enhancement values at all levels of the liver in portal phase (PP) and equilibrium phase (EP) were significantly higher in group B than in group A (p<0.01). Aortic enhancement values at two levels of the liver (middle and lower) in early hepatic arterial phase (EAP) were significantly higher in group B than in group A (p<0.05), however, there was no significant difference between groups A and B in aortic enhancement during the delayed hepatic arterial phase (DAP). Portal and hepatic venous enhancement values in PP and EP were significantly higher in group B than in group A (p<0.01).

CONCLUSIONS

On multiphasic dynamic CT, the use of a higher iodine concentration of contrast material results in higher hepatic parenchymal enhancement and aortic enhancement, as well as higher portal and hepatic venous enhancement.

Previous studies from this laboratory have demonstrated immunostainable beta-endorphin-like material (beta-EP) in Leydig cells and epithelia of the epididymis, seminal vesicle and vas deferens of the rat. These observations would be strengthened if it could be demonstrated that they were not a peculiarity of the rat. Accordingly, we now present immunocytochemical evidence for the presence of beta-EP in the Leydig cells of mouse, hamster, guinea pig and rabbit. No immunoreactive material was identified in Sertoli, myoid, endothelial or germ cells of any of the species examined. Immunostainable beta-EP was also demonstrated in the epididymides of mouse, guinea pig, rabbit, and rat, but not hamster. Immunostainable material was also present in the epithelia of the vas deferens and seminal vesicles of mouse and rat, the only two species thus far examined. Since beta-EP was present in Leydig cells, we wondered whether this peptide could be identified in other steroid-producing tissues. When rat ovaries and adrenals were reacted with anti-beta endorphin, staining was demonstrated in corpus luteum and adrenal cortex. No staining was observed in the adrenal medulla or other portions of the ovary. In order to determine whether the beta-EP detected in the testis and epididymis was derived from a pituitary source, animals were hypophysectomized and tissues examined 2 weeks later. Both the Leydig cells and the epididymal epithelium remained immunostainable. In summary, immunostainable beta-EP has been identified in Leydig cells of five species. Stainable material is also present in the epithelium of other portions of the male reproductive tract and in steroid-secreting cells of the ovary and the adrenal. Such beta-EP may have a paracrine function in the testis and other portions of the male reproductive tract.

Evidence strongly supports that low-level laser therapy (LLLT) is an effective physical modality for the treatment of pain associated with myofascial trigger points (MTrP). However, the effect of laser fluence (energy intensity in J/cm(2)) on biochemical regulation related to pain is unclear. To better understand the biochemical mechanisms modulated by high- and low-fluence LLLT at myofascial trigger spots (MTrSs; similar to human MTrPs) in skeletal muscles of rabbits, the levels of β-endorphin (β-ep), substance P (SP), tumor necrosis factor-α (TNF-α), and cyclooxygenase-2 (COX-2) were investigated in this study. New Zealand rabbits (2.5-3.0 kg in weight) were used in this study. High-fluence LLLT (27 J/cm(2)), low-fluence LLLT (4.5 J/cm(2)), or sham operations were applied on MTrSs of biceps femoris of rabbits for five sessions (one session per day). Effects of LLLT at two different fluences on biceps femoris, dorsal root ganglion (DRG), and serum were determined by β-ep, SP, TNF-α, and COX-2 immunoassays. LLLT irradiation with fluences of 4.5 and 27 J/cm(2) at MTrSs can significantly reduce SP level in DRG. LLLT with lower fluence of 4.5 J/cm(2) exerted lower levels of TNF-α and COX-2 expression in laser-treated muscle, but LLLT with higher fluence of 27 J/cm(2) elevated the levels of β-ep in serum, DRG, and muscle. This study demonstrated fluence-dependent biochemical effects of LLLT in an animal model on management of myofascial pain. The findings can contribute to the development of dosage guideline for LLLT for treating MTrP-induced pain.

Immunotherapy with interleukin-2 (IL-2) and lymphokine-activated killer (LAK) cells results in significant tumor regression in patients with advanced cancer. We have investigated the kinetics of circulating erythroid (BFU-E) and granulocytic-macrophage (CFU-GM) progenitors after IL-2 therapy in 11 cancer patients, mainly affected by metastatic melanoma and renal cell carcinoma. Administration of IL-2 from day 1 through day 5 constantly induced a dramatic decrease of the number of circulating BFU-E and CFU-GM, which then showed a striking rebound (up to values fourfold and sevenfold higher, respectively, than the pretherapy levels) on discontinuation of IL-2, ie, from day 5 through day 10. A similar kinetic pattern was observed during and after the second cycle of IL-2 administration. 3[H]-thymidine killing experiments showed that the cycling activity of the progenitors was virtually unmodified in the rebound phases. To explore the mechanism(s) underlying this kinetic pattern, we have analyzed the plasma concentration of several hematopoietic growth factors, including IL-1 beta, IL-3, IL-4, IL-6, granulocyte-macrophage colony-stimulating factor (GM-CSF), G-CSF, and erythropoietin (Ep). No modifications in the levels of IL-3, GM-CSF, or IL-1 beta were observed, whereas a pronounced increase of IL-6 and G-CSF concentration was monitored, starting at day 3 and peaking at day 5 of treatment (a parallel, but modest, increase of Ep level was also observed). The elevation of IL-6 and G-CSF concentration is directly correlated with and may, at least in part, underlie the subsequent rebound of circulating hematopoietic progenitors. Furthermore, the increase in IL-4 level observed at day 10 of therapy may mediate the eosinophilia gradually starting at this stage of treatment.

OBJECTIVE

The purpose of this study was to determine which imaging characteristics can be used as prognostic indicators in conjunction with beta-human chorionic gonadotropin (beta-hCG) levels in the treatment of ectopic pregnancy (EP) with single-dose methotrexate (MTX).

METHODS

A retrospective study was performed on 62 patients (age range, 16-47 years; mean, 29 years) treated with MTX for EP from November 2000 to August 2003. The transvaginal sonographic findings in each case were analyzed for the presence and size of an extraovarian mass or a pseudogestational sac, amount of free fluid, presence of a yolk sac, and fetal heart motion. Patient age and beta-hCG level were also noted. Success of treatment was defined as a single dose of MTX that resulted in appropriate lowering of beta-hCG levels.

RESULTS

Of 62 patients, 17 (27%) had single-dose MTX treatment failure. A yolk sac was identified in 15 (88%) of the 17 treatment failures and in none of the cases in which treatment was successful (positive predictive value, 100%). The average beta-hCG level in the cohort of patients who had single-dose treatment failure was 3282 mIU/mL compared with 1544 mIU/mL in the treatment success cohort. The presence of fetal heart motion was seen in only 1 patient, and this patient had treatment failure. The age of the patient, size of the extraovarian mass, presence of a pseudogestational sac, and amount of free fluid did not correlate with outcome.

CONCLUSIONS

The presence of a yolk sac was always associated with treatment failure in single-dose MTX treatment of EP and was the most reliable predictor of failure among all features analyzed. The beta-hCG level was a useful adjunct. A prediction rule was created correlating the probability of treatment success with the beta-hCG level.

1. This study was undertaken to characterize pharmacologically the prostanoid receptor subtypes mediating contraction in human umbilical vein (HUV). 2. HUV rings were mounted in organ baths and concentration-response curves to U-46619 (TXA(2) mimetic) were constructed in the absence or presence of SQ-29548 or ICI-192,605 (TP receptor antagonists). U-46619 was a potent constrictor (pEC(50): 8.03). SQ-29548 and ICI-192,605 competitively antagonized responses to U-46619 with pK(B) values of 7.96 and 9.07, respectively. 3. Concentration-response curves to EP receptor agonists: PGE(2), misoprostol and 17-phenyl-trinor-PGE(2) gave pEC(50) values of 5.06, 5.25 and 5.32, respectively. Neither pEC(50) nor maximum of PGE(2) and 17-phenyl-trinor-PGE(2) concentration-response curves were modified by the DP/EP(1)/EP(2) receptor antagonist AH 6809 (1 micro M). However, ICI-192,605 produced a concentration-dependent antagonism of the responses to all the EP receptor agonists. The pA(2) estimated for ICI-192,605 against PGE(2) or misoprostol were 8.91 and 9.22, respectively. 4. Concentration-response curves to FP receptor agonists: PGF(2)(alpha) and fluprostenol gave pEC(50) values of 6.20 and 5.82, respectively. ICI-192,605 (100 nM) was completely ineffective against PGF(2)(alpha) or fluprostenol. In addition, lack of antagonistic effect of AH 6809 (1 micro M) against PGF(2)(alpha) was observed. 5. In conclusion, the findings obtained with TP-selective agonist and antagonists provide strong evidence of the involvement of TP receptors promoting vasoconstriction in HUV. Furthermore, the action of the natural and synthetic EP receptor agonists appears to be mediated via TP receptors. On the other hand, the results employing FP receptor agonists and antagonists of different prostanoid receptors suggest the presence of FP receptors mediating vasoconstriction in this vessel.

The concentration of human beta-endorphin (beta h-EP)) was measured in various hypothalamic nuclei, in extrahypothalamic brain regions and in the anterior and posterior lobes of the pituitary using a specific radioimmunoassay (RIA) The beta h-EP concentrations in the arcuate nucleus (169 +/- 35 pg/100 micrograms protein, n = 7) and median eminence (163 +/- 32 pg/100 micrograms protein, n = 6) were among the highest in the 17 brain areas examined. The immunoreactive beta h-EP in the hypothalamus corresponded to authentic beta h-EP, as determined by gel exclusion chromatography. By chromatography and RIA the beta h-EP concentrations in anterior (1.53 X 10(5) +/- 0.51 X 10(5) pg/100 micrograms protein, n = 3) and posterior (1.41 X 10(5) +/- 0.38 X 10(5) g/100 micrograms protein, n - 5) pituitary were found to be approximately 1,000-fold higher than in hypothalamus. Within the pituitary beta h-EP was localized throughout the anterior lobe, in the pars intermedia and in that part of the posterior lobe nearest the pars intermedia, as judged by immunocytochemistry. Dense immunocytochemical staining was found along the perimeter of many blood vessels. Beta h-EP and adrenocorticotropin (ACTH) were co-localized in the same pituitary cells. The present data represent the first unequivocal localization and quantitation of beta h-EP in human brain and in the separate lobes of the human pituitary.

Previous research investigating the response of plasma beta-endorphins (beta-EP) to resistance exercise has resulted in equivocal findings. To further examine the effects of resistance exercise on beta-endorphin immunoreactivity, six resistance-trained athletes participated in a three-set series of eight repetitions of isotonic exercise. All exercises were performed at 80% maximal effort. Blood was sampled from the group by venepuncture, both prior to and following the exercise bout, and beta-endorphin concentration was determined by radioimmunoassay. The results indicated that mean (+/- S.E.) plasma levels of beta-endorphins following exercise (18.04 +/- 3.4 pg beta-EP ml-1) were not significantly changed from pre-exercise (control) levels (19.59 +/- 2.4 pg beta-EP ml-1), although there was considerable inter-individual variability. Our results support previous research which has reported no significant changes in beta-endorphin immunoreactivity following resistance exercise, as well as reported findings of considerable variability in the beta-endorphin response to exercise.

Four exopolysaccharides (EPS) obtained from Botryosphaeria rhodina strains isolated from rotting tropical fruit (graviola, mango, pinha, and orange) grown on sucrose were purified on Sepharose CL-4B. Total acid hydrolysis of each EPS yielded only glucose. Data from methylation analysis and (13)C NMR spectroscopy indicated that the EPS from the graviola isolate consisted of a main chain of glucopyranosyl (1-->3) linkages substituted at O-6 as shown in the putative structure below: [carbohydrate structure: see text]. The EPS of the other fungal isolates consisted of a linear chain of (1-->6)-linked glucopyranosyl residues of the following structure: [carbohydrate structure: see text]. FTIR spectra showed one band at 891 cm(-1), and (13)C NMR spectroscopy showed that all glucosidic linkages were of the beta-configuration. Dye-inclusion studies with Congo Red indicated that each EPS existed in a triple-helix conformational state. beta-(1-->6)-d-Glucans produced as exocellular polysaccharides by fungi are uncommon.

The purpose of this study was to investigate the allodynic effect of <em>b</em>icuculline (BIC) given topically to the dorsal surface of the rat spinal cord, and to determine if spinal prostaglandins (PGs) mediate the allodynic state arising from spinal GABA(A)-receptor <em>b</em>lockade. Male Sprague-Dawley rats (325-400 g) were anaesthetized with halothane and maintained with urethane for the continuous monitoring of <em>b</em>lood pressure (MAP), heart rate (HR) and cortical electroencephalogram (EEG). A laminectomy was performed to expose the dorsal surface of the spinal cord. Unilateral application of BIC (0.1 microg in 0.1 microl) to the L5 or L6 spinal segment induced a highly localized allodynia (e.g. one or two digits) on the ipsilateral hind paw. Thus, hair deflection (<em>b</em>rushing the hair with a cotton-tipped applicator) in the presence, <em>b</em>ut not a<em>b</em>sence of BIC, evoked an increase in MAP and HR, a<em>b</em>rupt motor responses (MR; e.g. withdrawal of the hind leg, kicking, and/or scratching) on the affected side, and desynchrony of the EEG. BIC-allodynia was dose-dependent, yielding ED(50)'s (95% CI's) of 45 ng (31-65) for MAP; 68 ng (46-101) for HR and 76 ng (60-97) for MR. Allodynia was sustained for up to 2 h with repeated BIC application without any detecta<em>b</em>le change in the location or area of peripheral sensitization. Pretreatment with either the <em>EP</em>(1)- receptor antagonist, SC-51322, the cyclooxygenase (COX)-2 selective inhi<em>b</em>itor, NS-398, or the NMDA-receptor antagonist, AP-7, inhi<em>b</em>ited BIC-allodynia in a dose-dependent manner. The results demonstrate: (a) BIC, applied to the dorsal surface of the spinal cord, induces highly localized allodynia; (<em>b</em>) this effect can <em>b</em>e sustained with repeated BIC application; (c) it is evoked <em>b</em>y NMDA-dependent afferent input; (d) spinal PGs are synthesized <em>b</em>y constitutive COX-2 during BIC-allodynia; and (e) spinal PGs contri<em>b</em>ute to the a<em>b</em>normal processing of tactile input via spinal <em>EP</em>1-receptors.

OBJECTIVE

Is annexin A2 involved in the reduced phagocytic ability of macrophages in endometriosis?

CONCLUSIONS

Data from women with endometriosis and a murine model of the disease show that expression of annexin A2 in peritoneal macrophages is inhibited by prostaglandin E2 (PGE2) and this impairs the phagocytic ability of macrophages.

BACKGROUND

Endometriosis is a chronic inflammatory disease that recruits many immune cells, especially macrophages, to the peritoneal cavity. The phagocytic ability of peritoneal macrophages isolated from women with endometriosis is reduced.

METHODS

A laboratory study. Thirty-five patients (20 with and 15 without endometriosis) of reproductive age with normal menstrual cycles were recruited.

METHODS

Peritoneal macrophages isolated from women with or without endometriosis were cultured and treated with vehicle, PGE2 and different EP receptor agonists, and the expression of annexin A2 was quantified by RT-PCR and western blotting. Annexin A2 was knocked down (by small interfering RNA) in normal macrophages or overexpressed (by treatment with recombinant protein) in endometriotic macrophages and their phagocytic ability was measured by flow cytometry. Peritoneal macrophages were isolated from a mouse model of endometriosis and treated with PGE2 or cyclo-oxygenase (COX) inhibitors, and annexin A2 mRNA was quantified.

RESULTS

Levels of annexin A2 were markedly reduced in peritoneal macrophages from women with endometriosis versus controls (mRNA: P < 0.01). The level of annexin A2 mRNA in the macrophages was reduced by PGE2 (P < 0.01/P < 0.05 in women without/with endometriosis versus control) via the <em>EP</em>2/<em>EP</em>4 receptor-dependent signaling pathway. Treatment with PGE2 or knockdown of annexin A2 inhibited the phagocytic ability of macrophages (P < 0.05 versus control), while treatment with annexin A2 recombinant protein enhanced phagocytosis. Autologous transplantation animal studies further confirmed that levels of annexin A2 in peritoneal macrophages were markedly reduced in mice treated with PGE2 (P < 0.01 versus control). In contrast, treatment with COX inhibitors to inhibit PGE2 production enhanced annexin A2 expression in peritoneal macrophages (P < 0.05 versus control).

CONCLUSIONS

We have provided no direct demonstration that phagocytic activity is indeed decreased in peritoneal cells from patients with endometriosis or that their endometriotic fluid contains increased amounts of PGE2 when compared with control subjects.

CONCLUSIONS

Inhibiting PGE2 signaling, in order to restore or enhance the phagocytic capability of macrophages, may represent a new direction of thinking in developing novel strategies against endometriosis.

BACKGROUND

This work was supported by grants from National Science Council of Taiwan, Republic of China (NSC97-2314-B-006-020-MY3) to M.-H.W. and (NSC98-2320-B-006-026-MY3) to S.-J.T., and grants from the Chang Gung Memorial Hospital, Taiwan, Republic of China (CMRPG891432 and CMRPG8A0531) to P.-C.C. None of the authors have any conflicts of interest.

Studies were performed to determine whether the isolated ovine anterior and intermediate pituitary might rhythmically secrete three POMC peptides, ACTH, ir-beta-endorphin (ir-beta-EP), and ir-alpha-melanocyte stimulating hormone (ir-alpha-MSH) in vivo. When blood was taken at 10-min intervals from four ewes with hypothalamo-pituitary-disconnection (HPD), a distinct POMC-peptide and cortisol ultradian rhythm was noted. A comparison of the four HPD ewes with five nonstressed hypothalamopituitary-intact (HPI) ewes revealed that the mean plasma levels of the three POMC-peptides and cortisol were increased, the mean ACTH and ir-alpha-MSH pulse amplitudes were increased, and the mean ir-beta-EP and ir-alpha-MSH interpulse intervals were decreased. When four HPI ewes were subjected to a mild stress, plasma POMC-peptide and cortisol levels increased significantly when compared with the five unstressed HPI animals. In addition, the ACTH and cortisol pulse amplitudes increased and the ir-beta-EP and ir-alpha-MSH interpulse intervals decreased. Although plasma ACTH levels in the stressed HPI and HPD ewes were comparable, mean plasma cortisol levels were 2-fold greater in the stressed HPI animals. To determine whether the ACTH hypersecretion in the HPD ewe might reflect a net reduction in hypothalamic inhibitory influence over ACTH secretion, we examined the effects of dopamine (DA), somatostatin (SS-14), and rat atrial natriuretic peptide [rANF(1-28)] on the secretion of ACTH from cultured ovine anterior pituitary cells. DA and SS-14 did not exert a discernible effect on basal, CRF-, or arginine vasopressin (AVP)-stimulated ACTH secretion. Although basal ACTH secretion was unaffected by rANF(1-28) (10(-12)-10(-8) M), a significant inhibition of CRF- and AVP-stimulated ACTH release was observed.(ABSTRACT TRUNCATED AT 400 WORDS)

Ia specificity 23 is a "combinatorial" antigen generated on a hybrid I region molecule, formed by the noncovalent binding of a 26,000- to 28,000-dalton beta polypeptide chain (Ae) coded by a gene in the I-A subregion with a 32,000- to 35,000-dalton alpha chain (E alpha) coded by a gene in the I-E subregion of the mouse H-2 gene complex. For expression of Ia.23, the Ae chain must be derived from the H-2d haplotype (I-Ad), while the E alpha can be provided by I-Ed, I-Ek, I-Ep, I-Er, I-Ev, and I-Ew3, but not I-Eb, I-Ef, I-Eq, I-Es, and I-Eu. With the exception of H-2u haplotype, all Ia.7 (I-E)-positive haplotypes can provide the permissive E alpha chain for generating Ia.23 by trans-complementation. In the H-2d haplotype, Ia.23 is generated by cis-complementation of Ad with Ed. Lymphocytes of F1 animals expressed two I-E subregion coded hybrid Ia specificities; one formed by cis-complementation and another by trans-complementation. It is postulated that such hybrid determinants are involved in the recognition and generation of immune response to antigens such as GL-Phe and cytochrome C where dual Ir gene control has been demonstrated. It is also suggested that there are two types of Ia specificities: (1) allotypic Ia specificities expressed on the alpha or beta chains (these could aid in the binding between the alpha and beta chains such as Ia.7); and (2) hybrid Ia specificities which are unique interaction determinants formed by the specific association of the alpha and beta chains (e.g., Ia.22,23). These interaction gene products may be involved in antigen recognition and presentation.

We studied the bacterial characteristics and incidence of invasive infections caused by group B streptococci (GBS) in adults in Iceland in 1975-2014. A total of 145 isolates were characterized by serotyping, antimicrobial susceptibility, multilocus sequence typing and surface protein gene profiling. Disease incidence increased during the studied period (p <0.001), reaching 2.17 cases/100 000 person-years in 2013-14. Overall, serotype Ia was the most frequently found (23%), but serotypes Ib, II, III and V showed similar prevalence (14%-17%). Although there were notable changes in the proportion of most serotypes during the study period, only the decline of serotype III was statistically supported (p = 0.003) and was reflected in a decrease of clonal complexes CC17 and CC19 that included most serotype III isolates (p <0.04). On the other hand, the increase in frequency of CC1 was caused by two lineages expressing distinct serotypes: ST1/V/alp3 and ST196/IV/eps. Underlying the relative stability of serotype Ia were major changes in the lineages expressing this serotype, with an increase in the relative importance of CC23, including both ST23/Ia/eps and ST24/Ia/bca lineages, and a decrease in CC7. Nine cases of invasive GBS disease were caused by ST7, of possible zoonotic origin. All isolates were susceptible to penicillin. Rates of erythromycin and clindamycin resistance were 8.3% and 9.7%, respectively. An over-representation of resistance solely to clindamycin was associated with the unusual lsaC gene and serotype III ST19/rib lineage (p <0.001).

We have tested the peptide specificity of positive selection using three transgenic alphabetaTCRs, originally selected on class II MHC (A(b)) covalently bound with one peptide Ealpha (52-68) (Ep). The transgenic TCR specific for the cytochrome c-derived (43-58) peptide was selected on A(b) bound with different arrays of endogenous peptides or the analogue of Ep covalently bound to A(b), but not on the original A(b)Ep complex. In contrast, transgenic TCRs specific for two different analogues of the Ep peptide and A(b) did not mature as CD4(+) T cells in various thymic environments, including the A(b)EpIi(-) mice. These results show that TCRs can be promiscuous or specific for the selecting MHC/peptide complex, and suggest that in mice described in this study transgenic expression of the TCR changes the original requirements for the positively selecting MHC/peptide complex. Future studies will determine whether the latter phenomenon is general or specific for this system.

The plasma patterns of ACTH, beta-lipotropin (beta LPH) and beta-endorphin (beta EP), in addition to those of cortisol and dehydroepiandrosterone sulfate (DHAS), were studied in 139 prepubertal children (subdivided into different age groups) and 38 adolescents (subdivided according to Tanner's pubertal stages) aged 10-16 yr. The adult control group was composed of 23 females and 12 males aged 17-40 yr. No sex differences were found in ACTH, beta LPH, beta EP, and cortisol plasma levels. ACTH plasma levels were slightly lower in the 1- to 3-yr-old groups than in males at 4-5 yr and females at 8-9 yr. No further significant differences were observed in any of the age or pubertal groups, the concentrations being constantly in the adult range. beta LPH and beta EP plasma levels were lowest at 1-3 yr in both males (beta LPH: 2.1 +/- 0.25, beta EP: 1.85 +/- 0.59 fmol/ml, mean +/- SE) and females (beta LPH: 2.8 +/- 0.31; beta EP: 2.41 +/- 0.41 fmol/ml); plasma levels of both hormones increased progressively in both sexes until Puberty 1 stage of sexual maturation, at which time levels were 7.3 +/- 0.78 and 8.69 +/- 1.0 fmol/ml in males and 7.1 +/- 0.34 and 6.76 +/- 0.13 fmol/ml in females; these levels are similar to adult values. A highly significant linear correlation was found between both beta LPH and beta EP concentrations and the age of the subjects; this was not true for ACTH plasma levels. Cortisol plasma levels were similar in all groups. DHAS plasma levels increased progressively from 1-3 yr to the end of sexual maturation when adult values were reached. During prepuberty, DHAS levels were significantly correlated with both beta LPH and beta EP, but not ACTH. These data indicate that plasma beta LPH and beta EP concentrations, in contrast to ACTH levels, increase progressively throughout prepuberty and suggest that the processing of the parent proopiocortin molecule or secretion of the processed peptides from the anterior pituitary (or other sources) may change from early infancy to adulthood. Furthermore, the correlation between both beta LPH and beta EP with DHAS plasma levels in prepuberty suggests a role of proopiocortin-related peptides in adrenarche.

We have previously demonstrated that anoxic preconditioning (APC) protects against a subsequent otherwise 'lethal' anoxic insult in hippocampal slices. Tested here are two hypotheses: (a) APC requires calcium to improve electrical recovery in hippocampal slices; and (b) mild excitation promotes preconditioning neuroprotection. Control hippocampal slices were given a single 'test' anoxic insult followed by reoxygenation. Experimental slices were preconditioned by three short anoxic insults of 1 min separated by 10 min of reoxygenation. At 30 min after the third 'conditioning' insult, slices underwent a 'test' anoxic insult [1 min of anoxic depolarization (AD)], and then slices were reoxygenated. Evoked potentials (EPs) were recorded throughout the experiment. In other slices, APC was emulated by inducing spreading depression (as determined by a negative DC shift) with KCL or by inducing increased neuronal excitability with the excitatory agent 8-cyclopentyl-1, 3-dipropylxanthine (DPCPX) (an adenosine A1 receptor blocker). 'Test' anoxic insults lasted 2 min of AD in these groups. To determine the role of calcium during APC, extracellular CaCl2 was decreased to 0.5 mM but only during the APC episodes ('test' anoxia, 1 min of AD). EP amplitudes recovered significantly better after anoxia in preconditioned slices, and in KCl- and DPCPX-treated slices (147.2+/-33.3, n=8, **p<0.01, 71.7+/-13.5, n=7, **p<0.01, and 117.8+/-37.3, n=5, ***p<0.001, respectively) compared to controls. Decreases in extracellular CaCl2 during APC blocked the recovery of EPs after 'test' anoxia (80.6+/-23.0, n=8). These data confirm that increases in excitability can emulate APC. These data also demonstrate that calcium influx during preconditioning is required for the induction of tolerance during APC.

When cauliflower plants (Brassica oleraceae) were misted with bacterial suspensions of Xanthomonas campestris pv. campestris (causal agent of black rot of cruciferous plants), two separate populations of the pathogen were associated with the leaves. Initially, bacteria removable by sonication and sensitive to sodium hypochlorite treatment predominated (easily removable epiphytic bacteria, EREB). However, after 2 weeks, bacteria not removable by sonication and insensitive to sodium hypochlorite treatment were dominant. Although the exact location of this second population of the pathogen was not determined, evidence is presented to support its location in protected sites on the leaf surface, pigB of this pathogen is required for production of extracellular polysaccharide (EPS), xanthomonadin pigments, and the diffusible signal molecule, DF (diffusible factor). DF can extracellularly restore EPS and xanthomonadin production to pigB mutant strains. Parent strain B-24 and pigB mutant strain BBB-24, Tn3HoHo1 pigB insertion mutation strain BBB mutation strain BBBB-24, BBBB populations of between approximately 3 and 6 (log [1 + CFU per g of fresh weight]) over a 3-week period, whereas BBB-24, BBBBBB population levels did not explain the observed differences in host infection frequencies, and the results indicated that strain BBB-24 and BBBBB-24 applied alone and BBB is required for epiphytic survival and natural host infection under the experimental conditions tested, and suggest that DF, xanthomonadins, and EPS could all be important for survival of this pathogen on the leaf surface, and/or for host infection.

Nine young (26.1 +/- 2.52 years) (M +/- SD) and seven elderly (66.0 +/- 5.85 years) untrained men performed a graded exercise to peak VO2 to compare whether serum beta-endorphin (B-EP) would respond similarly with age. B-EP levels were not different at rest (31.8 +/- 3.3 and 25.3 +/- 4.1 pg/ml in the old and young groups, respectively), and both groups displayed similar responses to exercise. Specifically, we noted no change in B-EP during the final stage of exercise followed by a significant rise for the recovery period (40.0 +/- 3.7 and 38.1 +/- 5.0 pg/ml in the aged and young groups, respectively). Post-exercise reports of anxiety and depression were unchanged in either group, and only minor changes in hostility were noted, as measured by the Multiple Affect Adjective Checklist. These results demonstrate a similar serum B-EP response in older men compared to young men following graded maximal exercise and support the contention that circulatory changes in B-EP may not be related to changes in specific affective states.

Mouse bone marrow cells infected in vitro with the anemia strain of Friend leukemia virus from large clusters (bursts) of erythroblasts after 5 days in culture in methylcellulose medium. Two types of erythroblast populations can be isolated from bursts of infected cells by manipulation of the culture conditions. One type of erythroblast, which is obtained when erythropoietin (EP) is added to the culture, has proliferated and undergoes differentiation to become an erythrocyte. The second type of erythroblast, which is obtained when no EP is added to the culture, is the product of extensive proliferation, but it fails to undergo the terminal stages of erythroblast differentiation. Comparisons of these two types of erythroblasts demonstrate that specific EP effects include changes in the nucleus, cytoplasm, and membrane of the treated cells. Those events of erythroid differentiation shown to be directed by EP were extrusion of the nucleus from the erythroblast, induction of uroporphyrinogen I synthetase activity, increased iron incorporation into protoporphyrin, synthesis of alpha- and beta-globin polypeptides due largely to increased mRNA production, and synthesis and incorporation of spectrin into the cell membrane. In this system, EP promotes these effects without observable stimulation of progenitor proliferation in addition to that caused by the virus alone. Thus, the role of EP in terminal erythrocyte differentiation is not simply that of an erythroid-specific mitogen.