Limited evidence of autocorrelation signaling upcoming affective episodes: a 12-month e-diary study in patients with bipolar disorder.

BACKGROUND: Increased autocorrelation (AR) of system-specific measures has been suggested as a predictor for critical transitions in complex systems. Increased AR of mood scores has been reported to anticipate depressive episodes in major depressive disorder, while other studies found AR increases to be associated with depressive episodes themselves. Data on AR in patients with bipolar disorders (BD) is limited and inconclusive. METHODS: Patients with BD reported their current mood via daily e-diaries for 12 months. Current affective status (euthymic, prodromal, depressed, (hypo)manic) was assessed in 26 bi-weekly expert interviews. Exploratory analyses tested whether self-reported current mood and AR of the same item could differentiate between prodromal phases or affective episodes and euthymia. RESULTS: A total of 29 depressive and 20 (hypo)manic episodes were observed in 29 participants with BD. Self-reported current mood was significantly decreased during the two weeks prior to a depressive episode (early prodromal, late prodromal), but not changed prior to manic episodes. The AR was neither a significant predictor for the early or late prodromal phase of depression nor for the early prodromal phase of (hypo)mania. Decreased AR was found in the late prodromal phase of (hypo)mania. Increased AR was mainly found during depressive episodes. CONCLUSIONS: AR changes might not be better at predicting depressive episodes than simple self-report measures on current mood in patients with BD. Increased AR was mostly found during depressive episodes. Potentially, changes in AR might anticipate (hypo)manic episodes.

[Ambulatory monitoring and digital phenotyping in the diagnostics and treatment of bipolar disorders]. / Ambulantes Monitoring und digitale Phänotypisierung in Diagnostik und Therapie bipolarer Erkrankungen.

BACKGROUND: Reliable and valid diagnostics and treatment of bipolar disorders and affective episodes are subject to extensive, especially methodological limitations in the clinical practice. OBJECTIVE: The use of smartphones and mobile sensor technology for improvement in diagnostics and treatment of bipolar disorders. METHODS: Critical discussion of current research on the use of ambulatory monitoring and digital phenotyping with bipolar disorders. RESULTS: In many studies the observation periods were too short and the sensors applied were too inaccurate to enable reliable and valid detection of behavioral changes in the context of affective episodes. CONCLUSION: The clarification and operationalization of psychopathological constructs to allow for the measurement of objectively observable and ascertainable behavioral changes during depressive and (hypo)manic states are essential for the successful application of modern mobile technologies in the diagnostics and treatment of bipolar disorders.

Clock gene expression in the liver of streptozotocin-induced and spontaneous type 1 diabetic rats.

Several investigations have shown a relation between diabetes and alterations of the liver circadian clock. We investigated the diurnal expression of clock genes and clock-controlled genes (CCGs) in 3-hour intervals for a 24-h period in the livers of male streptozotocin (STZ)-treated rats, male spontaneous type 1 diabetic LEW.1AR1-iddm (Iddm) rats, and Iddm rats treated for 10 days with insulin. Hepatic mRNA was extracted, and the relative expression of clock genes (Per1, Per2, Bmal1, Clock, Cry1), as well as CCGs (Dbp, E4bp4, RevErbα, Rorα, Pparγ), was analyzed by reverse transcription followed by real-time polymerase chain reaction. Diabetic STZ and Iddm rats, as well as insulin-substituted Iddm rats, exhibited a significant diurnal expression pattern of clock genes as determined by Cosinor analysis; however, the MESOR (midline estimating statistic of rhythm) of Bmal1, Per2, and Clock transcript expression was altered in Iddm and insulin-substituted Iddm rats. The hepatic expression of the CCGs Dbp and RevErbα revealed a diurnal rhythm in all investigated groups. Insulin administration to Iddm rats normalized the enhanced MESOR in the expression of Dbp, RevErbα, and E4bp4 to the levels of normoglycemic controls. Cosinor analysis indicated no diurnal rhythm of Pparγ expression in the livers of diabetic STZ or Iddm rats or in those of insulin-substituted Iddm rats. Also, insulin substitution could not reverse the decreased MESOR of Pparγ expression in Iddm rats. In consequence of the diabetic disease, changes in the expression of clock genes and CCGs suggest alterations in the hepatic peripheral clock mechanism.

GLUT4 in the endocrine pancreas--indicating an impact in pancreatic islet cell physiology?

The glucose transporter GLUT4 is well known to facilitate the transport of blood glucose into insulin-sensitive muscle and adipose tissue. In this study, molecular, immunohistochemical, and Western blot investigations revealed evidence that GLUT4 is also located in the mouse, rat, and human endocrine pancreas. In addition, high glucose decreased and insulin elevated the GLUT4 expression in pancreatic α-cells. In contrast, high glucose increased GLUT4 expression, whereas insulin led to a reduced expression level of the glucose transporter in pancreatic ß-cells. In vivo experiments showed that in pancreatic tissue of type 2 diabetic rats as well as type 2 diabetic patients, the GLUT4 expression is significantly increased compared to the nondiabetic control group. Furthermore, type 1 diabetic rats exhibited reduced GLUT4 transcript levels in pancreatic tissue, whereas insulin treatment of type 1 diabetic animals enhanced the GLUT4 expression back to control levels. These data provide evidence for the existence of GLUT4 in the endocrine pancreas and indicate a physiological relevance of this glucose transporter as well as characteristic changes in diabetic disease.

Catecholamines are the key for explaining the biological relevance of insulin-melatonin antagonisms in type 1 and type 2 diabetes.

In this paper, we analyze the biological relevance of melatonin in diabetogenesis. As has recently been demonstrated, melatonin decreases insulin secretion via specific melatonin receptor isoforms (MT1 and MT2) in the pancreatic ß-cells. In addition, type 2 diabetic rats, as well as patients, exhibit decreased melatonin levels, whereas the levels in type 1 diabetic rats are increased. The latter effects were normalized by insulin substitution, which signifies that a specific receptor-mediated insulin-melatonin antagonism exists. These results are in agreement with several recent genome-wide association studies, which have identified a number of single nucleotide polymorphisms in the MTNR1B gene, encoding the MT2 receptor, that were closely associated with a higher prognostic risk of developing type 2 diabetes. We hypothesize that catecholamines, which decrease insulin levels and stimulate melatonin synthesis, control insulin-melatonin interactions. The present results support this assertion as we show that catecholamines are increased in type 1 but are diminished in type 2 diabetes. Another important line of inquiry involves the fact that melatonin protects the ß-cells against functional overcharge and, consequently, hinders the development of type 2 diabetes. In this context, it is striking that at advanced ages, melatonin levels are reduced and the incidence of type 2 diabetes is increased. Thus, melatonin appears to have a protective biological role. Here, we strongly repudiate misconceptions, resulting from observations that melatonin reduces the plasma insulin level, that the blockage of melatonin receptors would be of benefit in the treatment of type 2 diabetes.

The insulin-melatonin antagonism: studies in the LEW.1AR1-iddm rat (an animal model of human type 1 diabetes mellitus).

AIMS/HYPOTHESIS: It is well documented that melatonin influences insulin secretion mediated by G-protein-coupled melatonin receptor isoforms MT1 and MT2, which are present in rat and human pancreatic islets, as well as in rat insulinoma cells. Recent investigations have proven that hyperinsulinaemic Goto-Kakizaki (GK) rats, which are a rat model of type 2 diabetic rats, and humans have decreased melatonin plasma levels, whereas a streptozotocin-induced rat model of diabetes developed reduced insulin levels combined with increased melatonin levels. METHODS: Plasma levels of glucose, insulin and melatonin as well as RNA expression of pineal Aanat, Hiomt (also known as Asmt), insulin receptor, adrenoceptor ß1 and the clock genes Per1 and Bmal1 (also known as Arntl) were determined in male and female LEW.1AR1-iddm rats as well as in insulin-substituted LEW.1AR1-iddm rats. RESULTS: Severe hypoinsulinaemia in diabetic LEW.1AR1-iddm rats was associated with decreased body weight and increased melatonin plasma levels combined with mainly elevated expression of Aanat, Hiomt, pineal insulin receptor and adrenoceptor ß1. The changes were normalised by insulin substitution. Diurnal profiles of plasma melatonin and of antagonistic clock genes Per1 and Bmal1 were maintained in diabetic and insulin-substituted rats. CONCLUSIONS/INTERPRETATION: The assumed causal relation between elevated melatonin and reduced insulin levels in LEW.1AR1-iddm rats is supported by the observation that insulin substitution normalised these changes. Further support for this interpretation comes from the observation that in GK rats an increase of plasma insulin was combined with a decrease of plasma noradrenaline (norepinephrine), the most important activator of melatonin synthesis. These relationships between the noradrenergic and insulin pathway support the existence of melatonin-insulin antagonism.

Transcripts of calcium/calmodulin-dependent kinases are changed after forskolin- or IBMX-induced insulin secretion due to melatonin treatment of rat insulinoma beta-cells (INS-1).

The objective of the present study was to examine the effects of melatonin on transcripts of isoforms of calcium/calmodulin-dependent protein kinases in rat insulinoma beta-cells INS-1. Investigations show that calcium/calmodulin-dependent kinase IV and calcium/calmodulin-dependent kinase 2d are expressed in human and rat pancreatic islets and INS-1 cells. By application of either forskolin or 3-isobutyl-1-methylxanthine for 6 hours, calcium spiking was evoked and the release of insulin was increased. The expression of the calcium/calmodulin-dependent kinase IV and calcium/calmodulin-dependent kinase 2d transcripts was significantly increased due to forskolin or 3-isobutyl-1-methylxanthine. Acute melatonin treatment (6 h) in the presence of either forskolin or 3-isobutyl-1-methylxanthine caused a significant decrease in insulin release and induced significant downregulation of calcium/calmodulin-dependent kinase IV and calcium/calmodulin-dependent kinase 2d transcripts in INS-1 batch cultures. The attenuating effect of melatonin on transcripts could be almost completely reversed by preincubation with the melatonin receptor antagonist luzindole. Thus, the insulin-inhibiting effect of melatonin in INS-1 cells is associated with significant changes in transcripts of calcium-signaling components suggesting that melatonin influences gene expression of components, which are known to be involved in insulin secretion or insulin gene expression.

Did the gradual loss of GLUT2 cause a shift to diabetic disorders in the New Zealand obese mouse (NZO/Hl)?

The New Zealand obese mouse (NZO/Hl) is characterised by hereditary obesity and type-2 diabetes, including insulin resistance, hyperinsulinaemia, and glucose intolerance. In other diabetic models, it has been revealed that the proper functioning of the glucose transporter isoform 2 (GLUT2) is essential for adequate secretion of insulin. The aim of this study was to compare the distribution of islet cells and GLUT2, as well as the expression of GLUT2-mRNA, in the pancreas of NZO mice and metabolically unimpaired NMRI (Naval Medical Research Institute) mice. Pancreas tissue was obtained from different stages of development. For molecular determination of the expression level of GLUT2-mRNA, total-RNA was extracted from the pancreas and analysed by quantitative real-time RT-PCR. All investigated NZO mice displayed increased weight, elevated hyperinsulinaemia, and slightly enhanced blood glucose levels compared with the NMRI control mice. By means of immunofluorescence microscopy drastically reduced insulin levels were detected, which might be compensated by the observed islet cell hyperplasia and hypertrophy. Furthermore, the normally peripheral localisation of the alpha-cells within islets was disturbed. By contrast, there were no changes in somatostatin cell distribution. However, considerable differences appeared with regard to GLUT2: whereas the beta-cells of NMRI mice showed dense immunostaining of the GLUT2 transporter on the cell surface, in all age groups of NZO mice, GLUT2 on the plasma membranes was reduced and dispersed in the cytoplasm. These findings agree with the molecular biological results, which displayed decreased mRNA-expression of GLUT2. In summary, the observed alteration of islet morphology and of GLUT2 expression in diabetic mice complements our previous results from a superfusion protocol and further clarifies the mechanisms of diabetogenesis in NZO mice.

Indication of circadian oscillations in the rat pancreas.

The central circadian oscillator of the suprachiasmatic nucleus controls diurnal rhythmicity of the body with light as its dominant zeitgeber. Recently, peripheral oscillators have been detected in liver and heart, which follow as yet unidentified cues. In this study real-time reverse transcription-polymerase chain reaction (RT-PCR) was used in analysis of the expression of the major clock genes Per1, Per2, Bmal1, Cry1, Tim (timeless) and Clock, as well as of the output genes Dbp and Rev-erbalpha in the pancreatic tissue of rats. The results presented here indicate a robust circadian expression of clock genes (e.g. Per1 and Bmal1) and the probable existence of a peripheral oscillator in the pancreas. Whether this oscillator regulates the diverse functions of the islets of Langerhans remains to be elucidated.

The natural beta-carbolines facilitate inositol phosphate accumulation by activating small G-proteins in human neuroblastoma cells (SH-SY5Y).

The naturally occurring beta-carbolines exert psychotropic actions in humans and have numerous behavioral effects in animals. The known in vitro activities of these substances do not provide a satisfactory explanation for their in vivo effects. The present study was undertaken to explore the possibility of a specific signal transduction pathway. The human neuroblastoma cell line SH-SY5Y was used as a model system. High-affinity binding sites for [3H]norharman (synonymous: beta-carboline) were detected. Pharmacological characterization revealed displacement of the ligand by beta-carbolines, to a weaker extent by indoleamines, but not by opioids, muscarinic receptor agonists, metabotropic glutamate receptor agonists or several peptide neurotransmitters. Inositol phosphate accumulation was only slightly affected by the beta-carbolines. However, the action of carbachol was clearly facilitated in a dose-dependent and pertussis toxin-insensitive manner. Pretreatment of the cells with Clostridium difficile toxin B blocked the facilitating effect of the beta-carbolines by concentrations which did not affect the action of carbachol alone. This suggests that low molecular weight GTP-binding proteins are involved in the facilitating action of the beta-carbolines. This mechanism was further supported by experiments measuring the concentrations of phosphatidylinositol phosphates after various activating compounds. In conclusion, the facilitating effect of beta-carbolines on inositol phosphate accumulation could play a part in the actions of beta-carbolines and may be produced by stimulating the generation of phosphatidylinositol-4,5-bisphosphate (PtdIns-4,5-P2), the key component in the activation of phosphoinositide-phospholipase C.

Characterization of growth hormone gene expression in the pituitary and plasma growth hormone concentrations during posthatch development in the chicken.

In this study both sexes of two strains of chicken with genetically different growth potentials (broiler- and laying-type) were used to investigate growth hormone (GH) gene expression during posthatch development from day 7 (D7) to D56 by using the in situ hybridization technique and Northern analysis. In pituitaries of both strains a high GH mRNA signal was found as early as D7 by in situ hybridization, showing clear differences in the pattern of gene expression between the two strains. By Northern hybridization sex differences were detectable in all age groups of broilers, with higher levels throughout in males. In layers, however, females showed consistently higher levels compared with males until D21. While signal intensities decreased in the broiler strain during the investigation period, the layer-type strain seemed to express GH mRNA more continuously, reaching significantly (P < 0.01) higher GH mRNA levels than broilers at D56. Plasma GH concentrations ran parallel to GH mRNA in early stages but showed a peak earlier at D14 and decreased after D35 in both sexes and strains. Determination of growth as weekly weight gains, however, proved that a period of rapid growth (at a higher level in both sexes of the broiler strain) at D7 was followed by a strong decrease from D14 to D21. A plateau of constant growth was reached until the end of the observation period with similar rates in both strains and sexes. Analysis of plasma thyroid hormones tri-iodothyronine/thyroxine (T3/T4) showed an increase in T3 concentrations in both strains and sexes in early stages and a decrease thereafter. No clear strain differences were measured. T4 plasma concentrations increased from D7 to D14 in broilers and D21 in layers when a plateau was reached. From the results we conclude that generally there is a good correlation between GH mRNA and plasma GH concentrations in both strains investigated. Neither parameter, however, is coupled directly with the growth rate. Thus the early rapid growth corresponds to relatively low levels of GH mRNA and plasma GH concentrations, but high T3 levels. Later, decreased growth rates are linked to increasing amounts of GH mRNA as well as increasing plasma GH concentrations in both layers and broilers. Towards the end of the observation period there was a strain divergence visible with increased amounts of GH mRNA in layers but a strong reduction in broilers. Moreover, plasma GH concentrations decreased more slowly in layers than in broilers.

Ontogeny of growth hormone and LH beta-, FSH beta- and alpha-subunit mRNA levels in the porcine fetal and neonatal anterior pituitary.

The mechanism underlying the ontogenetic increase in plasma growth hormone (GH), luteinizing hormone (LH) and follicle stimulating hormone (FSH) concentration during fetal life in mammalian species and the prenatal sex difference in these hormones in some species is not fully understood. To this end anterior pituitaries were collected from German Landrace fetuses and piglets at day (d) 50, 65, 80, 95, 110 pc and d 6 pp and pituitary GH, LH beta, FSH beta and alpha-subunit mRNA levels were determined by measuring Northern blot hybridization signals. GH mRNA was detected in both sexes as early as d 50 pc. The mRNA level markedly increased with age in both sexes (males > females, P < or = 0.05) reaching its maximum at d 95/110 pc. LH beta mRNA signals were first detected at d 50 pc in females and at d 65 pc in males increasing thereafter to a maximum at d 6 pp in both sexes (P < or = 0.05). In males the augmentation in LH beta mRNA was delayed compared to females (P < or = 0.01). Before d 80 pc no FSH beta mRNA hybridization signals were apparent. Thereafter the mRNA level continuously increased with age (P < or = 0.01) in both sexes reaching its maximum at d 6 pp. The FSH beta mRNA level in females was always higher than in males (P < or = 0.01). As early as d 50 pc the alpha-subunit mRNA level was high in both sexes and further increased without sex difference to d 6 pp (P < or = 0.05). In conclusion, the mRNA levels of GH, LH beta and FSH beta are age and sex dependent during fetal development. We suggest that the fetal increase in plasma GH concentration can be accounted for by changes in GH mRNA levels, while the dramatic perinatal decrease in plasma GH concentration seems to be primarily controlled at the posttranslational and/or secretion level. The fetal sex difference and the increase in plasma LH and FSH concentrations seems to be primarily dependent on the cellular concentration of the gonadotropin beta-subunit mRNAs and/or number of gonadotrophs.

Arginine vasotocin gene expression and hormone synthesis during ontogeny of the chicken embryo and the newborn chick.

Chicken embryos at different developmental stages (embryonal day (E) 6 to 21) and chicks at posthatch day 1 (D1) were monitored for the development of their hypothalamo-neurohypophysial system as indicated by the kinetics of arginine vasotocin (AVT) gene expression via mRNA concentration and brain AVT content. Our data concerning the onset of gene expression support previous results from our laboratory and others about an early activation of the AVT gene transcriptional and translational activity around E6. We could detect measurable amounts of AVT in chicken embryo brains at E6 and an exponential increase during further development until D1. Dot blots of hypothalamic RNA extracts indicated that AVT gene transcript concentrations rose between E12 and E17 and slightly dropped thereafter. Northern hybridization showed that this drop was caused by a decrease of full length message and an increase of smaller transcripts during late embryonal and D1 stages, probably an AVT mRNA specific degradation phenomenon. The dissociation between the increase of AVT concentration and AVT mRNA concentration visible at the D1 stage might be due to accumulation and storage of AVT in the magnocellular neurons, preferentially in their axon terminals in the neurohypophysis. Blood samples taken from E14 onwards revealed a constant increase in plasma osmolality and plasma AVT concentration. Our data suggest that, in the chicken, AVT seems to be required early during embryonal development, either for osmoregulatory or further unknown functions.

Impaired immunoglobulin M production by incubation of hybridoma cells with ethanol.

Several reports have presented results that demonstrate suppression of the immune system by ethanol. Using a hybridoma cell model, we studied the effects of ethanol on cell proliferation and on the production of immunoglobulin M (IgM) antibodies. The number of cells decreased while incubated with as little as 25 mM ethanol but not in a clonal subline incapable of IgM production, indicative of an increased vulnerability associated with the antibody-producing machinery. Levels of antibodies in cell culture supernatants were monitored by -heavy-chain-specific and -light-chain-specific enzyme-linked immunosorbent assays. We found a significant decrease in antibody concentration at 200 mM ethanol compared with findings for nonexposed cells. In addition, lower -chain compared with -chain values were monitored at ethanol concentrations of 50 mM and higher. This difference suggests irregular composition of the antibodies in the supernatant. Determination of IgM levels within the hybridoma cells revealed a linear increase in antibody concentrations by as much as three times the control levels with increasing ethanol concentrations when correlated with cell numbers. Analysis of the mRNA levels of two ethanol-inducible stress proteins, the 78-kilodalton glucose-regulated protein (GRP78) and the 70-kilodalton heat-shock protein (HSC70), by quantitative Northern hybridization yielded increased mRNA in a nonlinear fashion. The results demonstrate that ethanol impairs IgM composition, whereas antibody production within hybridoma cells is increased and the assembling machinery is activated, indicating compensating processes.

The hypothalamo-neurohypophyseal system in birds.

The hypothalamus is the central integrative structure for the neuroendocrine regulation of body fluid homeostasis. Despite many different modulatory endocrine and peptidergic inputs there are three main neuroendocrine 'channels' involved in osmoregulation: 1. corticotropin releasing hormone (CRH)-adrenocorticotropic hormone (ACTH)-adrenal steroids (aldosterone, corticosterone); 2. renin-angiotensin; 3. arginine-vasotocin (AVT). In this paper the important role of the AVT synthesizing hypothalamo-neurohypophysial system (HNS) in the control of osmoregulation is described. Although the onset of AVT gene expression and peptide synthesis is as early as around day 6 of embryonal life in the chick, osmoregulatory function may not occur before the end of the second week of incubation. Rapid maturational processes during the last week of embryonal development lead to a 'close-to-mature' response of the AVT secretory system to physiological (osmotic) stimulation in the one-day-old chicken. Stable AVT plasma concentrations imply continuous secretion and/or degradation of this peptide in the blood of the chicken. Osmotic challenge causes not only an increase in plasma osmolality, plasma sodium and AVT concentration, but also AVT gene expression is upregulated. The AVT system participates in adaptational] processes during the perihatching period and its secretory activity depends on modulatory effects caused by suboptimal humidity during incubation.

Circadian and age-dependent expression patterns of GLUT2 and glucokinase in the pancreatic beta-cell of diabetic and nondiabetic rats.

Alterations in glucose sensing are well-known in both humans and animal models of non-insulin-dependent diabetes mellitus. However, the circadian- and age-dependent expression of glucose-sensing genes has not previously been investigated in vivo. In the present paper, we show a progressive loss of beta-cell GLUT2-mRNA and, by immunocytochemistry, a gain of soluble, cytoplasmic GLUT2-protein in Goto-Kakizaki rat islets. We report that GLUT2-mRNA shows significant diurnal variation, which is stronger in metabolically healthy rats. We also demonstrate the significant diurnal variation of glucokinase-mRNA, with higher levels in the pancreas of 6-week-old Goto-Kakizaki rats than in Wistar rats. This leads to a maximum glucose phosphorylation capacity in-phase with food intake, enhanced glucose-stimulated insulin secretion, and prevents postprandial hyperglycemia. Perfusion experiments showed a reduction in glucose-stimulated insulin secretion in Goto-Kakizaki rat islets with an impaired first phase. Hyperglycemia and hypoinsulinemia in newborn and up to 3-week-old Goto-Kakizaki rats are thus probably due to reduced pancreatic beta-cell content, reduced beta-cell insulin content and impaired glucose sensing. The de-compensation of the metabolic situation in 42-week-old Goto-Kakizaki rats is likely to be caused by beta-cell destruction accompanied by negligible accumulation of GLUT2 in the cell membrane and further reduction of glucokinase expression.

Circadian changes of ether-a-go-go-related-gene (Erg) potassium channel transcripts in the rat pancreas and beta-cell.

Evidence has previously been presented that circadian rhythms play a role in islet hormone secretion. Here, RT-PCR was used to monitor the circadian expression of ether-a-go-go-related gene (Erg) potassium channel isoforms and Erg1 splice variants. Immunohistochemistry was used to identify the pancreatic distribution patterns of ERG1a and ERG1b, as well as ERG2 and ERG3. The influence of ERG on insulin secretion was monitored by perfusion of rat INS-1 beta-cells with the blockers E-4031 and rBeKm-1. We identified Erg1a, Erg1b, Erg2 and Erg3 transcripts in islets and INS-1 cells. Immunohistochemistry showed differential expression of ERG isoforms in the islet. Ca(2+) imaging and electrophysiological recordings of INS-1 cells during ERG blocking by E-4031 indicated functional ERG channels. Serum shock treatment of INS-1 cells elicited a time-dependent expression response for Erg transcripts. These results add to the current understanding of the function of ERG channels in beta-cells and the circadian secretion processes of insulin.

Differences in the target specificity of the transactivating factors MHBs(t) and HBx of hepatitis B virus.

Transient transfections of tissue culture cells with plasmids encoding the transactivating factors MHBs(t) and HBx of hepatitis B virus result in transcriptional stimulation of multiple target genes. Our experiments show that the NF-kappaB-binding enhancer element of simian virus 40 (SV40) and the AP-1-binding enhancer element of the human metallothionein IIA gene mediate the transactivating function of MHBs(t) and HBx. In contrast, the elements GT(IIC + I) and Sph(II + I) of the SV40 enhancer, that, as a common feature, require binding of transcription factor TEF-1 for activity, efficiently mediate transactivation only by HBx but not by MHBs(t). This finding suggests that at least one regulatory pathway exists that can only be activated by HBx but not by MHBs(t).

Distribution of AVT gene expressing neurons in the prosencephalon of japanese quail and chicken.

Detailed anatomical studies about the distribution of arginine-vasotocin (AVT) gene expressing neurons in the avian brain are so far non-existent. Most of the investigations are limited to discrete brain nuclei in the chicken and concern particular experimental conditions. Since no data on AVT gene expression are up to now available in Japanese quail, we report here the distribution of AVT mRNA containing neurons within the prosencephalon of this bird and a comparison with the same system of the chicken. The AVT mRNA containing perikarya were detected with a specific AVT cDNA probe directed toward the distal 3' glycopeptide encoding part of the chicken AVT gene. The specificity of the probe was tested by Northern and Southern blot analysis. The AVT gene expressing neurons showed the same pattern of distribution in both species; however, a larger number of labelled neurons was observed in the chicken by visual inspection of the sections. AVT gene expressing neurons were distributed within the hypothalamus (in the medial and ventral aspects of the rostral preoptic area, in the lateral forebrain bundle, close to the ependyma of the third ventricle and in several periventricular hypothalamic nuclei) and in some extrahypothalamic regions (around the nucleus rotundus, close to the nucleus commissurae pallii and in a region probably corresponding to the nucleus strial terminalis). AVT gene expressing neurons were observed close to the organum septi laterale of the chicken. Taken together, these results confirm the specificity of the employed probe and its suitability for further studies on the modulation of AVT gene expression in quail.

[Plasmid profiles of different Salmonella serovars from poultry flocks in Germany]. / Plasmidprofile verschiedener Salmonella-Serovare aus Geflügelbeständen in Deutschland.

151 salmonella isolates belonging to 6 serotypes and originating from diverse material collected in broiler chicken farms, hatcheries and poultry slaughteries in Northern Germany during 1984 and 1990 were investigated for their plasmid DNA content. This is the first report describing plasmids of S. saint-paul, isolated from poultry lines. The different plasmids of 22 Md, 3.1 Md, 2.4 Md and 1.1 Md molecular weight can be used for further epidemiological studies of salmonellosis in poultry. The 62 Md plasmid of S. typhimurium and the 36 Md plasmid of S. enteritidis have been found to be serovar specific. The smaller plasmids (m. w. less than 5.0 Md), found in S. virchow and S. blockley can be used for further epidemiological studies.