Tidal interaction in binary-black-hole inspiral.

Tidal torque may lead to important effects in the strong field interaction at the end point of the inspiral of two rapidly rotating black holes. We point out here that in such a case the exchange of spin and orbital angular momentum may significantly affect the energy and waveform of late stage gravitational waves, and may play an important role in the shedding of excess angular momentum. We argue that numerical relativity, the only tool for determining the importance of tidal torque, should be more specifically focused on binary configurations with aligned, large, angular momenta.

A splice variant of estrogen receptor beta missing exon 3 displays altered subnuclear localization and capacity for transcriptional activation.

There are two separate estrogen receptors (ERs), ERalpha and ERbeta. The ERbeta gene is variably spliced, and in some cases variant expression is high. Besides the full-length ERbeta (equivalent to ERbeta1), splice variants can encode proteins bearing an insert within the ligand-binding domain (beta2), a deletion of exon 3 (ERbeta1delta3) disrupting the DNA-binding domain, or both (ERbeta2delta3). Here we examine the intracellular localization and transcriptional properties of each of the ERbeta splice variants heterologously expressed in cultured cells. In accordance with ERalpha, ERbeta1 and ERbeta2 are both distributed in a reticular pattern within the nucleus after exposure to ligand. In contrast, ERbeta1delta3 and ERbeta2delta3 localize to discrete spots within the nucleus in the presence of ER agonists. In the presence of ER antagonists, the delta3 variants are distributed diffusely within the nucleus. We also show that the spots are stable nuclear structures to which the delta3 variants localize in a ligand-dependent manner. Coactivator proteins of ER colocalize with delta3 variants in the spots in the presence of agonists. The delta3 variants of ERbeta can activate luciferase reporter constructs containing an activator protein complex-1 site, but not an estrogen response element (ERE). These data suggest that without an intact DNA-binding domain, ERbeta is functionally altered, allowing localization to discrete nuclear spots and activation from activator protein-1-containing reporter genes.

Differential expression of estrogen receptor beta splice variants in rat brain: identification and characterization of a novel variant missing exon 4.

Estrogen receptor beta (ER-beta) mRNA is found in abundance in rat brain. The distribution of ER-beta mRNA in brain differs from that of ER-alpha suggesting they subserve different functions. ER-beta mRNA has been reported to be variably spliced, in contrast to ER-alpha, resulting in numerous isoforms that possess different functional properties. The present study was undertaken to determine whether the isoforms of ER-beta mRNA are differentially distributed in different brain regions. In order to assess the range of transcript forms expressed in various brain regions in the same assay, a micropunch dissection technique was combined with semiquantitative RT-PCR. The relative abundance of each ER-beta isoform (beta1>beta2>beta1delta3>beta2delta3) was similar in all ER-beta positive brain regions with the exception of the hippocampus, which contained low levels of most isoforms and a fifth ER-beta isoform, which we are calling ER-beta1delta4. Based on its sequence, ER-beta1delta4 encodes an ER-beta that is missing exon 4. Initial characterization studies of this showed that it did not bind estrogen, and that, unlike ER-beta1, it localized to the cytoplasm when expressed in cultured cells. The distribution of ER-beta1delta4 was different from that of the other isoforms in that it was expressed at high levels in the hippocampus, where the other isoforms were low, and that it was nearly undetectable in the brain regions that expressed the highest levels of the other ER-beta splice variants. These data suggest that a highly complex pattern of estrogen signaling can occur in a region specific manner in the rat brain.

Expression of estrogen receptor-beta protein and mRNA in the cerebellum of the rat.

Estrogen has been shown to play a modulatory role in cerebellar neuronal signaling. Recent reports have also shown that estrogen receptor beta (ER-beta) mRNA is expressed in cerebellum. The purpose of the present study was to identify and map ER-beta protein expression, and to determine the identity of the major splice variants of ER-beta mRNA in the cerebellum. Polyclonal antibodies raised against the NH(3)- and COOH-termini of rat ER-beta were used for immunohistochemistry. Mapping of ER-beta immunoreactivity was compared with the distribution of ER-beta mRNA using in situ hybridization. We also determined, using RT-PCR, whether the ER-beta mRNA was variably spliced in cerebellum. Our results show that in all cases the distribution of ER-beta protein was identical to the distribution of ER-beta mRNA. Both Purkinje cells and scattered cells in the granule cell layer, perhaps golgi cells, robustly expressed ER-beta. Reverse transcription-polymerase chain reaction analysis showed that three splice variants in addition to wild type ER-beta are expressed in rat cerebellum. However, wild type ER-beta was the predominant form. These observations provide anatomical evidence that neurons in the cerebellum express ER-beta and thus may be targets of estrogen action.

Estrogen and progesterone regulation of human fibroblast-like synoviocyte function in vitro: implications in rheumatoid arthritis.

OBJECTIVE: Despite increasing evidence regarding the significance of sex hormones in rheumatoid arthritis (RA), their etiopathological role and potential longterm effect on joint destruction remain unclear. We hypothesized that estrogen receptors (ER-alpha) are present in fibroblast-like synoviocytes, and 17beta-estradiol can modulate the production and activity of matrix degrading enzymes produced by these cells. Thus, depending on the endocrine balance, fibroblast-like synoviocyte activity can be suppressed or enhanced, leading to amelioration or exacerbation of the disease process, respectively. METHODS: By utilizing an in vitro cartilage invasion model, in combination with the molecular analyses of hormone receptors, matrix metalloproteinases (MMP) and their respective inhibitors, we investigated the effect of hormones (i.e., estrogen and progesterone) on fibroblast-like synoviocyte phenotypic changes, with particular emphasis on their functional interactions with cartilage. RESULTS: Our studies reveal the presence of functional ER-alpha in fibroblast-like synoviocytes. The findings indicate that estrogen exerts a stimulatory effect, while progesterone has an inhibitory effect on the expression of MMP, their tissue inhibitors (TIMP), and enzymatic activity of MMP produced by these cells. Furthermore, transfection of fibroblast-like synoviocytes with the ER-alpha gene resulted in the increased degradation and invasion of cartilage. CONCLUSION: We identified the presence of functional ER-alpha in fibroblast-like synoviocytes. This renders fibroblast-like synoviocytes as target cells for hormonal regulation. The regulatory effect of estrogen is partly targeted to the MMP and their respective inhibitors associated with fibroblast-like synoviocytes. Such studies provide a link between hormonal status and disease activity in RA and open new venues for future therapeutic intervention to combat this debilitating disease.

Two years after a job loss: long-term impact of the JOBS program on reemployment and mental health.

Analyses of data from a randomized field experiment with 1,801 participants (A.D. Vinokur, R.H. Price. & Y.Schul, 1995) examined the long-term effects of a job-search workshop (JOBS) and the independent effects of demographic and psychological factors on reemployment and mental health outcomes. Two years after the JOBS workshop, the experimental group had significantly higher levels of reemployment and monthly income, lower levels of depressive symptoms, lower likelihood of experiencing a major depressive episode in the last year, and better role and emotional functioning compared with the control group. Baseline job-search motivation and sense of mastery had both direct and interactive effects (with experimental condition) on reemployment and mental health outcomes, respectively. The interactive effects demonstrated larger benefits for those who had initial low levels of job-search motivation and mastery.

Estrogen receptor-beta messenger ribonucleic acid expression in the pituitary gland.

Estrogen plays a key role in the regulation of many pituitary hormones. The presence of estrogen receptor-beta (ER beta) messenger RNA (mRNA) has been demonstrated in the adult anterior pituitary by RT-PCR to be at a level much greater than that of ER beta mRNA. Because the number of ERs has been shown to change during development, in this study we examined the distribution of pituitary ER beta mRNA in adult and prepubertal rats. Using RT-PCR, we confirmed that ER beta mRNA expression is less than that of ER alpha mRNA in adult females. In contrast, in prepubertal female pituitaries, ER beta mRNA levels are much greater than those of ER alpha mRNA. Film densitometric analysis of whole pituitaries, similarly showed that ER beta mRNA is greater in prepubertal pituitaries than in adult pituitaries. However, after emulsion autoradiography, cell counts confirmed that prepubertal and adult pituitaries differ, not in the level of ER beta mRNA expression, but in the number of cells expressing ER beta mRNA. In postnatal day 15 pituitaries, there were twice as many cells per mm2 as in adults. A comparison between prepubertal males and females showed that females exhibited a 2-fold greater level of ER beta mRNA expression. To determine which cell types express ER beta mRNA, we performed in situ hybridization for ER beta mRNA coupled with immunohistochemistry for FSH or PRL. In prepubertal pituitaries, 84.5 +/- 2.3% of FSH-immunoreactive cells also express ER beta. Nearly all of the PRL-immunoreactive cells lacked ER beta mRNA. These data demonstrate sex- and age-related differences in ER beta mRNA expression in the anterior pituitary. Furthermore, these data suggest that ER beta is not the specific mediator of estrogen action in lactotrophs, whereas ER beta may be the prime mediator of estrogen action in FSH-containing gonadotrophs.

In praise of a cumulative prevention science.

Durlak and Wells (1997) provide a pivotal appraisal of prevention research on children and adolescents. Their meta-analytic approach has the advantages of reducing scientific misjudgments based on single studies, and providing a more balanced evaluation of impact of various interventions; it provides an opportunity for hypothesis finding helps set methodological standards, allows assessment of working classifications in the field, and an evaluation of the maturity of the prevention field itself. New developmental tasks for the field include incorporating and pursuing the leads produced by these findings, conducting similar research syntheses with other populations and outcomes, and using the results as an impetus to increased operational precision and parsimony.

National research agenda for prevention research. The National Institute of Mental Health Report.

This article reports some of the most promising ideas to emerge from a review conducted by the National Institute of Mental Health of the achievements and prospects for research on the prevention of mental disorders. These ideas are organized around 3 conceptual hubs: the development and transformation of biological and social risk and protective factors across the life span, classifying and relating various approaches to preventive interventions in a single logical system, and concepts about community contexts in which prevention trials are executed. These conceptual hubs clarify the relationship between 3 forms of research in prevention: longitudinal studies of risk, randomized preventive intervention trials, and the implementation of successful interventions as part of routine community practice.

Hard times and hurtful partners: how financial strain affects depression and relationship satisfaction of unemployed persons and their spouses.

The process linking unemployment and economic hardship to depression and marital or relationship satisfaction in couples was examined. Using structural equation modeling, the authors tested models in which financial strain and partners' symptoms of depression influence the behavioral exchange of the couples in terms of social support and social undermining and, in turn, the effects of support and undermining on relationship satisfaction and depressive symptoms. The analyses were based on longitudinal data from 815 recently unemployed job seekers and their spouses or partners. The results demonstrated that financial strain had significant effects on depressive symptoms of both partners, which in turn led the partner to withdraw social support and increase social undermining. Reduced supportive and increase undermining behaviors had additional adverse effects on satisfaction with the relationship and on depressive symptoms.

Impact of the JOBS intervention on unemployed workers varying in risk for depression.

Reports the results of the JOBS II randomized field experiment that included a sample of 1,801 recent job losers, 671 of which participated in a modified version of the JOBS I intervention for unemployed workers (Caplan, Vinokur, Price, & van Ryn, 1989). The intervention focused on enhancing the sense of mastery through the acquisition of job-search and problem-solving skills, and on inoculation against setbacks. JOBS II was intended to prevent poor mental health and to promote high quality reemployment. The study tested whether the efficacy of the intervention could be increased by screening and oversampling respondents who were at higher risk for a significant increase in depressive symptoms. Results demonstrated that the intervention primarily benefited the reemployment and mental health outcomes of the high-risk respondents. This suggests the feasibility of enhancing the efficacy of this preventive intervention by targeting it for high-risk unemployed workers who could be identified prospectively.

Immunocytochemical evidence that quisqualate is selectively internalized into a subset of hippocampal neurons.

Quisqualic acid (QUIS) has been shown to interact with several glutamate receptor subtypes and uptake sites. We have previously demonstrated that a brief exposure of hippocampal cells to QUIS sensitizes them to depolarization by the alpha-amino-omega-phosphonate analogues of glutamate, AP4, AP5, and AP6. This QUIS-induced sensitization is accompanied by the active uptake of QUIS into hippocampal slices. In order to localize the sites of QUIS uptake into rat hippocampal slices, a polyclonal antibody against QUIS was raised in rabbits. Utilizing immunocytochemical techniques, we have identified immunoreactive axons and dendrites after brief exposure times to QUIS, and perikarya after longer exposure times to QUIS. The intensity of the QUIS immunoreactivity increased as the exposure time to QUIS increased. QUIS immunoreactivity was primarily found in stratum oriens and stratum radiatum, of regions CA1, CA2, and CA3 of the hippocampus as well as in the hilus and molecular layer of the dentate gyrus. The distribution and morphology of QUIS immunoreactive cells appeared to be similar to those of GABAergic interneurons. Glial fibrillary acidic protein (GFAP) did not co-localize with the QUIS-internalizing cells suggesting that they are not glia. Ultrastructural analysis revealed QUIS immunoreactive profiles within the stratum radiatum. Immunostained profiles at both the light and EM levels appeared, in many cases, to be swollen and showed signs of degeneration. Such changes were only evident in tissue exposed to QUIS. These data demonstrate that QUIS is taken up by a select group of neurons in the rat hippocampus.

Distribution of nitric oxide synthase-immunoreactive interneurons in the spinal trigeminal nucleus.

The spinal trigeminal nucleus is involved in the transmission of orofacial sensory information. Neither the distribution of the neuromessenger, nitric oxide, within the trigeminal system nor the possible relationship of this simple gas with trigeminothalamic neurons has been carefully studied. Using immunocytochemical (against nitric oxide synthase) and histochemical (NADPH-diaphorase staining) techniques, we have found that nitric oxide neurons and processes are more prominent in the nucleus caudalis and the dorsomedial aspect of the nucleus oralis than in other spinal trigeminal regions. To study the relationship of nitric oxide to trigeminothalamic neurons and intertrigeminal interneurons of the spinal trigeminal nucleus, spinal trigeminal neurons were retrogradely labeled with fluorogold by thalamic injections or by injections into the junction of the nucleus interpolaris and nucleus caudalis. Medullary sections were subsequently processed with NADPH-diaphorase histochemistry. None of the diaphorase-stained neurons in the spinal trigeminal nucleus was found to contain fluorogold; however, some diaphorase-stained processes were found in close proximity to trigeminothalamic neurons. Following spinal trigeminal nucleus injections, many diaphorase-stained neurons were found to contain fluorogold, especially in the nucleus caudalis, suggesting that nitric oxide-containing neurons in the spinal trigeminal nucleus are intertrigeminal interneurons. Collectively, these data indicate that nitric oxide is most prominent in interneurons located in nucleus caudalis and that these interneurons give rise to processes that appose trigeminothalamic neurons, raising the possibility that they may indirectly influence orofacial nociceptive processing at the level of the spinal trigeminal nucleus via nitric oxide production.

Primary prevention of secondary disorders: a proposal and agenda.

This paper calls for consideration of a new class of preventive interventions designed explicitly to prevent comorbidity of psychiatric disorders. Epidemiologic data show that successful interventions of this type could be extremely valuable, as up to half of lifetime psychiatric disorders and an even larger percent of chronic and seriously impairing disorders occur to people with a prior history of some other disorder. Furthermore, a review of etiologic hypotheses concerning the causes of comorbidity suggests that interventions aimed at primary prevention of secondary disorders might be feasible. However, more basic risk factor research is needed on the causes of comorbidity before we can make a clear assessment of feasibility and discover promising intervention targets. A number of methodological problems arise in carrying out this type of formative research. These problems are reviewed and suggestions are offered for solutions involving innovations in measurement, design, and data analysis.

Cerebellar excitatory and inhibitory amino acid receptors in multiple system atrophy.

We studied excitatory and inhibitory amino acid binding sites autoradiographically in control and multiple system atrophy (MSA) cerebella. Within the dentate nucleus (DN) of MSA specimens, we found a significant increase in the level of GABAA, benzodiazepine, and metabotropic binding sites compared with controls. In the granule cell layer, kainate, N-methyl-D-aspartate, and GABAA binding sites were all decreased significantly in MSA specimens compared with controls. In the molecular layer of MSA cerebellum, alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate binding sites were decreased significantly compared with controls. Cerebellar cortical binding site decreases are likely due to Purkinje and granule cell loss. The increase of binding site levels in DN of MSA specimens may represent receptor up-regulation reflecting loss of descending inhibitory Purkinje cell and ascending excitatory afferents to the DN.

Nitric oxide synthase neurons in rat brain express more NMDA receptor mRNA than non-NOS neurons.

Nitric oxide, the gaseous neuronal messenger, is produced by nitric oxide synthase (NOS) in a Ca2+ dependent manner. Indirect evidence suggests that glutamate, acting through the NMDA subtype of excitatory amino acid receptor, is the principle activator signal for NOS in the brain. To date there is a lack of direct evidence demonstrating NOS and NMDA receptors, or receptor message, in the same neurons. A double labeling technique was developed which uses NOS immunocytochemistry in combination with in situ hybridization for NMDA NR1 receptor mRNA. Quantitative analysis of the silver grain labeling resulting from the hybridization procedure revealed that the majority of NOS positive cells in the cerebral cortex, striatum and midbrain contained a significantly greater amount of NR1 receptor mRNA than non-NOS neurons in the same regions. The amount of NR1 mRNA per cell varies in the cortical and striatal NOS cells, with some cells showing no NR1 expression. These results indicate that NOS containing neurons do not reflect a single phenotype of mRNA expression and further suggest that some NOS cells may be activated by a non-NMDA receptor mechanism.

Nitric oxide synthase is found in some spinothalamic neurons and in neuronal processes that appose spinal neurons that express Fos induced by noxious stimulation.

To determine if nitric oxide (NO) and Fos immunoreactivity induced by noxious stimulation were colocalized in spinothalamic neurons, double-staining immunocytochemical techniques were combined with retrograde neuroanatomical tracing procedures. Initial studies on three rats demonstrated that Fos and nitric oxide synthase (NOS), the synthesizing enzyme for nitric oxide, did not coexist in spinothalamic tract neurons. However, some spinothalamic neurons were found to contain NOS and some NOS immunoreactive processes were found to appose Fos containing neurons. Thus the remainder of the study: (1) analyzed the relationship of NOS positive neuronal processes with Fos stained neurons using a Fos immunocytochemical technique in combination with either NOS immunofluorescence or NADPH-diaphorase histochemistry; and (2) quantitated the number of NOS containing cells that project to the thalamus using a combined immunofluorescent-retrograde tracing procedure. Both NOS-like immunoreactive (NOS IR) neuronal processes and NADPH-diaphorase positive neuronal processes in the dorsal horn of the lumbar spinal cord were found to appose Fos positive neurons located in laminae I and II of the dorsal horn. Approximately 40% of Fos-labeled cells in these superficial laminae were found to be in apposition to or in close proximity to NOS labeled neuronal processes. Examination of spinal cord sections for NOS-containing spinothalamic tract neurons revealed that lamina X was the only spinal cord region containing such double-labeled neurons. Further quantification revealed that approximately 10% of NOS positive neurons in lamina X were double-labeled with Fluorogold. These findings support the hypothesis that nitric oxide is involved in nociceptive events occurring in the spinal cord in response to a peripheral noxious stimulus and further indicate that nitric oxide may contribute to the central transmission of spinothalamic information.

Excitatory amino acid receptor regulation after subthalamic nucleus lesions in the rat.

The subthalamic nucleus plays a pivotal role in the regulation of basal ganglia output. Recent electrophysiologic, lesion and immunocytochemical studies suggest that the subthalamic nucleus uses an excitatory amino acid as a neurotransmitter. After complete ablation of the subthalamic nucleus, we have examined the NMDA, AMPA, kainate and metabotropic subtypes of excitatory amino acid receptors in two major subthalamic projection areas (globus pallidus and substantia nigra pars reticulata) with quantitative autoradiography. Two weeks after ablation, binding sites for [3H]AMPA and [3H]kainate increased in substantia nigra pars reticulata ipsilateral to the lesion. In globus pallidus on the lesioned side, [3H]glutamate binding to the NMDA recognition site decreased. The results suggest that glutamate receptors regulate after interruption of subthalamic nucleus output.