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Background: Suicide rates and narcotic overdose have doubled since 2000. At least 30 percent of people with major depression are Treatment-Resistant (TR) and require novel therapeutics. ketamine at low doses has been shown in clinical trials to induce a rapid, short-lived anti-suicide and anti-depressant effect. Objective: To review the potential mechanism of action of ketamines’ alleviation of depressive symptoms from both animal and available human literature. Methods: This is a synthesis of information from papers listed in PUBMED Central. Although not exhaustive, this review highlights the most compelling work in the field related to this remarkable clinical rapid anti-depressant effect. Results: While there have been several theories and with some scientific evidence to date, the conclusion here is that currently, an exact and acceptable mechanism of action (MOA) for ketamines’ rapid anti-depressant effect is not apparent. The MOA of this compound with psychoactive abuse potential at a higher dosage and acute antidepressive effect in the most resistant patients is unknown. Discussion: Possible MOAs reviewed, include dopamine receptor modulation through epigenetic neuroadaptation via specific D1/D2 antagonism, D1 activation using optogenetic stimulation, and the role of D2/D3 availability in the ketamine therapeutic action. Conclusion: Unraveling MOA could guide the development of other unique Psychoplastogens capable of rapidly promoting structural and functional neural plasticity in cases of TR Major Depressive Episodes (MDE) and unipolar Major Depression Disorder (MDD).

To assess the performance of a symptom index (SI) and multivariate biomarker panel in the identification of ovarian cancer in women presenting for surgery with an adnexal mass.Prospective study of patients seen at a tertiary medical center. Following consent, patients completed an SI and preoperative serum was collected for individual markers (CA 125) and a second-generation FDA-cleared biomarker test (MIA2G). Results for the SI and MIA2G were correlated with operative findings and surgical pathology. Logistic regression modeling was performed to assess the interaction of the SI with MIA2G to determine the risk of malignancy (ROM).Of the 218 patients enrolled, the mean age was 53.6 years (range 18-86). One-hundred and forty-seven patients (67.4%) were postmenopausal. Sixty-four patients (29.4%) had epithelial ovarian cancer or fallopian tube cancer (EOC/FTC) and 17 (7.8%) had borderline ovarian tumors. A positive SI or MIA2G correctly identified 96.1% of patients with EOC/FTC. Using logistic regression, we found that both SI and MIA2G score were significantly associated with ROM (p 0.001). In a simulation with disease prevalence set at 5%, patients with a negative SI and a MIA2G score of 6 had a ROM of 1.8% whereas patients with the same MIA2G and positive SI had a 10.5% ROM, nearly a 6-fold higher risk.The combination of a patient-reported symptom index and refined biomarker panel allows for improved accuracy in the assessment for ovarian cancer in patients with an adnexal mass. This strategy could offer a personalized approach to addressing ROM to triage patients with an adnexal mass to appropriate care.

Small interfering RNAs (siRNA) and microRNAs (miRNA) are gaining considerable attention in the pharmaceutical and biotechnology industries, as research has revealed their likely clinical and agricultural applications. The capacity of siRNAs to dramatically and specifically reduce the expression of targeted genes has spawned multiple clinical trials to establish the therapeutic potential of small RNAs targeting viral, cancer and other disease-related genes. The successful application of siRNAs will enable the development of therapeutic applications based on miRNAs that have been observed to contribute to a variety of human diseases. This article reviews advances that have been made to apply small RNAs as therapeutics.

electron displays and lighting elements. [13,14] We strongly believe that one of the most prolific areas of their applications will be in biomedicine, where compact, strong, and high-performance devices can be engineered. These devices will exploit the properties of SWNTs and will compete with existing products. The novel technologies of diagnostics and therapeutics can be based on SWNT composites and individual tubes. Along these lines, SWNTs have been demonstrated as potential sensing materials of biological systems, [15–19] which are typically considered for the use in ex vivo modality. The potential use of SWNT-based structures for the purpose of healing neurological and brain-related injuries represents one of the major scientific and practical interests. The high mechanical strength and electrical properties possessed by SWNTs makes these materials perfect candidates for various prosthetic devices, including bone and joint repair. It is important to realize, however, that successful utilization of SWNT-based devices in biomedicine is hinged on the ability of such materials to interface with living cells, support their growth, and at the same time preserve their viability. [20–24] These factors are not well understood for any SWNT structures, which limits the development of in vivo, that is, implantable devices from such materials. The actual processes and techniques used for the preparation of macroscopic objects from SWNTs will play a significant role in determining cellular effects of SWNT composites. Substrates prepared from multi-walled carbon nanotubes (MWNTs) as well as SWNTs have been reported to be biocompatible platforms for neuronal growth and differentiation. [25–27] The use of carbon nanofiber composites as devices for neural- and bone-tissue-implant integration has also been described. [28] Molecular engineering of any SWNT-based composite should have a great effect on how the material performs during long-term contact with tissue. The layer-by-layer (LBL) approach to prepare SWNT structures can be particularly useful in this respect because it allows one to exert control over the structure of the SWNT/polymer systems from angstrom to nanometer and micrometer scale, which is necessity for the engineering of the cell/SWNT interface. [29] Recently, we demonstrated that SWNT LBL films can support the growth, viability, and differentiation of neuronal NG10815 neuroblastoma/glioma hybrid cells. [30] The first example of free-standing SWNT/polymer thin-film membranes that can be mechanically compatible with tissues and can be used as implants and repair devices for neurological- or brain-related injuries has been demonstrated. [30] The free-standing SWNT/ polyelectrolyte multilayers can serve as a prototype for a variety of nanotube-based implants.

Nanostructured thin films fabricated from semiconductor nanoparticles (NPs) are of great interest for biomedical applications, but NP materials based on heavy metals can be cytotoxic. In this work, the preparation of semiconductor NPs followed the protocol of layer-by-layer (LBL) assembly, which alleviates this problem. Collagen/poly(acrylic acid) bilayers were added to CdTe/polycation LBL films to produce porous collagen bilayers. Such stratified multilayer systems showed successful cell attachment and survival while native NP films were strongly cytotoxic.

The focus of our work on rapid actions of estrogens has been on the immuno-identification of a membrane version of the estrogen receptor-α (mERα) and the correlation of the presence of this receptor to the rapid secretion of prolactin in pituitary tumor cells. We demonstrated the mERα by both fluorescence and immuno-enzyme-cytochemistry and with both conventional and confocal microscopy in the cell line GH3/B6 and its sublines. Its presence on cells (including recently subcloned ones) is very heterogenous, unlike the nuclear ERα, which is present in every cell. An impeded ligand (estradiol covalently linked to BSA) binds to mERα and elicits the same response. A total of eight antibodies to ERα recognize mERα, making it likely that the membrane and nuclear proteins are highly related. Immuno-identification techniques have also been used to identify mERα on the MCF-7 human breast cancer cell line. Estradiol at very low concentrations elicits prolactin release from GH3/B6 cells within a few minutes of application. This response is bimodal, with effective concentrations in both the picomolar and nanomolar ranges. Prolactin release is also elicited or inhibited by ERα-specific antibodies. The characteristics of mERα and the membrane receptor for glucocorticoids have many similarities, suggesting that this mode of subcellular location/function alternative might be used by other members of the gene family.

Autopsy studies of patients with AIDS dementia have shown neuronal loss consistent with a neurotoxic component of this disease. In vitro studies suggest that viral products or cytokines from HIV-infected macrophages (Mphi) may modulate or directly mediate excitotoxic cell death of neurons. Mphi differentiated from peripheral mononuclear blood cultures were infected with HIV, and conditioned media (CM) were harvested from these cultures. Exposure of SK-N-MC (neuroblastoma) cells to CM from HIV-infected Mphi for 4, 24 or > or = 48 h resulted in a mean suppression of 12-34% of the glutamate transport Vmax with no appreciable change in transport Km. An astrocytoma tumor cell, U373MG, showed similar CM-mediated glutamate uptake suppression. Changes were evident in total and Na+-dependent glutamate uptake, with significantly more suppression of Na+-dependent uptake. Similar effects were seen with the nonmetabolizable transporter agonist D-aspartate, indicating that the effect was on transport and not metabolism. No suppression was seen with CM from uninfected Mphi or Mphi infected with heat-inactivated HIV. The magnitude of uptake suppression was not correlated with CM p24 values, and removal of CM virions by ultracentrifugation and immunoprecipitation did not alter the uptake-suppressive properties of infected Mphi CM. Uptake suppression was seen when Mphi were infected with Mphi-tropic strains HIV(SF162), HIV(JR-CSF), HIV(NFN-SX) and a Mphi-tropic patient isolate, but not the lymphotropic strain HIV(LAI). HIV-infected Mphi may produce substances which suppress neuronal and glial glutamate neurotransmitter uptake, resulting in higher extracellular glutamate levels and leading possibly to deficits in cell signaling and neurotoxicity.

Dopamine (DA), at concentrations greater than 100 microM, has previously been demonstrated to be toxic to mesencephalic, striatal and dorsal root ganglion cell cultures. Pharmacological experiments suggest that DA also may play a role in the cortical neurotoxicity caused by systemic administration of N-methyl-D-aspartate receptor antagonists such as phencyclidine and MK-801. In this study, the potential toxicity of DA in primary cortical cell cultures was determined in vitro. Using calcein and ethidium homodimer fluorescence as a marker for live and dead cells, respectively, we observed that a 24 h treatment with 10-100 microM DA produced a concentration-dependent increase in the number of ethidium homodimer-labelled cells. The cell death induced by 10 microM DA was dramatically reduced by co-administration of either superoxide dismutase and catalase or deferoxamine mesylate, an iron chelator. To verify this observation, the effects of 10 microM DA on the release of cytoplasmic lactate dehydrogenase (LDH) was measured. DA increased LDH release in a manner that was inhibited by both superoxide dismutase/catalase and deferoxamine. Nomifensine potentiated the effect of DA on LDH release, suggesting a protective role for DA uptake in this system. On the other hand, neither D1 nor D2 antagonists were able to prevent DA-induced LDH release. These data suggest that relatively low concentrations of DA can be injurious to cortical neurons through a mechanism that likely involves DA autooxidation and the formation of reactive oxygen species such as superoxide anion and hydroxyl radical. This mechanism may be important in the toxic effects of psychomotor stimulants such as amphetamine. However, the failure of DA receptor antagonists to affect DA-induced injury argues that the effect of DA on cortical neurons in culture does not model the toxic effect of phencyclidine and MK-801 observed in vivo.

Circulating hyaluronan (HA) levels were investigated as a function of age and diet in Fischer 344 male rats. A biphasic pattern of age-related changes was observed in rats fed ad libitum a diet in which the protein source was soya/fish meal. HA levels in 3- to 6- and 22- to 29-mo-old rats were not statistically different. However, HA levels in 12- to 20-mo-old rats were 10-29% of the levels in younger or aged adults. HA levels were also measured in rats fed ad libitum a semisynthetic diet in which the protein source was hydrolyzed casein. Whereas the two colonies exhibited similar biphasic age-related changes, HA levels differed 4- to 20-fold at every age examined. Caloric restriction affected HA levels in 19-mo-old casein-fed rats; HA levels were 2.3 times higher than age-matched controls and were not statistically different from young or aged animals. Serum and plasma HA levels were identical in the same individuals at all ages tested. These data suggest that HA turnover and metabolism in the rat are affected by age, dietary composition, and caloric intake.

We immunoselected GH(3)/B6 cells for a membrane estrogen receptor (mER) using antibodies generated against the rat intracellular ER (iER). Immunocytochemistry with anti-ER antibodies revealed bright fluorescence distributed in patches over the surface of mER-enriched cells, while cells immuno-depleted for mER showed only low-level membrane immunofluorescence. Quantitation via digital image analysis confirmed that immunoenriched populations show increases in both stained cell numbers and intensity of staining. Short-term culturing with serum reversibly decreased the intensity of immunostaining in mER-enriched cells to immuno-depleted cell levels. The mER-enriched populations initially contained ∼85% immunopositive cells in defined medium, but when cultured continuously with serum gradually decline to ∼22% immunopositive cells by 10 weeks. Cells enriched for mER showed a significant increase in rapid (after 2 or 5 min) prolactin release when treated with 17β-estradiol, while mER-depleted cells lacked this response. Immunoprecipitabie membrane proteins isolated from mER-enriched cells were 60,000, 74,000 and ∼ 200,000 MW, compared to an iER size of 67,000. Therefore, the presence and level of an mER that is antigenically related to iER is correlated with the ability of GH(3)/B6 cells to mediate a rapid action of estrogen.

For 179 male and female college and high school students, and 26 female eating-disordered patients body-image distortion was measured using a computer-based image-analysis of redrawn images of standardized human figures. Statistical analysis indicated that body-image distortion was the same for all groups. Body-image distortion was significantly and negatively related to weight:heigbt ratio as a function of a simple polynomial. These results suggest this evaluation of distortion of body-image yields a quantitative measure reliably related to weight status but also suggests the technique, and possibly measurement of body-image distortion in general, may not be a valid discriminator between eating-disordered and normal persons.

Cephalopods generally are thought to have only static iridophores, but this report provides qualitative and quantitative evidence for active control of certain iridescent cells in the dermis of the squid Lolliguncula brevis. In vivo observations indicate the expression of iridescence to be linked to agonistic or reproductive behavior. The neuromodulator acetylcholine (ACh) induced dramatic opitcla changes in active iridophores in vitro, whereas ACh had little effect on passive iridophores elsewhere in the mantle skin. Bath application of physiological concentrations of ACh (10(-7)M to 10(-6)M) to excised dermal skin layers transformed the active iridophores from a non-reflective diffuse blue to brightly iridescent colors, and this reaction was reversible and repeatable. The speed of change to iridescent in vitro corresponded well to the speed of changes in the living animal. Pharmacological results indicate the presence of muscarinic receptors in this system and that Ca++ is a mediator for the observed changes. Although ACh is present in physiological quantities in the dermal iridophore layer, it is possible that ACh release is not controlled directly by the nervous system because electrophysiological stimulation of major nerves in the periphery resulted in no iridescence in L. brevis; nor did silver staining or transmission electron microscopy reveal neuronal elements in the iridophore layer. Thus, active iridophores may be controlled by ACh acting as a hormone.

The remarkable optical and electrical properties of nanostructured materials are considered now as a source for a variety of biomaterials, biosensing, and cell interface applications. In this study, we report the first example of hybrid bionanodevice where absorption of light by thin films of quantum confined semiconductor nanoparticles of HgTe produced by the layer-by-layer assembly stimulate adherent neural cells via a sequence of photochemical and charge-transfer reactions. We also demonstrate an example of nanoscale engineering of the material driven by biological functionalities.

In this work, we demonstrate the use of Two-Photon Fluorescence Lifetime Imaging Microscopy (TP-FLIM) for intracellular calcium ([Ca 2+ ]i) measurement with a calcium sensitive fluorescent dye in a neuronal cell model and brain tissue. Calcium perturbations were induced via chemical stimulation and our results signify the potential of TP-FLIM for quantitative ([Ca 2+ ]i) measurement within the physiological range of intracellular [Ca 2+ ].

5556 Background: Many women with ovarian cancer receive non-guideline care. Failure to refer to a gynecologic oncologist is one likely cause. To improve specificity of Multivariate Index Assay (MIA...

5561 Background: The FDA cleared in vitro diagnostic multivariate index assay OVA1, with its intended utility to assist in referral of high risk patients to gynecologic oncologists, was by design t...

Single-walled carbon nanotubes (SWNTs) have unique mechanical, electrical, and optical properties and can be easily chemically modified; features that make them excellent candidate materials for applications as sensors and stimulators in neuronal tissue engineering. The purpose of this study was to demonstrate that SWNTs can support neuronal attachment and growth, that simple chemical modifications can be employed to control cell growth, that SWNTs do not interfere with ongoing neuronal function, and that neurons can be electrically coupled to SWNTs. Growth and attachment of the neuroblastoma*glioma NG108, a model neuronal cell, was assessed on unmodified SWNT substrates or substrates from SWNTs modified with 4-benzoic acid or 4-tert-butylphenyl functional groups using a simple functionalization method. SWNT films support cell growth, but at a reduced level compared to tissue culture-treated polystyrene. The order of viability and cell attachment was tissue culture treated polystyrene > SWNTs > 4-tert-butylphenyl-functionalized SWNTs > 4-benzoic acid-functionalized SWNTs. Decreased cell growth after culture on untreated (non adherent) polystyrene suggested that cell attachment was a critical determinant of proliferation and cell growth on SWNTs. Fluorescence and scanning electron microscopy revealed decreased neurite outgrowth in NG108 grown on SWNT substrates. We are also among the first groups to demonstrate electrical coupling of SWNTs and neurons by demonstrating that NG108 and rat primary peripheral neurons showed robust voltage-activated currents when electrically stimulated through transparent, conductive SWNT films. Our data suggest that SWNTs are flexible resource materials for tissue engineering application involving electrically excitable tissues such as muscles and nerves.

Whole-body hyperthermia (WBH) promotes cardiac protection against ischemia/reperfusion injury, in part by up-regulation of heat shock proteins (HSP). Whether heat stress also promotes up-regulation of angiogenic factors or induces endothelial cell proliferation is unknown. We studied the effects of heat stress on up-regulation of vascular endothelial growth factor (VEGF) and growth of new blood vessels following WBH. Anesthetized rats were subjected to WBH at 42 degrees C for 15 min. The control (n=23) and heated (n=55) groups were allowed to recover for 4, 12, 24, 48, or 72 h prior to harvesting the heart for Western Blot and immunohistochemical assessment of VEGF, HSP70, and platelet endothelial cell adhesion molecular-1 (PECAM-1). A significant increase in VEGF and HSP70 expression was observed as early as 4 h post-heating. The Western Blot analysis revealed a close temporal correlation between up-regulation of HSP70 and VEGF. Maximum VEGF and HSP70 expression occurred at 12 and 24 h post-heating in the left and right ventricles, respectively. The right ventricle showed the greatest expression of both VEGF and HSP70. Immunostaining revealed that VEGF was focally increased in the endothelial cells of capillaries, small arteries, and in interstitium. At 48 and 72 h post-heating, multiple areas of extensive capillary proliferation occurred in the epicardial region of the right ventricle. These observations were verified by quantitative analysis of the density of blood vessels as determined by PECAM-1 staining. Our experiments show that sublethal heat stress can lead to upregulation of both VEGF and HSP70 in cardiac tissue and promote focal endothelial proliferation in the heart.

Rattlesnakes, copperheads, and other pit vipers have highly sensitive heat detectors known as pit organs, which are used to sense and strike at prey. However, it is not currently known how temperature change triggers cellular and molecular events that activate neurons supplying the pit organ. We dissociated and cultured neurons from the trigeminal ganglia (TG) innervating the pit organs of the Western Diamondback rattlesnake ( Crotalus atrox) and the copperhead ( Agkistrodon contortix) to investigate electrophysiological responses to thermal stimuli. Whole cell voltage-clamp recordings indicated that 75% of the TG neurons from C. atrox and 74% of the TG neurons from A. contortix showed a unique temperature-activated inward current ( IΔT). We also found an IΔT-like current in 15% of TG neurons from the common garter snake, a species that does not have a specialized heat-sensing organ. A steep rise in the current-temperature relationship of IΔTstarted just below 18°C, and cooling temperature-responsive TG neurons from 20°C resulted in an outward current, suggesting that IΔTis on at relatively low temperatures. Ion substitution and Ca2+imaging experiments indicated that IΔTis primarily a monovalent cation current. IΔTwas not sensitive to capsaicin or amiloride, suggesting that the current did not show similar pharmacology to other mammalian heat-sensitive membrane proteins. Our findings indicate that a novel temperature-sensitive conductance with unique ion permeability and low-temperature threshold is expressed in TG neurons and may be involved in highly sensitive heat detection in snakes.

Summary Physiological and pathological aging of the central nervous system (CNS) is characterized by functional neuronal impairments which may lead to perturbed cell homeostasis and eventually to neuronal death. Many toxic events may underlie age-related neurodegeneration. These include the effects of beta amyloid, Tau and mutated presenilin proteins, free radicals and oxidative stress, pro-inflammatory cytokines and lack of growth factor support, which can be individually or collectively involved. Taken individually, these toxicants can induce very diverse cell responses, thus requiring individually targeted corrective interventions upstream of common cell death (apoptotic) pathways. Recent preliminary evidence suggests that the pro-inflammatory cytokine tumour necrosis factor alpha (TNFα) and growth factor withdrawal can both activate a common apoptotic pathway in nerve growth factor (NGF)-responsive PC12 cells involving caspase 3, albeit through very distinct upstream pathways: the former through active signalling and the latter through passive or lack of survival signalling. Here, we show that NGF can rescue PC12 cells from both growth factor withdrawal- and TNFα-promoted cell death. However, NGF rescue from growth factor withdrawal requires NGF signalling through the high-affinity tyrosine kinase receptor (TrkA), while NGF rescue from TNFα-promoted cell death requires NGF signalling through the low-affinity p75NTR receptor. These results strengthen the idea that prevention of age- or pathology-associated neurodegeneration may require varied molecular approaches reflecting the diversity of the toxicants involved, possibly acting simultaneously.

Macrophages and CD4+lymphocytes are the principal target cells for human immunodeficiency virus type 1 (HIV-1) infection, but the molecular details of infection may differ between these cell types. During studies to identify cellular molecules that could be involved in macrophage infection, we observed inhibition of HIV-1 infection of macrophages by monoclonal antibody (MAb) to the tetraspan transmembrane glycoprotein CD63. Pretreatment of primary macrophages with anti-CD63 MAb, but not MAbs to other macrophage cell surface tetraspanins (CD9, CD81, and CD82), was shown to inhibit infection by several R5 and dualtropic strains, but not by X4 isolates. The block to productive infection was postfusion, as assessed by macrophage cell-cell fusion assays, but was prior to reverse transcription, as determined by quantitative PCR assay for new viral DNA formation. The inhibitory effects of anti-CD63 in primary macrophages could not be explained by changes in the levels of CD4, CCR5, or β-chemokines. Infections of peripheral blood lymphocytes and certain cell lines were unaffected by treatment with anti-CD63, suggesting that the role of CD63 in HIV-1 infection may be specific for macrophages.

Age-associated neurodegenerative diseases such as Alzheimer's disease are characterised by neuronal impairment that leads to cognitive deficits. As certain affected neurons depend on trophic factors such as neurotrophins (NTs), impairment in NT function has been suggested to be a component of neuronal damage associated with such disorders. Age-related neurodegenerative diseases are also characterised by high levels of proinflammatory cytokines such as tumour necrosis factor alpha (TNFalpha) in the CNS. Because TNFalpha receptors and certain NT receptors share a high degree of homology and are capable of activating similar signalling pathways, one possibility is that altered cytokine levels may affect NT function in the aged or diseased CNS. Here we wish briefly to review the evidence suggesting a role for cytokine and NT in the onset of age-associated neurodegenerative diseases. We propose that cytokine/NT interactions may alter neuronal homeostasis, thus possibly contributing to some of the neuronal degeneration occurring during such age-associated CNS diseases.

CD8-positive T cells are thought to play an important role in the control of infection by human immunodeficiency virus (HIV) as a result of their cytotoxic activity and by releasing soluble factors. In AIDS patients, the absolute number of CD8+ T lymphocytes is decreased in peripheral blood and their turnover rate is increased, suggesting that there is more cell renewal and cell death occurring. Anti-retroviral therapy raises CD8+ T-cell counts in HIV-infected patients. Here we report that the death rate of CD8+ T cells by apoptosis increased markedly during HIV infection of peripheral blood mononuclear cells in vitro. Apoptosis is induced in a dose-dependent manner by recombinant envelope glycoprotein gp120 from HIV strain X4, or by stromal-derived factor-1 (SDF-1), the physiological ligand of the chemokine receptor CXCR4. Apoptosis is mediated by the interaction between tumour-necrosis factor-alpha bound to the membrane of macrophages (mbTNF) and a receptor on CD8+ T cells (TNF-receptor II, or TNFRII). The expression of both of these cell-surface proteins is upregulated by HIV infection or by treatment with recombinant gp120 or SDF-1. Apoptosis of CD8+ T cells isolated from HIV-infected patients is also mediated by macrophages through the interaction between mbTNF and TNFRII. These results indicate that the increased turnover of CD8+ T cells in HIV-infected subjects is mediated by the HIV envelope protein through the CXCR4 chemokine receptor.

GH3/B6 rat pituitary tumor cells exhibit rapid prolactin release (within 5 min) when treated with nanomolar amounts of estrogen. However, the putative protein mediator of this nongenomic action has not been described. Using antibodies directed against a peptide representing the hinge region of the intracellular estrogen receptor (iER), we have demonstrated that these cells contain a membrane ER (mER). We now report that confocal scanning laser microscopy of cells labeled live with the anti-peptide antibody further supports a membrane localization of ER. The monoclonal antibodies H226 and H222 and a polyclonal antibody, ER21, each recognizing a unique epitope on iER (NH2 terminal to the DNA-binding region, within the steroid binding region, and the NH2-terminal end, respectively), also immunohistochemically label membrane proteins of immuno-selected GH3/B6 cells. These cells also specifically bind a fluorescent estrogen-BSA conjugate. Coincubation of cells with anti-ER antibody and the fluorescent estrogen-BSA conjugate reveals that these labels colocalize on cells. These results suggest that mER may be structurally similar to iER.

Body-size distortion has been considered a central symptom of eating disorders. We studied 35 female eating-disordered patients and 85 controls using a computer-based body-size estimation technique. We have found almost identical linear relationships between body-size distortion and weight:height ratios in both groups. In the clinical group, distortion scores were not correlated with scores on the Eating Attitudes Test or Beck Depression Inventory but were negatively correlated with body dissatisfaction as measured on the Eating Disorder Inventory. These results raise further questions about the role of body-size distortion both as a diagnostic criterion and as a complicating phenomenon in eating disorders.

Environmental or nutritional estrogenic toxicants are thought to mediate developmental and carcinogenic pathologies. Estrogen receptor (ER) measurements are currently used to predict hormonal responsiveness; therefore all ER subpopulations should be considered. We have been involved in the immunoidentification and characterization of membrane steroid receptors in several systems and have recently shown that binding of estradiol (E2) to a subpopulation of ERs (mER) residing in the plasma membrane of GH3 pituitary tumor cells mediates the rapid release of prolactin (PRL). Here we review these findings and present other important characterizations of these receptors such as trypsin and serum susceptibility, movement in the membrane, confocal localization to the membrane, binding to and function of impeded ligands, and immunoseparation of cells bearing mER. We plan to use this system as a model for both the physiological and pathological nongenomic effects of estrogens and estrogenic xenobiotics. Specifically, it should be useful as an in vitro assay system for the ability of estrogenic xenobiotics to cause rapid PRL release as an example of nongenomic estrogen effects. Images Figure 4. A Figure 4. B Figure 4. C Figure 4. D Figure 4. E Figure 4. F Figure 5. A Figure 5. B Figure 6. A Figure 6. B Figure 6. C Figure 7. A Figure 7. B Figure 7. C Figure 7. D

An observational study of chamois behavior during the rut was conducted at the Los Angeles Zoo. The herd consisted of an adult male, two adult females, two subadult females, and two kids (1.1). During the 13-week study, observers collected scan data on all animals and recorded all occurrences of selected social behaviors during interactions which involved the adult male. Scan observations showed that open areas of the enclosure close to the public were the least preferred, and areas with good rest places were used most. Overall the herd spent 73% of their time stationary, 13% foraging/feeding, 6% moving, and less than 5% each in object investigation, maintenance, play, and social behavior. The adult male rested and fed less, and stood vigilant and engaged in social behavior more frequently than the others. Nearest neighbor data showed clear patterns of kin association beyond the mother-last kid bond. The adult male interacted most often with the regular keepers, the subordinate female and the oldest subadult female. Interactions with the females consisted mostly of herding and sexual behaviors, while those with keepers were almost exclusively competitive/aggressive. While most findings paralleled those from studies of wild chamois, no study has previously reported long-lasting associations among kin.