Online Screening and Personalized Education to Identify Post-Deployment Mental Health Need and Facilitate Access to Care.

INTRODUCTION: Reserve and National Guard (RNG) service members face increased risks for psychological and behavioral problems and are unlikely to seek mental health (MH) care after returning from military deployments. This article examines an online intervention (Web-Ed) with regard to participation, screening results, satisfaction, and intent to seek follow-up MH care, with comparisons by gender and post-deployment MH care receipt. MATERIALS AND METHODS: This was a cross-sectional study of 414 RNG service members (214 women and 200 men), who returned from deployments to or in support of the Iraq or Afghanistan wars within the prior 36 months. Participants completed Web-Ed, which includes screening, personalized education, and links to Veterans Health Administration health care. RESULTS: Positive post-deployment screening proportions, Web-Ed satisfaction, and intent to seek follow-up care were similar for men and women. Few had received MH care (33% women; 24% men), yet most screened positive on at least one screen (69% women; 72% men). Most indicated that they would recommend Web-Ed to other veterans (71% women; 67% men) and that they received useful information they may not have received otherwise (52% women; 53% men) regardless of gender or prior MH care. Almost half (40% women; 48% men) planned to seek further assessment from a health care provider as a direct result of Web-Ed. CONCLUSIONS: Efforts to facilitate access to MH care among post-deployed RNG veterans should be ongoing, regardless of the length of time since deployment, Veterans Affairs enrollment status, prior MH care receipt, or gender. Online screening and personalized education engage veterans who have not sought MH care and provide new information to those who have.

Factors Associated with U.S. Military Women Keeping Guns or Weapons Nearby for Personal Security Following Deployment.

Background: The relationship between postdeployment health characteristics and U.S. military women and women Veteran's gun/weapons use for personal safety outside of military is not well understood. The purpose of this exploratory study was to determine if Operation Enduring and Iraqi Freedom era military women and women Veterans keep guns/weapons nearby for personal security outside of military duties postdeployment and factors associated with this. Methods: A Midwestern community sample of US Army and Air Force currently serving Military women and women Veterans (N = 978) who had deployed to Iraq/Afghanistan (I/A) or outside of the United States completed telephone interviews (March 2010 to December 2011) querying sociodemographic and military characteristics, combat and gender-based trauma, and guns/weapons use postdeployment. Data were analyzed in June 2019 with chi-square, Fisher's Exact test, and odds ratios. A classification tree analysis identified subgroups with the greatest proportion of keeping guns/weapons nearby for security. Results: One-fifth of participants reported having guns/weapons nearby to feel secure. Women more likely to report this were younger, patrolled their homes for security (age adjusted odds ratio [aOR] 7.0); experienced combat (aOR 3.0-4.9) or gender-based traumas (aOR 1.9-2.0); self-reported mental health conditions (aOR 1.5-4.3), including post-traumatic stress disorder (PTSD; aOR 4.3); or relied on friends/family for housing (aOR 4.8). Most had seen a provider in the preceding year. The classification tree found women patrolling their homes, PTSD positive, and injured/wounded in I/A had the largest proportion of women keeping guns/weapons nearby for security. Conclusions: Keeping gun/weapons nearby for personal self-defense is a potential marker for complex postdeployment readjustment conditions and an overlooked public health concern. Provider recognition and assessment of women's postdeployment fears and safety-related activities are essential to address military women and women Veterans and their families' safety in this high-risk population.

Insertion of T4-lysozyme (T4L) can be a useful tool for studying olfactory-related GPCRs.

The detergents used to solubilize GPCRs can make crystal growth the rate-limiting step in determining their structure. The Kobilka laboratory showed that insertion of T4-lysozyme (T4L) in the 3rd intracellular loop is a promising strategy towards increasing the solvent-exposed receptor area, and hence the number of possible lattice-forming contacts. The potential to use T4L with the olfactory-related receptors hOR17-4 and hVN1R1 was thus tested. The structure and function of native and T4L-variants were compared. Both receptors localized to the cell membrane, and could initiate ligand-activated signaling. Purified receptors not only had the predicted alpha-helical structures, but also bound their ligands canthoxal (M(W) = 178.23) and myrtenal (M(W) = 150.22). Interestingly, the T4L variants had higher percentages of soluble monomers compared to protein aggregates, effectively increasing the protein yield that could be used for structural and function studies. They also bound their ligands for longer times, suggesting higher receptor stability. Our results indicate that a T4L insertion may be a general method for obtaining GPCRs suitable for structural studies.

Mechanoregulation of proliferation.

The proliferation of all nontransformed adherent cells is dependent upon the development of mechanical tension within the cell; however, little is known about the mechanisms by which signals regulated by mechanical tension are integrated with those regulated by growth factors. We show here that Skp2, a component of a ubiquitin ligase complex that mediates the degradation of several proteins that inhibit proliferation, is upregulated when increased mechanical tension develops in intact smooth muscle and that its upregulation is critical for the smooth muscle proliferative response to increased mechanical tension. Notably, whereas growth factors regulate Skp2 at the level of protein stability, we found that mechanical tension regulates Skp2 at the transcriptional level. Importantly, we demonstrate that the calcium-regulated transcription factor NFATc1 is a critical mediator of the effect of increased mechanical tension on Skp2 transcription. These findings identify Skp2 as a node at which signals from mechanical tension and growth factors are integrated to regulate proliferation, and they define calcium-NFAT-Skp2 signaling as a critical pathway in the mechanoregulation of proliferation.

Large-scale production and study of a synthetic G protein-coupled receptor: human olfactory receptor 17-4.

Although understanding of the olfactory system has progressed at the level of downstream receptor signaling and the wiring of olfactory neurons, the system remains poorly understood at the molecular level of the receptors and their interaction with and recognition of odorant ligands. The structure and functional mechanisms of these receptors still remain a tantalizing enigma, because numerous previous attempts at the large-scale production of functional olfactory receptors (ORs) have not been successful to date. To investigate the elusive biochemistry and molecular mechanisms of olfaction, we have developed a mammalian expression system for the large-scale production and purification of a functional OR protein in milligram quantities. Here, we report the study of human OR17-4 (hOR17-4) purified from a HEK293S tetracycline-inducible system. Scale-up of production yield was achieved through suspension culture in a bioreactor, which enabled the preparation of >10 mg of monomeric hOR17-4 receptor after immunoaffinity and size exclusion chromatography, with expression yields reaching 3 mg/L of culture medium. Several key post-translational modifications were identified using MS, and CD spectroscopy showed the receptor to be approximately 50% alpha-helix, similar to other recently determined G protein-coupled receptor structures. Detergent-solubilized hOR17-4 specifically bound its known activating odorants lilial and floralozone in vitro, as measured by surface plasmon resonance. The hOR17-4 also recognized specific odorants in heterologous cells as determined by calcium ion mobilization. Our system is feasible for the production of large quantities of OR necessary for structural and functional analyses and research into OR biosensor devices.

Study of a synthetic human olfactory receptor 17-4: expression and purification from an inducible mammalian cell line.

In order to begin to study the structural and functional mechanisms of olfactory receptors, methods for milligram-scale purification are required. Here we demonstrate the production and expression of a synthetically engineered human olfactory receptor hOR17-4 gene in a stable tetracycline-inducible mammalian cell line (HEK293S). The olfactory receptor gene was fabricated from scratch using PCR-based gene-assembly, which facilitated codon optimization and attachment of a 9-residue bovine rhodopsin affinity tag for detection and purification. Induction of adherent cultures with tetracycline together with sodium butyrate led to hOR17-4 expression levels of approximately 30 microg per 150 mm tissue culture plate. Fos-choline-based detergents proved highly capable of extracting the receptors, and fos-choline-14 (N-tetradecylphosphocholine) was selected for optimal solubilization and subsequent purification. Analysis by SDS-PAGE revealed both monomeric and dimeric receptor forms, as well as higher MW oligomeric species. A two-step purification method of immunoaffinity and size exclusion chromatography was optimized which enabled 0.13 milligrams of hOR17-4 monomer to be obtained at >90% purity. This high purity of hOR17-4 is not only suitable for secondary structural and functional analyses but also for subsequent crystallization trials. Thus, this system demonstrates the feasibility of purifying milligram quantities of the GPCR membrane protein hOR17-4 for fabrication of olfactory receptor-based bionic sensing device.

T-cell activation after chronic ethanol ingestion in mice.

Chronic excessive consumption of ethanol causes immunodeficiency in human beings and in mice. Immunologic changes have been described in both species, including T-cell and innate immune system cell activation, among others. The features of chronic ethanol-induced activation have similarities in the two species, including an increased effector subset in both CD4+ and CD8+ T cells. There are also features of activation observed in the splenic macrophages of mice consuming ethanol chronically, including increased up-regulation of CD80 and CD86. Because these molecules are involved in T-cell-antigen-presenting cell interactions in vivo, it is of interest to ask whether these and other pathways of interaction are important in the T-cell activation and cytokine skewing described in chronic ethanol abuse. Preliminary findings from comparisons of wild-type, CD40 ligand knock-out, and CD28 knock-out C57BL/6 mice strongly support the suggestion of a critical role for T-cell-antigen-presenting cell interactions in the immune alterations observed in chronic ethanol abuse.

Inhibition of mitogenic signaling and induction of apoptosis in human bladder smooth muscle cells treated with doxazosin.

PURPOSE: The alpha1 antagonist doxazosin is used to treat lower urinary tract symptoms and is believed to function primarily as a smooth muscle relaxant. However, doxazosin has been shown to inhibit proliferation and induce apoptosis in nonbladder smooth muscle. Consequently, we examined the effects of doxazosin on human bladder smooth muscle cell (SMC) proliferation and apoptosis. MATERIALS AND METHODS: Primary human bladder SMCs were cultured in M199 with 10% fetal bovine serum (FBS) until they reached 65% confluency and then they were made quiescent by serum starvation in M199 with 0.4% FBS for 24 hours. The quiescent bladder SMCs were pretreated for 30 minutes with doxazosin or vehicle (dimethyl sulfoxide) and then stimulated with 10% FBS for 24 hours. Measurement of 5'-bromo-2'-deoxyuridine (BrdU) uptake by flow cytometry was used to determine the effect of doxazosin on cell cycle progression. Western immunoblot was used to examine cell cycle protein expression and phosphorylation of the retinoblastoma protein (Rb) and cyclin A, both of which regulate cell cycle progression. RESULTS: Cellular proliferation was inhibited in a dose dependent manner by doxazosin. There was nearly a 50% decrease in BrdU uptake at 10 microM doxazosin and an approximately 90% decrease in BrdU at 25 microM doxazosin. Notably, doxazosin inhibited phosphorylation of Rb and expression of cyclin A, both of which are necessary for cell cycle progression. At concentrations of 25 microM doxazosin or greater apoptosis was induced in the bladder SMCs, as indicated by an increase in subG1 DNA content. CONCLUSIONS: Our study demonstrates that doxazosin inhibits mitogen induced proliferation of human bladder SMC by blocking cell cycle progression at the of G1/S border. Doxazosin induced cell cycle inhibition appears to be at least in part due to an inhibition of mitogen induced Rb phosphorylation and cyclin A expression. At higher concentrations doxazosin induces apoptosis in human bladder SMCs.

Factors associated with women's risk of rape in the military environment.

BACKGROUND: Health hazards specific to women workers have not been adequately documented. This study assessed military environmental factors associated with rape occurring during military service, while controlling for pre-military trauma experiences. METHODS: A national cross-sectional survey of 558 women veterans serving in Vietnam or in subsequent eras was obtained through structured telephone interviews. RESULTS: Rape was reported by 30% (n = 151) of participants, with consistent rates found across eras [corrected]. Military environmental factors were associated with increased likelihood of rape, including: sexual harassment allowed by officers (P < 0.0001), unwanted sexual advances on-duty (P < 0.0001) and in sleeping quarters (P < 0.0001). CONCLUSION: Violence towards military women has identifiable risk factors. Work and living environments where unwanted sexual behaviors occurred were associated with increased odds of rape. Officer leadership played an important role in the military environment and safety of women. Assailant alcohol and/or drug abuse at time of rape was notable. Interventions and policies based on modifiable environmental risk factors are needed to increase protection for women in the workplace.

Bcl-xL deamidation is a critical switch in the regulation of the response to DNA damage.

The therapeutic value of DNA-damaging antineoplastic agents is dependent upon their ability to induce tumor cell apoptosis while sparing most normal tissues. Here, we show that a component of the apoptotic response to these agents in several different types of tumor cells is the deamidation of two asparagines in the unstructured loop of Bcl-xL, and we demonstrate that deamidation of these asparagines imports susceptibility to apoptosis by disrupting the ability of Bcl-xL to block the proapoptotic activity of BH3 domain-only proteins. Conversely, Bcl-xL deamidation is actively suppressed in fibroblasts, and suppression of deamidation is an essential component of their resistance to DNA damage-induced apoptosis. Our results suggest that the regulation of Bcl-xL deamidation has a critical role in the tumor-specific activity of DNA-damaging antineoplastic agents.