Administration of glucocorticoids induces ocular hypertension in some patients. If untreated, these patients can develop a secondary glaucoma that resembles primary open-angle glaucoma (POAG). The underlying pathology of glucocorticoid-induced glaucoma is not fully understood, due in part to lack of an appropriate animal model. Here, we developed a murine model of glucocorticoid-induced glaucoma that exhibits glaucoma features that are observed in patients. Treatment of WT mice with topical ocular 0.1% dexamethasone led to elevation of intraocular pressure (IOP), functional and structural loss of retinal ganglion cells, and axonal degeneration, resembling glucocorticoid-induced glaucoma in human patients. Furthermore, dexamethasoneinduced ocular hypertension was associated with chronic ER stress of the trabecular meshwork (TM). Similar to patients, withdrawal of dexamethasone treatment reduced elevated IOP and ER stress in this animal model. Dexamethasone induced the transcriptional factor CHOP, a marker for chronic ER stress, in the anterior segment tissues, and Chop deletion reduced ER stress in these tissues and prevented dexamethasone-induced ocular hypertension. Furthermore, reduction of ER stress in the TM with sodium 4-phenylbutyrate prevented dexamethasone-induced ocular hypertension in WT mice. Our data indicate that ER stress contributes to glucocorticoid-induced ocular hypertension and suggest that reducing ER stress has potential as a therapeutic strategy for treating glucocorticoid-induced glaucoma.

Purpose Trial**** was a phase III randomized trial designed to determine the optimal duration of androgen deprivation therapy (ADT) when combined with definitive radiation therapy (RT) in the treatment of locally advanced non-metastatic adenocarcinoma of the prostate. Long-term follow-up results of this study now available are relevant to the management of this disease. Materials and Methods Men (N=1,554) with adenocarcinoma of the prostate (cT2c-T4, N0- Nx) with a prostate specific antigen (PSA) <150ng/ml and no evidence of distant metastasis were randomized (June 1992 to April 1995) to short term ADT (STAD: 4 months of flutamide 250mg three times per day and goserelin 3.6mg per month) and definitive RT verses long term ADT (LTAD: STAD with definitive RT plus an additional 24 months of monthly goserelin). Results Among 1,520 protocol eligible and evaluable patients, median follow up for this analysis was 19.6 years. In analysis adjusted for prognostic covariates, LTAD improved disease free survival (29% relative reduction in failure rate, p<0.0001), local progression (46% relative reduction, p=0.02), distant metastases (36% relative reduction, p<0.0001), disease specific survival (30% relative reduction, p=0.003), and overall survival (12% relative reduction, p=0.03). Other cause (non-prostate cancer) mortality did not differ (5% relative reduction, p=0.48). Conclusions LTAD and RT is superior to STAD and RT for the treatment of locally advanced non-metastatic adenocarcinoma of the prostate and should be considered the standard of care.

G9a is a mammalian histone methyltransferase that contributes to the epigenetic silencing of tumor suppressor genes. Evidence suggests that G9a is required to maintain the malignant phenotype, but little documentation show the role of G9a function in mediating tumor growth. We retrospectively analyzed the protein of G9a and monomethylated histone H3 lysine 9 (H3K9 me1), and dimethylated histone H3 lysine 9 (H3K9 me2) in 175 cases of gastric carcinoma by immunohistochemistry. RNAi-based inhibition of G9a in MGC803 cancer cell line was studied. G9a depletion was done by transient transfection using Lipofectamine 2000. Depletion efficiency of G9a was tested using real-time PCR and western blot analysis. Cell apoptosis and proliferation were detected by TUNEL assay and MTT, respectively. The proteins of H3K9 me1, me2, trimethylation of H3K9 (H3K9 me3), monomethylated histone H3 lysine 27 (H3K27 me1), dimethylated histone H3 lysine 27 (H3K27 me2) and histone acetylated H3, apoptotic proteins were studied by western blot analysis. G9a and H3K9 me2 expression was higher in gastric cancer cells compared to the control (p<0.05). Both G9a and H3K9 me2 were positively correlated with the degree of differentiation, depth of infiltration, lymphatic invasions and tumor-node-metastasis stage in gastric carcinoma, (p<0.05). RNAi-mediated knockdown of G9a induced cell apoptosis and inhibited cell proliferation. Depletion of G9a reduced the levels of H3K9 me1 and me2, H3K27 me1 and me2. Nonetheless, it did not activate acetylation of H3 and H3K9 me3. These data suggest that G9a is required in tumorigenesis, and correlated with prognosis. Furthermore, G9a plays a critical role in regulating epigenetics. Depletion of G9a inhibits cell growth and induces cells apoptosis in gastric cancer. It might be of therapeutic benefit in gastric cancers.

Ecological momentary assessment studies usually produce intensively measured longitudinal data with large numbers of observations per unit, and research interest is often centered around understanding the changes in variation of people's thoughts, emotions and behaviors. Hedeker et al developed a 2-level mixed effects location scale model that allows observed covariates as well as unobserved variables to influence both the mean and the within-subjects variance, for a 2-level data structure where observations are nested within subjects. In some ecological momentary assessment studies, subjects are measured at multiple waves, and within each wave, subjects are measured over time. Li and Hedeker extended the original 2-level model to a 3-level data structure where observations are nested within days and days are then nested within subjects, by including a random location and scale intercept at the intermediate wave level. However, the 3-level random intercept model assumes constant response change rate for both the mean and variance. To account for changes in variance across waves, as well as clustering attributable to waves, we propose a more comprehensive location scale model that allows subject heterogeneity at baseline as well as across different waves, for a 3-level data structure where observations are nested within waves and waves are then further nested within subjects. The model parameters are estimated using Markov chain Monte Carlo methods. We provide details on the Bayesian estimation approach and demonstrate how the Stan statistical software can be used to sample from the desired distributions and achieve consistent estimates. The proposed model is validated via a series of simulation studies. Data from an adolescent smoking study are analyzed to demonstrate this approach. The analyses clearly favor the proposed model and show significant subject heterogeneity at baseline as well as change over time, for both mood mean and variance. The proposed 3-level location scale model can be widely applied to areas of research where the interest lies in the consistency in addition to the mean level of the responses.

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Objective Cancer diagnosis in adolescents and young adults (AYAs) coincides with the developmental initiation of substance use and emergence of affective disturbance. We examined substance use behaviors and risk‐stratified associations with mental and physical health, as well as objective indicators of tobacco and cannabis use and concordance with self‐report and medical records. Methods AYAs were 15 to 39 years at cancer diagnosis and ≥18 years and ≥6 months postdiagnosis at study enrollment. Risk‐stratified groups included nonsmoker/nondrinker, nonsmoker/drinker, smoker/drinker. Assessments included demographics, past year tobacco, alcohol, and cannabis use, depression, anxiety, sleep, and physical activity. Urine analysis provided biochemical verification of tobacco and cannabis use. Results Participants included 100 AYAs (60% male) with primarily hematological cancers (88%). Past year alcohol, tobacco, and cannabis use prevalence rates were 80%, 15%, and 33%, respectively. A minority (non‐users) refrained from both alcohol and tobacco (20%), while most were exclusively alcohol users (65%) or alcohol and tobacco co‐users (15%). Relative to other sub‐groups, co‐users reported more depressive and anxious symptoms, while non‐users reported more physical activity. More frequent tobacco and cannabis use were associated with more depressive and anxious symptoms, while more frequent alcohol use was associated with lower physical activity. There were no group differences or associations with sleep quality. There was considerable discordance between tobacco use self‐report, biochemical verification, and medical record documentation. Conclusions Substance use among AYAs is common and detrimental to mental and physical health, especially among more frequent users and co‐users, highlighting the need for early assessment and intervention.

T-cell activation is a critical part of the adaptive immune system, enabling responses to foreign cells and external stimulus. In this process, T cell antigen receptor (TCR) activation stimulates translocation of the downstream kinase PKCθ to the membrane, leading to NF-κB activation and thus transcription of relevant genes. However, the details of how PKCθ is recruited to the membrane remain enigmatic. It is known that Annexin A5 (ANXA5), a calcium-dependent membrane-binding protein, has been reported to mediate PKCδ activation by interaction with PKCδ, a homologous of PKCθ, which implicates a potential role of ANXA5 involved in PKCθ signaling. Here we demonstrate that ANXA5 does play a critical role in the recruitment of PKCθ to the membrane during T-cell activation. ANXA5 knockout in Jurkat T cells substantially inhibited the membrane translocation of PKCθ upon TCR engagement and blocked the recruitment of CARMA1-BCL10-MALT1 (CBM) signalosome, which provides a platform for the catalytic activation of IKKs and subsequent activation of canonical NF- κB signaling in activated T cells. As a result, NF-κB activation was impaired in ANXA5-KO T cells. T-cell activation was also suppressed by ANAX5 knockdown in primary T cells. These results demonstrated a novel role of ANXA5 in PKC translocation and PKC signaling during T-cell activation.

ABSTRACT. Mutations in the CYP1B1 gene were detected in primary openangle glaucoma (POAG) patients. However, the association between these mutations and the incidence of POAG remains to be elucidated. Here, we have conducted a meta-analysis to analyze this correlation, using relevant studies obtained from an extensive search of various electronic databases, including EMBase, Web of Science, and PubMed. The extracted studies were selected for the meta-analysis based on the inclusion and exclusion criteria. The quality of each included study was assessed by the Newcastle- Ottawa scale (NOS), and the I 2 value was calculated to evaluate the heterogeneity between studies. The combined effect size was presented as the odds ratio (OR), and confidence intervals (CI) were used to assess the association between POAG and CYP1B1 mutations. Eight studies, each with a high NOS score, were included in the analysis. Compared to the mutant allele, the wild-type allele of the rs180040 polymorphism in POAG patients showed a 12% decrease in OR (OR = 0.88, 95%CI = 0.76-1.00); also, the wild-type allele of rs1056827 showed a 23% decrease in OR of the incidence of POAG (OR = 0.77, 95%CI = 0.60-0.99). However, the latter result was controversial. Polymorphisms at rs1056836, rs10012, and rs1056837 were not correlated with the incidence of POAG (using any evaluation model). In conclusion, three of the tested SNPs in the CYP1B1 gene were correlated with POAG; however, the SNPs rs180040 and rs1056827 showed an association with risk of POAG. These results must be further validated with larger-scale evaluations.