The role of AKT isoforms in glioblastoma: AKT3 delays tumor progression.

The growth factor receptor/PI3K/AKT pathway is an important drug target in many cancers including Glioblastoma. AKT, a key node in the pathway, has 3 isoforms, AKT1, AKT2 and AKT3. Here we investigate their role in GBM. We find each activated, ser473 phosphorylated isoform is present in some GBMs but expression patterns vary. There is a direct relationship between human GBM patient outcome and both AKT1 and AKT2 mRNA levels, but an inverse relationship with AKT3 mRNA. Furthermore, AKT3 mRNA levels were high in a less aggressive GBM subtype. Overexpressing AKT3 improves survival in a rodent model of GBM and decreases colony forming efficiency, but not growth rate, in glioma cells. Silencing AKT3 slows cell cycle progression in one cell line and increases apoptosis in another. Our studies of AKT3 substrates indicate (1) silencing both AKT2 and AKT3 reduces GSK3 phosphorylation (2) only AKT2 silencing reduces S6 phosphorylation. Since S6 phosphorylation is a marker of mTORC1 activity this indicates that AKT2 activates mTORC1, but AKT3 does not. Our results indicate AKT isoforms have different roles and downstream substrates in GBM. Unexpectedly, they indicate AKT3 delays tumor progression. Therefore strategies that inhibit AKT3 may be unhelpful in some GBM patients.

Factors associated with high-risk behaviour among migrants in the state of maharashtra, India.

Studies among migrants show that they are more susceptible to HIV infection than the general population and thereby spread the epidemic from high prevalence to low prevalence areas. It is therefore critical to enhance the body of knowledge on factors associated with condom use among migrants. This study, conducted in 2009 in the State of Maharashtra, covers 4595 single in-migrants aged 15-49 years and aims at understanding the factors associated with non-use of condoms consistently. Information was collected using a Structured Interview Schedule covering demographic, socioeconomic profile, sexual history, knowledge, behaviour and stigma and discrimination indicators. Logistic regression analysis was used to understand the association between unprotected sex and various socio-demographic and environmental factors. The models were run using the Enter method. The goodness-of-fit of the model was assessed using Hosmer and Lemeshow chi-squared statistics. A significant association is observed between sex with sex workers and older migrants (>24 years), the literate, those who are mobile, unmarried, employed in the textile, quarry and construction industries, who often consume alcohol and who watch pornographic films. The factors associated with unprotected sex are age between 30 and 34 years and no literacy. Migrants who are mobile and consume alcohol show a significant association with unprotected sex. The findings suggest a need for a comprehensive HIV prevention programme including strategies to address the stressful work conditions. The prevention programmes should focus not only on skills for safer sex practices, but also on alcohol use reduction.

Family intervention improves outcomes for patients with delirium: Systematic review and meta-analysis.

OBJECTIVE: To determine if family caregiver involvement in interventions with patients with delirium improves patient outcomes. METHODS: A search of three databases (Medline-Ovid, CINAHL and Embase) was conducted. Eligibility criteria included adult patients and involvement of family caregivers in any delirium intervention. Data were extracted from each study (determined by PEDro scale) using a customised form. A meta-analysis was undertaken which compared the length of hospital stay and duration of delirium. PROSPERO registration number is CRD42017077650. RESULTS: Five studies involving 505 participants published over a 5-year period were suitable for inclusion. Low-level evidence demonstrated family caregiver involvement may reduce caregiver's anxiety and hospital staff viewed administration of education to family caregivers as efficient. Meta-analysis suggested family interventions reduce length of hospital stay for patients with delirium. It remains unclear if it affects the duration of delirium. CONCLUSION: Family caregivers providing interventions to patients with delirium can improve patient outcomes.

Communicative Competence: Responding to Residents' Health Changes in Assisted Living.

BACKGROUND AND OBJECTIVES: Frail and disabled individuals, including assisted living (AL) residents, are embedded in care convoys composed of dynamic networks of formal and informal care partners. Yet, little is known about how care convoys operate over time, especially when health changes occur. Thus, our aim was to provide an in-depth understanding of care convoy communication during times of residents' health changes in AL. RESEARCH DESIGN AND METHODS: Data for this analysis come from a Grounded Theory study that involved 50 residents and their care convoy members (n = 169) from 8 diverse AL communities followed over 2 years. Researchers conducted formal and informal interviewing, participant observation, and record review. RESULTS: We identified "communicative competence" as an explanatory framework in reference to a resident's or care partner's ability, knowledge, and action pertaining to communication and health change. Individual and collective competencies were consequential to timely and appropriate care. Communication involved: identifying; assessing significance; informing, consulting or collaborating with others; and responding to the change. Variability in communication process and properties (e.g., pace and timing; sequencing, timing, content, and mode of communication) depended on multiple factors, including the nature of the change and resident, informal and formal caregiver, convoy, AL community, and regulatory influences. DISCUSSION AND IMPLICATIONS: Formal and informal care partners need support to establish, enhance, and maintain communicative competence in response to health changes. Findings reinforce the need for timely communication, effective systems, and well-documented accessible health care directives and have implications that are applicable to AL and other care settings.

Evaluating the Introduction of an Allied Health Clinical Research Office at a Health Service: Effects on Research Participation, Interest, and Experience of Allied Health Professionals.

Following the introduction of an allied health clinical research office at a large metropolitan health service, we aimed to measure change in self-reported research participation, interest and experience of allied health professionals. METHODS: Allied health professionals were surveyed using the Research Spider tool in 2015 (n=245), and the results were compared to a similar survey completed in 2007 at the same health service (n=132). RESULTS: Overall, allied health professionals rated themselves as having "some research interest" and "little research experience," with no significant difference from 2007 to 2015. Allied health professionals with at least some research interest reported increased experience in critically reviewing literature (p=0.045) and finding relevant literature (p=0.009) and a trend to increased experience of publishing research (p=0.059) in 2015 compared with 2007. The proportion of allied health professionals who classified themselves as participating in research had increased from 41% in 2007 to 51% in 2015 (p=0.028). CONCLUSIONS: The introduction of an allied health clinical research office has been associated with increased participation in research with some improvements in research experience for those with at least some interest in research. Despite these positive changes, most allied health professionals at this health service still report little research experience and only some interest in research.

Glioma cells on the run - the migratory transcriptome of 10 human glioma cell lines.

BACKGROUND: Glioblastoma multiforme (GBM) is the most common primary intracranial tumor and despite recent advances in treatment regimens, prognosis for affected patients remains poor. Active cell migration and invasion of GBM cells ultimately lead to ubiquitous tumor recurrence and patient death. To further understand the genetic mechanisms underlying the ability of glioma cells to migrate, we compared the matched transcriptional profiles of migratory and stationary populations of human glioma cells. Using a monolayer radial migration assay, motile and stationary cell populations from seven human long term glioma cell lines and three primary GBM cultures were isolated and prepared for expression analysis. RESULTS: Gene expression signatures of stationary and migratory populations across all cell lines were identified using a pattern recognition approach that integrates a priori knowledge with expression data. Principal component analysis (PCA) revealed two discriminating patterns between migrating and stationary glioma cells: i) global down-regulation and ii) global up-regulation profiles that were used in a proband-based rule function implemented in GABRIEL to find subsets of genes having similar expression patterns. Genes with up-regulation pattern in migrating glioma cells were found to be overexpressed in 75% of human GBM biopsy specimens compared to normal brain. A 22 gene signature capable of classifying glioma cultures based on their migration rate was developed. Fidelity of this discovery algorithm was assessed by validation of the invasion candidate gene, connective tissue growth factor (CTGF). siRNA mediated knockdown yielded reduced in vitro migration and ex vivo invasion; immunohistochemistry on glioma invasion tissue microarray confirmed up-regulation of CTGF in invasive glioma cells. CONCLUSION: Gene expression profiling of migratory glioma cells induced to disperse in vitro affords discovery of genomic signatures; selected candidates were validated clinically at the transcriptional and translational levels as well as through functional assays thereby underscoring the fidelity of the discovery algorithm.

Effect of Physical Violence on Sexually Transmitted Infections and Treatment Seeking Behaviour among Female Sex Workers in Thane District, Maharashtra, India.

BACKGROUND: Violence against sex workers can heighten their vulnerability to HIV and other sexually transmitted infections (STIs). Evidence suggests the risk of acquiring STI/HIV infections among female sex workers (FSWs) who have experienced violence to be almost three-times higher than FSWs, who have not experienced violence. Moreover, an experience of physical and sexual violence makes it difficult for them to negotiate safer sex with their partners and often act as a barrier to utilization of prevention services. METHODS: This study utilizes data from 2785 FSWs aged 18 years and above who participated in a cross-sectional behavioural study conducted during 2013-14 in Thane district, Maharashtra. A probability-based two-stage cluster sampling method was used for data collection. This study assesses the effect of physical violence on self-reported STI symptoms (any STI and multiple STIs) and treatment seeking for the last STI symptom using propensity score matching method. RESULTS: About 18% of sampled FSWs reported physical violence at the time of the survey. The likelihood of experiencing such violence was significantly higher among FSWs who solicited clients at public places, engaged in other economic activities apart from sex work, had savings, and reported high client volume per week. FSWs experiencing violence were also inconsistent condom users while engaging in sex with regular partners and clients. The average adjusted effect of violence clearly depicted an increase in the risk of any STI (11%, p<0.05) and multiple STIs (8%, p<0.10) and reduction in treatment seeking (10%, p<0.05). CONCLUSIONS: This study demonstrates a significant effect of physical violence on reporting of any STI symptom and treatment seeking. Findings call for the immediate inclusion of strategies aimed to address violence related challenges in HIV prevention program currently being provided at Thane district. Such strategies would further help in enhancing the access to tailored STI prevention and care services among FSWs in the district.

Use of a conformational switching aptamer for rapid and specific ex vivo identification of central nervous system lymphoma in a xenograft model.

Improved tools for providing specific intraoperative diagnoses could improve patient care. In neurosurgery, intraoperatively differentiating non-operative lesions such as CNS B-cell lymphoma from operative lesions can be challenging, often necessitating immunohistochemical (IHC) procedures which require up to 24-48 hours. Here, we evaluate the feasibility of generating rapid ex vivo specific labeling using a novel lymphoma-specific fluorescent switchable aptamer. Our B-cell lymphoma-specific switchable aptamer produced only low-level fluorescence in its unbound conformation and generated an 8-fold increase in fluorescence once bound to its target on CD20-positive lymphoma cells. The aptamer demonstrated strong binding to B-cell lymphoma cells within 15 minutes of incubation as observed by flow cytometry. We applied the switchable aptamer to ex vivo xenograft tissue harboring B-cell lymphoma and astrocytoma, and within one hour specific visual identification of lymphoma was routinely possible. In this proof-of-concept study in human cell culture and orthotopic xenografts, we conclude that a fluorescent switchable aptamer can provide rapid and specific labeling of B-cell lymphoma, and that developing aptamer-based labeling approaches could simplify tissue staining and drastically reduce time to histopathological diagnoses compared with IHC-based methods. We propose that switchable aptamers could enhance expeditious, accurate intraoperative decision-making.

AKT pathway genes define 5 prognostic subgroups in glioblastoma.

Activity of GFR/PI3K/AKT pathway inhibitors in glioblastoma clinical trials has not been robust. We hypothesized variations in the pathway between tumors contribute to poor response. We clustered GBM based on AKT pathway genes and discovered new subtypes then characterized their clinical and molecular features. There are at least 5 GBM AKT subtypes having distinct DNA copy number alterations, enrichment in oncogenes and tumor suppressor genes and patterns of expression for PI3K/AKT/mTOR signaling components. Gene Ontology terms indicate a different cell of origin or dominant phenotype for each subgroup. Evidence suggests one subtype is very sensitive to BCNU or CCNU (median survival 5.8 vs. 1.5 years; BCNU/CCNU vs other treatments; respectively). AKT subtyping advances previous approaches by revealing additional subgroups with unique clinical and molecular features. Evidence indicates it is a predictive marker for response to BCNU or CCNU and PI3K/AKT/mTOR pathway inhibitors. We anticipate Akt subtyping may help stratify patients for clinical trials and augment discovery of class-specific therapeutic targets.

Migrating glioma cells activate the PI3-K pathway and display decreased susceptibility to apoptosis.

Glioma cells that migrate out of the main tumor mass into normal brain tissue contribute to the failure of most gliomas to respond to treatment. Treatments that target migratory glioma cells may enhance the therapeutic response. Multiple lines of evidence suggest that suppression of apoptosis accompanies activation of the migratory phenotype. Here, we determine whether migration and apoptosis are consistently linked in glioma cells and whether manipulation of migration influences cytotoxic therapy-induced apoptosis. Camptothecin and Trail-induced apoptosis were decreased 2-5-fold in actively migrating glioma cells relative to migration-restricted cells. Consistent with a mechanistic link between migration and apoptosis, the dose-response for stimulation of migration on laminin was inversely proportional to apoptosis induction. Treatment of glioma cells with migration inhibitors alone had little effect on basal rates of apoptosis and had little effect on Trail-induced or camptothecin-induced apoptosis in migration-restricted cells. By contrast, migration inhibitors increased camptothecin and Trail-induced apoptosis in actively migrating glioma cells. Migrating glioma cells have increased amounts of phosphorylated Akt and its downstream substrate glycogen synthase kinase-3 relative to migration restricted cells. Treatment of migrating cells with a specific inhibitor of phosphoinositide 3-kinase (PI3-K), LY294002, blocked the phosphorylation of Akt and increased the sensitivity to apoptosis. LY294002 had no effect on the migration of restricted cells. This suggests that migrating glioma cells activate the PI3-K survival pathway, protecting migrating cells from apoptosis. Taken together, these data provide support for a link between migration and apoptosis in glioma cells. In addition, evidence indicates that treatment with migration inhibitors, while not affecting apoptosis-induction in migration-restricted cells, can sensitize migrating glioma cells to cytotoxic agents.