PET-CT-guided characterisation of progressive, preclinical tuberculosis infection and its association with low-level circulating Mycobacterium tuberculosis DNA in household contacts in Leicester, UK: a prospective cohort study.

BACKGROUND: Incipient tuberculosis, a progressive state of Mycobacterium tuberculosis infection with an increased risk of developing into tuberculosis disease, remains poorly characterised. Animal models suggest an association of progressive infection with bacteraemia. Circulating M tuberculosis DNA has previously been detected in pulmonary tuberculosis by use of Actiphage, a bacteriophage-based real-time PCR assay. We aimed to investigate whether serial [18F]fluorodeoxyglucose ([18F]FDG)-PET-CT could be used to characterise the state and progressive trajectory of incipient tuberculosis, and examine whether these PET-CT findings are associated with Actiphage-based detection of circulating M tuberculosis DNA. METHODS: We did a prospective 12-month cohort study in healthy, asymptomatic adults (aged ≥16 years) who were household contacts of patients with pulmonary tuberculosis, and who had a clinical phenotype of latent tuberculosis infection, in Leicester, UK. Actiphage testing of participants' blood samples was done at baseline, and [18F]FDG PET-CT at baseline and after 3 months. Baseline PET-CT features were classified as positive, indeterminate, or negative, on the basis of the quantitation (maximum standardised uptake value [SUVmax]) and distribution of [18F]FDG uptake. Microbiological sampling was done at amenable sites of [18F]FDG uptake. Changes in [18F]FDG uptake after 3 months were quantitatively categorised as progressive, stable, or resolving. Participants received treatment if features of incipient tuberculosis, defined as microbiological detection of M tuberculosis or progressive PET-CT change, were identified. FINDINGS: 20 contacts were recruited between Aug 5 and Nov 5, 2020; 16 of these participants had a positive result on IFNγ release assay (QuantiFERON-TB Gold Plus [QFT]) indicating tuberculosis infection. Baseline PET-CT scans were positive in ten contacts (all QFT positive), indeterminate in six contacts (three QFT positive), and negative in four contacts (three QFT positive). Four of eight PET-CT-positive contacts sampled had M tuberculosis identified (three through culture, one through Xpert MTB/RIF Ultra test) from intrathoracic lymph nodes or bronchial wash and received full antituberculosis treatment. Two further unsampled PET-CT-positive contacts were also treated: one with [18F]FDG uptake in the lung (SUVmax 9·4) received empirical antituberculosis treatment and one who showed progressive [18F]FDG uptake received preventive treatment. The ten untreated contacts with [18F]FDG uptake at baseline (seven QFT positive) had stable or resolving changes at follow-up and remained free of tuberculosis disease after 12 months. A positive baseline Actiphage test was associated with the presence of features of incipient tuberculosis requiring treatment (p=0·018). INTERPRETATION: Microbiological and inflammatory features of incipient tuberculosis can be visualised on PET-CT and are associated with M tuberculosis detection in the blood, supporting the development of pathogen-directed blood biomarkers of tuberculosis risk. FUNDING: MRC Confidence in Concept.

Optimization of Class I Histone Deacetylase PROTACs Reveals that HDAC1/2 Degradation is Critical to Induce Apoptosis and Cell Arrest in Cancer Cells.

Class I histone deacetylase (HDAC) enzymes 1, 2, and 3 organize chromatin as the catalytic subunits within seven distinct multiprotein corepressor complexes and are established drug targets. We report optimization studies of benzamide-based Von Hippel-Lindau (VHL) E3-ligase proteolysis targeting chimeras (PROTACs) and for the first time describe transcriptome perturbations resulting from these degraders. By modifying the linker and VHL ligand, we identified PROTACs 7, 9, and 22 with submicromolar DC50 values for HDAC1 and/or HDAC3 in HCT116 cells. A hook effect was observed for HDAC3 that could be negated by modifying the position of attachment of the VHL ligand to the linker. The more potent HDAC1/2 degraders correlated with greater total differentially expressed genes and enhanced apoptosis in HCT116 cells. We demonstrate that HDAC1/2 degradation by PROTACs correlates with enhanced global gene expression and apoptosis, important for the development of more efficacious HDAC therapeutics with reduced side effects.

Using a Food Bank as a Platform for Educating Communities during the COVID-19 Pandemic.

PURPOSE: To describe a partnership between a public university and a regional foodbank aimed to promote health in food insecure communities, and to support nursing students' learning experiences in community health. DESIGN: A single setting case study. METHODS: A comprehensive health needs assessment conducted through a windshield survey and key informant interviews was used to identify the community's priorities. FINDINGS: Nursing interventions were tailored to the community's needs by creating healthy recipes and providing education on food safety, lower back injury prevention, and chronic disease prevention and 15 management. CONCLUSION: During pandemics, nursing program partnerships with food banks could play pivotal roles in community health promotion. CLINICAL EVIDENCE: Food banks can serve as clinical platforms for nursing education and community wellness activities.

American Muslim Adolescent Daughters' Perception of Maternal Relationships and the Influence on their Health Behaviors: A Conceptual Model.

Objectives: The goal of this qualitative research study was to better understand of how the mother-daughter relationship shaped by different ecologies in a Muslim community in the United States (US) influences their daughters' health behaviors. Methods: Using a criterion sampling strategy, 11 immigrant Muslim mothers and their American Muslim adolescent daughters aged 12-18 years who were born and also raised in the US were recruited (N=22) and interviewed. The interviews were transcribed verbatim, coded, and analyzed following phenomenological research methods. Results: Mothers in this sample explained that to share their health values with their daughters, they needed to be close, supportive, open-minded, and good listeners to them. The results revealed that daughters who perceived that their mothers' values were shaped by 3 factors - religion, culture of origin, and acculturation were more likely to follow healthy behaviors. Conclusion: The findings and the conceptual model will help explain how these maternal factors can work together to shape American Muslim adolescent daughters' health behaviors.

Theorizing the Process of Leaving a Violent Marriage and Getting a Divorce in Tehran.

This study utilized qualitative methods to develop a theory regarding the process used by Tehranian women who leave violent marriages to get a divorce. Findings from semistructured, in-depth interviews with nine women in Tehran who left their abusive husbands suggested that there are six stages in this process: "denial," "using cognitive and internal strategies to save marriage," "using behavioral and external strategies to save marriage," "seeking power to end violence," "preparation to leave marriage," and "termination" stage. The implications of this study help therapists working with women in Tehran who are living in violent marriages and provide effective prevention and intervention services that are appropriately targeted to the specific needs of Iranian women.

Outrage and the emotional labour associated with environmental activism among nurses.

AIM: To investigate the emotions associated with environmental activism among nurses. BACKGROUND: Nursing has a long history of political activism in relation to social justice issues. Some nurses are environmental activists. DESIGN: Ethnography. METHODS: Focus groups and individual interviews were triangulated with additional observational data. The 40 participants (USA = 23, UK = 17) were nurses actively engaged with environmental/sustainability issues. Data collection was from March-October 2017. Data were analysed using discourse analysis. RESULTS: The trigger for nurses' environmental activism was a strong emotional response to recognizing risks to human health. Their activism results in an additional emotion burden above which they routinely experience in caring for patients. Many feel over-whelmed by the magnitude of the issues. Nurses in America feel more supported and empowered to be activists than UK nurses. CONCLUSION: This study reveals for the first time that advocating for the environment is emotional labour, additional to the previously recognized emotional labour associated with caring for patients. Encouragement and support from others, particularly fellow nurses, helps nurse environmental activists continue to fight for the health of communities and the planet. IMPACT: Outrage at threats to human health triggers nurses' environmental activism. Environmental activism creates an additional emotional burden above that routinely experienced in caring for patients. Nurses in America feel more supported and empowered to be activists than UK. Nurses whose practice embraces environmental activism need practical and emotional support from other nurses.

Becoming and being an environmentally 'woke' nurse: A phenomenological study.

BACKGROUND: Increasing numbers of nurses view environmental activism as part of their role. No studies have explored the reasons. AIM: This study aimed to identify what awakened and shaped US and UK nurses' environmental activism. DESIGN: A Gadamerian hermaneutic phenomenological study. METHODS: Forty nurses were recruited in 2017 using purposive sampling (USA n = 23, UK n = 17). Four focus groups were followed by individual interviews. Iterative, fully immersive phenomenological analysis was conducted with corroboration of themes and a final "shared horizon." FINDINGS: Participants described "lightbulb" events whereby they recognized environmental threats to people and the planet. Their nursing-centered values of social justice, generational fairness, and alleviating suffering underpinned a shared belief in their ability to help mitigate those threats. They believed it was their duty as nurses to take action on behalf of individuals, communities, and the planet. DISCUSSION: The nursing skill set of these nurses makes for effective leadership and environmental activism. Environmentally "woke" nurses have the skills to create solutions and systems approaches that increase sustainability, improve health and make a difference to people's lives, now and for future generations.

"I Am Not A Victim. I Am A Survivor": Resilience as a Journey for Female Survivors of Child Sexual Abuse.

This study was designed to expand our understanding of the positive aspects of coping and resilience in female survivors of child sexual abuse. Research questions focused on women's lived experiences of being survivors of child sexual abuse and how they have experienced resilience, developed healthy intimate relationships, and viewed themselves as sexual beings. Using a qualitative research lens of phenomenology, we captured the essence of survivors' experiences of resilience. Although each woman's experience was unique, similar patterns of processes and outcomes emerged as meaningful in their development of resilient and healthy sexuality and relationship functioning. However, participants emphasized resilience as a process or journey-recovery from trauma, reconceptualization of self, and development of healthy sexuality included deliberate efforts occurring over time. Implications for future research and practice using a positive lens of resilience are provided.

Preventable Injuries/Fatalities Due to Distracted Driving: A Call for a Coordinated Action.

In the past decade, distracted driving has emerged as one of the most prominent global road safety concerns. The singular and combined efforts of many organizations, such as Drop It And Drive (DIAD) and the Traffic Injury Research Foundation (TIRF), have identified the need for partnerships, shared information, and access to interpretable research findings to guide decision making and to identify effective strategies to reduce, if not eliminate, preventable injuries/fatalities from distracted driving, including whiplash-associated disorders. This editorial describes activities in 5 target areas identified in our environmental scan in Canada: (1) provincial/territorial government strategies, (2) enforcement, (3) data collection and measurement, (4) awareness campaigns, and (5) legislation. We will also reinforce the potential benefits of seeking and establishing meaningful interdisciplinary partnerships to reduce distracted driving across the globe. Finally, this editorial provides an example of how these interdisciplinary partnerships can help reduce preventable, life-altering injuries/deaths caused by distracted driving. J Orthop Sports Phys Ther 2016;46(10):818-821. doi:10.2519/jospt.2016.0113.

Comet assay measures of DNA damage as biomarkers of irinotecan response in colorectal cancer in vitro and in vivo.

The use of irinotecan to treat metastatic colorectal cancer (CRC) is limited by unpredictable response and variable toxicity; however, no reliable clinical biomarkers are available. Here, we report a study to ascertain whether irinotecan-induced DNA damage measures are suitable/superior biomarkers of irinotecan effect. CRC-cell lines (HCT-116 and HT-29) were treated in vitro with irinotecan and peripheral blood lymphocytes (PBL) were isolated from patients before and after receiving irinotecan-based chemotherapy. Levels of in vitro-, in vivo-, and ex vivo-induced DNA damage were measured using the Comet assay; correlations between damage levels with in vitro cell survival and follow-up clinical data were investigated. Irinotecan-induced DNA damage was detectable in both CRC cell-lines in vitro, with higher levels of immediate and residual damage noted for the more sensitive HT-29 cells. DNA damage was not detected in vivo, but was measurable in PBLs upon mitogenic stimulation prior to ex vivo SN-38 treatment. Results showed that, following corrections for experimental error, those patients whose PBLs demonstrated higher levels of DNA damage following 10 h of SN-38 exposure ex vivo had significantly longer times to progression than those with lower damage levels (median 291 vs. 173 days, P = 0.014). To conclude, higher levels of irinotecan-induced initial and residual damage correlated with greater cell kill in vitro and a better clinical response. Consequently, DNA damage measures may represent superior biomarkers of irinotecan effect compared to the more often-studied genetic assays for differential drug metabolism.

Grief and risk of depression in context: the emotional outcomes of bereaved cancer caregivers.

We investigated the relationships of grief and depression to cancer caregiving in early bereavement. We began with three expectations: (a) each outcome would reflect different situational predictors, (b) grief would be more directly related to such predictors, and (c) components of grief would relate differently to the caregiving context and depressed mood. We conducted telephone interviews with family caregivers of incurable cancer patients from two hospitals. A total of 199 family caregivers were interviewed at the time of the patient's diagnosis and reinterviewed 3 months after the patient's death. Results showed grief severity was predicted by caregiving circumstances, but bereavement depressed mood was largely unrelated to caregiving. Grief was the main predictor of depressed mood and mediated almost all other effects. We conclude that while grief may trigger depression, the dissimilar connection to context means that the two emotional states should not be equated based purely on similarity of expression.

Gender differences in the experiences of HIV/AIDS-related stigma: a qualitative study in Ghana.

Globally more women have been diagnosed with HIV/AIDS and are more likely to be stigmatized than men, especially in male-dominant societies. Gender differences in the experience of HIV-related stigma, however, have not been extensively explored. Researchers investigate the gender differences in HIV/AIDS-related stigma experiences here. Interviews were conducted with eight HIV patients and their nine discordant family members in Ghana. Our findings include gender differences in disclosure and response to HIV/AIDS diagnosis. The negative impact of HIV-related stigma was found to be more extensive for women than for men. Our findings may be used to facilitate an awareness and understanding through which supportive interventions can be implemented.

Comet assay measures of DNA damage are predictive of bladder cancer cell treatment sensitivity in vitro and outcome in vivo.

Bladder cancer patients suffer significant treatment failure, including high rates of recurrence and poor outcomes for advanced disease. If mechanisms to improve tumour cell treatment sensitivity could be identified and/or if tumour response could be predicted, it should be possible to improve local-control and survival. Previously, we have shown that radiation-induced DNA damage, measured by alkaline Comet assay (ACA), correlates bladder cancer cell radiosensitivity in vitro. In this study we first show that modified-ACA measures of cisplatin and mitomycin-C-induced damage also correlate bladder cancer cell chemosensitivity in vitro, with essentially the same rank order for chemosensitivity as for radiosensitivity. Furthermore, ACA studies of radiation-induced damage in different cell-DNA substrates (nuclei, nucleoids and intact parent cells) suggest that it is a feature retained in the prepared nucleoids that is responsible for the relative damage sensitivity of bladder cancer cells, suggestive of differences in the organisation of DNA within resistant vs. sensitive cells. Second, we show that ACA analysis of biopsies from bladder tumours reveal that reduced DNA damage sensitivity associates with poorer treatment outcomes, notably that tumours with a reduced damage response show a significant association with local recurrence of non-invasive disease and that reduced damage response was a better predictor of recurrence than the presence of high-risk histology in this cohort. In conclusion, this study demonstrates that mechanisms governing treatment-induced DNA damage are both central to and predictive of bladder cancer cell treatment sensitivity and exemplifies a link between DNA damage resistance and both treatment response and tumour aggression.

Simplified method for the collection, storage, and comet assay analysis of DNA damage in whole blood.

Single-cell gel electrophoresis (comet assay) is one of the most common methods used to measure oxidatively damaged DNA in peripheral blood mononuclear cells (PBMC), as a biomarker of oxidative stress in vivo. However, storage, extraction, and assay workup of blood samples are associated with a risk of artifactual formation of damage. Previous reports using this approach to study DNA damage in PBMC have, for the most part, required the isolation of PBMC before immediate analysis or freezing in cryopreservative. This is very time-consuming and a significant drain on human resources. Here, we report the successful storage of whole blood in ~250 µl volumes, at -80°C, without cryopreservative, for up to 1 month without artifactual formation of DNA damage. Furthermore, this blood is amenable for direct use in both the alkaline and the enzyme-modified comet assay, without the need for prior isolation of PBMC. In contrast, storage of larger volumes (e.g., 5 ml) of whole blood leads to an increase in damage with longer term storage even at -80°C, unless a cryopreservative is present. Our "small volume" approach may be suitable for archived blood samples, facilitating analysis of biobanks when prior isolation of PBMC has not been performed.

Relationships and emotional wellbeing among African American and White advanced cancer caregivers.

OBJECTIVE: Advanced cancer family caregivers who have good relationships with other family members and with patient's health care providers (PHCPs) have less emotional distress than caregivers with poor relationships. Given a history of different experiences in medical settings among Whites and African Americans, we examined moderation effects by race. METHODS: Baseline data from an ongoing study were collected via telephone interviews with 397 family caregivers of advanced cancer patients at two cancer clinics. Depressed mood and anxiety were measured with the 14-item Profile of Mood States. RESULTS: Caregivers reporting good relationships with family (p<.001) and PHCPs (p<.001) had lower anxiety and less depressed mood (family, p<.01; PHCP, p<.001). Caregiver race moderated relationship quality: Whites with good PHCP relationships felt less depressed mood (p<.01) and anxiety (p<.01). African Americans with good family relationships showed less depressed mood (p<.05), but no association with anxiety. CONCLUSION: Good relationships are important for caregivers, but PHCPs may have more influence on the wellbeing of White than of African American caregivers. PRACTICE IMPLICATIONS: Developing relationships with caregivers of advanced cancer patients may improve wellbeing for caregivers. In addition, creating strategies to support family relationships may be a useful intervention, especially for African American advanced cancer caregivers.

DNA strand breaks and hypoxia response inhibition mediate the radiosensitisation effect of nitric oxide donors on prostate cancer under varying oxygen conditions.

Prostate cancer cells can exist in a hypoxic microenvironment, causing radioresistance. Nitric oxide (NO) is a radiosensitiser of mammalian cells. NO-NSAIDs are a potential means of delivering NO to prostate cancer cells. This study aimed to determine the effect and mechanism of action of NO-sulindac and radiation, on prostate cancer cells and stroma, under normoxia (21% oxygen) and chronic hypoxia (0.2% oxygen). Using clonogenic assays, at a surviving fraction of 10% the sensitisation enhancement ratios of radiation plus NO-sulindac over radiation alone on PC-3 cells were 1.22 and 1.42 under normoxia and hypoxia, respectively. 3D culture of PC-3 cells revealed significantly reduced sphere diameter in irradiated spheres treated with NO-sulindac. Neither NO-sulindac nor sulindac radiosensitised prostate stromal cells under normoxia or hypoxia. HIF-1α protein levels were reduced by NO-sulindac exposure and radiation at 21 and 0.2% oxygen. Alkaline Comet assay analysis suggested an increased rate of single strand DNA breaks and slower repair of these lesions in PC-3 cells treated with NO-sulindac prior to irradiation. There was a higher level of γ-H2AX production and hence double strand DNA breaks following irradiation of NO-sulindac treated PC-3 cells. At all radiation doses and oxygen levels tested, treatment of 2D and 3D cultures of PC-3 cells with NO-sulindac prior to irradiation radiosensitised PC-3, with minimal effect on stromal cells. Hypoxia response inhibition and increased DNA double strand breaks are potential mechanisms of action. Neoadjuvent and concurrent use of NO-NSAIDs have the potential to improve radiotherapy treatment of prostate cancer under normoxia and hypoxia.

Pediatric brain tumor cancer stem cells: cell cycle dynamics, DNA repair, and etoposide extrusion.

Reliable model systems are needed to elucidate the role cancer stem cells (CSCs) play in pediatric brain tumor drug resistance. The majority of studies to date have focused on clinically distinct adult tumors and restricted tumor types. Here, the CSC component of 7 newly established primary pediatric cell lines (2 ependymomas, 2 medulloblastomas, 2 gliomas, and a CNS primitive neuroectodermal tumor) was thoroughly characterized. Comparison of DNA copy number with the original corresponding tumor demonstrated that genomic changes present in the original tumor, typical of that particular tumor type, were retained in culture. In each case, the CSC component was approximately 3-4-fold enriched in neurosphere culture compared with monolayer culture, and a higher capacity for multilineage differentiation was observed for neurosphere-derived cells. DNA content profiles of neurosphere-derived cells expressing the CSC marker nestin demonstrated the presence of cells in all phases of the cell cycle, indicating that not all CSCs are quiescent. Furthermore, neurosphere-derived cells demonstrated an increased resistance to etoposide compared with monolayer-derived cells, having lower initial DNA damage, potentially due to a combination of increased drug extrusion by ATP-binding cassette multidrug transporters and enhanced rates of DNA repair. Finally, orthotopic xenograft models reflecting the tumor of origin were established from these cell lines. In summary, these cell lines and the approach taken provide a robust model system that can be used to develop our understanding of the biology of CSCs in pediatric brain tumors and other cancer types and to preclinically test therapeutic agents.

Trepidation: a family member's response to shift change in the intensive care unit.

Every shift change in the intensive care unit presents an opportunity for uncertainty among family members regarding rules and information exchange. High levels of uncertainty maintain a presence in the intensive care unit environment, and feelings of trepidation can increase with uncertainty. Critical care nurses in the intensive care unit play a pivotal role in mediating family members' feelings of trepidation by what they do or do not do. Further research into family members' responses to shift change and into inconsistencies in nursing decisions regarding rules and information exchange in the intensive care unit is needed to better understand the feelings that family members experience in the intensive care unit environment and to motivate meaningful changes that will contribute to a clearer sense of purpose for family and their loved ones.

Primary care physicians' involvement in the cancer care of older long-term survivors.

OBJECTIVE: This study investigated survivors' reports of primary care physicians' (PCPs) involvement in three key cancer survivorship activities: discussing cancer history, whether the PCP initiated discussions, and whether discussions led to tests/procedures. METHOD: The sample included 215 older survivors whose health care was maintained in primary care. Logistic regression explored predictors of the three activities, including demographics, cancer characteristics, survivor/PCP association characteristics, health characteristics, and psychosocial well-being. RESULTS: Nearly two thirds of survivors indicated discussing cancer history; most said discussions were PCP initiated and nearly half said discussions resulted in tests/procedures. Predictors of discussing cancer history were African American race and more comorbid conditions. PCP-initiated discussions were related to older age, surviving breast cancer, more years in the PCP's practice, and having less general health worry. The tests/procedures model was not significant. CONCLUSIONS: As older survivors focused more on other health concerns, PCPs remained attentive to cancer issues, prompting discussions about history and ordering tests.