Designing cytochrome P450 enzymes for use in cancer gene therapy.

Cancer is a significant global socioeconomic burden, as millions of new cases and deaths occur annually. In 2020, almost 10 million cancer deaths were recorded worldwide. Advancements in cancer gene therapy have revolutionized the landscape of cancer treatment. An approach with promising potential for cancer gene therapy is introducing genes to cancer cells that encode for chemotherapy prodrug metabolizing enzymes, such as Cytochrome P450 (CYP) enzymes, which can contribute to the effective elimination of cancer cells. This can be achieved through gene-directed enzyme prodrug therapy (GDEPT). CYP enzymes can be genetically engineered to improve anticancer prodrug conversion to its active metabolites and to minimize chemotherapy side effects by reducing the prodrug dosage. Rational design, directed evolution, and phylogenetic methods are some approaches to developing tailored CYP enzymes for cancer therapy. Here, we provide a compilation of genetic modifications performed on CYP enzymes aiming to build highly efficient therapeutic genes capable of bio-activating different chemotherapeutic prodrugs. Additionally, this review summarizes promising preclinical and clinical trials highlighting engineered CYP enzymes' potential in GDEPT. Finally, the challenges, limitations, and future directions of using CYP enzymes for GDEPT in cancer gene therapy are discussed.

Do patients actually understand? An evaluation of the informed consent process for endoscopic procedures in rural Uganda.

INTRODUCTION: Improving surgical access in low- and middle-income countries is vital for the 5 billion people who lack safe surgical care. Tailoring a culturally sensitive approach to consent is essential for patient comprehension and comfort, thereby alleviating the effects of resource constraints and advancing equitable care. This study examines the consenting process for endoscopy at Kyabirwa Surgical Center in Kyabirwa, Jinja, Uganda, to assess patients' knowledge and attitudes as a potential barrier to participating in endoscopic procedures. METHODS: All adult upper endoscopy (EGD) and colonoscopy patients were recruited to participate in a survey of their demographics, knowledge, and attitudes toward their procedure. All patients received a standard consultation explaining the procedure and its risks and benefits. RESULTS: 75 patients were included; median age was 54 years and 56% (n = 42) were women. 92% (n = 69) of patients had never had an endoscopy before and 73% (n = 55) of patients were scheduled for an EGD while the remaining 27% (n = 20) were scheduled for a colonoscopy. Most patients 80% (n = 60) had a basic understanding of what an endoscopy is and 87% (n = 65) its diagnostic purpose. Few patients 15% (n = 11) knew of the most common side effects or if they would have a surgical scar 27% (n = 20). Overall, 46.7% (n = 35) of patients were moderately or severely fearful of getting an endoscopy. Additionally, 45.3% (n = 34) of patients were moderately or severely fearful of receiving anesthesia during their endoscopic procedure. Despite this fear, most patients 85.3% (n = 64) stated that they understood the benefits of the procedure either very well or extremely well. CONCLUSIONS: Most patients understood the role that an endoscopic procedure plays in their care and its potential benefits. Despite this, many patients continued to have high levels of fear associated with both the endoscopic procedure and with receiving anesthesia during their procedure. Future patient education should focus on addressing patients' fears and the risks of undergoing an endoscopy, which may improve the utilization of surgical services.

Ethnic-Racial Identity and Attitude Change: Assessments of Outgroup and Diversity Attitudes among Adolescents in Sweden.

Outgroup and diversity attitudes are important components of intercultural understanding and well-being. Despite the potential of ethnic-racial identity development as a means to foster positive outgroup and diversity attitudes, little is known about its effectiveness in rapidly diversifying contexts such as Sweden. This pre-registered study filled this gap by examining if adolescents taking part in an intervention focused on ethnic-racial identity exploration, the Identity Project, also reported change in outgroup and diversity attitudes, and whether migration background, education type, and ethnic-racial identity development predicted such change. Twenty-three tenth-grade classes in Sweden (N = 509; Mage = 16.28; SDage = 0.80; 66% female; 51% migration background) participated in the intervention and were assessed in four waves over a period of 26 weeks. Whereas ethnic-racial identity exploration and resolution increased for the intervention group, the adolescents reported no change in outgroup and diversity attitudes when compared to a control group. Increases in ethnic-racial identity exploration and resolution co-varied with increases in attitudes, but only at Time 3. The results do not provide support for the link between ethnic-racial identity development and positive outgroup and diversity attitudes, and challenge the notion of attitude change as a cascading effect of the Identity Project intervention in non-US sociocultural contexts. All aspects of the study were pre-registered on the Open Science Framework platform ( https://osf.io/f5896 ).

Collaborative international partnership: Enabling the development of a state-of-the-art minimally invasive surgical center in rural Uganda through telementoring.

BACKGROUND: Access to minimally invasive surgery (MIS) is limited in Sub-Saharan African countries. In 2019, the Mount Sinai Department of Surgery in New York collaborated with local Ugandans to construct the Kyabirwa Surgical Center (KSC), an independent, replicable, self-sustaining ambulatory surgical center in Uganda. We developed a focused MIS training program using a combination of in-person training and supervised telementoring. We present the results of our initial MIS telementoring experience. METHODS: We worked jointly with Ugandan staff to construct the KSC in the rural province of Jinja. A solar-powered backup battery system ensured continuous power availability. Underground fiber optic cables were installed to provide stable high-speed Internet. The local Ugandan general surgeon (JOD) underwent a mini-fellowship in MIS and then trained extensively using the Fundamentals of Laparoscopic Surgery program. After a weeklong in-person session to train the Ugandan OR team, JOD performed laparoscopic cases with telementoring, which was conducted remotely by surgeons in New York via audiovisual feeds from the KSC OR. RESULTS: From October 2021 to February 2024, JOD performed 61 telementored laparoscopic operations at KSC including 37 appendectomies and 24 cholecystectomies. Feedback was provided regarding patient positioning, port placement, surgical technique, instrument use, and critical steps of the operation. There were no intra-operative complications. Postoperatively, field medical workers visited patients at home to collect follow-up information. Two superficial wound infections (3.3%) were reported in the short-term follow-up. CONCLUSION: Telementoring can be safely implemented to assist surgeons in previously underserved areas to provide advanced laparoscopic surgical care to the local patient population.

Promoting Adolescents' Heritage Cultural Identity Development: Exploring the Role of Autonomy and Relatedness Satisfaction in School-Based Interventions.

Given the significance of heritage cultural identity for optimal adolescent development, it is imperative to investigate factors influencing the efficacy of interventions aimed at promoting heritage cultural identity. Using latent profile cluster analysis and multinomial logistic regressions, this longitudinal study examined how autonomy and relatedness need satisfaction at school (1) related to heritage cultural identity development trajectories, and (2) moderated effects of a school-based intervention. The study included N = 198 adolescents (Mage = 12.86 years, SDage = 0.75, 52% female, 41% immigrant descent, 49% intervention group) in Germany. Teacher-student relationships played an important role in facilitating intervention effects on identity development trajectories, emphasizing the importance of the relational context when implementing school-based interventions to promote heritage cultural identity development.

Vesicular monoamine transporter-2 inhibitor JPC-141 prevents methamphetamine-induced dopamine toxicity and blocks methamphetamine self-administration in rats.

Previous research has demonstrated therapeutic potential for VMAT2 inhibitors in rat models of methamphetamine use disorder. Here, we report on the neurochemical and behavioral effects of 1-(2-methoxyphenethyl)-4-phenethypiperazine (JPC-141), a novel analog of lobelane. JPC-141 potently inhibited (Ki = 52 nM) [3H]dopamine uptake by VMAT2 in striatal vesicles with 50 to 250-fold greater selectivity for VMAT2 over dopamine, norepinephrine and serotonin plasmalemma transporters. Also, JPC-141 was 57-fold more selective for inhibiting VMAT2 over [3H]dofetilide binding to hERG channels expressed by HEK293, suggesting relatively low potential for cardiotoxicity. When administered in vivo to rats, JPC-141 prevented the METH-induced reduction in striatal dopamine content when given either prior to or after a high dose of METH, suggesting a reduction in METH-induced dopaminergic neurotoxicity. In behavioral assays, JPC-141 decreased METH-stimulated locomotor activity in METH-sensitized rats at doses of JPC-141 which did not alter locomotor activity in the saline control group. Moreover, JPC-141 specifically decreased iv METH self-administration at doses that had no effect on food-maintained responding. These findings support the further development of VMAT2 inhibitors as pharmacotherapies for individuals with methamphetamine use disorder.

The Use of the Acute Stroke Management Questionnaire to Measure Acute Care Nurses' Stroke Knowledge.

ABSTRACT: BACKGROUND: Nurses are key in identifying and treating in-hospital strokes (IHSs). Delayed treatment times and poor patient outcomes are associated with IHSs. Information is needed on nurses' stroke knowledge and the objective measurement of stroke knowledge using a validated tool. The Acute Stroke Management Questionnaire (ASMaQ) was recently developed to test stroke knowledge of healthcare professionals but has not been used on a nursing-specific population. METHODS: Through online surveying and use of ASMaQ, we will measure stroke knowledge of nurses caring for adult, hospitalized patients in an urban, southeast US health system. RESULTS: Total N is 196. Most participants (74.5%, n = 146) never worked on a stroke floor; however, almost all (95.9%, n = 188) cared for a stroke patient in the past. Most participants (65.3%, n = 128) reported receiving prelicensure stroke education, and 98.5% (n = 193) received postlicensure stroke education. Acute Stroke Management Questionnaire total scores ranged from 93 to 133 (mean [SD], 117.35 [8.15]). Most participants scored in the good stroke knowledge range for all 3 ASMaQ domains and total ASMaQ score. CONCLUSION: The online delivery of the ASMaQ was successful in testing nurses' stroke knowledge, and nurses were shown to have good stroke knowledge. Future initiatives should focus on discerning whether certain nurse characteristics predict higher or lower levels of stroke knowledge to help inform educational initiatives to improve IHS outcomes.

Increase in Full-Length Dystrophin by Exon Skipping in Duchenne Muscular Dystrophy Patients with Single Exon Duplications: An Open-label Study.

Single exon duplications account for disease in a minority of Duchenne muscular dystrophy patients. Exon skipping in these patients has the potential to be highly therapeutic through restoration of full-length dystrophin expression. We conducted a 48-week open label study of casimersen and golodirsen in 3 subjects with an exon 45 or 53 duplication. Two subjects (aged 18 and 23 years) were non-ambulatory at baseline. Upper limb, pulmonary, and cardiac function appeared stable in the 2 subjects in whom they could be evaluated. Dystrophin expression increased from 0.94 % ±0.59% (mean±SD) of normal to 5.1% ±2.9% by western blot. Percent dystrophin positive fibers also rose from 14% ±17% at baseline to 50% ±42% . Our results provide initial evidence that the use of exon-skipping drugs may increase dystrophin levels in patients with single-exon duplications.

[Coumarin or DOAC in frail elderly patients with atrial fibrillation?] / Cumarine of DOAC bij kwetsbare ouderen met atriumfibrilleren?

OBJECTIVE: To investigate whether frail elderly people with atrial fibrillation (AF) who are currently using a vitamin K antagonist (VKA) should be switched to a direct-acting oral anticoagulant (DOAC). DESIGN: Randomized clinical trial. METHODS: 662 frail elderly AF patients were switched to a DOAC, and 661 patients continued their VKA. The primary endpoint was a major or clinically relevant non-major bleeding during 1 year of follow-up. Secondary endpoints included thrombo-embolic events. RESULTS: The mean age of the included patients was 83 years. In the 'switch to DOAC arm', 101 bleeding events (15.3%) occurred and in the 'continue with VKA arm', 62 bleeding events (9.4%); an increase of 69% more bleeding events (P-value 0.001). The number of thrombo-embolic events was not significantly different between both groups. CONCLUSION: Switching from a VKA to a DOAC in frail elderly people with AF leads to 69% more bleeding, without a difference in thrombo-embolic events.

Defining Roles and Responsibilities of the Health Workforce to Respond to the Climate Crisis.

Importance: The adverse effects of climate change are now apparent, disproportionately affecting marginalized and vulnerable populations and resulting in urgent worldwide calls to action. Health professionals occupy a critical position in the response to climate change, including in climate mitigation and adaptation, and their professional expertise and roles as health messengers are currently underused in the society-wide response to this crisis. Observations: Clinical and public health professionals have important roles and responsibilities, some of which are shared, that they must fill for society to successfully mitigate the root causes of climate change and build a health system that can reduce morbidity and mortality impacts from climate-related hazards. When viewed through a preventive framework, the unique and synergizing roles and responsibilities provide a blueprint for investment in climate change-related prevention (primary, secondary, and tertiary), capacity building, education, and training of the health workforce. Substantial investment in increasing the competence and collaboration of health professionals is required, which must be undertaken in an urgent, coordinated, and deliberate manner. Conclusions and Relevance: Exceptional collaboration, knowledge sharing, and workforce capacity building are essential to tackle the complex ways in which climate change threatens health. This framework serves as a guide for health system leaders, education institutions, policy planners, and others seeking to create a more resilient and just health system.

Feasibility and utility of in-home body weight support harness system use in young children treated for spinal muscular atrophy: A single-arm prospective cohort study.

PURPOSE: This single-arm prospective cohort study aimed to evaluate the feasibility and utility of in-home body weight support harness system (BWSS) use in children treated for spinal muscular atrophy (SMA). METHODS: Individuals with 2 or 3 copies of SMN2 who received pharmacotherapeutic treatment, had head control, and weight <50lbs were enrolled. Families were provided a BWSS and documented use. Motor outcome assessments were completed at baseline, month 3 and month 6. Families provided feedback in an end of study survey. RESULTS: All 32 participants (2.9 (SD 1.9) yrs), improved or remained stable on all outcomes. Average reported frequency of use was 4.1(2.3) hrs/week. Controlling for other covariates, frequency of use explained over 70% of the variability in change scores. Family feedback was overwhelmingly positive. CONCLUSION: Use of in-home BWSS is a safe, feasible and useful option to increase exercise dosage after treatment in SMA and may help optimize motor abilities. TRIAL REGISTRATION: Study registered with: Clinicaltrials.gov Clinicaltrials.gov identifier: NCT05715749.

Gene therapy with bidridistrogene xeboparvovec for limb-girdle muscular dystrophy type 2E/R4: phase 1/2 trial results.

Limb-girdle muscular dystrophy 2E/R4 is caused by mutations in the ß-sarcoglycan (SGCB) gene, leading to SGCB deficiency and consequent muscle loss. We developed a gene therapy approach based on functional replacement of the deficient SCB protein. Here we report interim results from a first-in-human, open-label, nonrandomized, phase 1/2 trial evaluating the safety and efficacy of bidridistrogene xeboparvovec, an adeno-associated virus-based gene therapy containing a codon-optimized, full-length human SGCB transgene. Patients aged 4-15 years with confirmed SGCB mutations at both alleles received one intravenous infusion of either 1.85 × 1013 vector genome copies kg-1 (Cohort 1, n = 3) or 7.41 × 1013 vector gene copies kg-1 (Cohort 2, n = 3). Primary endpoint was safety, and secondary endpoint was change in SGCB expression in skeletal muscle from baseline to Day 60. We report interim Year 2 results (trial ongoing). The most frequent treatment-related adverse events were vomiting (four of six patients) and gamma-glutamyl transferase increase (three of six patients). Serious adverse events resolved with standard therapies. Robust SGCB expression was observed: Day 60 mean (s.d.) percentage of normal expression 36.2% (2.7%) in Cohort 1 and 62.1% (8.7%) in Cohort 2. Post hoc exploratory analysis showed preliminary motor improvements using the North Star Assessment for Limb-girdle Type Muscular Dystrophies maintained through Year 2. The 2-year safety and efficacy of bidridistrogene xeboparvovec support clinical development advancement. Further studies are necessary to confirm the long-term safety and efficacy of this gene therapy. ClinicalTrials.gov registration: NCT03652259 .

Safety of Switching From a Vitamin K Antagonist to a Non-Vitamin K Antagonist Oral Anticoagulant in Frail Older Patients With Atrial Fibrillation: Results of the FRAIL-AF Randomized Controlled Trial.

BACKGROUND: There is ambiguity whether frail patients with atrial fibrillation managed with vitamin K antagonists (VKAs) should be switched to a non-vitamin K oral anticoagulant (NOAC). METHODS: We conducted a pragmatic, multicenter, open-label, randomized controlled superiority trial. Older patients with atrial fibrillation living with frailty (≥75 years of age plus a Groningen Frailty Indicator score ≥3) were randomly assigned to switch from international normalized ratio-guided VKA treatment to an NOAC or to continued VKA treatment. Patients with a glomerular filtration rate <30 mL·min-1·1.73 m-2 or with valvular atrial fibrillation were excluded. Follow-up was 12 months. The cause-specific hazard ratio was calculated for occurrence of the primary outcome that was a major or clinically relevant nonmajor bleeding complication, whichever came first, accounting for death as a competing risk. Analyses followed the intention-to-treat principle. Secondary outcomes included thromboembolic events. RESULTS: Between January 2018 and June 2022, a total of 2621 patients were screened for eligibility and 1330 patients were randomly assigned (mean age 83 years, median Groningen Frailty Indicator score 4). After randomization, 6 patients in the switch-to-NOAC arm and 1 patient in the continue-with-VKA arm were excluded due to the presence of exclusion criteria, leaving 662 patients switched from a VKA to an NOAC and 661 patients continued VKAs in the intention-to-treat population. After 163 primary outcome events (101 in the switch arm, 62 in the continue arm), the trial was stopped for futility according to a prespecified futility analysis. The hazard ratio for our primary outcome was 1.69 (95% CI, 1.23-2.32). The hazard ratio for thromboembolic events was 1.26 (95% CI, 0.60-2.61). CONCLUSIONS: Switching international normalized ratio-guided VKA treatment to an NOAC in frail older patients with atrial fibrillation was associated with more bleeding complications compared with continuing VKA treatment, without an associated reduction in thromboembolic complications. REGISTRATION: URL: https://eudract.ema.europa.eu; Unique identifier: 2017-000393-11. URL: https://eudract.ema.europa.eu; Unique identifier: 6721 (FRAIL-AF study).

Health Screening Among a Rural, Hourly Workforce: North Carolina SERVIRE Project.

INTRODUCTION: This study demonstrates the feasibility of a novel, business-partnered, and worksite-based approach to healthcare access to facilitate chronic disease screening and diagnosis among rural hourly workers. The prevalence of undiagnosed and untreated diabetes and hypertension among screening participants was determined. METHODS: From February 2021 to June 2023, investigators partnered with 29 businesses to screen 1,114 workers. Health screenings included a demographic questionnaire, A1c testing for prediabetes (A1c of 5.7-6.4) and diabetes (A1c≥6.5), hypertension (Stage 1: systolic blood pressure of 130-139 mmHg; Stage 2: systolic blood pressure ≥140 mmHg), kidney disease (estimated glomerular filtration rate <60; urine protein ≥1+), and questionnaire assessment of stroke (CHA2DS2-VASc) and sleep apnea (STOP-bang) risk. RESULTS: Of the 1,114 individuals screened (n=632, 56.7% male; n=497, 44.6% Black)), 388 (36%) screened positive for prediabetes or diabetes. Diabetes was previously undiagnosed in 273 (70.4%) of these participants. More than half of the participants (n=680, 62.4%) had an elevated blood pressure reading during the screening, and the majority of these participants (n=445, 65.4%) had not been previously diagnosed with hypertension. In addition, 241 (21.6%) participants were at an increased risk of stroke (CHA2DS2-VASc≥2), and 182 (23.7%) had a STOP-Bang score ≥4, indicating an increased risk of obstructive sleep apnea. CONCLUSIONS: By partnering with local businesses to deliver worksite-based health screenings, high rates of undiagnosed and uncontrolled diabetes and hypertension were identified among the rural, hourly workforce. This worksite-based approach to healthcare access could facilitate early detection of chronic disease, improve patient engagement in the healthcare system, and ultimately yield better long-term public health outcomes.

Critical, active, and well adapted: Antecedents and consequences of adolescents' critical consciousness profiles.

This preregistered study aimed to identify antecedents and consequences of adolescents' critical consciousness (CC) profiles with person-centered approaches based on data from 663 ethnically diverse German adolescents collected from 2017 to 2019 (Mage = 12.91, 50% male, 50% female). Latent profile analyses of adolescents' critical reflection and interpersonal and structural critical action intentions yielded three profiles: "uncritical," "armchair activists," and "actionists." Discrimination experiences, but not CC classroom climate, predicted a higher likelihood of being in the armchair activist or actionist profiles. The actionist profile showed better, but the armchair activist profile worse socioemotional and academic adaptation cross-sectionally and over time (vs. the other profiles). The results highlight the potential of person-centered approaches and of fostering developmentally appropriate forms of critical action among adolescents in novel contexts.