Generation Rx: evaluation of medication misuse prevention education for older adults.

Objective: Increased medication misuse over the last two decades has prompted extensive discussion about the lack of evidence-based and evidence-informed prevention education programs targeting the topic. As older adults are high utilizers of medications, this is an important population to reach with such educational programming. This study was designed to assess the change in knowledge and behavioral intentions of older adult participants after attending an educational session focused on safe medication use utilizing the Generation Rx Older Adult Toolkit (GROAT) resources. Methods: The Generation Rx team at The Ohio State University College of Pharmacy (OSU COP) partnered with The Ohio State University Extension offices (OSU Extension) across the state of Ohio to provide GROAT educational programming in their communities. OSU Extension Educators were trained via the standardized virtual training program, Generation Rx Ambassadors. Program participants were surveyed immediately before and after the educational events. Pre- and post-survey data was then analyzed to assess knowledge gain and behavioral intentions about safe medication practices, as well as program perception and program satisfaction. Results: Programming occurred between May 2022 and September 2022. In total, OSU Extension Educators collectively engaged 843 individuals in a prevention education program utilizing the GROAT materials. After excluding participants under 50 years of age, there were 297 pre surveys and 245 post surveys included in the data analysis. Knowledge gains from pre- to post-survey showed a significant increase in correct responses in seven of the eight questions asked regarding safe medication practices. All five questions evaluating behavioral intentions demonstrated positive results after the programming (p < 0.001). Participants' perceptions and program satisfaction were also favorable. Conclusion: This study found through pre- and post-survey results that the Generation Rx Older Adult Toolkit programming delivered by Generation Rx trained OSU Extension Educators significantly increased older adult participants' knowledge and favorably impacted behavioral intentions around safe medication use practices.

N-substituted-4-(pyridin-4-ylalkyl)piperazine-1-carboxamides and related compounds as Leishmania CYP51 and CYP5122A1 inhibitors.

CYP5122A1, an enzyme involved in sterol biosynthesis in Leishmania, was recently characterized as a sterol C4-methyl oxidase. Screening of a library of compounds against CYP5122A1 and CYP51 from Leishmania resulted in the identification of two structurally related classes of inhibitors of these enzymes. Analogs of screening hit N-(3,5-dimethylphenyl)-4-(pyridin-4-ylmethyl)piperazine-1-carboxamide (4a) were generally strong inhibitors of CYP51 but were less potent against CYP5122A1 and typically displayed weak inhibition of L. donovani promastigote growth. Analogs of screening hit N-(4-(benzyloxy)phenyl)-4-(2-(pyridin-4-yl)ethyl)piperazine-1-carboxamide (18a) were stronger inhibitors of both CYP5122A1 and L. donovani promastigote proliferation but also remained selective for inhibition of CYP51. Two compounds in this series, N-(4-((3,5-bis(trifluoromethyl)benzyl)oxy)phenyl)-4-(2-(pyridin-4-yl)ethyl)piperazine-1-carboxamide (18e) and N-(4-((3,5-di-tert-butylbenzyl)oxy)phenyl)-4-(2-(pyridin-4-yl)ethyl)piperazine-1-carboxamide (18i) showed modest selectivity for inhibiting L. donovani promastigote proliferation compared to J774 macrophages and were effective against intracellular L. donovani with EC50 values in the low micromolar range. Replacement of the 4-pyridyl ring present in 18e with imidazole resulted in a compound (4-(2-(1H-imidazol-1-yl)ethyl)-N-(4-((3,5-bis(trifluoromethyl)benzyl)oxy)phenyl)piperazine-1-carboxamide, 18p) with approximately fourfold selectivity for CYP5122A1 over CYP51 that inhibited both enzymes with IC50 values ≤ 1 µM, although selective potency against L. donovani promastigotes was lost. Compound 18p also inhibited the proliferation of L. major promastigotes and caused the accumulation of 4-methylated sterols in L. major membranes, indicating that this compound blocks sterol demethylation at the 4-position in Leishmania parasites. The molecules described here may therefore be useful for the future identification of dual inhibitors of CYP51 and CYP5122A1 as potential antileishmanial drug candidates and as probes to shed further light on sterol biosynthesis in Leishmania and related parasites.

Genetic Alteration of p18INK4C and its Expression in Peripheral Blood Mononuclear Cells Were Not Associated With Progression-free Survival of Multiple Myeloma Patients After Autologous Stem Cell Transplant.

BACKGROUND/AIM: The tumor suppressor CDKN2C (also designed as p18INK4C or p18) is deleted in approximately 7% to 40% of multiple myeloma (MM) patients, indicative of its potential association with myeloma pathogenesis. MATERIALS AND METHODS: A quantitative real-time PCR-based assay was developed to determine p18 deletion in DNA from peripheral blood mononuclear cells (PBMCs) in a cohort of 108 multiple myeloma patients after autologous stem cell transplant (ASCT). Additionally, the p18 mRNA expression level in PBMCs was quantitatively assessed using Taqman® gene expression assays. RESULTS: p18 was homozygously and hemizygously deleted in PBMCs from 3 (2.8%) and 5 (4.6%) patients, respectively. Neither the p18 deletion nor the p18 mRNA levels in PBMCs were associated with progression-free survival (PFS), 90-day response, and the occurrence of severe mucositis in these patients (all p-values >0.20). CONCLUSION: Neither p18 deletion nor p18 mRNA expression in PBMCs can be used as a prognostic biomarker in MM patients.

Evaluation of Cocurricular Learning and the Influence of Student Choice.

OBJECTIVE: The primary objective of this study was to evaluate the students' choice of activities and perception of a cocurricular program (CCP). Attitude and skill development and areas for program improvement were also assessed. METHODS: Data were evaluated from 2 sources: a student survey administered to all Doctor of Pharmacy students to determine student perceptions of the CCP and barriers to success and student activity reporting data from postexperience submissions to evaluate the CCP. Data were stratified by student demographics to identify trends. A comparison of data was conducted from both sources on the students' perception of learning value. RESULTS: Data for 405 students were available for analysis. The highest preference overall from students for hosts of activities were professional student organizations; the highest preference of location was online, asynchronous. The most meaningful benefits were learning information about a new subject (n = 258, 63.7%), ability to extend learning from the classroom to real life (n = 247, 61%), and networking opportunities (n = 218, 53.8%). The top barriers for completion of the program included scheduling challenges (n = 296, 73.1%), lack of time (n = 249, 61.5%), and lack of interest (n = 187, 46.2%). Subpopulations identified different benefits and barriers to completion. CONCLUSION: Students have overall positive perceptions of the CCP and its value for skill development. Findings were applied to adjust the CCP at the institution, including a reduction in total hours required. The continuous evaluation of CCPs is important to optimize student learning and address curricular overload.

Impact of point-of-care gonorrhea and chlamydia testing in the emergency department on reducing overtreatment rates.

BACKGROUND: Chlamydia trachomatis (CT) and Neisseria gonorrhoeae (NG) infections continue to increase in the United States. Advancement in technology with point-of-care (POC) testing can improve the overall treatment of sexually transmitted infections (STI) in the emergency department (ED) by shortening the time to test result and administration of accurate treatment. The purpose of this study was to assess if the POC test reduced the rate of overtreatment for CT and/or NG compared to the standard-of-care (SOC) test. METHODS: This retrospective cohort study included adult patients tested for CT and NG at two urban EDs between August 2020 and October 2022. This cohort excluded hospital admissions, elopement, pregnancy, rectal and oral samples, victims of sexual assault, and diagnoses for which antimicrobial treatment overlapped that of CT/NG. The primary outcome assessed overtreatment, defined as receiving treatment in the ED or a prescription prior to discharge for patients who tested negative for CT and/or NG. Secondary outcomes included undertreatment rates, overtreatment rates in select populations, test turnaround time, and ED length of stay (LOS). RESULTS: Of 327 patients screened, 97 patients were included in the SOC group and 100 in POC. Overtreatment for CT was provided in zero POC patients and 29 (29.9%) SOC patients (p < 0.001). NG was overtreated in 1 (1%) POC and 23 (23.7%) SOC (p < 0.001). POC was associated with undertreatment of CT and/or NG in two patients, compared to four patients tested with SOC. Overall, treatment was deemed inappropriate for 5 (5%) of those tested with POC, compared to 35 (36%) tested with SOC (p < 0.001). There was no difference in ED LOS (2.7 vs 3.01 h, p = 0.41). CONCLUSIONS: POC testing facilitated the return of results prior to patients being discharged from the ED. Compared to standard testing, POC improved appropriateness of CT and NG treatment by reducing the rates of overtreatment.

Improving Self-Perceived Competencies of Second-Year Pharmacy Students Through an Introductory Medication Reconciliation Rotation.

OBJECTIVE: Medication reconciliation (MedRec) is an essential health care function that is particularly relevant to pharmacists' expertise and a learning opportunity for pharmacy students. Our objective was to assess change across clinical competence, confidence, and communication skills after the completion of a MedRec rotation by second-year pharmacy students. METHODS: A retrospective post-then-pre-survey including 29 questions was developed/delivered to students after the completion of required MedRec hours. The primary end point was the change in 3 domains via summed scores from individual questions. Cohen's difference (d) was used to determine group effect size change. The secondary end points included individual question change, perceived patient impact, and subgroup analyses. RESULTS: Of 115 second-year pharmacy students, 81.7% (n = 94) participated in the study. Students self-reported increases on the Likert scale (0-10) of 2.49 ± 1.90 in clinical competency domain, 3.57 ± 2.13 in confidence domain , and 3.12 ± 2.15 in communication skills domain, representing statistically significant and large group effect changes across all 3. A total of 21 of the 22 individual questions had large group effect changes; 1 question (nursing communications) had a moderate group effect change. Student perception of MedRec impact on patient care (Likert scale 0-10) was positive: post-rotation score 7.39 ± 1.57. CONCLUSION: To the best of our knowledge, this is the first larger-scale study that examines student-evaluated outcomes of a MedRec-based rotation. Students self-reported high levels of post-rotation competency across all domains; students from ethnic minorities and with less work/MedRec experience increased their lower pre-rotation scores to statistically similar post-rotation scores, compared with non-minority and more experienced peers. Further study of the model and outcomes is advised.

Impact of Clinical Pharmacists in the Inflammatory Bowel Disease Clinic.

Background: Limited evidence exists regarding pharmacist involvement and impact in inflammatory bowel disease (IBD) interdisciplinary clinic care models. The purpose is to describe pharmacist utilization in an interdisciplinary IBD clinic and evaluate clinical impact on patient quality of life. Methods: This was a retrospective cohort study comparing outcomes in patients with Crohn's disease initiated on therapy when the implementation of pharmacy services began (Early Phase) to the expansion of pharmacy services (Recent Phase). The primary outcome compared the proportion of patients referred to a pharmacist and those achieving a Harvey-Bradshaw Index (HBI) reduction of ≥3 points after therapy initiation. Results: 50 patients were included in the Early Phase and 43 patients in the Recent Phase. Utilization in pharmacy referrals increased from 48% (n = 24) in the Early Phase to 72% (n = 31) in the Recent Phase (P = 0.03). The proportion of patients achieving a HBI reduction of ≥3 points increased from 35% (n = 14) in the Early Phase to 51% (n = 18) in the Recent Phase (P = 0.23). Results also found a greater proportion of patients remaining steroid free in the Recent Phase compared to the Early Phase (50% vs 63%; P = 0.01) and C-reactive protein (CRP) improved significantly in the Recent Phase (-11) compared to (-3) in the Early Phase (P = 0.006). Conclusion: The utilization of pharmacists in an interdisciplinary IBD clinic increased and showed to impact patient care through improving symptom relief as seen by the achievement rate of the HBI score reduction, reducing steroid use after therapy initiation, and making clinically significant interventions.

A case of gastric granulosa cell tumor resected by endoscopic submucosal dissection.

Granular cell tumors, uncommon soft tissue growths, predominantly manifest in the subcutaneous and tongue areas, while those in the gastrointestinal tract are scarce and develop slowly. Patients typically show no distinct clinical symptoms and are hard to differentiate from gastrointestinal mesenchymal tumors, smooth muscle tumors, neural sheath tumors, and rhabdomyosarcomas using endoscopy. This paper details a case of a granular cell tumor in the stomach addressed through endoscopic submucosal dissection, focusing on its endoscopic attributes and clinicopathological traits.

Evaluation of a Student Pharmacist-Driven Fall-Prevention Program for Older People.

There is limited research on the impact of fall prevention education for older community-living people led by student pharmacists, which includes a medication review to identify Fall Risk-Increasing Drugs (FRIDs). Study objectives were to first assess the knowledge and behavioral intentions of older people after attending a student pharmacist-led fall-prevention program (FPP) and secondly to quantify the number of FRIDs identified during a medication review. Between October 2022 and April 2023, four independent-living facilities and two senior centers served as programming locations. Events began with a fall prevention-focused presentation provided by student pharmacists. Attendees voluntarily filled out surveys to assess their knowledge and behavioral intentions regarding fall prevention. Optional medication reviews were offered. Additional survey questions were asked of medication review participants. If FRIDs were identified, the individual was provided documentation to share with their prescriber. Fall prevention bingo was offered at select events to review educational content and engage those waiting for a medication review. Eighty-six older people attended the presentations; 45 people completed medication reviews across six sites. Survey information was available for 65 presentation attendees and 29 medication review participants. After programming, 64 out of 65 participants stated they felt comfortable speaking to their pharmacist or provider about falls and their medications. Most survey respondents correctly selected which medications increase fall risk. Twenty-two of 29 medication review participants were taking at least one FRID. The FPP described showed positive results through a post-survey evaluation. Participants demonstrated knowledge of fall hazards including medications and a willingness to discuss falls and FRIDs with health professionals. These factors may lead to concrete interventions to avoid falls and their associated health consequences for older people.

Circumvention of Topoisomerase IIα Intron 19 Intronic Polyadenylation in Acquired Etoposide-Resistant Human Leukemia K562 Cells.

DNA topoisomerase IIα (TOP2α; 170 kDa, TOP2α/170) is an essential enzyme for proper chromosome dysjunction by producing transient DNA double-stranded breaks and is an important target for DNA damage-stabilizing anticancer agents, such as etoposide. Therapeutic effects of TOP2α poisons can be limited due to acquired drug resistance. We previously demonstrated decreased TOP2α/170 levels in an etoposide-resistant human leukemia K562 subline, designated K/VP.5, accompanied by increased expression of a C-terminal truncated TOP2α isoform (90 kDa; TOP2α/90), which heterodimerized with TOP2α/170 and was a determinant of resistance by exhibiting dominant-negative effects against etoposide activity. Based on 3'-rapid amplification of cDNA ends, we confirmed TOP2α/90 as the translation product of a TOP2α mRNA in which a cryptic polyadenylation site (PAS) harbored in intron 19 (I19) was used. In this report, we investigated whether the resultant intronic polyadenylation (IPA) would be attenuated by blocking or mutating the I19 PAS, thereby circumventing acquired drug resistance. An antisense morpholino oligonucleotide was used to hybridize/block the PAS in TOP2α pre-mRNA in K/VP.5 cells, resulting in decreased TOP2α/90 mRNA/protein levels in K/VP.5 cells and partially circumventing drug resistance. Subsequently, CRISPR/CRISPR-associated protein 9 with homology-directed repair was used to mutate the cryptic I19 PAS (AATAAA→ACCCAA) to prevent IPA. Gene-edited clones exhibited increased TOP2α/170 and decreased TOP2α/90 mRNA/protein and demonstrated restored sensitivity to etoposide and other TOP2α-targeted drugs. Together, results indicated that blocking/mutating a cryptic I19 PAS in K/VP.5 cells reduced IPA and restored sensitivity to TOP2α-targeting drugs. SIGNIFICANCE STATEMENT: The results presented in this study indicate that CRISPR/CRISPR-associated protein 9 gene editing of a cryptic polyadenylation site (PAS) within I19 of the TOP2α gene results in the reversal of acquired resistance to etoposide and other TOP2-targeted drugs. An antisense morpholino oligonucleotide targeting the PAS also partially circumvented resistance.

Benefit of clinical pharmacists in neurology clinics at an academic medical center.

BACKGROUND: Patients with neurologic diseases have complex medical needs and may benefit from the addition of clinical pharmacists in their care. OBJECTIVES: This study aimed to describe integration and benefit of clinical pharmacists in neuroimmunology and neuromuscular clinics at an academic medical center. METHODS: This retrospective chart review evaluated patients initiated on a neurology medication for a neuroimmunology or neuromuscular disease state before and after pharmacist integration in neurology clinics. The primary outcome measured access to an initially prescribed neuroimmunology or neuromuscular medication within 90 days of prescription. Secondary outcomes included access to an initially prescribed or alternative neurology medication owing to insurance requirements within 90 days, time from initial prescription to start, and description of pharmacist involvement. RESULTS: There were 101 patients in the pregroup and 101 patients in the postgroup. The percentage of patients with confirmed initially prescribed medication access at 90 days increased in the postgroup compared with the pregroup (87.1% vs. 72.5%, respectively, P = 0.014). For secondary outcomes, the percentage of patients who started on an initially prescribed or alternative neuroimmunology or neuromuscular medication within 90 days also increased in the postgroup compared with the pregroup (90.0% vs. 73.3%, respectively, P = 0.004). Additional pharmacist involvement occurred in 64 patients (63.4%) in the postgroup and included prior authorization approval assistance, drug information support, and medication liaison interventions, with an average of 4.7 pharmacist interventions at each pharmacy-led encounter. CONCLUSION: The addition of pharmacists into neuroimmunology and neuromuscular clinics improved operational access to medications for neuroimmunology and neuromuscular conditions. In addition, pharmacists were able to assist with multiple areas of patient care including medication education, monitoring, and serving as a medication liaison. This study supports continuing to offer clinical pharmacy services in neuroimmunology and neuromuscular departments and may support the addition of clinical pharmacists into neurology services at other institutions.

Vincristine Disposition and Neurotoxicity Are Unchanged in Humanized CYP3A5 Mice.

Previous studies have suggested that the incidence of vincristine-induced peripheral neuropathy (VIPN) is potentially linked with cytochrome P450 (CYP)3A5, a polymorphic enzyme that metabolizes vincristine in vitro, and with concurrent use of azole antifungals such as ketoconazole. The assumed mechanism for these interactions is through modulation of CYP3A-mediated metabolism, leading to decreased vincristine clearance and increased susceptibility to VIPN. Given the controversy surrounding the contribution of these mechanisms, we directly tested these hypotheses in genetically engineered mouse models with a deficiency of the entire murine Cyp3a locus [Cyp3a(-/-) mice] and in humanized transgenic animals with hepatic expression of functional and nonfunctional human CYP3A5 variants. Compared with wild-type mice, the systemic exposure to vincristine was increased by only 1.15-fold (95% confidence interval, 0.84-1.58) in Cyp3a(-/-) mice, suggesting that the clearance of vincristine in mice is largely independent of hepatic Cyp3a function. In line with these observations, we found that Cyp3a deficiency or pretreatment with the CYP3A inhibitors ketoconazole or nilotinib did not influence the severity and time course of VIPN and that exposure to vincristine was not substantially altered in humanized CYP3A5*3 mice or humanized CYP3A5*1 mice compared with Cyp3a(-/-) mice. Our study suggests that the contribution of CYP3A5-mediated metabolism to vincristine elimination and the associated drug-drug interaction potential is limited and that plasma levels of vincristine are unlikely to be strongly predictive of VIPN. SIGNIFICANCE STATEMENT: The current study suggests that CYP3A5 genotype status does not substantially influence vincristine disposition and neurotoxicity in translationally relevant murine models. These findings raise concerns about the causality of previously reported relationships between variant CYP3A5 genotypes or concomitant azole use with the incidence of vincristine neurotoxicity.

Evaluation of gender-affirming care experiences of transgender and gender diverse patients within a LGBTQ+ community pharmacy.

BACKGROUND: Transgender and gender diverse (TGD) populations require personalized care. Lived experiences and needs TGD populations express, compounded by limited care access, negatively shape health care involvement. Manifestations from these barriers may present as health care avoidance, identity concealment, or preventive care hinderance. Community pharmacies remain engagement points for TGD patients, but gender diverse services remain limited. What remains unknown is how TGD pharmacy perceptions and behaviors are influenced with gender-affirming care (GAC) accessibility. OBJECTIVES: The primary objective is to assess how TGD patient perceptions and behaviors toward community pharmacy experiences are affected through a lesbian, gay, bisexual, transgender, queer/questioning, and others (LGBTQ+) community-based health system. METHODS: A cross-sectional, multisite, reflective survey was conducted at 4 LGBTQ+ community pharmacies in central and southwest Ohio. Nine 5-point Likert-item questions and one ordinal question were used to analyze perception and behavior. Participants responded for LGBTQ+ and external pharmacy experiences respectively. Data were analyzed through descriptive methods, paired Student's t test, and Fisher's exact test or c2 test where appropriate. RESULTS: In total, 267 surveys were completed with 96 TGD submissions qualifying for analysis. Perceptions toward pharmacy experience saw statistically significant differences among all evaluations of perception. Behavioral assessment demonstrated statistically significant improvements in pharmacy outreach except for seeking medications from outside sources. Respondents indicated more involvement with the LGBTQ+ pharmacies versus external pharmacies in discussing medications (96.9% vs. 60.4%), care plans (64.6% vs. 41.6%), disclosure of pronouns or gender (97.9% vs. 43.8%), and feeling needs were understood (96.8% vs. 51%). CONCLUSION: Inclusive community pharmacies may positively affect pharmacy perceptions and behaviors of TGD patients. These findings call attention to barriers in the provision of care for TGD patients while highlighting the change community pharmacies can have when providing these services. Community pharmacies should be encouraged to incorporate inclusive environments to improve TGD patient care involvement and access.

The effects of organic food on human health: a systematic review and meta-analysis of population-based studies.

CONTEXT: Although the nutritional composition of organic food has been thoroughly researched, there is a dearth of published data relating to its impact on human health. OBJECTIVE: This systematic review aimed to examine the association between organic food intake and health effects, including changes in in vivo biomarkers, disease prevalence, and functional changes. DATA SOURCES: PubMed, EMBASE, Web of Science, the Cochrane Library, and ClinicalTrials.gov were searched from inception through Nov 13, 2022. DATA EXTRACTION: Both observational and interventional studies conducted in human populations were included, and association between level of organic food intake and each outcome was quantified as "no association," "inconsistent," "beneficial correlation/harmful correlation," or "insufficient". For outcomes with sufficient data reported by at least 3 studies, meta-analyses were conducted, using random-effects models to calculate standardized mean differences. DATA ANALYSIS: Based on the included 23 observational and 27 interventional studies, the association between levels of organic food intake and (i) pesticide exposure biomarker was assessed as "beneficial correlation," (ii) toxic metals and carotenoids in the plasma was assessed as "no association," (iii) fatty acids in human milk was assessed as "insufficient," (iv) phenolics was assessed as "beneficial", and serum parameters and antioxidant status was assessed as "inconsistent". For diseases and functional changes, there was an overall "beneficial" association with organic food intake, and there were similar findings for obesity and body mass index. However, evidence for association of organic food intake with other single diseases was assessed as "insufficient" due to the limited number and extent of studies. CONCLUSION: Organic food intake was found to have a beneficial impact in terms of reducing pesticide exposure, and the general effect on disease and functional changes (body mass index, male sperm quality) was appreciable. More long-term studies are required, especially for single diseases. SYSTEMATIC REVIEW REGISTRATION: PROSPERO registration no. CRD42022350175.

Oxycodone Extended-Release Capsule Utilization for Pain Management in a Cancer Palliative Care Clinic: A Retrospective Review.

Xtampza ER™, an oxycodone extended-release capsule (OERC), was the first long-acting opioid to feature abuse-deterrent properties and various routes of administration without pharmacokinetic alterations. The primary objective of this study was to evaluate changes in reported pain scores after initiation of or rotation to OERC from a previous opioid.  Baseline scores were from patients' outpatient visits immediately before starting OERC and were compared to those at the next two follow-up visits. Secondary objectives identified variables that influenced pain scores. Methods included screening for cancer patients with outpatient OERC prescriptions seen in the palliative care clinic. Eighty-two charts were reviewed with 66 included. Overall mean pain scores at both follow-ups were lower than those at baseline (-0.7 ± 2.1; -1.1 ± 2.4). Results were statistically significant between first and second-reported pain scores versus baseline (p = 0.009; 0.012) but clinically insignificant, defined as a ≥ 2-point change in numeric pain scores. Most patients discontinued OERC at the first or second follow-up (35; 53%), and 12.1% of patients who started OERC were prescribed OERC at the end of the study. There were no significant variables identified to influence pain scores either statistically or clinically. Further studies are needed to determine the long-term efficacy and safety in cancer palliative-care patients.

CYP5122A1 encodes an essential sterol C4-methyl oxidase in Leishmania donovani and determines the antileishmanial activity of antifungal azoles.

Visceral leishmaniasis, caused by Leishmania donovani, is a life-threatening parasitic disease, but current antileishmanial drugs are limited and have severe drawbacks. There have been efforts to repurpose antifungal azole drugs for the treatment of Leishmania infection. Antifungal azoles are known to potently inhibit the activity of cytochrome P450 (CYP) 51 enzymes which are responsible for removing the C14α-methyl group of lanosterol, a key step in ergosterol biosynthesis in Leishmania. However, they exhibit varying degrees of antileishmanial activities in culture, suggesting the existence of unrecognized molecular targets for these compounds. Our previous study reveals that, in Leishmania, lanosterol undergoes parallel C4- and C14-demethylation reactions to form 4α,14α-dimethylzymosterol and T-MAS, respectively. In the current study, CYP5122A1 is identified as a sterol C4-methyl oxidase that catalyzes the sequential oxidation of lanosterol to form C4-oxidation metabolites. CYP5122A1 is essential for both L. donovani promastigotes in culture and intracellular amastigotes in infected mice. Overexpression of CYP5122A1 results in growth delay, differentiation defects, increased tolerance to stress, and altered expression of lipophosphoglycan and proteophosphoglycan. CYP5122A1 also helps to determine the antileishmanial effect of antifungal azoles in vitro. Dual inhibitors of CYP51 and CYP5122A1, e.g., clotrimazole and posaconazole, possess superior antileishmanial activity against L. donovani promastigotes whereas CYP51-selective inhibitors, e.g., fluconazole and voriconazole, have little effect on promastigote growth. Our findings uncover the critical biochemical and biological role of CYP5122A1 in L. donovani and provide an important foundation for developing new antileishmanial drugs by targeting both CYP enzymes.

CDK4/6 and autophagy inhibitors synergize to suppress the growth of human head and neck squamous cell carcinomas.

Head and neck squamous cell carcinoma (HNSCC) accounts for over 10,000 deaths in the United States annually. Approximately 80% of HNSCC are human papillomavirus (HPV)-negative which have an overall poorer prognosis compared to the HPV-positive disease. Treatment options are mainly nontargeted chemotherapy, radiation, and surgery. The cyclin-d-CDK4/6-RB pathway, which regulates cell cycle progression, is often deregulated in HNSCC, making it an attractive therapeutic target. In the current study, we investigated the therapeutic effects of cyclin-dependent kinase 4/6 (CDK4/6) inhibitors in preclinical models of HNSCCs. Our results show that the specific CDK4/6 inhibitor, abemaciclib, inhibited cell growth, and induced apoptosis in HNSCC cell lines. We also demonstrated that both the pro-survival autophagy pathway and the ERK pathway in HNSCC cells were activated with abemaciclib treatment through the generation of reactive oxygen species (ROS). Coinhibition of CDK4/6 and autophagy synergistically decreased cell viability, induced apoptosis, and inhibited tumor growth in both in vitro and in vivo preclinical HNSCC models. These results reveal a potential therapeutic strategy that supports the rationale for further clinical development of a combination of CDK4/6 and autophagy inhibitors in HNSCC.

PARP1 and POLD2 as prognostic biomarkers for multiple myeloma in autologous stem cell transplant.

Multiple Myeloma (MM) is an incurable plasma cell malignancy often treated by autologous stem cell transplant (ASCT). Clinical response to ASCT has been associated with DNA repair efficiency. Here we interrogated the role of the base excision DNA repair (BER) pathway in MM response to ASCT. Across 450 clinical samples and six disease stages, expression levels of genes in the BER pathway were found to be highly upregulated during the development of MM. In a separate cohort of 559 patients with MM treated with ASCT, expression of BER pathway members MPG and PARP3 was positively associated with overall survival (OS) while expression of PARP1, POLD1, and POLD2 was negatively associated with OS. In a validation cohort of 356 patients with MM treated with ASCT, PARP1 and POLD2 findings were replicated. In patients with MM who never received ASCT (n=319), PARP1 and POLD2 were not associated with OS, suggesting that the prognostic effect of these genes may be treatment-dependent. In preclinical models of MM, synergy was observed in anti-tumor activity when poly (ADPribose) polymerase (PARP) inhibitors (olaparib, talazoparib) were used in combination with melphalan. The negative prognosis associated with PARP1 and POLD2 expression along with the apparent melphalan-sensitizing effect of PARP inhibition may suggest this pathway as a potential biomarker in patients with MM in the setting of ASCT. Further understanding of the role of the BER pathway in MM is vital to improve therapeutic strategies related to ASCT.

The Impact of a Diabetes Transitions of Care Clinic on Hospital Utilization and Patient Care.

BACKGROUND: There is currently limited guidance from the American Diabetes Association regarding transitions of care for patients with diabetes. OBJECTIVE: This study's aim was to determine the impact of a diabetes-specific transitions of care clinic (TOCC) on hospital utilization and patient outcomes in recently discharged patients with diabetes. METHODS: This retrospective study evaluated patients seen by TOCC as compared with similar patients discharged from the study institution the year prior. The primary outcome was a composite of the number of unique patients with readmissions/emergency department (ED) visits within 30 days of discharge. Secondary outcomes included a subcomponent analysis of readmissions/ED visits, index hospital length of stay (LOS), and to describe clinical interventions made in clinic. This study was approved by the institutional review board of the Office of Responsible Research Practice at the Ohio State University Wexner Medical Center. RESULTS: There were 165 patients in the TOCC group and 157 in the control group based on the matching criteria. There was a statistically significant decrease in the primary outcome in the TOCC group versus the control group (18% vs 36%, P < 0.001). In evaluation of its subcomponents, there was a statically significant decrease in patients with readmissions (11% vs 26%, P < 0.001) but not ED visits (10% vs 17%, P = 0.096). The LOS for the TOCC group was shorter at 4 days versus 5 days in the control group (P = 0.055). CONCLUSIONS AND RELEVANCE: The implementation of a diabetes-specific TOCC can decrease both readmissions and ED visits and may impact hospital LOS. In addition, a TOCC can be used to identify gaps in preventive care. The results from this study may help support the creation of similar TOCC at other institutions.

Effects of hsa-miR-9-3p and hsa-miR-9-5p on Topoisomerase IIß Expression in Human Leukemia K562 Cells with Acquired Resistance to Etoposide.

DNA topoisomerase IIα (TOP2α/170; 170 kDa) and topoisomerase IIß (TOP2ß/180; 180 kDa) are targets for a number of anticancer drugs, whose clinical efficacy is attenuated by chemoresistance. Our laboratory selected for an etoposide-resistant K562 clonal subline designated K/VP.5. These cells exhibited decreased TOP2α/170 and TOP2ß/180 expression. We previously demonstrated that a microRNA-9 (miR-9)-mediated posttranscriptional mechanism plays a role in drug resistance via reduced TOP2α/170 protein in K/VP.5 cells. Here, it is hypothesized that a similar miR-9 mechanism is responsible for decreased TOP2ß/180 levels in K/VP.5 cells. Both miR-9-3p and miR-9-5p are overexpressed in K/VP.5 compared with K562 cells, demonstrated by microRNA (miRNA) sequencing and quantitative polymerase chain reaction. The 3'-untranslated region (3'-UTR) of TOP2ß/180 contains miRNA recognition elements (MRE) for both miRNAs. Cotransfection of K562 cells with a luciferase reporter plasmid harboring TOP2ß/180 3'-UTR plus miR-9-3p or miR-9-5p mimics resulted in statistically significant decreased luciferase expression. miR-9-3p and miR-9-5p MRE mutations prevented this decrease, validating direct interaction between these miRNAs and TOP2ß/180 mRNA. Transfection of K562 cells with miR-9-3p/5p mimics led to decreased TOP2ß protein levels without a change in TOP2ß/180 mRNA and resulted in reduced TOP2ß-specific XK469-induced DNA damage. Conversely, K/VP.5 cells transfected with miR-9-3p/5p inhibitors led to increased TOP2ß/180 protein without a change in TOP2ß/180 mRNA and resulted in enhancement of XK469-induced DNA damage. Taken together, these results strongly suggest that TOP2ß/180 mRNA is translationally repressed by miR-9-3p/5p, that these miRNAs play a role in acquired resistance to etoposide, and that they are potential targets for circumvention of resistance to TOP2-targeted agents. SIGNIFICANCE STATEMENT: Results presented here indicate that miR-9-3p and miR-9-5p play a role in acquired resistance to etoposide via decreased DNA topoisomerase IIß 180 kDa protein levels. These findings contribute further information about and potential strategies for circumvention of drug resistance by modulation of microRNA levels. In addition, miR-9-3p and miR-9-5p overexpression in cancer chemoresistance may lead to future validation as biomarkers of responsiveness to DNA topoisomerase II-targeted therapy.