The E-liquid flavoring vanillin alters energy and autophagic pathways in human proximal tubule (HK-2) epithelial cells.

The use of flavored e-liquids in electronic nicotine delivery systems (ENDS) has become very popular in recent years, but effects of these products have not been well characterized outside the lung. In this study, acute exposure to the popular flavoring vanillin (VAN) was performed on human proximal tubule (HK-2) kidney cells. Cells were exposed to 0-1000 µM VAN for 24 or 48 h and cellular stress responses were determined. Mitochondrial viability using MTT assay showed a significant decrease between the control and 1000 µM group by 48 h. Seahorse XFp analysis showed significantly increased basal respiration, ATP production, and proton leak after 24 h exposure. By 48 h exposure, these parameters remained significantly increased in addition to non-mitochondrial respiration and maximal respiration. Glycolytic activity after 24 h exposure showed significant decreases in glycolysis, glycolytic capacity, glycolytic reserve, and non-glycolytic acidification. The autophagy markers microtubule-associated protein 1A/1B light chain 3 (LC3B-I and LC3B-II) were probed via western blotting. The ratio of LC3B-II/LC3B-I was significantly increased after 24 h exposure to VAN, but by 48 h this ratio significantly decreased. The mitophagy marker PINK1 showed an increasing trend at 24 h, and its downstream target Parkin was significantly increased between the control and 750 µM group only. Finally, the oxidative stress marker 4-HNE was significantly decreased after 48 h exposure to VAN. These results indicate that acute exposure to VAN in the kidney HK-2 model can induce energy and autophagic changes within the cell.

The e-liquid flavoring cinnamaldehyde induces cellular stress responses in human proximal tubule (HK-2) kidney cells.

Flavored e-liquid use has become popular among e-cigarette users recently, but the effects of such products outside the lung are not well characterized. In this work, acute exposure to the popular flavoring cinnamaldehyde (CIN) was performed on human proximal tubule (HK-2) kidney cells. Cells were exposed to 0-100 µM CIN for 24-48 h and cellular stress responses were assessed. Mitochondrial viability via MTT assay was significantly decreased at 20 µM for 24 and 48 h exposure. Seahorse XFp analysis showed significantly decreased mitochondrial energy output at 20 µM by 24 h exposure, in addition to significantly reduced ATP Synthase expression. Seahorse analysis also revealed significantly decreased glycolytic function at 20 µM by 24 h exposure, suggesting inability of glycolytic processes to compensate for reduced mitochondrial energy output. Cleaved caspase-3 expression, a mediator of apoptosis, was significantly increased at the 24 h mark. C/EBP homologous protein (CHOP) expression, a mediator of ER-induced apoptosis, was induced by 48 h and subsequently lost at the highest concentration of 100 µM. This decrease was accompanied by a simultaneous decrease in its downstream target cleaved caspase-3 at the 48 h mark. The autophagy marker microtubule-associated protein 1 A/1B light chain 3 (LC3B-I and LC3B-II) expression was significantly increased at 100 µM by 24 h. Autophagy-related 7 (ATG7) protein and mitophagy-related proteins PTEN-induced putative kinase 1 (PINK1) and PARKIN expression were significantly reduced at 24 and 48 h exposure. These results indicate acute exposure to CIN in the kidney HK-2 model induces mitochondrial dysfunction and cellular stress responses.

Gender disparity within the Canadian Urological Association A comparison with the Quebec Urological Association.

INTRODUCTION: The aim of this study was to examine gender diversity within the Canadian Urological Association (CUA) and to compare it with the Quebec Urological Association (QUA). METHODS: A retrospective review of women's representation regarding membership, committees' composition, awards, grants, and conferences between 2012 and 2022 was performed. Data and gender were extracted from databases and annual meeting programs provided by the CUA and the QUA. RESULTS: In 2022, females accounted for 18% (256/1431) of the membership at the CUA and 23% (52/228) at the QUA. The female proportion of committee members at the CUA increased from 9% (63/676) from 2012-2016 to 14% (177/1230) from 2017-2022 (p≤0.0001). In 2022, the QUA had a significantly higher proportion of female committee members than the CUA, with 39% (15/38) vs. 22% (50/225) women (p=0.0226), respectively. Moreover, from 2012-2022, 11% (5/46) of the CUA awards were given to women, whereas 38% (13/34) of the award winners at the QUA were women over the same time period (p=0.0038). Between 2012 and 2022, there were 16% (20/126) female CUA grant recipients and 44% (14/32) at the QUA (p=0.0095). The proportion of grants awarded to women at the CUA increased from 13% (5/39) in 2012-2016 to 17% (15/87) in 2017-2022. Two percent (1/53) of the plenary invited speakers at the CUA annual meetings from 2012-2016 were women, compared with 21% (14/66) from 2017-2022 (p=0.0016). In 2022, 53% (9/17) of invited plenary faculty were women at the QUA annual conference, compared to 23% (3/13) at the CUA annual meeting (p=0.0980). CONCLUSIONS: Over the past 10 years, there has been an increase in women's representation at the CUA and the QUA; however, data show that the increase in female representation at the QUA has outpaced that of the CUA.

An Improved Protocol for the Matrigel Duplex Assay: A Method to Measure Retinal Angiogenesis.

Neovascular diseases of the retina, such as diabetic retinopathy (DR) and age-related macular degeneration (AMD), are proliferative retinopathies involving the growth of new blood vessels on the retina, which in turn causes impairment and potential loss of vision. A drawback of conventional angiogenesis assays is that they are not representative of the angiogenic processes in the retina. In the retina, the new blood vessels grow (from pre-existing blood vessels) and migrate into a non-perfused region of the eye including the inner limiting membrane of the retina and the vitreous, both of which contribute to vision loss. The Matrigel Duplex Assay (MDA) measures the migration of angiogenic capillaries from a primary Matrigel layer to a secondary Matrigel layer, which resembles the pathological angiogenesis in AMD and DR. The methodology of MDA is comprised of two steps. In the first step, the human retinal microvascular endothelial cells (HRMECs) are mixed with phenol red-containing Matrigel (in a 1:1 ratio) and seeded in the center of an 8-well chamber slide. After 24 h, a second layer of phenol red-free Matrigel is overlaid over the first layer. Over the course of the next 24 h, the HRMECs invade from the primary Matrigel layer to the secondary layer. Subsequently, the angiogenic sprouts are visualized by brightfield phase contrast microscopy and quantified by ImageJ software. The present manuscript measures the angiogenesis-inhibitory activity of the Src kinase inhibitor PP2 in primary HRMECs using the MDA. The MDA may be used for multiple applications like screening anti-angiogenic drugs, measuring the pro-angiogenic activity of growth factors, and elucidating signaling pathways underlying retinal angiogenesis in normal and disease states.

Investigating Ecological Momentary Assessed Physical Activity and Core Executive Functions in 18- to 24-Year-Old Undergraduate Students.

Although evidence for young children (<10) and older adults (>64) highlights an association between physical activity (PA) and executive functions (EFs), there is a paucity of research on adolescents aged 18-24 years. Thus, this study examined the associations between PA and EF and the difference in EF between individuals who achieve the moderate-to-vigorous (MVPA) guidelines and those who do not. Forty-seven participants engaged in a Stroop task, a reverse Corsi-block test, and a task-switching test, to measure inhibition, working memory, and cognitive flexibility, respectively. An ecological momentary assessment (EMA) was used to determine the participant's MVPA and step count, through the "Pathverse" app. Multiple regressions were run to predict the task-switch cost, the Stroop effect, and the backward Corsi span from time spent in MVPA. A two-way ANCOVA examined the effects of achieving the MVPA guidelines on EF. MVPA and step count did not significantly predict EF. There were no significant differences in EF between participants achieving the MVPA guidelines and those that did not. Time spent in MVPA and step count were not significantly associated with working memory, cognitive flexibility, or inhibition in adolescents. Further research is warranted to understand other factors that may significantly affect EF, within and outside an individual's control.

Anticancer Activity of Region B Capsaicin Analogs.

The heterocyclic vanilloid compound capsaicin is responsible for the spicy and pungent flavor of chili peppers. Several convergent studies have shown that capsaicin suppresses the growth of multiple human cancers. Apart from capsaicin, natural and synthetic capsaicin-like compounds display growth suppressive activity in human cancers. The pharmacophore of capsaicin is comprised of three regions, namely region A (the aromatic ring), region B (the amide bond), and region C (the side chain). The present manuscript describes the isolation and synthesis of capsaicin analogs which have structural modifications in region B of the molecule. Furthermore, the pharmacokinetic properties, anticancer activity of region B capsaicin analogs, as well as the signaling pathways (underlying the growth-inhibitory effects of region B capsaicin analogs) have also been described. The discovery of novel, second-generation region B capsaicin analogs may foster the hope of innovative nutrition-based combination therapies in human cancers.

Examining the impact of postoperative opioid use on length of hospital stay following radical cystectomy.

INTRODUCTION: Radical cystectomy (RC) is associated with high rates of morbidity, prolonged hospital stay, and increased opioid use for postoperative pain management; however, the relationship between postoperative opioid use and length of stay (LOS ) remains uncharacterized. This study serves to investigate the association between postoperative opioid use and length of hospital stay after RC. The relationship between patient and surgical factors on LOS was also characterized. METHODS: We retrospectively reviewed all patients between 2009 and 2019 who underwent RC at our institution. Patient and perioperative variables were analyzed to determine the relationship between postoperative opioid use and LOS using multivariable linear regression analysis. RESULTS: We identified 240 patients for study inclusion with a median age of 70.0 years. Median LOS was 10.0 days, with median daily mg morphine equivalent use of 57.5 for patients. Daily mg morphine equivalent use was significantly associated with an increased LOS, as were previous pelvic radiation, postoperative ileus, and higher Clavien-Dindo grade complication during admission (all p<0.05). Median LOS increased by one day for each increase of 13.2 daily mg morphine equivalents received. CONCLUSIONS: Increased daily opioid use was associated with increased length of hospital stay after RC. Non-opioid-based pain management approaches may be effective in reducing LOS after RC.

Clinical utility of routine postoperative serial hemoglobin measurements in patients undergoing radical cystectomy for urothelial carcinoma.

INTRODUCTION: Routine measurements of serum hemoglobin (sHgb) are common after abdominal surgery; however, prolonged measurements may be associated with patient anxiety, increased costs, and longer hospitalization without clinical benefit. The objective of this study was to determine the utility of routine sHgb measurements after radical cystectomy (RC) and factors associated with transfusion of packed red blood cell (pRB C) beyond postoperative day (POD ) 2. METHODS: We retrospectively reviewed patients who underwent RC between 2009 and 2019 at a single academic tertiary care center. The number of sHgb measurements for each patient was examined and pRB C transfusion rates were calculated. Multivariable logistic regression was used to determine factors associated with transfusion beyond POD 2. RESULTS: The median number of sHgb measurements per patient during admission was nine (interquartile range [IQR] 7, 25). Overall, 69/240 (28.7%) patients received a postoperative transfusion, including 46/240 (19.2%) patients receiving a transfusion beyond POD 2. Among patients with a sHgb ≥100 g/L on POD 2, 7/85 (8.2%) went on to receive a transfusion beyond this day compared with 39/155 (25.2%) patients with sHgb <100 g/L. On multivariable analysis, risk factors associated with pRB C transfusion beyond POD 2 included older age, lower sHgb on POD 2, and longer length of stay in hospital. CONCLUSIONS: Transfusion of pRB Cs beyond POD 2 was found to be common; however, patients with sHgb ≥100 g/L on POD 2 were at low risk of requiring subsequent transfusion. Discontinuing further routine sHgb checks in these patients may serve to decrease patient anxiety, healthcare costs, and delays in hospital discharge.

The Feasibility and Acceptability of an Online CPD Programme to Enhance PE Teachers' Knowledge of Muscular Fitness Activity.

Schools provide an opportunity to enhance muscular fitness of English youth during physical education (PE). Continued professional development (CPD) among teachers may improve muscular fitness activity delivery in schools. This study sought to assess the feasibility and acceptability of an online CPD programme to enhance PE teachers' knowledge of muscular fitness activity. METHODS: Co-production of an online CPD platform was undertaken with five secondary school PE teachers. A quasi-experimental pre-post control group design for the CPD was adopted (CPD group n = 76, control n = 32). Upon CPD completion feedback was solicited for qualitative analysis. RESULTS: Pre-and-post CPD knowledge quiz data were available from 65 participants (55.4% male). The median knowledge quiz change score was significantly higher in the CPD group than in the wait list control group (CPD score vs. control score; U =37, z = -5.96, p < 0.01). Three primary themes reflecting factors associated with the acceptability and feasibility were (1) practical application, (2) support and resources, and (3) knowledge and confidence. CONCLUSIONS: Co-production of an online CPD programme can improve knowledge and affect practice. PE teachers that completed the CPD reported the online platform was beneficial to overcoming the limitations of face-to-face CPD such as time and financial constraints and suggested the content covered was beneficial and appropriate to their teaching. Future work is required to establish links between teachers' learning following CPD, the translation into PE practice and student MF outcomes.

The NXDC-MEN-301 Study on 5-ALA for Meningiomas Surgery: An Innovative Study Design for the Assessing the Benefit of Intra-Operative Fluorescence Imaging.

BACKGROUND: 5-aminolevulinic acid (5-ALA; GleolanTM, NX Development Corps., Lexington, USA) is approved for fluorescence-guided resections of suspected malignant gliomas. Experience has demonstrated that meningiomas also show fluorescence, which may be a useful surgical adjunct. We present an innovative design for a multi-center, prospective study to determine the clinical safety and potential benefit of fluorescence-guided resection of meningiomas with utmost bias reduction. METHODS: All patients with suspected meningioma (all grades) receive GleolanTM 20 mg/kg 2-4 h prior to surgery supported by fluorescence excitation from a blue light source (Blue400, Zeiss Meditech, Oberkochen, Germany; FL400, Leica Microsystems, Heerbrugg, Switzerland). Surgeons are asked whether a residual tumor can be observed to fluoresce under blue light (BL) after the tumor is no longer recognizable using conventional illumination at the end of surgery. In addition, when faced with tissues of uncertain tissue type (so-called "indeterminate" tissue), this study records how often surgeons make a correct decision based on fluorescence and how this influences surgical strategy. The primary endpoint is the percentage of patients in whom one of these two benefits are observed. Other endpoints include the diagnostic accuracy of fluorescence compared to white light (WL) versus correlative histology. For bias reduction, pertinent data are derived from surgical videos reviewed by independent reviewers blinded to surgeons' assessments of tissue type and fluorescence status. Data will be included from approximately 100 study participants completing the study at approximately 15 centers in the United States, Germany, and Austria. RESULTS: As of May 2022, 88 patients have completed the study. No adverse safety signal has been detected. CONCLUSIONS: Preliminary data confirm the feasibility of our study design. Accrual is targeted for completion in the third quarter of 2022.

The Physical Behaviour Intensity Spectrum and Body Mass Index in School-Aged Youth: A Compositional Analysis of Pooled Individual Participant Data.

We examined the compositional associations between the intensity spectrum derived from incremental acceleration intensity bands and the body mass index (BMI) z-score in youth, and investigated the estimated differences in BMI z-score following time reallocations between intensity bands. School-aged youth from 63 schools wore wrist accelerometers, and data of 1453 participants (57.5% girls) were analysed. Nine acceleration intensity bands (range: 0−50 mg to ≥700 mg) were used to generate time-use compositions. Multivariate regression assessed the associations between intensity band compositions and BMI z-scores. Compositional isotemporal substitution estimated the differences in BMI z-score following time reallocations between intensity bands. The ≥700 mg intensity bandwas strongly and inversely associated with BMI z-score (p < 0.001). The estimated differences in BMI z-score when 5 min were reallocated to and from the ≥700 mg band and reallocated equally among the remaining bands were −0.28 and 0.44, respectively (boys), and −0.39 and 1.06, respectively (girls). The time in the ≥700 mg intensity band was significantly associated with BMI z-score, irrespective of sex. When even modest durations of time in this band were reallocated, the asymmetrical estimated differences in BMI z-score were clinically meaningful. The findings highlight the utility of the full physical activity intensity spectrum over a priori-determined absolute intensity cut-point approaches.

Working With Survivors of Human Trafficking: Results From a Needs Assessment in a Midwestern State, 2019.

OBJECTIVES: While the needs of human trafficking survivors have been documented in the academic discourse, the saliency of such needs is understudied. This study aimed to reveal the critical needs of service provision for human trafficking survivors in a Midwestern state as perceived by multidisciplinary service providers. METHODS: Targeting health care, social service, law enforcement, public health, and other service providers involved in anti-human trafficking service delivery, we disseminated a survey using purposive and snowball sampling. Drawing from survey responses collected in 2019 from 107 service providers working with 422 survivors of human trafficking in the previous 12 months in a Midwestern state, we examined the saliency of needs from the perspective of service providers in the health care, criminal justice, and social service sectors. RESULTS: Respondents indicated on a Likert scale (1-5) the level of need for 37 social, health care, and legal services in their communities. The top-indicated needs statewide were mental health care, shelter/housing, peer mentorship programs, legal services, transportation, and provider training in juvenile courts, schools, law enforcement, and health care settings on human trafficking. CONCLUSION: The results suggest the need for statewide responses, such as legal assistance networks, telemental health options, and enhanced training collaborations between interpersonal violence and anti-human trafficking service providers. The study findings are generalizable beyond the study site in 3 ways that can guide strategic action: (1) they provide a framework for state-level analysis and strategic planning that parallels the results; (2) they show that regional variation is possible and should be accounted for in state-level study design, analysis, and strategic planning; and (3) the implications for housing assistance, legal assistance, mental health/substance use disorder-related health care, and training are scalable.

Utility of Postoperative Serial Renal Function Monitoring in Patients Undergoing Radical Cystectomy for Urothelial Carcinoma.

PURPOSE: The aim of this study is to examine the rates of clinical actions (CAs) as a result of serial creatinine (SCr) values and to examine postoperative SCr trends to assess the utility of SCr measurements in radical cystectomy (RC) patients. METHODS: We performed a retrospective chart review using our institution's database on all patients who underwent radical cystectomy for urothelial carcinoma (UC) from 2009 to 2019. Preoperative and all postoperative inpatient creatinine values were recorded. Minor CAs included fluid boluses, fluid rate changes, and medication changes. Major CAs included OR take-backs for stent repositioning, nephrostomy tube placement, nephrology consultation, and hemodialysis. RESULTS: A total of 238 RCs were performed with a resultant 2952 SCr measurements. The median number of SCr measurements per patient was 9 (interquartile range, IQR 7) with median length of hospital stay of 10 days (IQR 9 days). There were 92 minor and 12 major CAs as a result of 3% and 0.44% of SCr measurements, respectively. All major CAs were seen in patients experiencing complicated postoperative course. The median postoperative day with the highest creatinine was day 2. Predictors of postoperative CAs included preoperative renal dysfunction and obesity. CONCLUSION: SCr measurements remain a clinically valuable tool in postoperative management. Nonetheless, this present study suggests that prolonged SCr monitoring is of limited clinical utility. As such, discontinuing SCr checks after postoperative day 3 in patients experiencing uncomplicated postoperative course is safe and may lead to both cost savings and decreased patient discomfort.

Anti-cancer activity of sustained release capsaicin formulations.

Capsaicin (trans-8-methyl-N-vanillyl-6-noneamide) is a hydrophobic, lipophilic vanilloid phytochemical abundantly found in chili peppers and pepper extracts. Several convergent studies show that capsaicin displays robust cancer activity, suppressing the growth, angiogenesis and metastasis of several human cancers. Despite its potent cancer-suppressing activity, the clinical applications of capsaicin as a viable anti-cancer drug have remained problematic due to its poor bioavailability and aqueous solubility properties. In addition, the administration of capsaicin is associated with adverse side effects like gastrointestinal cramps, stomach pain, nausea and diarrhea and vomiting. All these hurdles may be circumvented by encapsulation of capsaicin in sustained release drug delivery systems. Most of the capsaicin-based the sustained release drugs have been tested for their pain-relieving activity. Only a few of these formulations have been investigated as anti-cancer agents. The present review describes the physicochemical properties, bioavailability, and anti-cancer activity of capsaicin-sustained release agents. The asset of such continuous release capsaicin formulations is that they display better solubility, stability, bioavailability, and growth-suppressive activity than the free drug. The encapsulation of capsaicin in sustained release carriers minimizes the adverse side effects of capsaicin. In summary, these capsaicin-based sustained release drug delivery systems have the potential to function as novel chemotherapies, unique diagnostic imaging probes and innovative chemosensitization agents in human cancers.