Minimal Protocol Research Articles

Essential(ly forgotten) workers: Latine youth farmworkers during the COVID-19 pandemic.

Essential workers across multiple industries faced a disproportionate burden of morbidity and mortality during the COVID-19 pandemic. The effects were especially severe for agricultural and food processing workers, many of whom are members of the structurally vulnerable Latine community. Under current U.S. federal laws, children under 12 years old can legally work in agriculture, one of the most hazardous U.S. industries. Many of these working youth are Latine and experience health and educational inequities. Using a community-based participatory research approach and a qualitative design, we conducted semi-structured in-depth interviews in North Carolina in 2022, with service providers in health, education, and advocacy fields (n = 10) and Latine youth farmworkers aged 10-17 (n = 24). We used participatory qualitative analysis methods and a reflexive thematic analysis to understand and describe the work experiences of these youth during the first 2 years of the pandemic. Amidst precarious economic and school situations exacerbated by the COVID-19 pandemic, many youths described an increased need to work to support their families. While aware and sometimes fearful of the added occupational health risks of COVID-19, youth and their families felt they had few other options. Service providers and youth described minimal employer-provided safety protocols or equipment, yet some workers organized their own attempts at safety protocols. Youth narratives imply limited knowledge of basic workplace safety requirements. Study findings emphasize the urgent need to address structural vulnerabilities shaping workplace policies and norms to protect Latine youth farmworkers to support their healthy development.

A Minimal Training Protocol Improves Strength And Aerobic Fitness In Trained And Untrained Populations

A Minimal Training Protocol Improves Strength And Aerobic Fitness In Trained And Untrained Populations

The Minimal assessment Protocol for Cerebral Stroke 2020 (PMIC2020): a multicenter feasibility study in post-stroke inpatient rehabilitation.

In 2008, a Working Group of the Italian Society of Physical and Rehabilitation Medicine (SIMFER) published the first minimum protocol for assessing stroke patients (PMIC) to define functional needs and outcomes. The recent PMIC revision (PMIC2020) introduces a document for all rehabilitation settings, incorporating updated measurement tools. The aim of this study was to investigate the PMIC2020 feasibility and administration time (AT) in post-stroke inpatients and to examine the influence of demographic and clinical variables on AT. Multicenter prospective observational study. Eight Italian rehabilitation centers for post-acute inpatients. Adult patients consecutively admitted to rehabilitation after ischemic/hemorrhagic stroke, reporting the first event or recurrence, with a modified Barthel Index (mBI)<75 points, without cognitive impairment and clinical instability. PMIC2020 was administered at admission (T0) and discharge (T1), recording AT of each section/ tool. A feasibility questionnaire was administered to assessors. Univariate and multivariate analyses were conducted to investigate the effect of demographics and clinical variables on AT. One hundred fifty-one subjects were enrolled at T0 and 139 at T1; the mean±SD AT (seconds) was 1634±401 at T0 and 1087±360 at T1 (P<0.001). National Institute of Health-Stroke Scale and Mini-Mental State Examination required the highest AT. All but two scales had significantly lower AT at T1 (P<0.05). Severe disability (as measured by mBI) was associated with higher AT than either complete or minimal/absent disability. The feasibility questionnaire showed good PMIC2020 appraisal by assessors without relevant critical issues. PMIC2020 was feasible in post-acute inpatient rehabilitation settings. No relevant critical issue was raised by users. Even though more comprehensive than PMIC, PMIC2020 required only slightly more AT (27 minutes at T0 and 18 minutes at T1, on average); more AT was needed to assess patients with severe disability. The study has immediate transferability for the National Health Service, as PMIC2020 can be routinely implemented in clinical practice and research to assess stroke patients' needs and outcomes. The updated measures allow more immediate comparisons with international data on stroke rehabilitation. Future research should investigate the PMIC2020 feasibility in other rehabilitation settings and its relevance in predicting stroke rehabilitation needs and outcomes.

Minimal dose resistance training enhances strength without affecting cardiac autonomic modulation in menopausal women: a randomized clinical trial

The proposition of a minimal dose of resistance training (RT) to elicit health benefits, encompassing physiological and psychological aspects, has garnered attention. While empirical investigations have demonstrated the efficacy of low-volume RT protocols in inducing adaptations such as enhanced strength and functional capacity, further exploration of the effects of this paradigm across a broader spectrum of variables is warranted. Thus, this study aimed to investigate the effects of minimal dose RT on strength and functional capacity, cardiac autonomic modulation, and hemodynamic parameters in menopausal women. Twenty-six women were randomly assigned to the training (TG: 63.2 ± 9.3 years) or control group (CG: 59.3 ± 7.6 years). Anthropometric measurements, strength and functional performance tests, cardiac autonomic assessment, and hemodynamic parameters were performed before and after four weeks of intervention. The TG performed the minimum dose RT twice weekly for four weeks (2 sets of 8–12 repetitions in three dynamic exercises, plus three 1-min isometric planks), and the CG had a weekly meeting with lectures and stretching. Two-way ANOVA with repeated measures was applied to each variable. Regarding time comparisons, there was a significant increase for LniRR (F = 4.78; ω2 = 0.046; p = 0.04), one repetition maximum (1RM) bench press (F = 8.06; ω2 = 0,013; p = 0.01), and 1RM leg press (F = 17.3; ω2 = 0,098; p < 0.01). There was a group*time interaction only for the index LnRMSSD (F = 5.11; ω2 = 0.042; p = 0.03), and 1RM bench press (F = 9.52; ω2 = 0,016; p = 0.01). No between-group main effect for any variable was found. The minimal dose RT protocol improved muscle strength, while cardiac autonomic and hemodynamic variables, as well as functional capacity, remained stable over 4 weeks in menopausal women.

A modular cell-free protein biosensor platform using split T7 RNA polymerase.

Conventional laboratory protein detection techniques are not suitable for point-of-care (POC) use because they require expensive equipment and laborious protocols, and existing POC assays suffer from long development timescales. Here, we describe a modular cell-free biosensing platform for generalizable protein detection that we call TLISA (T7 RNA polymerase-Linked ImmunoSensing Assay), designed for extreme flexibility and equipment-free use. TLISA uses a split T7 RNA polymerase fused to affinity domains against a protein. The target antigen drives polymerase reassembly, inducing reporter expression. We characterize the platform, then demonstrate its modularity by using 16 affinity domains against four different antigens with minimal protocol optimization. We show TLISA is suitable for POC use by sensing human biomarkers in serum and saliva with a colorimetric readout within one hour and by demonstrating functionality after lyophilization. Altogether, this technology could have potentially revolutionary impacts, enabling truly rapid, reconfigurable, equipment-free detection of virtually any protein.

Noneffectiveness of cryostorage duration on clinical and neonatal outcomes after single vitrified-warmed blastocyst transfers

BackgroundIn the current era of assisted reproductive technology, the strategy of single vitrified-warmed blastocyst transfer (SVBT) is acknowledged for positively impacting clinical outcomes and preventing the risk of multiple conceptions. Previous studies have indicated that blastocyst grade and the day of blastocyst positively correlate with outcomes. Moreover, storage duration has no significant effect on survival rates, clinical outcomes, and neonatal outcomes. However, some researchers express controversial opinions on cryostorage duration, and their findings suggest a negative impact on clinical outcomes. These results remain subject to controversy, and limited studies exist regarding the outcomes after SVBT. Therefore, our study aims to investigate the impact of the day of blastocyst, blastocyst grade, and blastocyst cryostorage duration on clinical and neonatal outcomes following SVBT in patients who underwent clomiphene-citrate-based minimal stimulation.Material and methodsIn this study, retrospective cohort study data collected from June 2015 to April 2023 included 2107 patients with first SVBT cycles who underwent a clomiphene-based minimal ovarian stimulation protocol or a drug-free natural protocol at the Ojinmed IVF Center. Patients were categorized into four groups based on blastocyst cryostorage duration: group 1 (< 2 months, n = 882), group 2 (3–6 months, n = 794), group 3 (7–12 months, n = 187), group 4 (13–24 months, n = 126), and group 5 (25–81 months, n = 118). The patient’s clinical and neonatal outcomes were compared with cryostorage duration after the propensity score matched.ResultMultivariable logistic regression analysis revealed that prolonged cryostorage duration insignificantly correlated with clinical outcomes. Additionally, neonatal outcomes are not correlated with cryostorage duration.ConclusionsThe patient must consider several parameters when selecting embryos for transfer, including the duration of cryostorage. Our study results show that for the first single vitrified-warmed blastocyst transfer of patients who underwent clomiphene citrate-based minimal stimulation, cryostorage duration does not affect outcomes.

O-198 The decline in semen quality parameters enhances mosaicism in trophectoderm biopsy of human blastocysts

Abstract Study question Do poor sperm parameters impact chromosomal analysis compared to sperm with normal parameters? Summary answer Fertilization with more than two abnormal patterns, including severe oligoasthenoteratozoospermia sperm, may increase the rate of embryo mosaicism. What is known already Sperm cells constitute the genetic half of the embryo, and the male factor plays a crucial role in embryo chromosome integrity. While PGT-A is widely used to enhance IVF outcomes, the influence of male factors on embryo genetics remains underexplored. We aimed to determine the effect of semen profile on the outcome of PGT-A cycles performed at the blastocyst stage. Study design, size, duration A single-center retrospective study conducted between January 2019 and July 2023 analyzed 609 blastocysts with Next Generation Sequencing (NGS) of trophectoderm (TE) biopsies. Four study groups were formed based on sperm parameters: normozoospermia (N), moderate male factor (MMF), severe oligoasthenoteratozoospermia (s-OAT), obstructive azoospermia (OA). Participants/materials, setting, methods Patients treated with a minimal ovarian stimulation protocol. PGT-A cycles were divided into three paternal age groups. OUTCOMES: Euploidy, aneuploidy, mosaicism rates; blast formation time; good-quality blastocyst rates. METHODS: Multivariate logistic analysis, Chi-squared tests, and STATA 12.1 statistical software. Main results and the role of chance In the 609 TE biopsied blastocysts, euploidy rate was 49.9%, aneuploidy rate was 31.5%, and mosaicism rate was 18.6%. Paternal age ≥43 increased aneuploidy risk compared with &amp;lt;35 (40.1% vs 26.1%, p &amp;lt; 0.05). The euploidy rate and aneuploidy rate were no different in MMF, s-OAT, and OA compared with N (44.8%, 40.6%, and 42.1% vs. 54.2%; 36.8%, 25.0%, and 31.6% vs. 30.5%). Mosaicism rates were significantly difference between sperm parameter groups (N 17.1%, MMF 18.4%, s-OAT 34.4% and OA 26.3%, p &amp;lt; 0.05). In logistic regression analysis adjusted for confounders, severe oligoasthenoteratozoospermia showed 2.5 times higher mosaicism rate (aOR 2.58; 95% CI 1.08-5.08, p = 0.027). The good-quality blastocyst rate was significantly reduced in the OA compared with N, MMF, and s-OAT (21.1% vs 43.4%, 39.2% and 37.5%, p &amp;lt; 0.05). No impact of semen parameters on blastocyst formation time and vitrification time outcomes was observed. Limitations, reasons for caution Retrospective, single-center study. Excludes DNA fragmentation index and TESE results, impacting sperm quality evaluation. Wider implications of the findings Severe oligoasthenoteratozoospermia is positively associated with mosaicism rate. Poor sperm parameters lead to mosaic embryos. Couples undergoing PGT-A for male factors should discuss potential outcomes. Patients with abnormal semen parameters should consider pretreatment for improved semen quality. Results useful for genetic counseling and clinical decision-making. Trial registration number None

P-114 The timing from post-insemination to blastocyst vitrification correlates with ploidy and gender

Abstract Study question Which morphology parameter of blastocysts has the strongest correlation with the euploid rate? Summary answer The vitrification times post-insemination and trophectoderm grade (TE) are more predictive parameters for euploid blastocysts, and female blastocysts require less time for vitrification. What is known already In current ART, the use of preimplantation genetic tests for aneuploidy (PGT-A) has increased by 2 to 25 percent in different countries. PGT-A analyses have helped patients achieve an increased pregnancy rate, choose the gender of the embryo and prevent miscarriages. Moreover, the potential for blastocyst euploid is strongly correlated with the developmental speed and morphology grades. However, for patients without PGT-A, the criteria for selecting transferred blastocysts are mostly based on morphology and the day of the blastocyst. Therefore, it is essential to clarify which parameter is more predictive of euploid embryos during elective FET cycles. Study design, size, duration This retrospective analysis included 713 blastocysts from 417 patients treated at the Ojinmed IVF Center, Mongolia, between May 2019 and June 2023. The morphology parameters (morphology grade, blastocyst formation time, vitrification time and inner diameter) were compared between the PGT-A results and the control results. Blastocyst quality was evaluated with the Gardner and Schoolcraft grading system, and the inner diameter of the blastocyst was measured. Participants/materials, setting, methods All patients underwent a clomiphene-based minimal ovarian stimulation protocol or drug-free natural cycle of IVF treatment. When the blastocyst diameter exceeded 160 μm, a trophectoderm (TE) biopsy was taken, and the blastocyst was immediately vitrified using the Cryotop method. The time of vitrification and formation of the blastocyst were recorded from the insemination. The patients were analyzed using high-resolution NGS PGT-A (10 Mb resolution for copy number variations and 30%-70% for mosaicism). Main results and the role of chance The PGT-A analysis reported euploid, aneuploid, and mosaic rates. Euploid, aneuploid, and mosaic rates were significantly correlated with blastocyst formation time (114.78 ± 4.49 h, 116.12 ± 6.42 h and 116.33 ± 6.87 h, p = 0.003) and vitrification blastocyst time (126.65 ± 9.8 h, 131.57 ± 10.8 h and 130.44 ± 11.19 h, p &amp;lt; 0.001), along with women’s age (34.9 ± 4.32, 38.14 ± 3.77, and 35.84 ± 4.5, p &amp;lt; 0.001). In logistic regression analysis, aneuploid rates were significantly lower in blastocysts with A and B grade of ICM (OR 0.36, CI 95% 0.24-0.32, p &amp;lt; 0.01 and OR 0.45, CI 95% 0.32-0.63, p &amp;lt; 0.01) and A and B grade of TE (OR 0.20, CI 95% 0.11-0.37, p &amp;lt; 0.01 and OR 0.20, CI 95% 0.25-0.48, p &amp;lt; 0.01) compared to C grades, respectively. The AUCs for prediction of euploid were maternal age 0.66 (95% CI 0.62-0.70), TE grade 0.64 (95% CI 0.57-0.64), blastocyst vitrification time 0.62 (95% CI 0.58-0.66), ICM grade 0.60 (95% CI 0.61-0.68), blastocyst formation time 0.55 (95% CI 0.51-0.59) and inner diameter 0.46 (95% CI 0.42-0.5). Although blastocyst vitrification time is correlated with blastocyst gender (female blastocyst 127.9 ± 10.25h vs male blastocyst 129.68 ± 10.84h, p = 0.02). Limitations, reasons for caution The results of the retrospective study are limited to data from a single IVF center and conventional incubators were used in this study. Wider implications of the findings These findings might help non PGT-A patients in deciding on embryo selection for transfer. The vitrification time and TE grade of blastocysts are strongly correlated with euploid rate than the ICM, inner diameter and formation time. Also, female blastocysts take less time to vitrification time than male blastocysts. Trial registration number *

Enhanced hippocampal LTP but normal NMDA receptor and AMPA receptor function in a rat model of CDKL5 deficiency disorder

BackgroundMutations in the X-linked gene cyclin-dependent kinase-like 5 (CDKL5) cause a severe neurological disorder characterised by early-onset epileptic seizures, autism and intellectual disability (ID). Impaired hippocampal function has been implicated in other models of monogenic forms of autism spectrum disorders and ID and is often linked to epilepsy and behavioural abnormalities. Many individuals with CDKL5 deficiency disorder (CDD) have null mutations and complete loss of CDKL5 protein, therefore in the current study we used a Cdkl5−/y rat model to elucidate the impact of CDKL5 loss on cellular excitability and synaptic function of CA1 pyramidal cells (PCs). We hypothesised abnormal pre and/or post synaptic function and plasticity would be observed in the hippocampus of Cdkl5−/y rats.MethodsTo allow cross-species comparisons of phenotypes associated with the loss of CDKL5, we generated a loss of function mutation in exon 8 of the rat Cdkl5 gene and assessed the impact of the loss of CDLK5 using a combination of extracellular and whole-cell electrophysiological recordings, biochemistry, and histology.ResultsOur results indicate that CA1 hippocampal long-term potentiation (LTP) is enhanced in slices prepared from juvenile, but not adult, Cdkl5−/y rats. Enhanced LTP does not result from changes in NMDA receptor function or subunit expression as these remain unaltered throughout development. Furthermore, Ca2+ permeable AMPA receptor mediated currents are unchanged in Cdkl5−/y rats. We observe reduced mEPSC frequency accompanied by increased spine density in basal dendrites of CA1 PCs, however we find no evidence supporting an increase in silent synapses when assessed using a minimal stimulation protocol in slices. Additionally, we found no change in paired-pulse ratio, consistent with normal release probability at Schaffer collateral to CA1 PC synapses.ConclusionsOur data indicate a role for CDKL5 in hippocampal synaptic function and raise the possibility that altered intracellular signalling rather than synaptic deficits contribute to the altered plasticity.LimitationsThis study has focussed on the electrophysiological and anatomical properties of hippocampal CA1 PCs across early postnatal development. Studies involving other brain regions, older animals and behavioural phenotypes associated with the loss of CDKL5 are needed to understand the pathophysiology of CDD.

Premature luteinization and pregnancy outcomes in depot goserelin-downregulated assisted reproductive technology cycles: A cross-sectional study from Ethiopia.

To compare the rate of premature luteinization in depot goserelin-downregulated invitro fertilization (IVF) cycles with other IVF protocols at a teaching hospital in Ethiopia. We conducted a cross-sectional study on the effects of premature luteinization on IVF outcomes at St. Paul's Hospital Millennium Medical College, a tertiary teaching hospital in Ethiopia. Patients who had IVF at this hospital between 2019 and 2020 were included in the study. Patient records were reviewed and collected on Open Data Kit. We used Stata release 15 to analyze the data. A simple descriptive analysis and bivariate analysis were performed as appropriate. A P-value less than 0.05 was considered asstatistically significant. A total of 305 patients (40 cases with premature luteinization and 265 without premature luteinization) were included. There was no difference in the rate of premature luteinization in the depot goserelin long protocol cycles (6.4%), compared to minimal stimulation (14.1%) and antagonist protocols (16.7%), P = 0.19. Embryo transfer was carried out in 27 (67.5%) patients in the premature luteinization group, which was lower than the 86.0% (228/265) in the non-premature luteinization group, P = 0.003. There was no difference in the median number of oocytes retrieved (8.5 [interquartile range 5.0, 13.0] per cycle in the premature luteinization group vs 5.0 [interquartile range 3.0, 10.0] in those without premature luteinization, P = 0.10). A depot goserelin-downregulated long protocol for IVF is a cost-effective and convenient option for controlled ovarian hyperstimulation without increased risk of having premature luteinization compared to antagonist and minimal stimulation protocols.

Comparison In Vitro Fertilization Outcomes between DouStim and Minimal Stimulation Protocols in Poor Ovarian Responders: A Randomized Clinical Trial.

Various protocols have been approved to improve the response rate leading to successful fertilization in poor ovarian responders (PORs). The application of double ovarian stimulation (DuoStim) in the follicular and luteal phases of the same ovarian cycle has been shown as an intriguing option to achieve more oocyte retrievals in the shortest time. The aim of the current study is to compare the outcomes of different protocols, minimal stimulation (MS) and Duostim. This randomized clinical trial was performed on 42 in vitro fertilization (IVF) candidates with POR diagnosis. Patients were classified into two equal groups and treated with the DuoStim protocol and MS protocol. The IVF outcomes, including retrieved follicles, oocytes, metaphase II (MII) oocytes and embryos, were compared between these groups. The patients' characteristics including age, anti-mullerian hormone (AMH), follicle stimulating hormone (FSH), luteinizing hormone (LH), and antral follicle count (AFC) were collected and compared. It showed there was no significant difference between the two groups baseline characteristics (P>0.05). We observed that the DuoStim protocol resulted in a significantly higher score in comparison with the MS protocols , including the number of follicles (6.23 ± 2.93 vs. 1.77 ± 1.66, P<0.001), retrieved oocytes (3.86 ± 2.57 vs. 1.68 ± 1.58, P=0.002), MII oocytes (3.36 ± 2.42 vs. 1.27 ± 1.27, P=0.001) and obtained embryos (2.04 ± 1.64 vs. 0.77 ± 0.86, P=0.003). The DuoStim protocol is a favourable and time saving plan that is associated with more oocytes in a single stimulation cycle. The DuoStim protocol significantly can result in more frequent MII oocytes and embryos. We figured that the higher number of oocytes and embryos might have led to a higher rate of pregnancy (registration number: IRCT20200804048303N1).

Clinical and neonatal outcomes of complete zona pellucida removal by laser-assisted hatching after single vitrified-warmed blastocyst transfer.

Laser-assisted hatching (LAH) is a widely used and unavoidable technique in assisted reproductive technology after frozen embryo transfer (FET). We aimed to investigate the safety and efficacy of completely ZP (zona pellucida) removed by an assisted laser group compared to an intact group after FET. This retrospective study involved 320 SVBT single vitrified-warmed blastocyst transfers of 213 patients who underwent clomiphene citrate (CC) based minimal stimulation protocol. This study compared 160 SVBT with ZP that were completely removed using laser-assisted (laser beams of 10-15, a wavelength of 1.48μm, and duration of 1.8-2.2ms) to 160 cases of intact SVBT. Fisher's exact test, chi-squared test, and logistic regression are used for statistical analysis. The women's age, cause of infertility, BMI, transferred embryo day, insemination methods, and blastocyst grade were not significantly different between two groups. The implantation rate (66.2% vs. 51.2%, P < 0.01), the clinical pregnancy rate (CPR, 52.5% vs. 39.3%, P = 0.01), and the live birth rate (LBR, 43.7% vs. 29.3%, P < 0.01) were significantly higher in the completely ZP removed than the intact group. In logistic regression analysis, the laser-assisted hatching group showed about 2 times higher implantation rate and CPR and LBR than the control group. But miscarriage, gestational weeks, gender, birth weight, and twin births were insignificant between two groups. Our study suggests ZP free embryo transfer by assisted laser is more effective and safer than intact embryo transfer.

Cultivation and Genomic DNA Extraction of Klebsiella pneumoniae.

Klebsiella pneumoniae is a Gram-negative, rod-shaped bacterium of medical significance. It typically exists as part of the normal flora of the human intestine but can cause severe infections in the healthcare setting due to its rapid acquisition of antibiotic resistance. Cultivating and extracting genomic DNA from this bacterium is crucial for downstream characterization and comparative analyses. To provide a standardized approach for growing K. pneumoniae in the laboratory setting, this collection of protocols provides step-by-step procedures for routine culturing, generating growth curves, storing bacteria, and extracting genomic DNA. © 2024 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Reviving K. pneumoniae from frozen stocks Basic Protocol 2: Cultivating K. pneumoniae in rich growth medium Alternate Protocol: Cultivating in minimal liquid growth medium Basic Protocol 3: Enumerating K. pneumoniae colony forming units Basic Protocol 4: Growth curves Basic Protocol 5: Genomic DNA extraction Basic Protocol 6: Characterizing K. pneumoniae strains based on genomic sequence Basic Protocol 7: Storage of K. pneumoniae frozen stocks in glycerol Basic Protocol 8: Storage of K. pneumoniae in agar stabs.

Beyond effectiveness in eHealth trials: Process evaluation of a stepped-care programme to support healthcare workers with psychological distress (RESPOND-HCWs).

This study presents the process evaluation of an effective stepped-care programme of eHealth interventions (Doing What Matters in Times of Stress [DWM] and Problem Management Plus [PM+]) for healthcare workers (HCWs) with psychological distress (RESPOND-HCWs trial) conducted in Spain. The aim is to analyse the context in which the programme was delivered, assess key implementation outcomes and explore mechanisms of action. We used mixed methods. Quantitative data came from routine randomised control trial monitoring and structured observation, and qualitative data were collected using semi-structured, in-depth interviews with trial participants (n = 12) and decision-makers (n = 7) and a focus group discussion with intervention providers (n = 7). We conducted a descriptive analysis of quantitative data using R software and a thematic analysis of qualitative data using NVivo. Context analysis revealed implementation barriers, including unrealistic expectations of participants about the programme and mental health-related stigma. The flexibility of interventions and the opportunity for mental health actions were enabling factors. Implementation outcomes showed that the trial was feasible, appropriate and timely, and that the intervention was delivered with minimal protocol deviations and good acceptance among participants. Mechanisms of action included confidence in the positive effect of the intervention, a good therapeutic relationship and specific intervention components. These results supplement the outcome evaluation and can help inform large-scale implementation in similar settings. Specific recommendations include increasing mental health awareness and reducing stigma in the implementation setting, including a short orientation session and ensuring flexibility in schedules and peer support. NCT04980326.

#287 : A Comparison of hCG Trigger Versus Dual Trigger for Final Oocyte Maturation in Poor Ovarian Responders Undergoing Minimal Stimulation

Background and Aims: Traditionally, hCG has been used for oocyte triggering. However, women with poor ovarian response manifest low oocyte yield and compromised occyte quality. More recently, a combination of hCG and GnRHa (dual trigger) has been shown to improve follcile collection yield and oocyte maturation. The aim of the study is tocompare the result of final oocyte maturation and the embryo formation between two groups of poor ovarian responders undergoing minimal stimulation after triggering by hCG alone versus dual trigger. Method: This randomized control trial was conducted at National Assisted Reproductive Center, National Hospital of Obstetrics and Gynecology. Patients defined as poor ovarian responders according to the Bologna criteria were equally divided into two groups: control (hCG trigger) and investigation (dual trigger) groups. The group received the hCG trigger (6500 IU) and the investigation group received the dual trigger (0.2 mg of Triptorelin plus 6500 IU of hCG). Minimal stimulation IVF protocols consist of 5 days of 100mg clomiphene citrate and small amounts of gonadotropin. Results: One hundred and two patients were included in the study, with 51 women in each treatment group. Dual triggering showed a significant higher number of retrieved oocytes (5.35 ± 3.13 vs 4.35 ± 2.44, P=0.038) and metaphase II oocytes (4.25 ± 3.12 vs 3.35 ± 2.02, P=0.043). Although not reaching statistical significances, increases in the number of fertilized oocytes, embryos, and number of top-quality embryos (TQE) were observed. Conclusion: Dual trigger for final oocyte maturation is associated with better IVF outcome in poor ovarian responders undergoing minimal stimulation. Acknowledgements: The authors are grateful to all physicians, administrative staff at the National Assisted Reproductive Technology Center, National Hospital of Obstetrics and Gynecology, Vietnam for allowing and supporting us to undertake this research.

Decreased Opioid Use and Equivalent Pain Score Outcomes After Robotic Urologic Surgery Using a Simplified Opioid Minimization Protocol: A Safety-Net Hospital Experience.

We aimed to implement a simplified opioid minimization (OM) protocol after robotic urologic surgery in a safety-net hospital to decrease opioid consumption without compromising patient-reported pain or satisfaction. Robotic urologic surgery was performed in 103 consecutive patients at a safety-net hospital. An opioid control (OC) cohort was established from January to May 2021, and the OM protocol was implemented from June to October 2021. On postoperative day (POD) 2 and POD7, a validated survey was used to assess pain and satisfaction. Opioid dispensation records were queried from the Prescription Monitoring Program. Outcomes were compared by univariate methods. There were no demographic differences between the OM (n = 45) and OC (n = 35) cohorts. Total opioids received within 30 days of surgery decreased by 68% in the OM vs OC cohort (median [IQR] 32.5 [7.5-65] vs 100 [30-173] morphine milligram equivalents, P < .001). The median amount of opioids prescribed at discharge for the OM cohort was 0 (IQR:0-0) vs 75 morphine milligram equivalents (IQR:0-112.5) for the OC cohort (P < .001). Pain severity did not differ between cohorts on POD2 (median [IQR]: OM=3/10 [2-5], OC=3.5/10 [2-6]; P = .5) or POD7 (median [IQR]: OM=2/10 [0-3], OC=1/10 [0-3]; P = .8), and POD7 satisfaction with pain management remained high for both cohorts (P = .8). Our simplified OM protocol decreased total opioid use after robotic urologic surgery by 68% without compromising pain or satisfaction.

Coconut oil versus standard of care for the prevention of diaper dermatitis in the neonatal intensive care unit: A randomized clinical trial

ObjectiveTo evaluate the effectiveness and safety of coconut oil (CO) for diaper dermatitis (DD) prevention. MethodsA prospective, 2-group, randomized trial compared organic CO applied at each diaper change to minimal standard of care (SOC) in 1 neonatal intensive care unit (NICU). DD characteristics were documented in diaper rash logs. Primary outcome was “time to rash” calculated using log-rank tests of Kaplan-Meier plots. Adverse events and parental satisfaction were secondary measures. ResultsInfants (N = 149, 40% female, median gestational age 35 weeks) were randomized to CO (n = 75) or SOC (n = 74). Time to rash plots showed no DD prevention for either group. Adverse events were documented in 29% of participants. Parental satisfaction scores were driven by rash outcome with favorable scores related to no DD. ConclusionCoconut oil and minimal SOC protocols did not prevent DD in NICU infants. Parents should be advised that coconut oil may not protect against DD.

A Perspective into "TEL"-Tenofovir, Emtricitabine and Lamivudine Antileprotic Activities by Drug Repurposing and Exploring the Possibility of Combination Chemotherapy with Drug Rescued Molecules for a Leprosy Free Mankind.

Since leprosy bacilli cannot grow in vitro, testing for antimicrobial resistance against Mycobacterium leprae or assessing the anti-leprosy activity of new drugs remains hard. Furthermore, developing a new leprosy drug through the traditional drug development process is not economically captivating for pharmaceutical companies. As a result, repurposing existing drugs/approved medications or their derivatives to test their anti-leprotic potency is a promising alternative. It is an accelerated method to uncover different medicinal and therapeutic properties in approved drug molecules. The study aims to explore the binding potential of anti-viral drugs such as Tenofovir, Emtricitabine, and Lamivudine (TEL) against Mycobacterium leprae using molecular docking. The current study evaluated and confirmed the possibility of repurposing antiviral drugs such as TEL (Tenofovir, Emtricitabine, and Lamivudine) by transferring the graphical window of the BIOVIA DS2017 with the Crystal Structure of a phosphoglycerate mutase gpm1 from Mycobacterium leprae (PDB ID: 4EO9). Utilizing the smart minimizer algorithm, the protein's energy was reduced in order to achieve a stable local minima conformation. The protein and molecule energy minimization protocol generated stable configuration energy molecules. The protein 4EO9 energy was reduced from 14264.5 kcal/mol to -17588.1 kcal/mol. The CHARMm algorithm-based CDOCKER run docked all three molecules (TEL) inside the 4EO9 protein binding pocket (Mycobacterium leprae). The interaction analysis revealed that tenofovir had a better binding molecule with a score of - 37.7297 kcal/mol than the other molecules.

Low dose tacrolimus exposure and early steroid withdrawal with strict body weight control can improve post kidney transplant glucose tolerance in Japanese patients.

The development of diabetes mellitus (DM) after living donor kidney transplantation (KT) is a risk factor for worsening transplant kidney function, cardiac disease, and cerebrovascular disease, which may affect prognosis after KT. At our institution, all patients' glucose tolerance is evaluated perioperatively by oral glucose tolerance tests (OGTTs) at pre-KT, and 3, 6, and 12 month (mo.) after KT. We analyzed the insulinogenic index (ISI) and homeostasis model assessment beta cell (HOMA-β) based on the immunoreactive insulin (IRI) levels to determine how glucose tolerance changed after KT in 214 patients who had not been diagnosed with DM before KT. In addition, we analyzed the body mass index (BMI) which may also influence glucose tolerance after KT. The concentration of tacrolimus (TAC) in blood was also measured as the area under the curve (AUC) to examine its effects at each sampling point. The preoperative-OGTTs showed that DM was newly diagnosed in 22 of 214 patients (10.3%) who had not been given a diagnosis of DM by the pre-KT fasting blood sugar (FBS) tests. The glucose tolerance was improved in 15 of 22 DM patients at 12 mo. after KT. ISI and IRI deteriorated only at 3 mo. after KT but improved over time. There was a trend of an inverse correlation between HOMA-β and TAC-AUC. We also found inverse correlations between IRI and an increase in BMI from 3 to 12 mo. after KT. Early corticosteroid withdrawal or the steroid minimization protocol with tacrolimus to maintain a low level of diabetogenic tacrolimus and BMI decrease after KT used by our hospital individualizes lifestyle interventions for each patient might contribute to an improvement in post-KT glucose tolerance.

Single-cell RNA sequencing of bronchoscopy specimens: development of a rapid minimal handling protocol.

Single-cell RNA sequencing (scRNA-seq) is an important tool for understanding disease pathophysiology, including airway diseases. Currently, the majority of scRNA-seq studies in airway diseases have used invasive methods (airway biopsy, surgical resection), which carry inherent risks and thus present a major limitation to scRNA-seq investigation of airway pathobiology. Bronchial brushing, where the airway mucosa is sampled using a cytological brush, is a viable, less invasive method of obtaining airway cells for scRNA-seq. Here we describe the development of a rapid and minimal handling protocol for preparing single-cell suspensions from bronchial brush specimens for scRNA-seq. Our optimized protocol maximizes cell recovery and cell quality and facilitates large-scale profiling of the airway transcriptome at single-cell resolution.