Analyzing efficiency of a lentiviral shRNA knockdown system in human enteroids using western blot and flow cytometry.

Enteroids are in vitro models to study gastrointestinal pathologies and test personalized therapeutics; however, the inherent complexity of enteroids often renders standard gene editing approaches ineffective. Here, we introduce a refined lentiviral transfection protocol, ensuring sufficient lentiviral engagement with enteroids while considering spatiotemporal growth variability throughout the extracellular matrix. Additionally, we highlight a selection process for transduced cells, introduce a protocol to accurately measure transduction efficiency, and explore methodologies to gauge effects of gene knockdown on biological processes.

Robotic resection of presacral tumors.

BACKGROUND: Presacral tumors are a rare entity typically treated with an open surgical approach. A limited number of minimally invasive resections have been described. The aim of the study is to evaluate the safety and efficacy of roboticresection of presacral tumors. METHODS: This is a retrospective single system analysis, conducted at a quaternary referral academic healthcare system, and included all patients who underwent a robotic excision of a presacral tumor between 2015 and 2023. Outcomes of interest were operative time, estimated blood loss, complications, length of stay, margin status, and recurrence rates. RESULTS: Sixteen patients (11 females and 5 males) were included. The median age of the cohort was 51 years (range 25-69 years). The median operative time was 197 min (range 98-802 min). The median estimated blood loss was 40 ml, ranging from 0 to 1800 ml, with one patient experiencing conversion to open surgery after uncontrolled hemorrhage. Urinary retention was the only postoperative complication that occurred in three patients (19%) and was solved within 30 days in all cases. The median length of stay was one day (range 1-6 days). The median follow-up was 6.7 months (range 1-110 months). All tumors were excised with appropriate margins, but one benign and one malignant tumor recurred (12.5%). Ten tumors were classified as congenital (one was malignant), two were mesenchymal (both malignant), and five were miscellaneous (one malignant). CONCLUSIONS: Robotic resection of select presacral pathology is feasible and safe. Further studies must be conducted to determine complication rates, outcomes, and long-term safety profiles.

Tablet Versus Quick Response Code: Difference in Research Participation Rates?

Conducting research with immunocompromised populations, especially within the context of a global pandemic, warrants consideration of alternative research methods and modes of administration to keep participants safe. Digital and internet-based research methods have been utilized to minimize the risk of harm with cancer patients; however, adolescents and young adults with cancer (AYAs) remain an under served and understudied population with high levels of unmet needs. The purpose of the current study was to examine differences in AYA research participation rates based on two digital survey administration methods (tablet versus QR code). AYAs were randomly assigned to complete an online survey using either a tablet or quick response (QR) code, and participation rates in each group were compared. The total participation rate was 22.9%, with 75% of completed surveys from the tablet group and 25% from the QR code group. While the use of a QR code allows for reduced costs for in-clinic recruitment and may be the most sanitary option during COVID-19, eligible patients in the current study showed trends of increased engagement using a sanitized tablet. It is important to consider how psychosocial research and electronic surveys are administered, as the method may impact recruitment and/or information obtained.

Smoking History and the Development of Incisional Umbilical Hernia After Laparoscopic and Laparoendoscopic Single-Site Cholecystectomy.

The aim of this study was to analyze umbilical hernia occurrences in patients who underwent laparoscopic or laparoendoscopic single-sight (LESS) cholecystectomy. Patients who underwent cholecystectomy by a single surgeon between 2015 and 2020 were surveyed. Data are presented as median (mean +/- standard deviation). Two hundred and fifty-three patients were sent the survey and 130 (51%) patients responded. The overall age was 57 (31 +/- 18) and the overall BMI was 30 (31 +/- 7). Twelve (9%) patients developed an umbilical hernia. Seventeen patients were active smokers and four (24%) developed an umbilical hernia. One hundred and thirteen patients were inactive smokers and eight (7%) developed an umbilical hernia. There was a statistical significance between umbilical hernia occurrence and smoking history (P < .05). Active smokers have a higher risk of developing an umbilical hernia following a minimally invasive cholecystectomy, regardless of operative approach. Elective cholecystectomy should be reconsidered for current smokers.

Adolescent and Young Adult Communication Preferences.

The purpose of this study was to examine adolescents and young adults preferred methods of communication by health care professionals and to identify whether this aligns/differs from overall preferred methods of social media use. Sixty-seven participants completed an electronic survey about their communication preferences. Forty-eight participants were patients and 19 were parents. Logistic regression and chi-square analysis were performed to predict social media use and communication preferences from predictors including age, gender, race, and treatment status. More research is needed to generalize these results, and future research could further evaluate the effectiveness of implementing health care interventions using preferred platforms.

The AYA Care Plan: Initial Evaluation of a Web-Based Psychosocial Intervention.

PURPOSE: Adolescents and young adults (AYA) with cancer do not fit neatly into pediatric or older adult oncology care settings. Recent efforts have led to the development of psychosocial interventions for AYAs, but studies show AYAs demonstrate low levels of engagement in psychosocial services. The AYA Care Plan is one of the only web-based tools providing a personalized, psychosocial resource that addresses unmet needs for AYAs in active treatment and post-treatment survivorship. The current study aims to assess the usability and utility of the AYA Care Plan and identify opportunities for improvement. METHODS: Clinic staff administered an online distress and needs assessment to AYA patients with cancer at outpatient oncology clinics. Personalized care plans were sent to participants on the basis of their responses. A total of 11 AYAs between the ages 18 and 39 years, with a mean age of 31.64 years, then completed qualitative interviews about their experiences. Thematic analysis was used to identify themes on the AYA Care Plan. RESULTS: A majority of participants reported positive usability features. Half of the participants reported using their care plan to make health care decisions. One person indicated not finding the resources helpful, and the other half of participants reported not engaging with the care plan. Participants also offered suggestions for improvement. CONCLUSION: The AYA Care Plan appears to be a useful psychosocial intervention for some AYAs with cancer. Future research should continue to examine the AYA Care Plan's usability and utility, and specify when, how, and for whom the AYA Care Plan is useful.

Academic Detailing to Reduce Sedative-Hypnotic Prescribing in Older Veterans.

BACKGROUND: Sedative-hypnotics, including benzodiazepines (BZDs) and benzodiazepine receptor agonists (BZD-RA), are considered potentially inappropriate medications (PIMs) in older adults. Academic detailing, an educational outreach delivered by trained clinicians to other clinicians to encourage evidence-based care, can promote deprescribing of PIMs. OBJECTIVE: To evaluate the impact of academic detailing on sedative-hypnotic prescribing to older veterans. METHODS: A retrospective analysis was performed to evaluate the impact of academic detailing on BZD and BZD-RA prescribing to veterans aged 75 years and older. Prescribing trends for primary care and mental health prescribers in the Veterans Health Administration (VA) Southeast Network were calculated for the 18 months before and after an initial academic detailing session for each prescriber. Pre-post interrupted time series analyses (ITSAs) were conducted, and period prevalence was calculated as the number of prescriptions per 1000 older veterans. RESULTS: A total of 155 prescribers were followed for 36 months. BZD prevalence declined by 23% (69.08-53.33 per 1000 population; P < .001) and by 15% for BZD-RA (18.07-15.38 per 1000 population; P < .001). New starts on BZD declined by 54% (2.36-1.09 per 1000 population; P < .001) and new starts on BZD-RA declined by 53% (1.02-0.48 per 1000 population; P < .001). Alternative medications for insomnia increased by 23% (39.98-49.27 per 1000 population; P < .001). Findings from the ITSA confirmed those of the pre-post analysis with sustained effects in the postintervention period. CONCLUSIONS: Academic detailing was associated with reduced sedative-hypnotic prescribing in the primary care and mental health setting.

Boundary spanning at the science-policy interface: the practitioners' perspectives.

Cultivating a more dynamic relationship between science and policy is essential for responding to complex social challenges such as sustainability. One approach to doing so is to "span the boundaries" between science and decision making and create a more comprehensive and inclusive knowledge exchange process. The exact definition and role of boundary spanning, however, can be nebulous. Indeed, boundary spanning often gets conflated and confused with other approaches to connecting science and policy, such as science communication, applied science, and advocacy, which can hinder progress in the field of boundary spanning. To help overcome this, in this perspective, we present the outcomes from a recent workshop of boundary-spanning practitioners gathered to (1) articulate a definition of what it means to work at this interface ("boundary spanning") and the types of activities it encompasses; (2) present a value proposition of these efforts to build better relationships between science and policy; and (3) identify opportunities to more effectively mainstream boundary-spanning activities. Drawing on our collective experiences, we suggest that boundary spanning has the potential to increase the efficiency by which useful research is produced, foster the capacity to absorb new evidence and perspectives into sustainability decision-making, enhance research relevance for societal challenges, and open new policy windows. We provide examples from our work that illustrate this potential. By offering these propositions for the value of boundary spanning, we hope to encourage a more robust discussion of how to achieve evidence-informed decision-making for sustainability.

Antihypertensive medication adherence and blood pressure control among central Alabama veterans.

Medication nonadherence is associated with adverse outcomes. To evaluate antihypertensive medication adherence and its association with blood pressure (BP) control, the authors described population adherence to prescribed antihypertensive medication (proportion of days covered ≥80%) and BP control (mean BP <140/90 mm Hg) among central Alabama veterans during the fiscal year 2015. Overall, 75.1% of patients receiving antihypertensive medication were considered adherent, and 66.1% had adequate BP control. Patients adherent to antihypertensive medication were more likely to have adequate BP control compared with patients classified as nonadherent (67.4% vs 62.0%; adjusted odds ratio 1.33; 95% confidence interval, 1.22-1.44 [P<.0001]). Among patients who had uncontrolled BP, 73.6% were considered adherent to medication. Adherence to antihypertensive medication was associated with adequate BP control; however, a substantial proportion of patients with inadequate BP control were also considered adherent. Interventions to increase BP control could address more aggressive medication management to achieve BP goals.

Investigating the validity of the species status of the false codling moth in South African deciduous fruit orchards using mating studies and mtDNA.

The false codling moth is a polyphagous pest of various kinds or fruit, and it has expanded its geographical distribution and host range. The expanding host range could result in subspecies requiring varied pest management options. Laboratory no-choice cross-mating tests were conducted to establish whether Thaumatotibia leucotreta individuals from six areas and three host species, in South Africa, share mating characteristics and belong to the same subspecies or strain. The no-choice cross-mating tests indicated that all individuals in self- and out-crosses readily mated within 24 h with those derived from different hosts and areas. The mtDNA results confirmed that all individuals formed one group or clade. Overall, the results indicate that T. leucotreta individuals from the six areas and three host species in the Western Cape Province and two other provinces in South Africa represent a single genetical species. The results imply that similar control options can be effective across host ranges and distribution areas.

Investigating population differentiation in a major African agricultural pest: evidence from geometric morphometrics and connectivity suggests high invasion potential.

The distribution, spatial pattern and population dynamics of a species can be influenced by differences in the environment across its range. Spatial variation in climatic conditions can cause local populations to undergo disruptive selection and ultimately result in local adaptation. However, local adaptation can be constrained by gene flow and may favour resident individuals over migrants-both are factors critical to the assessment of invasion potential. The Natal fruit fly (Ceratitis rosa) is a major agricultural pest in Africa with a history of island invasions, although its range is largely restricted to south east Africa. Across Africa, C. rosa is genetically structured into two clusters (R1 and R2), with these clusters occurring sympatrically in the north of South Africa. The spatial distribution of these genotypic clusters remains unexamined despite their importance for understanding the pest's invasion potential. Here, C. rosa, sampled from 22 South African locations, were genotyped at 11 polymorphic microsatellite loci and assessed morphologically using geometric morphometric wing shape analyses to investigate patterns of population structure and determine connectedness of pest-occupied sites. Our results show little to no intraspecific (population) differentiation, high population connectivity, high effective population sizes and only one morphological type (R2) within South Africa. The absence of the R1 morphotype at sites where it was previously found may be a consequence of differences in thermal niches of the two morphotypes. Overall, our results suggest high invasion potential of this species, that area-wide pest management should be undertaken on a country-wide scale, and that border control is critical to preventing further invasions.

Setting conservation management thresholds using a novel participatory modeling approach.

We devised a participatory modeling approach for setting management thresholds that show when management intervention is required to address undesirable ecosystem changes. This approach was designed to be used when management thresholds: must be set for environmental indicators in the face of multiple competing objectives; need to incorporate scientific understanding and value judgments; and will be set by participants with limited modeling experience. We applied our approach to a case study where management thresholds were set for a mat-forming brown alga, Hormosira banksii, in a protected area management context. Participants, including management staff and scientists, were involved in a workshop to test the approach, and set management thresholds to address the threat of trampling by visitors to an intertidal rocky reef. The approach involved trading off the environmental objective, to maintain the condition of intertidal reef communities, with social and economic objectives to ensure management intervention was cost-effective. Ecological scenarios, developed using scenario planning, were a key feature that provided the foundation for where to set management thresholds. The scenarios developed represented declines in percent cover of H. banksii that may occur under increased threatening processes. Participants defined 4 discrete management alternatives to address the threat of trampling and estimated the effect of these alternatives on the objectives under each ecological scenario. A weighted additive model was used to aggregate participants' consequence estimates. Model outputs (decision scores) clearly expressed uncertainty, which can be considered by decision makers and used to inform where to set management thresholds. This approach encourages a proactive form of conservation, where management thresholds and associated actions are defined a priori for ecological indicators, rather than reacting to unexpected ecosystem changes in the future.

Are we missing the boat? Current uses of long-term biological monitoring data in the evaluation and management of marine protected areas.

Protected area management agencies are increasingly using management effectiveness evaluation (MEE) to better understand, learn from and improve conservation efforts around the globe. Outcome assessment is the final stage of MEE, where conservation outcomes are measured to determine whether management objectives are being achieved. When quantitative monitoring data are available, best-practice examples of outcome assessments demonstrate that data should be assessed against quantitative condition categories. Such assessments enable more transparent and repeatable integration of monitoring data into MEE, which can promote evidence-based management and improve public accountability and reporting. We interviewed key informants from marine protected area (MPA) management agencies to investigate how scientific data sources, especially long-term biological monitoring data, are currently informing conservation management. Our study revealed that even when long-term monitoring results are available, management agencies are not using them for quantitative condition assessment in MEE. Instead, many agencies conduct qualitative condition assessments, where monitoring results are interpreted using expert judgment only. Whilst we found substantial evidence for the use of long-term monitoring data in the evidence-based management of MPAs, MEE is rarely the sole mechanism that facilitates the knowledge transfer of scientific evidence to management action. This suggests that the first goal of MEE (to enable environmental accountability and reporting) is being achieved, but the second and arguably more important goal of facilitating evidence-based management is not. Given that many MEE approaches are in their infancy, recommendations are made to assist management agencies realize the full potential of long-term quantitative monitoring data for protected area evaluation and evidence-based management.

The osmotic tissue expander: a 5-year experience.

Tissue expansion is a valuable technique in soft tissue reconstruction. Osmotic expanders are self-inflating and obviate the need for repeated injections. In doing so, they eliminate port site problems and may reduce the potential to introduce infection. The use of such expanders has become more common in recent years. We report on our experience with the Osmed™ osmotic expanders over the last 5-years.

The bionomics of an invasive species Sitona lepidus during its establishment in New Zealand.

The egg, larval, pupal and adult abundance of the clover root weevil Sitona lepidus Gyllenhal (Coleoptera: Curculionidae) was monitored at three sites for the first ten years following the discovery of this exotic pest in the Waikato region of New Zealand. The species went through an initial boom and bust cycle at two sites, with populations reaching up to 1800 larvae m-2. Thereafter, winter larval populations were relatively stable, ranging between 450-750 m-2. Unlike in the Northern Hemisphere, S. lepidus was found to have two generations a year in the Waikato region of New Zealand. Pasture white clover content at the time of peak adult numbers was positively related to the subsequent peak larval populations for each generation. The factors contributing to the emergence of S. lepidus as one of the most important pasture pests in New Zealand are discussed.

Elevated hepatic transaminases associated with the use of interferon alfacon-1 and ribavirin.

PURPOSE: The case of a patient with hepatitis C who developed elevated hepatic transaminase levels associated with the use of interferon alfacon-1 and ribavirin is described. SUMMARY: A 55-year-old Caucasian man arrived at a hepatitis C clinic to discuss alternative treatment options for his hepatitis C virus (genotype 1a) infection, which did not respond to a 48-week course of peginterferon and ribavirin therapy. He was subsequently treated with interferon alfacon-1 9 microg subcutaneously daily plus ribavirin 200 mg orally twice daily. During treatment with interferon alfacon-1, he developed elevated hepatic transaminase levels despite a decrease in viral load. His hepatic transaminase levels returned to baseline when interferon alfacon-1 was discontinued and rose again upon rechallenge. Ribavirin was not the likely cause of the increase in transaminases since the patient previously tolerated it in combination with peginterferon. While activation of autoimmune hepatitis is a potential cause of acute decompensation in patients treated with interferons, it was not believed to be the case in this patient. Interferon alfacon-1 was determined to be the probable cause of the rise in hepatic transaminase levels in this patient, since his levels declined when therapy was discontinued and rose dramatically once it was restarted. This case illustrates the importance of monitoring both viral loads and hepatic transaminase levels in patients with hepatitis C being treated with interferon therapy. CONCLUSION: A patient with hepatitis C developed elevated hepatic transaminase levels despite showing an improvement in viral load after receiving interferon alfacon-1 and ribavirin.

Shock outcome prediction before and after CPR: a comparative study of manual and automated active compression-decompression CPR.

We report on a study designed to compare the relative efficacy of manual CPR (M-CPR) and automated mechanical CPR (ACD-CPR) provided by an active compression-decompression (ACD) device. The ECG signals of out-of-hospital cardiac arrest patients of cardiac aetiology were analysed just prior to, and immediately after, cardiopulmonary resuscitation (CPR) to assess the likelihood of successful defibrillation at these time points. The cardioversion outcome prediction (COP) measure previously developed by our group was used to quantify the probability of return of spontaneous circulation (ROSC) after counter-shock and was used as a measure of the efficacy of CPR. An initial validation study using COP to predict shock outcome from the patient data set resulted in a performance of 60% specificity achieved at 100% sensitivity on a blind test of the data. This is comparable with previous studies and provided confidence in the robustness of the technique across hardware platforms. Significantly, the COP marker also displayed an ability to stratify according to outcomes: asystole, ventricular fibrillation (VF), pulseless electrical activity (PEA), normal sinus rhythm (NSR). We then used the validated COP marker to analyse the ECG data record just prior to and immediately after the chest compression segments. This was initially performed for 87 CPR segments where VF was both the pre- and post-CPR waveform. An increase in the mean COP values was found for both CPR types. A signed rank sum test found the increase due to manual CPR not to be significant (p>0.05) whereas the automated CPR was found to be significant (p<0.05). This increase was larger for the automated CPR (1.26, p=0.024) than for the manual CPR (0.99, p=0.124). These results indicate that the application of CPR does indeed provide beneficial preparation of the heart prior to defibrillation therapy whether manual or automated CPR is applied. The COP marker shows promise as a definitive, quantitative determinant of the immediate positive effect of both types of CPR regardless of the details of use. In work of a more exploratory nature we then used the validated COP marker to analyse the ECG pre- and post-CPR for all rhythm types (212 traces). We show a significant increase in the COP measure (p<0.001 in both cases) as indicated by a shift in the median COP marker distribution values. This increase was more pronounced for automated ACD-CPR than for manual CPR. However, a detailed statistical analysis carried out between the groups adjusted for pre-CPR value showed no significant difference between the two methods of CPR (p=0.20). Similarly, adjusting for length of CPR showed no significant difference between the groups. Secondary, subgroup analysis of the ECG according to the length of time for which CPR was performed showed that both types of CPR led to an increase in the likelihood of successful defibrillation after increasing durations of CPR, however results were less reliable after longer periods of continuous CPR.