Mapping the genetic landscape of DNA double-strand break repair.

Cells repair DNA double-strand breaks (DSBs) through a complex set of pathways critical for maintaining genomic integrity. To systematically map these pathways, we developed a high-throughput screening approach called Repair-seq that measures the effects of thousands of genetic perturbations on mutations introduced at targeted DNA lesions. Using Repair-seq, we profiled DSB repair products induced by two programmable nucleases (Cas9 and Cas12a) in the presence or absence of oligonucleotides for homology-directed repair (HDR) after knockdown of 476 genes involved in DSB repair or associated processes. The resulting data enabled principled, data-driven inference of DSB end joining and HDR pathways. Systematic interrogation of this data uncovered unexpected relationships among DSB repair genes and demonstrated that repair outcomes with superficially similar sequence architectures can have markedly different genetic dependencies. This work provides a foundation for mapping DNA repair pathways and for optimizing genome editing across diverse modalities.

Bicoid-Dependent Activation of the Target Gene hunchback Requires a Two-Motif Sequence Code in a Specific Basal Promoter.

In complex genetic loci, individual enhancers interact most often with specific basal promoters. Here we investigate the activation of the Bicoid target gene hunchback (hb), which contains two basal promoters (P1 and P2). Early in embryogenesis, P1 is silent, while P2 is strongly activated. In vivo deletion of P2 does not cause activation of P1, suggesting that P2 contains intrinsic sequence motifs required for activation. We show that a two-motif code (a Zelda binding site plus TATA) is required and sufficient for P2 activation. Zelda sites are present in the promoters of many embryonically expressed genes, but the combination of Zelda plus TATA does not seem to be a general code for early activation or Bicoid-specific activation per se. Because Zelda sites are also found in Bicoid-dependent enhancers, we propose that simultaneous binding to both enhancers and promoters independently synchronizes chromatin accessibility and facilitates correct enhancer-promoter interactions.

A feed-forward relay integrates the regulatory activities of Bicoid and Orthodenticle via sequential binding to suboptimal sites.

The K50 (lysine at amino acid position 50) homeodomain (HD) protein Orthodenticle (Otd) is critical for anterior patterning and brain and eye development in most metazoans. In Drosophila melanogaster, another K50HD protein, Bicoid (Bcd), has evolved to replace Otd's ancestral function in embryo patterning. Bcd is distributed as a long-range maternal gradient and activates transcription of a large number of target genes, including otd Otd and Bcd bind similar DNA sequences in vitro, but how their transcriptional activities are integrated to pattern anterior regions of the embryo is unknown. Here we define three major classes of enhancers that are differentially sensitive to binding and transcriptional activation by Bcd and Otd. Class 1 enhancers are initially activated by Bcd, and activation is transferred to Otd via a feed-forward relay (FFR) that involves sequential binding of the two proteins to the same DNA motif. Class 2 enhancers are activated by Bcd and maintained by an Otd-independent mechanism. Class 3 enhancers are never bound by Bcd, but Otd binds and activates them in a second wave of zygotic transcription. The specific activities of enhancers in each class are mediated by DNA motif variants preferentially bound by Bcd or Otd and the presence or absence of sites for cofactors that interact with these proteins. Our results define specific patterning roles for Bcd and Otd and provide mechanisms for coordinating the precise timing of gene expression patterns during embryonic development.

Blood-brain barrier dysfunction mediated by the EZH2-Claudin-5 axis drives stress-induced TNF-α infiltration and depression-like behaviors.

Growing evidence suggests that neurovascular dysfunction characterized by blood-brain barrier (BBB) breakdown underlies the development of psychiatric disorders, such as major depressive disorder (MDD). Tight junction (TJ) proteins are critical modulators of homeostasis and BBB integrity. TJ protein Claudin-5 is the most dominant BBB component and is downregulated in numerous depression models; however, the underlying mechanisms remain elusive. Here, we demonstrate a molecular basis of BBB breakdown that links stress and depression. We implemented an animal model of depression, chronic unpredictable mild stress (CUMS) in male C57BL/6 mice, and showed that hippocampal BBB breakdown was closely associated with stress vulnerability. Concomitantly, we found that dysregulated Cldn5 level coupled with repression of the histone methylation signature at its promoter contributed to stress-induced BBB dysfunction and depression. Moreover, histone methyltransferase enhancer of zeste homolog 2 (EZH2) knockdown improved Cldn5 expression and alleviated depression-like behaviors by suppressing the tri-methylation of lysine 27 on histone 3 (H3K27me3) in chronically stressed mice. Furthermore, the stress-induced excessive transfer of peripheral cytokine tumor necrosis factor-α (TNF-α) into the hippocampus was prevented by Claudin-5 overexpression and EZH2 knockdown. Interestingly, antidepressant treatment could inhibit H3K27me3 deposition at the Cldn5 promoter, reversing the loss of the encoded protein and BBB damage. Considered together, these findings reveal the importance of the hippocampal EZH2-Claudin-5 axis in regulating neurovascular function and MDD development, providing potential therapeutic targets for this psychiatric illness.

The use of a multi-disciplinary geriatric telemedicine service (TELEG) and its acceptance at a tertiary care centre in Malaysia.

BACKGROUND: The COVID-19 pandemic has fueled the widespread adoption of telemedicine in healthcare, particularly in Sarawak, Malaysia. This study investigates the use and acceptance of Sarawak's inaugural multidisciplinary geriatric telemedicine service, TELEG. METHODS: This cross-sectional study took place at the Sarawak Heart Centre's geriatric department from July 1, 2021, to April 30, 2022. Convenient sampling included all TELEG-enrolled patients during this period, to achieve minimum sample size of 148. TELEG's utilization was assessed in terms of medication therapy and treatment plan optimization, as well as enhanced healthcare accessibility. Participants' acceptance of TELEG was measured using the Service User Technology Acceptability Questionnaire (SUTAQ) administered through Google Forms. Descriptive statistics percentages illustrated the proportion of participants who found TELEG moderately to highly acceptable. Associations between baseline characteristics and overall acceptance were explored through bivariate analyses, including Pearson's correlation test, independent t-test, and ANOVA. The influence of six SUTAQ dimensions on overall acceptance, multivariable linear regression using enter method was employed. Statistical significance was determined by p-values less than 0.5. RESULTS: Among 180 geriatric patients enrolled in TELEG during the study period, 149 agreed to participate. TELEG led to medication therapy optimization for 88.6% of participants, primarily involving dose adjustment (44.7%), de-prescribing (31.8%), and prescribing (15.9%). Additionally, 53.8% received treatment plan optimization, predominantly in the form of self-care education (56.3%), referrals for further treatment (33.8%), additional laboratory investigations (29.6%), and increased monitoring (26.8%). Among those educated in self-care (n = 40), dietary intake (27.5%), lower limb exercise (25.0%), and COVID-19 vaccination (12.5%) were the most common topics. All participants expressed moderate to high acceptance of TELEG (mean = 4.9, SD = 0.65, on a scale of 1 to 6). Notably, care personnel concern (B = 0.256; p < 0.001) had the most significant impact on overall acceptance. CONCLUSION: This pioneering study evaluates the utilization and user acceptance of a geriatric telemedicine service in the region, providing valuable insights to support its expansion. Follow-up surveys or interviews to gain insights into users' experiences are crucial to further enhance acceptance.

Impact of Perceived Safety and Barriers on Physical Activity Levels in Community-Dwelling Older Adults During the COVID-19 Pandemic in Singapore: A Cross-Sectional Mixed Methods Study.

This descriptive cross-sectional mixed methods study conducted in Singapore aimed to describe community-dwelling older adults' differences in physical activity (PA) based on perceived safety to exercise, barriers to PA, and preferred modes of PA during a pandemic. Out of 268 older adults, 25.4% felt unsafe to exercise during the pandemic. More participants who felt unsafe were aged 75 years and older (72.1% vs. 57.0%, p = .028) and lacked formal education (54.4% vs. 37.0%, p = .040). Barriers included difficulties exercising with masks, family concerns, and exercise center closures. Those who felt unsafe were significantly more likely to exercise at home and had significantly shorter duration of exercise and walks per week (2.72 vs. 4.50 hr, p = .002). Perceived barriers and exercise preferences should be considered when developing programs to improve older adults' PA during pandemics.

Suboptimal Intermediates Underlie Evolution of the Bicoid Homeodomain.

Changes in regulatory networks generate materials for evolution to create phenotypic diversity. For transcription networks, multiple studies have shown that alterations in binding sites of cis-regulatory elements correlate well with the gain or loss of specific features of the body plan. Less is known about alterations in the amino acid sequences of the transcription factors (TFs) that bind these elements. Here we study the evolution of Bicoid (Bcd), a homeodomain (HD) protein that is critical for anterior embryo patterning in Drosophila. The ancestor of Bcd (AncBcd) emerged after a duplication of a Zerknullt (Zen)-like ancestral protein (AncZB) in a suborder of flies. AncBcd diverged from AncZB, gaining novel transcriptional and translational activities. We focus on the evolution of the HD of AncBcd, which binds to DNA and RNA, and is comprised of four subdomains: an N-terminal arm (NT) and three helices; H1, H2, and Recognition Helix (RH). Using chimeras of subdomains and gene rescue assays in Drosophila, we show that robust patterning activity of the Bcd HD (high frequency rescue to adulthood) is achieved only when amino acid substitutions in three separate subdomains (NT, H1, and RH) are combined. Other combinations of subdomains also yield full rescue, but with lower penetrance, suggesting alternative suboptimal activities. Our results suggest a multistep pathway for the evolution of the Bcd HD that involved intermediate HD sequences with suboptimal activities, which constrained and enabled further evolutionary changes. They also demonstrate critical epistatic forces that contribute to the robust function of a DNA-binding domain.

Norepinephrine promotes glioma cell migration through up-regulating the expression of Twist1.

BACKGROUND: Glioma cells are characterized by high migration ability, resulting in aggressive growth of the tumors and poor prognosis of patients. It has been reported that the stress-induced hormone norepinephrine (NE) contributes to tumor progression through mediating a number of important biological processes in various cancers. However, the role of NE in the regulation of glioma migration is still unclear. Epithelial-to-mesenchymal transition (EMT) is one of the most important steps for tumor migration and metastasis. Twist1, as a key regulator of EMT, has been found to be elevated during glioma migration. But it is still unknown whether Twist1 is involved in the effect of NE on the migration of glioma cells. METHODS: Wound healing assay and transwell assay were conducted to evaluate the migration of glioma cells upon different treatments. The mesenchymal-like phenotype and the expression of Twist1 after NE treatment were assessed by cell diameters, real-time PCR, western blot and immunofluorescence staining. The gain-and loss-of-function experiments were carried out to investigate the biological function of Twist1 in the migration induced by NE. Finally, the clinical significance of Twist1 was explored among three public glioma datasets. RESULTS: In this study, our finding revealed a facilitative effect of NE on glioma cell migration in a ß-adrenergic receptor (ADRB)-dependent way. Mechanistically, NE induced mesenchymal-like phenotype and the expression of Twist1. Twist1 overexpression promoted glioma cells migration, while knockdown of Twist1 abolished the discrepancy in the migration ability between NE treated glioma cells and control cells. In addition, the clinical analysis demonstrated that Twist1 was up-regulated in malignant gliomas and recurrent gliomas, and predicted a poor prognosis of glioma patients. CONCLUSIONS: NE enhanced the migration ability of glioma cells through elevating the expression of Twist1. Our finding may provide potential therapeutic target for protecting patients with glioma from the detrimental effects of stress biology on the tumor progression.

Factors influencing fear of falling in community-dwelling older adults in Singapore: a cross-sectional study.

BACKGROUND: Fear of falling (FoF) has far-reaching implications including activity restriction, functional decline and reduced quality of life. It is a common consequence of falls but may be present even in non-fallers. This study aimed to determine the factors associated with FoF in a segment of Singapore's community-dwelling older adults. METHODS: This descriptive cross-sectional study recruited a convenience sample of adults aged 65 and above from 4 primary care clinics from September 2020 to March 2021. Data were collected on demographic factors, clinical factors such as multi-morbidity, falls characteristics such as history of falls, injuries, and reasons for falls and frailty as determined by the Clinical Frailty Scale (CFS). FoF was measured using the Short Falls Efficacy Scale-International (Short FES-I), cut-off score of 14 and above indicated high FoF. Logistic regression was used to determine factors associated with high FoF. RESULTS: Out of 360 older adults, 78.1% were Chinese and 59.7% females. The mean age was 78.3 years and 76 (21.1%) had a history of falls in the past six months. Almost half (43.1%) were mildly to moderately frail and most (80.6%) had multi-morbidity. The mean FoF score was 15.5 (SD 5.97) and 60.8% reported high FoF. There were statistically significant differences in age, gender, ethnicity, marital status, educational level, use of walking aid, multi-morbidity, frailty status, history of falls within six months and reason for falls between patients who had high FoF versus those who had moderate or low FoF. Logistic regression found that Malay ethnicity (OR = 5.81, 95% CI 1.77-19.13), marital status, use of walking aids (OR = 3.67, 95% CI = 1.54-8.77) and frailty were significant factors associated with high FoF. Compared to those who were never married, the odds of high FoF were significantly higher in married older adults (OR = 6.75, 95% CI 1.39 to 32.76), those who were separated or divorced (OR 10.40, 95% CI 1.13 to 95.76) and those who were widowed (OR = 7.41, 95% CI 1.51 to 36.41). Compared to well older adults, the odds of high FoF were significantly higher in pre frail older adults (OR = 6.87, 95% CI = 2.66-17.37), mildly frail older adults (OR = 18.58, 95% CI = 4.88-70.34) and moderately frail older adults (OR = 144.78, 95% CI = 13.86-1512.60). CONCLUSIONS: The study found that pre frail to moderately frail older adults as determined by CFS have significantly higher risk of high FoF. The demographic factors such as marital status and ethnicity and falls characteristics associated with FoF in this study will be helpful to develop targeted and tailored interventions for FoF.

Preparation and Characterization of Polysaccharides from Turpiniae Folium and Its Antioxidative, Anti-Inflammatory Activities and Antiproliferative Effect on VSMCs.

Turpiniae Folium, the dried leaves of Turpinia arguta Seem., is a kind of historic traditional Chinese medicine. Here, based on our previous study, we extracted the Turpiniae Folium polysaccharides (TFP) and isolated three polysaccharide fractions from TFP. Then, TFP and one of the major polysaccharide fractions (TFP-1a) were identified through HPLC, HPGPC, and ATR-FTIR. Furthermore, the evaluations of their antioxidative, anti-inflammatory activities and inhibitory effect on angiotensin II-induced vascular smooth muscle cells (VSCMs) proliferation in vitro were conducted. Both TFP and TFP-1a showed strong hydroxyl radical scavenging, DPPH radical scavenging, and Fe2+ chelating activities, and exerted strong anti-inflammatory activity. Moreover, TFP and TFP-1a also possessed a strong inhibitory effect on Ang II-induced VSCMs proliferation. On these premises, we inferred that TFP and TFP-1a could be potential and promising natural antioxidants, anti-inflammatory agents, and implicated to treat cardiovascular disease.

Impacts of the ubiquitous factor Zelda on Bicoid-dependent DNA binding and transcription in Drosophila.

In vivo cross-linking studies suggest that the Drosophila transcription factor Bicoid (Bcd) binds to several thousand sites during early embryogenesis, but it is not clear how many of these binding events are functionally important. In contrast, reporter gene studies have identified >60 Bcd-dependent enhancers, all of which contain clusters of the consensus binding sequence TAATCC. These studies also identified clusters of TAATCC motifs (inactive fragments) that failed to drive Bcd-dependent activation. In general, active fragments showed higher levels of Bcd binding in vivo and were enriched in predicted binding sites for the ubiquitous maternal protein Zelda (Zld). Here we tested the role of Zld in Bcd-mediated binding and transcription. Removal of Zld function and mutations in Zld sites caused significant reductions in Bcd binding to known enhancers and variable effects on the activation and spatial positioning of Bcd-dependent expression patterns. Also, insertion of Zld sites converted one of six inactive fragments into a Bcd-responsive enhancer. Genome-wide binding experiments in zld mutants showed variable effects on Bcd-binding peaks, ranging from strong reductions to significantly enhanced levels of binding. Increases in Bcd binding caused the precocious Bcd-dependent activation of genes that are normally not expressed in early embryos, suggesting that Zld controls the genome-wide binding profile of Bcd at the qualitative level and is critical for selecting target genes for activation in the early embryo. These results underscore the importance of combinatorial binding in enhancer function and provide data that will help predict regulatory activities based on DNA sequence.

Factors Associated with Healing Outcomes in Primary Care Patients with Diabetic Foot Ulcers: A Retrospective Study in a Multiethnic Sample.

OBJECTIVE: To identify and determine patient- and ulcer-related factors associated with healing outcomes within 3 months for patients with diabetic foot ulcer (DFU) in a multiethnic primary care sample. METHODS: Retrospective data were collected over 3 months from 520 primary care patients with a DFU between April 1, 2016 and March 31, 2017. Multivariable prevalence ratios (PRs) were calculated using Poisson regression to find associations between patient- and ulcer-related factors and healing outcomes. RESULTS: Most patients were male (66%) and Chinese (49.8%) and had a diabetes mellitus duration longer than 5 years (81.8%). Toe ulcers (64%) were most common. Healing occurred for 33.9% of participants; 19.1% and 1.5% underwent minor and major amputation, respectively. Wound sizes between 1 and 10 cm2 (PR, 0.61; 95% confidence interval [CI], 0.46-0.76; P < .001) and over 10 cm2 (PR, 0.55; 95% CI, 0.33-0.76; P = .003), ulcer duration 6 months or longer (PR, 0.36; 95% CI, 0.19-0.53; P < .001), ischemic ulcers (PR, 0.54; 95% CI, 0.22-0.86; P = .044), and neuroischemic ulcers (PR, 0.73; 95% CI, 0.53-0.93; P = .027) were negatively associated with healing outcomes. Women were more likely to experience healing (PR, 1.18; 95% CI, 0.91-1.45; P = .157). CONCLUSIONS: Ulcer healing varied by sex and was affected by wound size, wound duration, and ischemic etiology, regardless of ethnicity. Prompt attention to these risk factors may reduce healing time. Further studies are warranted to elucidate the mechanism underlying sex differences in association with DFU healing.

Description and Utilization of Telewound Monitoring Services in Primary Care Patients with Acute Wounds in Singapore: A Retrospective Study.

OBJECTIVE: To describe an inaugural telewound monitoring service (TMS) designed for the remote monitoring of acute wounds to empower primary care patients, and identify factors associated with the utilization of the TMS. METHODS: Retrospective data were collected from 204 patients who participated in the TMS between June 19, 2016 and August 31, 2017 and analyzed using both descriptive and multiple regression analysis. RESULTS: The mean patient age was 27.9 years (SD, 12.4); wound area was 7.8 cm2 (SD, 21.2); and duration of healing was 11.7 days (SD, 6.9). A multiple regression model based on patients' demographics and wound factors predicted which patients were likely to have more telewound sessions than face-to-face sessions. The model was statistically significant (F = 2.093 (11, 124), P = .025) with 15.7% of variance explained by the variables. An increase in age (P = .043) and increased days to healing (P = .043) were associated with a reduction in the number of telewound sessions. CONCLUSIONS: The TMS is a valuable alternative to face-to-face wound care that enables patients with acute wounds to assume the roles of both patient and carer simultaneously. Age and healing duration are predictors for utilization of this service. Prompt attention to these predictors may improve service allocation and utilization.

[Establishment of a prostate cancer prognostic risk model based on the TCGA database and inflammation-related genes].

OBJECTIVE: To investigate the effect of inflammation-related genes on the prognosis of prostate cancer (PCa). METHODS: We downloaded PCa-related clinical data and mRNA sequencing data from the database Cancer Genome Atlas (TCGA) and inflammation-related pathway gene sets from MsigDB. Using univariate regression and LASSO regression analyses, we screened inflammation-related genes for the construction of a prognostic risk model and evaluated the performance of the model in predicting the prognosis of PCa by Kaplan-Meier and ROC analyses. Based on the nomogram, we calculated the risk scores of the patients, divided them into a high-risk and a low-risk group based on the median values of their risk scores, identified differentially expressed genes for enrichment analysis and verified the expression level of SPHK1 in the PCa tissue microarrays by immunohistochemical staining. RESULTS: Totally 19 inflammation-related genes were identified from 172 candidate genes for the construction of the prognostic risk model, including the risk genes CD14, PIK3R5, GABBR1, RELA, IRF7, SCARF1, MSR1, SPHK1, OSM and STAB1, and the protective genes AQP9, LPAR1, ATP2C1, NDP, CXCL6, P2RY2, DCBLD2, PCDH7, and IFNAR1. Kaplan-Meier analysis showed that the patients with high risk scores had a significantly lower recurrence-free survival and a worse prognosis than those with low risk scores. Differentially expressed genes were involved mainly in the activation of inflammatory response pathways. Immunohistochemical results indicated that the expression of SPHK1 was significantly higher in the tumorous than in the normal tissue and increased with the Gleason score. There was a correlation between the SPHK1 expression and envelope invasion. CONCLUSION: The prognostic risk model of inflammation-related genes constructed based on the TCGA database can effectively predict the prognosis of PCa.

The molecular mechanism underlying mitophagy-mediated hippocampal neuron apoptosis in diabetes-related depression.

Diabetes-related depression (DD) is a major complication of diabetes mellitus. Our previous studies indicated that glutamate (Glu) and hippocampal neuron apoptosis are key signal and direct factor leading to diabetes-related depression, respectively. However, the accurate pathogenesis remains to be unclear. We hypothesized that diabetes-related depression might be associated with the mitophagy-mediated hippocampal neuron apoptosis, triggered by aberrant Glu-glutamate receptor2 (GluR2)-Parkin pathway. To testify this hypothesis, here the rat model of DD in vivo and in vitro were both established so as to uncover the potential mechanism of DD based on mitophagy and apoptosis. We found that DD rats exhibit an elevated glutamate levels followed by monoamine neurotransmitter deficiency and depressive-like behaviour, and DD modelling promoted autophagosome formation and caused mitochondrial impairment, eventually leading to hippocampal neuron apoptosis via aberrant Glu-GluR2-Parkin pathway. Further, in vitro study demonstrated that the simulated DD conditions resulted in an abnormal glutamate and monoamine neurotransmitter levels followed by autophagic flux increment, mitochondrial membrane potential reduction and mitochondrial reactive oxygen species and lactic dehydrogenase elevation. Interestingly, both GluR2 and mammalian target of rapamycin (mTOR) receptor blocker aggravated mitophagy-induced hippocampal neuron apoptosis and abnormal expression of apoptotic protein. In contrast, both GluR2 and mTOR receptor agonist ameliorated those apoptosis in simulated DD conditions. Our findings revealed that mitophagy-mediated hippocampal neuron apoptosis, triggered by aberrant Glu-GluR2-Parkin pathway, is responsible for depressive-like behaviour and monoamine neurotransmitter deficiency in DD rats. This work provides promising molecular targets and strategy for the treatment of DD.

Comparison of outcomes between neoadjuvant imatinib and upfront surgery in patients with localized rectal GIST: An inverse probability of treatment weighting analysis.

BACKGROUND AND OBJECTIVES: This study aimed to compare outcomes between neoadjuvant imatinib and upfront surgery in patients with localized rectal gastrointestinal stromal tumors (GIST) patients. METHODS: Eighty-five patients with localized rectal GIST were divided into two groups: upfront surgery ± adjuvant imatinib (Group A, n = 33) and the neoadjuvant imatinib + surgery + adjuvant imatinib (Group B, n = 52). Baseline characteristics between groups were controlled for with inverse probability of treatment weighting (IPTW) adjusted analysis. RESULTS: The response rate to neoadjuvant imatinib was 65.9%. After the IPTW-adjusted analysis, patients who underwent neoadjuvant therapy had better distant recurrence-free survival (DRFS) and disease-specific survival (DSS) compared with those who underwent upfront surgery (5-year DRFS 97.8 vs. 71.9%, hazard ratio [HR], 0.15; 95% CI, 0.03-0.87; p = 0.03; 5-year DSS 100 vs. 77.1%; HR, 0.11; 95% CI, 0.01-0.92; p = 0.04). While no significant association was found between overall survival (OS) and treatment groups (p = 0.07), 5-year OS was higher for the neoadjuvant group than upfront surgery group (97.8% vs. 71.9%; HR, 0.2; 95% CI, 0.03-1.15). CONCLUSIONS: In patients with localized rectal GIST, neoadjuvant imatinib not only shrunk the tumor size but also decreased the risk of metastasis and tumor-related deaths when compared to upfront surgery and adjuvant imatinib alone.

Pain resolution and glucose control in pediatric patients with chronic pancreatitis after total pancreatectomy with islet auto-transplantation.

BACKGROUND: Chronic pancreatitis (CP) and acute recurrent pancreatitis (ARP) in pediatric patients are strongly associated with genetic mutations and lead to pan-parenchymal disease refractory to medical and endoscopic treatment. Our aim was to assess pain resolution and glucose control in patients with CP and ARP following total pancreatectomy with islet auto-transplantation (TPIAT). METHODS: We retrospectively analyzed prospectively collected clinical data of 12 children who developed CP and ARP and underwent TPIAT when 21 years old or younger at the University of Chicago between December 2009 and June 2020. Patients with recurrent or persistent abdominal pain attributed to acute or chronic pancreatic inflammation and a history of medical interventions attempted for the relief of pancreatic pain were selected by a multi-disciplinary team for TPIAT. We followed patients post-operatively and reported data for pre-TPIAT, post-operative day 75, and yearly post-TPIAT. RESULTS: All 12 patients experienced complete resolution of pancreatic pain. The overall insulin-independence rate after 1 year was 66% (8/12) and 50% (3/6) at 4 years. Shorter duration of CP/ARP pre-TPIAT, higher mass of islets infused, and lower BMI, BMI percentile, and BSA were associated with insulin-independence post-TPIAT. CONCLUSIONS: TPIAT is a viable treatment option for pediatric patients with CP and ARP. Pediatric patients undergoing TPIAT for CP achieved resolution of pancreatic-type pain and reduced opioid requirements. The majority were able to achieve insulin-independence which was associated with lower pre-TPIAT BMI and higher islet mass transplanted (i.e., over 2000 IEQ/kg), the latter of which can be achieved by earlier TPIAT. LEVEL OF EVIDENCE: Treatment study, Level IV.

Understanding nurse preceptors' experiences in a primary health care setting: A descriptive qualitative study.

AIM: This study sought to understand primary health care nurse preceptors' experiences of precepting junior nurses. BACKGROUND: Nurse preceptors provide clinical teaching to nurses and pre-registration nurses and play a supervisory role in their work. The demand for preceptors in the primary health care setting is on the rise, and there is a huge responsibility placed on them on top of their routine workload. METHODS: A descriptive qualitative approach was adopted, and purposive sampling was used to select the participants. 14 nursing preceptors across six polyclinics were interviewed. Thematic analysis was used to analyse the data. RESULTS: The analysis revealed three themes: (1) a vessel for transferring knowledge; (2) building a therapeutic relationship; and (3) overcoming organisational barriers. CONCLUSION: In light of the study's findings, strategies to address the insufficient knowledge of nurse preceptors, lack of recognition and preceptorship opportunities must be developed. IMPLICATIONS FOR NURSING MANAGEMENT: More attention and opportunities for nurse preceptors' professional development such as courses to enhance their knowledge on educational and research developments as well as teaching seminars to maintain and build effective relationships with their preceptees. In addition, recognizing the role of nurse preceptors as a pillar of guiding the future generation of nurses and research to support nurse preceptors in their training cannot be undermined.

"When nothing happens, nobody is afraid!" beliefs and perceptions around self-care and health-seeking behaviours: Voices of patients living with diabetic lower extremity amputation in primary care.

Self-management and self-care are the cornerstone of diabetes care and an essential part of successfully preventing or delaying diabetes complications. Yet, despite being armed with the required information and guidance for self-management, self-care and adherence to foot self-care recommendations and compliance to medication among patients with diabetic foot ulcer and diabetic lower extremity amputations remain low and suboptimal. This study reveals in-depth account of nine such patients' beliefs and perceptions around their illness, their self-care, and their health-seeking behaviours. Patients living with diabetic lower extremity amputation displayed profound lack of knowledge of self-care of diabetes and foot and passive health-related behaviours. The overarching sense that "when nothing happens, nobody is afraid," points to a lack of motivation in taking charge of one's own health, whether this is with reference to treatment or care adherence, following recommended self-care advice, or seeking timely treatment. The Health Beliefs Model provides the theoretical framework for probing into the factors for the participants' suboptimal self-care and passive health-seeking behaviours. Two themes emerged from data analysis: profound knowledge deficit and passive health-related behaviours. The beliefs and perceptions around self-care and health-seeking behaviours for patients with lower extremity amputation are interpreted as the "ignorant self" with passive health-seeking behaviours. Patients with diabetes and diabetic foot diseases may benefit from personalized education, motivational interviewing, and family support.

BETA-2 score is an early predictor of graft decline and loss of insulin independence after pancreatic islet allotransplantation.

This study aimed to evaluate whether the BETA-2 score is a reliable early predictor of graft decline and loss of insulin independence after islet allotransplantation. Islet transplant procedures were stratified into 3 groups according to clinical outcome: long-term insulin independence without islet graft decline (group 1, N = 9), initial insulin independence with subsequent islet graft decline and loss of insulin independence (group 2, N = 13), and no insulin independence (group 3, N = 13). BETA-2 was calculated on day 75 and multiple times afterwards for up to 145 months posttransplantation. A BETA-2 score cut-off of 17.4 on day 75 posttransplantation was discerned between group 1 and groups 2 and 3 (area under the receiver operating characteristic 0.769, P = .005) with a sensitivity and negative predictive value of 100%. Additionally, BETA-2 ≥ 17.4 at any timepoint during follow-up reflected islet function required for long-term insulin independence. While BETA-2 did not decline below 17.4 for each of the 9 cases from group 1, the score decreased below 17.4 for all transplants from group 2 with subsequent loss of insulin independence. The reduction of BETA-2 below 17.4 predicted 9 (1.5-21) months in advance subsequent islet graft decline and loss of insulin independence (P = .03). This finding has important implications for posttransplant monitoring and patient care.