G-quadruplex is critical to epigenetic activation of the lncRNA HOTAIR in cancer cells.

The cancer-promoting lncRNA HOTAIR has multiple isoforms. Which isoform of HOTAIR accounts for its expression and functions in cancer is unknown. Unlike HOTAIR's canonical intergenic isoform NR_003716 (HOTAIR-C), the novel isoform NR_047517 (HOTAIR-N) forms an overlapping antisense transcription locus with HOXC11. We identified HOTAIR-N as the dominant isoform that regulates the gene expression programs and networks for cell proliferation, survival, and death in cancer cells. The CpG island in the HOTAIR-N promoter was marked with epigenetic markers for active transcription. We identified a G-quadruplex (G4) motif rich region in the HOTAIR-N CpG island. Our findings indicate that G4s in HOTAIR-N CpG island is critical for expression of HOTAIR-N in cancer cells. Disruption of G4 may represent a novel therapeutic approach for cancer. The transcriptomes regulated by HOTAIR-N and Bloom in cancer cells as provided herein are important resources for the exploration of lncRNA, DNA helicases, and G4 in cancer.

Nursing harm reduction education and care for people who use drugs or who engage in sex work.

Harm reduction is an evidence-based approach that could promote health equity through the implementation of pragmatic strategies for people who use drugs or who engage in sex work. The purposes of this study were to describe whether nurses received harm reduction education in their nursing programs, whether nurses are practicing harm reduction, and strategies for integrating harm reduction within nursing curriculum. An online survey was delivered to nurses and an interview with a nurse harm reductionist was conducted. Among the 30 survey participants, only 17% (n = 5) stated they were taught about harm reduction in their nursing programs. On average, survey participants practiced harm reduction less than "half of the time" by meeting people where they are in their drug use or sex work. The nurse harm reductionist described how harm reduction should be taught as a skillset for evidence-based nursing practice. By incorporating harm reduction principles into nursing education and nursing practice, nurses can provide competent care for people who use drugs or who engage in sex work.

Overview of stab injuries in Far North Queensland: A new insight into the demographics, injury patterns and management.

BACKGROUND: Cairns Hospital is the northernmost tertiary referral hospital in Far North Queensland (FNQ) and manages trauma from a large catchment area. A large burden of stab injuries occurs in at-risk patient groups, such as Indigenous and mental health patients, in this region. This research aims to present an overview of the demographics, injury patterns, management and outcomes for stabbings injuries in FNQ. METHODS: A five-year retrospective single-centre study of all patients treated for neck, torso or junctional stab wounds in Far North Queensland was performed searching for all patients with a coded diagnosis of stabbing or knife injury from 1 March 2016 to 31 March 2021. RESULTS: 214 knife injuries were identified and 50.5% of those injured identified as Aboriginal and/or Torres Strait Islander. Stabbing injury locations were most commonly the abdomen/flank/pelvis (n = 81) and the chest/thorax (n = 77). Two-thirds of injuries that breached abdominal fascia had concurrent intra-abdominal injury. Hollow viscus injury commonly involved the small bowel (n = 8), colon (n = 5) and stomach (n = 2), whilst the liver was the most frequently injured solid organ (n = 6). There were 19 vascular injuries, excluding the extremities. 89.2% received diagnostic imaging in the emergency department. FAST scan had 76% sensitivity and 100% specificity for intra-abdominal injury at operation. Overall, 35% of patients required an operation. There were only two in-hospital deaths. CONCLUSION: Stab injuries annually in FNQ are comparable to other centres in Australia. Overall injury severity was low, with excellent survival rates and outcomes for patients who reached hospital. Operative intervention rates for abdominal stab wounds were low in FNQ compared to available data and imaging again appears protective against negative laparotomy rate.

Fistulating diverticular disease masquerading as a peri-anal abscess: a laparoscopic approach to management.

We present a rare case of complicated sigmoid diverticulitis presenting as a peri-anal abscess from an extra-sphincteric fistulous tract. This presentation of a colocutaneous peri-anal abscess is extremely rare, with only a handful of cases described in the literature. Most are managed with an open sigmoid colectomy, however, this case was successfully managed laparoscopically. It highlights the need to consider extra-levator causes of peri-anal abscess, such as pelvic sepsis causing fistulating disease, and to consider early magnetic resonance imaging if there is clinical suspicion of underlying pathology. It also demonstrates that a safe and potentially less morbid outcome is possible via laparoscopic approach when compared to traditional open surgical approach.

Papua New Guinea agri-food trade and household consumption trends point towards dietary change and increased overweight and obesity prevalence.

BACKGROUND: Papua New Guinea (PNG) experienced positive GDP growth at approximately 4.3% per year during the last decade. With increases in overall wealth within the country, PNG is facing a double burden of malnutrition: comparatively high child stunting rates and a growing overweight and obesity epidemic. We focus on the latter by evaluating trends in agri-food import data from 2001 to 2018 and household consumption data from 2018 and 2009/10. RESULTS: The analysis presented in this paper raises three red flags. First, international food import data suggest that the demand for ultra-processed, sugar-sweetened beverages and food have increased substantially over time in PNG. Sugar-sweetened beverages dominated the largest growth in processed food imports, increasing by 23% per capita per year between 2001 and 2018. Second, households across the country with a greater food expenditure on sugar-sweetened beverages have a higher probability of an overweight child (under 5 years old). Last, the probability of soft-drink consumption in PNG increases with greater income acquisition and improved market access. While the price of a soft drink is negatively correlated with the quantity consumed, analysis suggests that total household income has a quantitatively larger (and positive) association with soft drink consumption. CONCLUSIONS: Taxing (or increasing taxes on) sugar-sweetened beverages may not be a sufficient policy mechanism to curb overconsumption of soft drinks in PNG. Education and advocacy programs should be fostered that integrate improved dietary information on packaging, as well as greater access to and understanding of nutrition and diet information of common household consumption items. While increases in household income and market access are crucial to economic growth and transformation, PNG's economic transition must be dovetailed with programs that expand and enhance health and nutrition information and education to improve household consumption decisions and overall household wellbeing.

Impacts of COVID-19 induced income and rice price shocks on household welfare in Papua New Guinea: Household model estimates.

Concerns over the potential effects of the COVID-19 pandemic have led to trade restrictions by major rice exporters, contributing to an average 25% increase in Thai and Vietnamese rice export prices between December 2019 and March-September 2020. This article assesses the consequences of these rice price increases in Papua New Guinea (PNG), where 99% of rice is imported. Utilizing data from a PNG 2018 rural household survey along with earlier national household survey data, we examine rice consumption patterns in PNG and estimate demand parameters for urban and rural households. Model simulations indicate that a 25% rise in the world price of rice would reduce total rice consumption in PNG by 14% and reduce rice consumption of the poor (bottom 40% of total household expenditure distribution) by 15%. Including the effects of a possible 12% decrease in household incomes because of the COVID-19 related economic slowdown, rice consumption of the urban and rural poor fall by 20% and 17%, respectively. Maintaining functioning domestic supply chains of key staple goods is critical to mitigating the effects of global rice price increases, allowing urban households to increase their consumption of locally produced staples.

Unusual source of fever: the curious case of an infected arterial stent graft presenting as a ruptured popliteal artery aneurysm.

A 71-year-old man presented to the emergency department with a 1-week history of lethargy, general malaise and intermittent high fever. He had presented 18 months ago, and again 12 months earlier with similar symptoms. On this third presentation, the fever was accompanied by right thigh swelling and pain. The patient was referred to the vascular surgeons with concern regarding an infected vascular stent graft from previous treatment of popliteal artery aneurysms (PAA) 8 years earlier. CT angiogram demonstrated a collapsed right PAA sac with a large collection, consistent with ruptured PAA, and a single gas bubble around the indwelling stent graft. The patient recovered well after a course of antibiotics and surgical explantation of the graft. Infection is a potential complication of any implanted arterial advice, and needs to be considered even years after initial intervention and with no localising symptoms.

Limits to commercially iodized salt to address dietary iodine deficiency in rural Papua New Guinea.

BACKGROUND AND OBJECTIVES: Fortifying commercial table salt with iodine is the principal strategy used globally to prevent dietary iodine deficiency. However, the costs of providing fortified salt to remote communities may result in it not being locally available or too expensive for many households. This study shows that barriers to consuming adequately iodized salt remain significant for remote rural households in Papua New Guinea (PNG). METHODS AND STUDY DESIGN: Using data from a rural household survey conducted in four areas of PNG in 2018, two issues are examined. First, we contrast the characteristics of households that reported consuming or not consuming iodized table salt, respectively. Second, the adequacy of the iodine content of samples of table salt consumed was assessed in the laboratory. RESULTS: Nine percent of the 1,026 survey households reported not consuming iodized table salt. These households tend to live in remote communities, are among the poorest households, have received no formal education, and have experienced recent food insecurity. Second, 17 percent of the 778 salt samples tested had inadequate iodine. The brand of salt most commonly consumed had the highest share of samples with inadequate iodine levels. CONCLUSIONS: Particularly in remote communities, ensuring that individuals consume sufficient iodine will require going beyond salt iodization to use other approaches to iodine supplementation. To ensure that the iodine intake of those using commercial table salt is adequate, closer monitoring of the iodine content in table salt produced or imported into PNG and enforcement of salt iodization regulations is required.

A small molecule G6PD inhibitor reveals immune dependence on pentose phosphate pathway.

Glucose is catabolized by two fundamental pathways, glycolysis to make ATP and the oxidative pentose phosphate pathway to make reduced nicotinamide adenine dinucleotide phosphate (NADPH). The first step of the oxidative pentose phosphate pathway is catalyzed by the enzyme glucose-6-phosphate dehydrogenase (G6PD). Here we develop metabolite reporter and deuterium tracer assays to monitor cellular G6PD activity. Using these, we show that the most widely cited G6PD antagonist, dehydroepiandosterone, does not robustly inhibit G6PD in cells. We then identify a small molecule (G6PDi-1) that more effectively inhibits G6PD. Across a range of cultured cells, G6PDi-1 depletes NADPH most strongly in lymphocytes. In T cells but not macrophages, G6PDi-1 markedly decreases inflammatory cytokine production. In neutrophils, it suppresses respiratory burst. Thus, we provide a cell-active small molecule tool for oxidative pentose phosphate pathway inhibition, and use it to identify G6PD as a pharmacological target for modulating immune response.

Women's experience of induction of labor using PGE2 as an inpatient versus balloon catheter as an outpatient.

OBJECTIVE: Induction of labor (IOL) typically involves cervical priming in an inpatient setting. Outpatient cervical priming may be a safe and cost-effective alternative. However, little is known about women's preference and the impact of outpatient cervical priming on their healthcare experience. The objective was to compare women's healthcare experiences following IOL using a balloon catheter and going home, versus prostaglandin (PG) and remaining an inpatient. STUDY DESIGN: A randomized controlled trial was undertaken across eight Australian maternity hospitals. Between September 2015 and October 2018, 695 women with uncomplicated term singleton pregnancies were randomized. Of these, 215 and 233 women in the balloon-outpatient and PG-inpatient groups, respectively, received the allocated intervention. The PG group received Dinoprostone gel or controlled-release tape. The balloon group had a double-balloon catheter inserted and went home. Experiential and quality-of-life outcomes were measured via written questionnaire after birth. The primary outcome was a composite neonatal measure. Women's healthcare experience, health-state (EQ-5D-3 L) and pain scores are reported here. RESULTS: Questionnaire data were available for 366 (81.7 %) women enrolled who received their treatment allocation. More women in the balloon-outpatient group reported they would choose IOL next pregnancy (49.2 % vs 38.4 %; p = 0.037) and desire the same method (72.4 % vs 61.1 %; p = 0.022). The balloon-outpatient group experienced higher pain scores at the start of IOL (median (IQR) 3(2-5) vs 2(1-4); p = 0.002) but lower scores at time of rupture of membranes (3(1-5) vs 4(2-6); p = 0.007). The EQ-5D-3 L health-utility index did not differ significantly between the groups (0.77 vs 0.78; p = 0.899). CONCLUSIONS: Women report similar healthcare experiences following balloon-outpatient compared to PG-impatient IOL, but are more likely to desire the same method next pregnancy if IOL is required. If both options are available, then differences in experience should be shared with women, alongside differences in clinical outcomes as part of their decision-making process.

Delayed Respiratory Depression Secondary to Opioid Overdose After Multimodal Analgesia With Transversus Abdominis Plane Blocks in Postanesthesia Care Unit: A Case Report.

Pain control after open abdominal surgery often includes multimodal analgesia with thoracic epidural or transversus abdominis plane (TAP) block. After liposomal bupivacaine was approved for TAP blocks in 2015, it became an alternative to indwelling catheters. However, the pharmacokinetics and safety of its use during the perioperative period have not been thoroughly investigated, especially in conjunction with parenteral opioids. We present a case report of an elderly patient having urgent laparoscopic converted to open abdominal surgery, who experienced postoperative respiratory depression in the recovery room after multimodal therapy with liposomal bupivacaine TAP blocks, intravenous (IV) opioids, and ketorolac.

Rapid Thermostabilization of Bacillus thuringiensis Serovar Konkukian 97-27 Dehydroshikimate Dehydratase through a Structure-Based Enzyme Design and Whole Cell Activity Assay.

Thermostabilization of an enzyme with complete retention of catalytic efficiency was demonstrated on recombinant 3-dehydroshikimate dehydratase (DHSase or wtAsbF) from Bacillus thuringiensis serovar konkukian 97-27 (hereafter, B. thuringiensis 97-27). The wtAsbF is relatively unstable at 37 °C, in vitro (t1/237 = 15 min), in the absence of divalent metal. We adopted a structure-based design to identify stabilizing mutations and created a combinatorial library based upon predicted mutations at specific locations on the enzyme surface. A diversified asbF library (∼2000 variants) was expressed in E. coli harboring a green fluorescent protein (GFP) reporter system linked to the product of wtAsbF activity (3,4-dihydroxybenzoate, DHB). Mutations detrimental to DHSase function were rapidly eliminated using a high throughput fluorescence activated cell sorting (FACS) approach. After a single sorting round and heat screen at 50 °C, a triple AsbF mutant (Mut1), T61N, H135Y, and H257P, was isolated and characterized. The half-life of Mut1 at 37 °C was >10-fold higher than the wtAsbF (t1/237 = 169 min). Further, the second-order rate constants for both wtAsbF and Mut1 were approximately equal (9.9 × 105 M-1 s-1, 7.8 × 105 M-1 s-1, respectively), thus demonstrating protein thermostability did not come at the expense of enzyme thermophilicity. In addition, in vivo overexpression of Mut1 in E. coli resulted in a ∼60-fold increase in functional enzyme when compared to the wild-type enzyme under the identical expression conditions. Finally, overexpression of the thermostable AsbF resulted in an approximate 80-120% increase in DHB accumulation in the media relative to the wild-type enzyme.

Beyond Helper Phage: Using "Helper Cells" to Select Peptide Affinity Ligands.

Peptides are important affinity ligands for microscopy, biosensing, and targeted delivery. However, because they can have low affinity for their targets, their selection from large naïve libraries can be challenging. When selecting peptidic ligands from display libraries, it is important to: 1) ensure efficient display; 2) maximize the ability to select high affinity ligands; and 3) minimize the effect of the display context on binding. The "helper cell" packaging system has been described as a tool to produce filamentous phage particles based on phagemid constructs with varying display levels, while remaining free of helper phage contamination. Here we report on the first use of this system for peptide display, including the systematic characterization and optimization of helper cells, their inefficient use in antibody display and their use in creating and selecting from a set of phage display peptide libraries. Our libraries were analyzed with unprecedented precision by standard or deep sequencing, and shown to be superior in quality than commercial gold standards. Using our helper cell libraries, we have obtained ligands recognizing Yersinia pestis surface antigen F1V and L-glutamine-binding periplasmic protein QBP. In the latter case, unlike any of the peptide library selections described so far, we used a combination of phage and yeast display to select intriguing peptide ligands. Based on the success of our selections we believe that peptide libraries obtained with helper cells are not only suitable, but preferable to traditional phage display libraries for selection of peptidic ligands.

High-Capacity and Rapid Removal of Refractory NOM Using Nanoscale Anion Exchange Resin.

As human health concerns over disinfection byproducts (DBP) in drinking water increase, so does the need to develop new materials that remove them rapidly and at high capacity. Ion exchange (IEX) is an effective method for the removal of natural organic matter (NOM), especially anion exchange resins (AERs) with quaternary ammonium functional groups. However, capacity is limited in existing commercial resin materials because adsorbates can only interact with the outermost surface area, which makes these products inefficient on a mass basis. We have synthesized a novel "NanoResin" exploiting the enhanced NOM removal of the quaternary ammonium resin while utilizing the vast surface area of SWCNTs, which act as scaffolding for the resin. Our nanomaterials show increased adsorption capacity compared to commercially available adsorbents, in a fraction of the time. This NanoResin requires only about 10 s to reach ion-exchange equilibrium. Comparatively, commercial AERs only achieved partial removal after more than 30 min. High capacity adsorption of a low molecular weight (MW) surrogate has been measured. NOM removal was demonstrated in solutions of both low and high specific UV absorbance (SUVA) composition with these nanomaterials. Additionally, the NanoResin showed enhanced removal of a NOM concentrate sample taken from Myrtle Beach, SC, demonstrating NanoResin is an effective method of removal for refractory NOM in a natural aqueous environment. Synthesis and characterization of the polymers and nanomaterials are presented below. Adsorption capacity, adsorption kinetics, and the regeneration and reusability of these new materials for NOM removal are described. The open matrix microstructure precludes any intraparticle diffusion of adsorbates; thus, these nanomaterials act as a "contact resin".

Achalasia in Pregnancy: Botulinum Toxin A Injection of Lower Esophageal Sphincter.

Background. Achalasia, a rare esophageal motility disorder that may cause malnutrition during pregnancy, can result in fetal and maternal morbidity and mortality. Many medical treatment regimens are contraindicated or not tolerated during pregnancy, and surgery is generally avoided due to potential risks to the fetus. Case Report. Severe, medically refractory achalasia in a 23-year-old pregnant woman that caused malnutrition was successfully managed by administering a botulinum toxin A injection to the lower esophageal sphincter. The injection was performed at approximately 14 weeks' gestation and the patient reported clinically significant relief from dysphagia. She gained weight and ultimately delivered a healthy baby girl at term, but her symptoms returned a few months postpartum. She underwent a second treatment of botulinum toxin A injection, but it offered only one month of relief. Roughly eight months after delivery, the patient underwent a laparoscopic extended Heller myotomy and Dor fundoplication. The patient resumed a normal diet one week postoperatively, and her baby has had no complications. Conclusion. This is only the second reported case of botulinum toxin A injection being used to treat achalasia in pregnancy. This treatment proved to be a safe temporary alternative without the risks of surgery and anesthesia during pregnancy.

Concern for Another's Distress in Toddlers at High and Low Genetic Risk for Autism Spectrum Disorder.

We examined concern for others in 22-month-old toddlers with an older sibling with autism spectrum disorder (ASD) and low risk typically-developing toddlers with older siblings. Responses to a crying infant and an adult social partner who pretended to hurt her finger were coded. Children with a later diagnosis of ASD showed limited empathic concern in either context compared to low risk toddlers. High risk toddlers without a later diagnosis fell between the ASD and low risk groups. During the crying baby probe the low risk and high risk toddlers without a diagnosis engaged their parent more often than the toddlers with ASD. Low levels of empathic concern and engagement with parents may signal emerging ASD in toddlerhood.

CTLA-4 controls follicular helper T-cell differentiation by regulating the strength of CD28 engagement.

Cytotoxic T-lymphocyte-associated antigen-4 (CTLA-4) is an essential regulator of T-cell responses, and its absence precipitates lethal T-cell hyperactivity. However, whether CTLA-4 acts simply to veto the activation of certain clones or plays a more nuanced role in shaping the quality of T-cell responses is not clear. Here we report that T cells in CTLA-4-deficient mice show spontaneous T-follicular helper (T(FH)) differentiation in vivo, and this is accompanied by the appearance of large germinal centers (GCs). Remarkably, short-term blockade with anti-CTLA-4 antibody in wild-type mice is sufficient to elicit T(FH) generation and GC development. The latter occurs in a CD28-dependent manner, consistent with the known role of CTLA-4 in regulating the CD28 pathway. CTLA-4 can act by down-regulating CD80 and CD86 on antigen presenting cells (APCs), thereby altering the level of CD28 engagement. To mimic reduced CD28 ligation, we used mice heterozygous for CD28, revealing that the magnitude of CD28 engagement is tightly linked to the propensity for T(FH) differentiation. In contrast, other parameters of T-cell activation, including CD62L down-regulation and Ki67 expression, were relatively insensitive to altered CD28 level. Altered T(FH) generation as a result of graded reduction in CD28 was associated with decreased numbers of GC B cells and a reduction in overall GC size. These data support a model in which CTLA-4 control of immunity goes beyond vetoing T-cell priming and encompasses the regulation of T(FH) differentiation by graded control of CD28 engagement.

Social engagement with parents in 11-month-old siblings at high and low genetic risk for autism spectrum disorder.

Infant siblings of children with an autism spectrum disorder are at heightened genetic risk to develop autism spectrum disorder. We observed high risk (n = 35) and low risk (n = 27) infants at 11 months during free play with a parent. Children were assessed for autism spectrum disorder in toddlerhood. High-risk infants with a later diagnosis (n = 10) were less socially engaged with their parents than were low-risk infants. Parent behavior during play did not vary by group. Within the high-risk group, ratings of social reciprocity at 11 months predicted Autism Diagnostic Observation Schedule severity scores at follow-up, suggesting that systematic observations of parent-infant play may be a useful addition to early assessments of emerging autism spectrum disorder.

A novel upstream enhancer of FOXP3, sensitive to methylation-induced silencing, exhibits dysregulated methylation in rheumatoid arthritis Treg cells.

Treg-cell function is compromised in rheumatoid arthritis (RA). As the master regulator of Treg cells, FOXP3 controls development and suppressive function. Stable Treg-cell FOXP3 expression is epigenetically regulated; constitutive expression requires a demethylated Treg-specific demethylated region. Here, we hypothesised that methylation of the FOXP3 locus is altered in Treg cells of established RA patients. Methylation analysis of key regulatory regions in the FOXP3 locus was performed on Treg cells from RA patients and healthy controls. The FOXP3 Treg-specific demethylated region and proximal promoter displayed comparable methylation profiles in RA and healthy-donor Treg cells. We identified a novel differentially methylated region (DMR) upstream of the FOXP3 promoter, with enhancer activity sensitive to methylation-induced silencing. In RA Treg cells we observed significantly reduced DMR methylation and lower DNA methyltransferase (DNMT1/3A) expression compared with healthy Treg cells. Furthermore, DMR methylation negatively correlated with FOXP3 mRNA expression, and Treg cells isolated from rheumatoid factor negative RA patients were found to express significantly higher levels of FOXP3 than Treg cells from RhF-positive patients, with an associated decrease in DMR methylation. In conclusion, the novel DMR is involved in the regulation of Treg-cell FOXP3 expression, but this regulation is lost post-transcriptionally in RA Treg cells.