RHAMM regulates MMTV-PyMT-induced lung metastasis by connecting STING-dependent DNA damage sensing to interferon/STAT1 pro-apoptosis signaling.

BACKGROUND: RHAMM is a multifunctional protein that is upregulated in breast tumors, and the presence of strongly RHAMM+ve cancer cell subsets associates with elevated risk of peripheral metastasis. Experimentally, RHAMM impacts cell cycle progression and cell migration. However, the RHAMM functions that contribute to breast cancer metastasis are poorly understood. METHODS: We interrogated the metastatic functions of RHAMM using a loss-of-function approach by crossing the MMTV-PyMT mouse model of breast cancer susceptibility with Rhamm-/- mice. In vitro analyses of known RHAMM functions were performed using primary tumor cell cultures and MMTV-PyMT cell lines. Somatic mutations were identified using a mouse genotyping array. RNA-seq was performed to identify transcriptome changes resulting from Rhamm-loss, and SiRNA and CRISPR/Cas9 gene editing was used to establish cause and effect of survival mechanisms in vitro. RESULTS: Rhamm-loss does not alter initiation or growth of MMTV-PyMT-induced primary tumors but unexpectedly increases lung metastasis. Increased metastatic propensity with Rhamm-loss is not associated with obvious alterations in proliferation, epithelial plasticity, migration, invasion or genomic stability. SNV analyses identify positive selection of Rhamm-/- primary tumor clones that are enriched in lung metastases. Rhamm-/- tumor clones are characterized by an increased ability to survive with ROS-mediated DNA damage, which associates with blunted expression of interferon pathway and target genes, particularly those implicated in DNA damage-resistance. Mechanistic analyses show that ablating RHAMM expression in breast tumor cells by siRNA knockdown or CRISPR-Cas9 gene editing blunts interferon signaling activation by STING agonists and reduces STING agonist-induced apoptosis. The metastasis-specific effect of RHAMM expression-loss is linked to microenvironmental factors unique to tumor-bearing lung tissue, notably high ROS and TGFB levels. These factors promote STING-induced apoptosis of RHAMM+ve tumor cells to a significantly greater extent than RHAMM-ve comparators. As predicted by these results, colony size of Wildtype lung metastases is inversely related to RHAMM expression. CONCLUSION: RHAMM expression-loss blunts STING-IFN signaling, which offers growth advantages under specific microenvironmental conditions of lung tissue. These results provide mechanistic insight into factors controlling clonal survival/expansion of metastatic colonies and has translational potential for RHAMM expression as a marker of sensitivity to interferon therapy.

Soil bacterial community dynamics in plots managed with cover crops and no-till farming in the Lower Mississippi Alluvial Valley, USA.

AIMS: Assess bacterial community changes over time in soybean (Glycine max) crop fields following cover crop (CC) and no-till (NT) implementation under natural abiotic stressors. METHOD AND RESULTS: Soil bacterial community composition was obtained by amplifying, sequencing, and analysing the V4 region of the 16S rRNA gene. Generalized linear mixed models were used to assess the effects of tillage, CC, and time on bacterial community response. The most abundant phyla present were Acidobacteria, Actinobacteria, Bacteroidetes, and Verrucomicrobia. Bacterial diversity increased in periods with abundant water. Reduced tillage (RT) increased overall bacterial diversity, but NT with a CC was not significantly different than RT treatments under drought conditions. CCs shifted abundances of Firmicutes and Bacteroidetes depending on abiotic conditions. CONCLUSIONS: In the Lower Mississippi Alluvial Valley (LMAV), USA, NT practices lower diversity and influence long-term community changes while cover crops enact a seasonal response to environmental conditions. NT and RT management affect soil bacterial communities differently than found in other regions of the country.

Can Pesticides Dissolved in Runoff and Exposed to Maturing Rice (Oryza sativa) Plants be Transferred to Seeds?

Agriculture's global challenge to feed an estimated 7.7 billion people is further exacerbated by less available cropland for production and rapidly changing climate patterns. Pesticides are often utilized to minimize crop losses due to pest infestations; however, problems arise when these chemicals are transported off production acreage, either by storm or irrigation events, and into nearby water bodies. Innovative management practices are needed to not only reduce the volume of runoff, but also to mitigate various pollutants, such as pesticides, within the runoff. One such practice being evaluated involves using rice (Oryza sativa) as a pesticide mitigation tool. While rice plants may serve as a mechanism for phytoremediation, whether the seeds harvested from exposed plants could then be utilized as a human food source is an unanswered question. Thirty round mesocosms (55 L volume; 56 cm diameter; six replicates per treatment) were established with rice and exposed to aqueous concentrations of the pesticides clomazone, propanil, or cyfluthrin, as well as a mixture of the three pesticides. Six replicates with rice and no pesticide exposure served as controls. Initial pesticide exposure took place 8 weeks post-planting and continued once a week for 5 weeks. Rice plants, unmilled seeds, and mesocosm sediment were collected from each mesocosm 2 weeks after seed formation began and analyzed for pesticide concentrations using gas chromatography. Concentrations of pesticides in unmilled seed were below detection for individual exposures of clomazone, propanil, and cyfluthrin. When rice was exposed to the pesticide mixture, the mean ± SE unmilled seed cyfluthrin concentration was 14.8 ± 1.25 µg kg-1. These small-scale, preliminary studies offer insight into the possibility of using immature rice plants as a phytoremediation tool, while harvesting its grain after plant maturation for human consumption. Further research is needed to address this question on a larger scale and with multiple pesticide mixtures.

Increased urokinase and consumption of α2 -antiplasmin as an explanation for the loss of benefit of tranexamic acid after treatment delay.

Essentials Delayed treatment with tranexamic acid results in loss of efficacy and poor outcomes. Increasing urokinase activity may account for adverse effects of late tranexamic acid treatment. Urokinase + tranexamic acid produces plasmin in plasma or blood and disrupts clotting. α2 -Antiplasmin consumption with ongoing fibrinolysis increases plasmin-induced coagulopathy. SUMMARY: Background Tranexamic acid (TXA) is an effective antifibrinolytic agent with a proven safety record. However, large clinical trials show TXA becomes ineffective or harmful if treatment is delayed beyond 3 h. The mechanism is unknown but urokinase plasminogen activator (uPA) has been implicated. Methods Inhibitory mechanisms of TXA were explored in a variety of clot lysis systems using plasma and whole blood. Lysis by tissue plasminogen activator (tPA), uPA and plasmin were investigated. Coagulopathy was investigated using ROTEM and activated partial thromboplastin time (APTT). Results IC50 values for antifibrinolytic activity of TXA varied from < 10 to > 1000 µmol L-1 depending on the system, but good fibrin protection was observed in the presence of tPA, uPA and plasmin. However, in plasma or blood, active plasmin was generated by TXA + uPA (but not tPA) and coagulopathy developed leading to no or poor clot formation. The extent of coagulopathy was sensitive to available α2 -antiplasmin. No clot formed with plasma containing 40% normal α2 -antiplasmin after short incubation with TXA + uPA. Adding purified α2 -antiplasmin progressively restored clotting. Plasmin could be inhibited by aprotinin, IC50 = 530 nmol L-1 , in plasma. Conclusions Tranexamic acid protects fibrin but stimulates uPA activity and slows inhibition of plasmin by α2 -antiplasmin. Plasmin proteolytic activity digests fibrinogen and disrupts coagulation, exacerbated when α2 -antiplasmin is consumed by ongoing fibrinolysis. Additional direct inhibition of plasmin by aprotinin may prevent development of coagulopathy and extend the useful time window of TXA treatment.

Advancing the Sustainability of US Agriculture through Long-Term Research.

Agriculture in the United States must respond to escalating demands for productivity and efficiency, as well as pressures to improve its stewardship of natural resources. Growing global population and changing diets, combined with a greater societal awareness of agriculture's role in delivering ecosystem services beyond food, feed, fiber, and energy production, require a comprehensive perspective on where and how US agriculture can be sustainably intensified, that is, made more productive without exacerbating local and off-site environmental concerns. The USDA's Long-Term Agroecosystem Research (LTAR) network is composed of 18 locations distributed across the contiguous United States working together to integrate national and local agricultural priorities and advance the sustainable intensification of US agriculture. We explore here the concept of sustainable intensification as a framework for defining strategies to enhance production, environmental, and rural prosperity outcomes from agricultural systems. We also elucidate the diversity of factors that have shaped the past and present conditions of cropland, rangeland, and pastureland agroecosystems represented by the LTAR network and identify priorities for research in the areas of production, resource conservation and environmental quality, and rural prosperity. Ultimately, integrated long-term research on sustainable intensification at the national scale is critical to developing practices and programs that can anticipate and address challenges before they become crises.

Can Rice (Oryza sativa) Mitigate Pesticides and Nutrients in Agricultural Runoff?

Phytoremediation of nutrients and pesticides in runoff is a growing conservation effort, particularly in agriculturally intensive areas such as the lower Mississippi River Valley. In the current study, rice (Oryza sativa) was examined for its mitigation capacity of nitrogen, phosphorus, diazinon, and permethrin. Twenty-two high density polyethylene circular containers (56 cm x 45 cm) were used as mesocosms, with 12 mesocosms planted with rice and 10 mesocosms remaining unvegetated. Mesocosms were hydraulically connected and arranged in a series of two, with each system providing a 4 h hydraulic retention time (HRT) for a total system retention time of 8 h. Two treatments (RICE/RICE and RICE/BARE) of four replicates each were utilized, with three replicates of controls (BARE/BARE). Systems with RICE/RICE (8 h HRT) significantly reduced diazinon (p = 0.0126), cis-permethrin (p = 0.0442), filtered orthophosphate (p = 0.0058), and total orthophosphate (p = 0.0123) compared to control systems. No significant differences were noted for trans-permethrin, nitrate, or ammonium. Results indicate promise in phytoremediation of agricultural runoff by rice. If further studies reveal contaminants are not transferred into seeds, then rice could potentially serve as both a remediation tool and food source in countries facing agricultural pollution challenges.

Salmonella Enteritidis with double deletion in phoP fliC and a competitive exclusion culture elicit substantial additive protective effects against Salmonella exposure in newly hatched chicks.

A live Salmonella Enteritidis vaccine (SE147N ΔphoP fliC), able to express both a homologous intestinal colonisation-inhibition effect and a systemic invasion-inhibition effect, was tested for its potential to generate a postulated additive protective effect in case of combined application with a competitive exclusion (CE) culture against Salmonella exposure in very young chicks. Both, SE147N ΔphoP fliC and the CE culture alone were highly protective against systemic and intestinal colonisation of the challenge strain in case of moderate Salmonella exposure, consequently, additive protective effects in combined use could not be detected. However, in case of high Salmonella Enteritidis challenge with 106 cfu/bird at day 3 of life the combination of the ΔphoP fliC vaccine and the CE culture resulted in a protective effect much more pronounced than either of the single preparations and most substantial compared to untreated control birds. The term additive protective effects reflects the recognition that exclusion effects by gut flora cultures and inhibition effects by Salmonella vaccines are caused by different mechanisms.

Application of Polyacrylamide (PAM) through Lay-Flat Polyethylene Tubing: Effects on Infiltration, Erosion, N and P Transport, and Corn Yield.

Polyacrylamides (PAMs), when applied as a soil amendment, purportedly improve soil infiltration, decrease erosion, and reduce offsite agrochemical transport. The effect of PAM on infiltration, erosion, agrochemical transport, and crop yield when applied in furrow to mid-southern US production systems has not been evaluated. The objective of this study was to assess PAM effects on infiltration, erosion, corn ( L.) grain yield, and nitrogen (N) and phosphorus (P) transport when applied at 10 mg L through lay-flat polyethylene tubing. A 2-yr field study was conducted at the Mississippi State Delta Research and Extension Center in Stoneville, MS, on a Dundee silt loam and a Forestdale silty clay loam. The experimental design was a randomized complete block with four replications of each treatment: irrigated plus no PAM (control) and irrigated plus PAM at 10 mg L. Each irrigation event delivered 102 mm of water at 18.9 L m per furrow, and runoff was captured in a holding tank on the lower end of each plot. Pooled over year and soil texture, PAM increased infiltration and corn grain yield by 6% ( ≤ 0.0398). Polyacrylamide effects on the offsite transport of sediment and N and P were inconsistent, varying across year and soil texture. Results indicate that PAM improves infiltration and corn grain yield on silt loam and silty clay loam textured soils; however, further research is required before PAM can be recommended as a best management practice for mitigating erosion and offsite agrochemical transport in mid-southern production systems.

Pyoderma gangrenosum with pathergy: A potentially significant complication following breast reconstruction.

The failure of postoperative surgical site infection to resolve after appropriate antibiotic therapy should alert the clinician to other diagnoses. Pyoderma gangrenosum (PG) is an inflammatory neutrophilic dermatosis that is typically characterized by necrotizing ulceration. PG can be exacerbated by minor trauma leading to exaggerated skin injury, a condition known as pathergy. We present a case series of PG arising after immediate reconstruction for breast oncological surgery from 1st January 2006 to 1st September 2014. 395 immediate breast reconstructions were performed in 335 patients. Three cases of post-surgical PG were identified (0.9%), all in the setting of mastectomy for breast cancer. Two cases underwent immediate reconstruction with pedicled transverse rectus abdominus myocutaneous flaps, and one underwent submuscular expander insertion. A mean delay of 6.3 days was observed from first presentation of symptoms to diagnosis of PG. Immunosuppressants commonly used included methylprednisolone, prednisone, and ciclosporin, with good success at halting disease progress. Significant scarring affected all three women. Once the disease was deemed quiescent, intravenous immunoglobulin used in the perioperative period for further surgical procedures provided favorable results. A diagnostic algorithm is suggested to guide surgeons in investigations and management when post-surgical PG is suspected.

A survey of dental practitioners in Wales to evaluate the management of deep carious lesions with vital pulp therapy in permanent teeth.

Objective To evaluate the management of deep carious lesions with vital pulp therapy in permanent teeth by dental practitioners within Wales.Design Postal questionnaire.Setting General practitioners (GDS), community (CDS) and hospital-based dentists (HDS) in Wales.Methods Community and hospital dental services with a remit for provision of restorative dentistry (CDS = 71; HDS = 46) and general dental practitioners (N = 510) were approached regarding their management of deep carious lesions with vital pulp therapy in permanent teeth. The postal questionnaire took the form of an anonymous survey. Questions covered usage parameters, training issues and reasons for material choice.Results The response rate was 29%. The majority of HDS (89%) used MTA or Biodentine for vital pulp therapy in contrast to GDS (41%) and CDS (32%). The main reasons cited for avoiding the use of MTA or Biodentine included cost, lack of training and difficulty in material handling.Conclusion Usage of MTA or Biodentine for vital pulp therapies is low in the general dental and community dental settings. Cost and lack of training are the main barriers for the uptake of these materials. Postgraduate training may be useful in addressing these barriers. Increasing their adoption would be advantageous as they have been shown to produce a more predictable outcome compared to traditional materials (for example, calcium hydroxide).

Using aquatic vegetation to remediate nitrate, ammonium, and soluble reactive phosphorus in simulated runoff.

Within the agriculturally-intensive Mississippi River Basin of the United States, significant conservation efforts have focused on management practices that reduce nutrient runoff into receiving aquatic ecosystems. Only a small fraction of those efforts have focused on phytoremediation techniques. Each of six different aquatic macrophytes were planted, in monoculture, in three replicate mesocosms (1.2 m × 0.15 m × 0.65 m). Three additional unvegetated mesocosms served as controls for a total number of 21 mesocosms. Over two years, mesocosms were amended once each summer with sodium nitrate, ammonium sulfate, and potassium phosphate dibasic to represent nitrogen and phosphorus in agricultural runoff. System retention was calculated using a simple aqueous mass balance approach. Ammonium retention in both years differed greatly, as Panicum hemitomon and Echinodorus cordifolius retentions were significantly greater than controls in the first year, while only Myriophyllum aquaticum and Typha latifolia were significantly greater than controls in the second year. Greater soluble reactive phosphorus retention was observed in T. latifolia compared to controls in both years. Several other significant differences were observed in either the first or second year, but not both years. In the first year's exposure, P. hemitomon was significantly more efficient than the control, Saururus cernuus, and T. latifolia for overall percent nitrate decrease. Results of this novel study highlight inherent variability within and among species for nutrient specific uptake and the temporal variations of species for nutrient retention. By examining this natural variability, scientists may design phytoremediation systems with greater impact on improving agricultural runoff water quality.

Drying and Storage Methods Affect Cyfluthrin Concentrations in Exposed Plant Samples.

Standard procedures do not exist for drying and storage of plant samples prior to chemical analyses. Since immediate analysis is not always possible, current research examined which plant drying and storage method yielded the highest cyfluthrin recovery rates compared to traditional mechanical freeze-drying methods. Fifteen mesocosms were planted with rice. Cyfluthrin (5 mg L(-1)) was amended into the water column of individual mesocosms. 48 h later, plant material in the water column was collected from each mesocosm. Control (mechanical freeze drying) recovery was significantly greater (p < 0.001) than all 14 combinations of drying and storage. Significant differences also existed between all 14 different combinations. Greatest cyfluthrin recoveries in non-control plants were from the freezer-greenhouse-freezer drying and storage method. Results offer evidence for the efficient plant drying and storage methods prior to cyfluthrin analysis. Future studies should perform comparable analyses on various pesticide classes to determine possible relationships.

Genomic copy number variation in Mus musculus.

BACKGROUND: Copy number variation is an important dimension of genetic diversity and has implications in development and disease. As an important model organism, the mouse is a prime candidate for copy number variant (CNV) characterization, but this has yet to be completed for a large sample size. Here we report CNV analysis of publicly available, high-density microarray data files for 351 mouse tail samples, including 290 mice that had not been characterized for CNVs previously. RESULTS: We found 9634 putative autosomal CNVs across the samples affecting 6.87% of the mouse reference genome. We find significant differences in the degree of CNV uniqueness (single sample occurrence) and the nature of CNV-gene overlap between wild-caught mice and classical laboratory strains. CNV-gene overlap was associated with lipid metabolism, pheromone response and olfaction compared to immunity, carbohydrate metabolism and amino-acid metabolism for wild-caught mice and classical laboratory strains, respectively. Using two subspecies of wild-caught Mus musculus, we identified putative CNVs unique to those subspecies and show this diversity is better captured by wild-derived laboratory strains than by the classical laboratory strains. A total of 9 genic copy number variable regions (CNVRs) were selected for experimental confirmation by droplet digital PCR (ddPCR). CONCLUSION: The analysis we present is a comprehensive, genome-wide analysis of CNVs in Mus musculus, which increases the number of known variants in the species and will accelerate the identification of novel variants in future studies.

Diazinon and permethrin mitigation across a grass-wetland buffer.

Vegetated buffers of different designs are often used as edge-of-field treatment practices to remove pesticides that may be entrained in agricultural runoff. However, buffer system efficacy in pesticide runoff mitigation varies widely due to a multitude of factors including, but not limited to, pesticide chemistry, vegetation composition, and hydrology. Two experimental systems, a control (no vegetation) and a grass-wetland buffer system, were evaluated for their ability to retain diazinon and permethrin associated with a simulated storm runoff. The two systems were equally inefficient at retaining diazinon (mean 9.6 % retention for control and buffer). Grass-wetland buffers retained 83 % and 85 % of cis- and trans-permethrin masses, respectively, while the control only retained 39 % and 44 % of cis- and trans-permethrin masses, respectively. Half-distances (the distance required to decrease pesticide concentration by one-half) for both permethrin isomers were 26 %-30 % shorter in grass buffers (22-23 m) than in the control (32 m). The current study demonstrates treatment efficacy was a function of pesticide properties with the more strongly sorbing permethrin retained to a greater degree. The study also demonstrates challenges in remediating multiple pesticides with a single management practice. By using suites of management practices, especially those employing vegetation, better mitigation of pesticide impacts may be accomplished.

Biological relevance of CNV calling methods using familial relatedness including monozygotic twins.

BACKGROUND: Studies involving the analysis of structural variation including Copy Number Variation (CNV) have recently exploded in the literature. Furthermore, CNVs have been associated with a number of complex diseases and neurodevelopmental disorders. Common methods for CNV detection use SNP, CNV, or CGH arrays, where the signal intensities of consecutive probes are used to define the number of copies associated with a given genomic region. These practices pose a number of challenges that interfere with the ability of available methods to accurately call CNVs. It has, therefore, become necessary to develop experimental protocols to test the reliability of CNV calling methods from microarray data so that researchers can properly discriminate biologically relevant data from noise. RESULTS: We have developed a workflow for the integration of data from multiple CNV calling algorithms using the same array results. It uses four CNV calling programs: PennCNV (PC), Affymetrix® Genotyping Console™ (AGC), Partek® Genomics Suite™ (PGS) and Golden Helix SVS™ (GH) to analyze CEL files from the Affymetrix® Human SNP 6.0 Array™. To assess the relative suitability of each program, we used individuals of known genetic relationships. We found significant differences in CNV calls obtained by different CNV calling programs. CONCLUSIONS: Although the programs showed variable patterns of CNVs in the same individuals, their distribution in individuals of different degrees of genetic relatedness has allowed us to offer two suggestions. The first involves the use of multiple algorithms for the detection of the largest possible number of CNVs, and the second suggests the use of PennCNV over all other methods when the use of only one software program is desirable.

Effect of storage method and associated holding time on nitrogen and phosphorus concentrations in surface water samples.

Assessments were conducted to determine the effect of sample storage method and associated holding time on surface water nutrient concentrations from field sites. Six surface water sites and two nutrient spiked, laboratory water samples were evaluated for nitrate, nitrite, ammonium, filtered orthophosphorus, and total orthophosphorus concentrations on four separate days throughout the period of 1 year. Samples stored at ambient temperature (23°C) for 24 h prior to nutrient analyses resulted in 18 % ± 2 % of results being significantly different from controls (which were analyzed immediately upon collection). Samples placed in the cooler (4°C) for 7 days prior to nutrient analyses resulted in 30 % ± 1 % of values being significantly different from controls. Samples placed in the freezer (-20°C) for 7 days prior to analyses resulted in 34 % ± 12 %, 44 % ± 10 %, and 28 % ± 5.7 % of ammonium, filtered orthophosphate, and total orthophosphate, respectively, values being significantly different from controls. This study highlights the challenges facing researchers in efficient collection, storage and nutrient analysis of samples, especially when sites are remote and difficult to access .

A survey of adoption of endodontic nickel-titanium rotary instrumentation part 1: general dental practitioners in Wales.

OBJECTIVE: To assess adoption of endodontic nickel-titanium (NiTi) rotary technology by general dental practitioners and identify factors influencing its uptake. DESIGN: Postal questionnaire. SETTING: General dental practitioners working in Wales. METHODS: General dental practitioners (n = 584) were approached regarding their usage or otherwise of nickel-titanium rotary instrumentation during root canal shaping. The postal questionnaire took the form of an anonymous survey comprising 13 questions. These questions covered usage parameters, satisfaction, training issues and reasons for avoidance of NiTi instruments. RESULTS: The response rate was 71%. Nickel-titanium rotary instruments were used routinely by 67% of those responding practitioners. Principle factors cited as being implicated in the decision to not adopt NiTi use included cost (65% of responses), lack of training and the perceived risk of instrument fracture. CONCLUSIONS: Over two thirds of dental practitioners in Wales use rotary NiTi endodontic technology with the majority having converted to such systems more than three years ago. There was, however, a significant disparity in NiTi usage between solely NHS practitioners (42%) and private practitioners (90%). Continued provision of high quality hands-on practical workshops may be of benefit in facilitating a positive initial NiTi experience in order to assist the transfer to these newer technologies.

A survey of adoption of endodontic nickel-titanium rotary instrumentation part 2: community and hospital dental practitioners in Wales.

OBJECTIVES: To assess adoption of endodontic nickel-titanium rotary technology (NiTi) by community and hospital dental clinicians within Wales and identify factors that may restrict uptake. DESIGN: Postal questionnaire. SETTING: Community and hospital-based dentists in Wales. METHODS: Community and hospital-based dentists with a remit for provision of restorative dentistry (community dental setting = 32; hospital dental setting = 36) were approached regarding their usage or otherwise of nickel-titanium rotary instrumentation for endodontic treatments. The postal questionnaire took the form of an anonymous survey comprising 12 questions. These questions covered usage parameters, satisfaction and training and broached reasons for NiTi avoidance. RESULTS: The response rate was 77%. NiTi rotary instruments were used routinely by an encouraging 82% of those in the hospital-based restorative dental services but only 13% of community staff. Factors cited as being implicated in the decision to avoid their use included cost (62% of responses) lack of training and the perceived lack of benefit. CONCLUSION: The adoption of rotary NiTi endodontic technology by the hospital dental practitioners of Wales is encouraging with the majority having converted to such systems in excess of three years prior to the survey. There was, however, a significant disparity in NiTi usage between community and hospital settings, the implications for which and possible solutions for increased training and uptake are discussed.