Phase Behavior and Rheological Properties of Size-Fractionated MXene (Ti3C2Tx) Dispersions.

Understanding the dispersion behavior of MXenes is interesting from a fundamental colloid science perspective and critical to enabling the fluid-phase manufacturing of MXene devices with controlled microstructures and properties. However, the polydispersity, irregular shape, and charged surfaces of MXenes result in a complex phase behavior that is difficult to predict through theoretical calculations. As two-dimensional (2D) nanomaterials, MXenes can form lyotropic liquid crystal phases, gels, and aggregates. This work aims to elucidate the effects of MXene (Ti3C2Tx) sheet size on their phase behavior and associated rheological properties. Aqueous dispersions of large sheets with an average lateral dimension of 3.0 µm, small sheets with an average lateral dimension of 0.3 µm, and a bimodal mixture of the two sizes were investigated by using cross-polarized optical microscopy and rheology. At low concentrations, the large MXene dispersions exhibited lyotropic liquid crystal behavior and extended aligned textures, but increasing concentration resulted in the formation of dense flocs. Dispersions of small sheets formed small birefringent domains with increasing concentration but lacked long-range ordering. A bimodal mixture of these sizes enabled the formation of liquid crystalline phases with extended aligned textures with less floc formation. These results provide insights into using polydispersity to tune dispersion microstructure and rheological properties that can be applied to designing dispersions for fluid-phase manufacturing methods, such as direct ink writing.

Short-Term Dorsal Genital Nerve Stimulation Increases Subjective Arousal in Women With and Without Spinal Cord Injury: A Preliminary Investigation.

OBJECTIVES: Female sexual dysfunction (FSD) affects an estimated 40% of women. Unfortunately, FSD is understudied, leading to limited treatment options for FSD. Neuromodulation has shown some success in alleviating FSD symptoms. We developed a pilot study to investigate the short-term effect of electrical stimulation of the dorsal genital nerve and tibial nerve on sexual arousal in healthy women, women with FSD, and women with spinal cord injury (SCI) and FSD. MATERIALS AND METHODS: This study comprises a randomized crossover design in three groups: women with SCI, women with non-neurogenic FSD, and women without FSD or SCI. The primary outcome measure was change in vaginal pulse amplitude (VPA) from baseline. Secondary outcome measures were changes in subjective arousal, heart rate, and mean arterial pressure from baseline. Participants attended one or two study sessions where they received either transcutaneous dorsal genital nerve stimulation (DGNS) or tibial nerve stimulation (TNS). At each session, a vaginal photoplethysmography sensor was used to measure VPA. Participants also rated their level of subjective arousal and were asked to report any pelvic sensations. RESULTS: We found that subjective arousal increased significantly from before to after stimulation in DGNS study sessions across all women. TNS had no effect on subjective arousal. There were significant differences in VPA between baseline and stimulation, baseline and recovery, and stimulation and recovery periods among participants, but there were no trends across groups or stimulation type. Two participants with complete SCIs experienced genital sensations. CONCLUSIONS: To our knowledge, this is the first study to measure sexual arousal in response to short-term neuromodulation in women. This study indicates that short-term DGNS but not TNS can increase subjective arousal, but the effect of stimulation on genital arousal is inconclusive. This study provides further support for DGNS as a treatment for FSD.

Primary and Secondary Organic Aerosol Formation from Asphalt Pavements.

Asphalt is ubiquitous across cities and a source of organic compounds spanning a wide range of volatility and may be an overlooked source of urban organic aerosols. The emission rate and composition depend strongly on temperature, but emissions have been observed at both application temperatures and surface temperatures during warm sunny days. Here we report primary organic aerosol (POA) emissions and secondary organic aerosol (SOA) production from asphalt. We reheated real-world asphalt samples to application-relevant temperatures (∼130 °C) and typical summertime road-surface temperatures (∼55 °C) and then flushed the emitted vapors into an environmental oxidation chamber containing ammonium sulfate seed particles. SOA was then formed following the photo-oxidation of emissions under high-NOx conditions typical of urban atmospheres. We find that POA only forms at application temperature as it does not require further oxidation, whereas SOA forms under both conditions; with the resulting POA and SOA both being semi-volatile. While total OA formation rates were substantially greater under the limited time spent under application conditions, SOA formation from passive asphalt heating presents a potential long-term source, as heating continues for the lifetime of the road surface. This suggests that persistent asphalt solar heating is likely a considerable and continued source of summertime SOA in urban environments.

Limited Secondary Organic Aerosol Production from Acyclic Oxygenated Volatile Chemical Products.

Volatile chemical products (VCPs) have recently been identified as potentially important unconventional sources of secondary organic aerosol (SOA), in part due to the mitigation of conventional emissions such as vehicle exhaust. Here, we report measurements of SOA production in an oxidation flow reactor from a series of common VCPs containing oxygenated functional groups and at least one oxygen within the molecular backbone. These include two oxygenated aromatic species (phenoxyethanol and 1-phenoxy-2-propanol), two esters (butyl butyrate and butyl acetate), and four glycol ethers (carbitol, methyl carbitol, butyl carbitol, and hexyl carbitol). We measured gas- and particle-phase products with a suite of mass spectrometers and particle-sizing instruments. Only the aromatic VCPs produce SOA with substantial yields. For the acyclic VCPs, ether and ester functionality promotes fragmentation and hinders autoxidation, whereas aromatic rings drive SOA formation in spite of the presence of ether groups. Therefore, our results suggest that a potential strategy to reduce urban SOA from VCPs would be to reformulate consumer products to include less oxygenated aromatic compounds.

Residual Effects of Cannabis Use on Effort-Based Decision-Making.

OBJECTIVE: Acute Δ9-tetrahydrocannabinol (THC) administration in humans (Lawn etal., 2016) and rats (Silveira, Adams, Morena, Hill, & Winstanley, 2016) has been associated with decreased effort allocation that may explain amotivation during acute cannabis intoxication. To date, however, whether residual effects of cannabis use on effort-based decision-making are present and observable in humans have not yet been determined. The goal of this study was to test whether prolonged cannabis use has residual effects on effort-based decision-making in 24-hr abstinent cannabis using adults. METHOD: We evaluated performance on the Effort Expenditure for Reward Task (EEfRT) in 41 adult cannabis users (mean age = 24.63 years, 21 males) and 45 nonusers (mean age = 23.90 years, 19 males). A mixed 2x3x3 ANOVA with age as a covariate was performed to examine the effect of group, probability of winning, and reward amount on EEfRT performance. EEfRT performance was operationalized as % of trials for which the hard (vs. easy) condition was chosen. Pearson's correlations were conducted to test the relationship between EEfRT performance and measures of cannabis use, anhedonia and motivation. RESULTS: We found that cannabis users selected hard trials significantly more than nonusers regardless of win probability or reward level. Frequency of cannabis use was positively correlated with amount of % hard trials chosen. There were no significant correlations between % hard trials chosen, self-reported anhedonia, or motivation. CONCLUSIONS: These results suggest that unlike acute effects, residual effects of cannabis following 24 hrs of abstinence are associated with greater effort allocation during effort-based decision-making.

Using NMR Spectroscopy To Measure Protein Binding Capacity on Gold Nanoparticles.

A simple one-dimensional 1H NMR experiment that quantifies protein bound to gold nanoparticles has been developed for upper-division biochemistry and physical chemistry students. This laboratory experiment teaches the basics of NMR techniques, which is a highly effective tool in protein studies and supports students to understand the concepts of NMR spectroscopy and nanoparticle-protein interactions. Understanding the interactions of gold nanoparticles (AuNPs) with biological macromolecules is becoming increasingly important as interest in the clinical use of nanoparticles has been on the rise. Applications in drug delivery, biosensing, diagnostics, and enhanced imaging are all tangible possibilities with a better understanding of AuNP-protein interactions. The ability to use AuNPs as biosensors for drug delivery methods in cellular uptake is dependent on the amount of protein that is able to bind to the surface of the nanoparticle. This laboratory experiment solidifies concepts such as quantitative NMR spectroscopy while reinforcing precision laboratory titrations. Students learn how 1H proton NMR spectra can be used to measure free protein in solution and protein bound to AuNPs. A simple formula is used to determine the binding capacity of the nanoparticle. This analysis helps students to understand the impact of nanoparticle-protein interactions, and it allows them to conceptualize macromolecular binding using NMR spectroscopy.

Adipose tissue-specific knockout of AMPKα1/α2 results in normal AICAR tolerance and glucose metabolism.

AMP-activated protein kinase (AMPK) is a member of Ser/Thr kinases that has been shown to regulate energy balance. Although recent studies have demonstrated the function of AMPK in adipose tissue using different fat-specific AMPK knockout mouse models, the results were somewhat inconsistent. For this study, we tested the hypothesis that AMPK in adipose tissue regulates whole body glucose metabolism. To determine the role of adipose tissue AMPK in vivo, we generated fat-specific AMPKα1/α2 knockout mice (AMPKFKO) using the Cre-loxP system. Body weights of AMPKFKO mice were not different between 8 and 27 weeks of age. Furthermore, tissue weights (liver, kidney, muscle, heart and white and brown adipose tissue) were similar to wild type littermates and DEXA scan analysis revealed no differences in percentages of body fat and lean mass. Knockout of AMPKα1/α2 in adipose tissue abolished basal and AICAR-stimulated phosphorylation of AMPK and Acetyl-CoA Carboxylase, a downstream of AMPK. Despite of the ablation of AICAR-stimulated AMPK phosphorylation, the blood glucose-lowering effect of AICAR injection (i.p.) was normal in AMPKFKO mice. In addition, AMPKFKO displayed normal fasting blood glucose concentration, glucose tolerance, insulin tolerance, and insulin signaling, indicating normal whole body glucose metabolism. These data demonstrate that adipose tissue AMPK plays a minimum role in whole body glucose metabolism on a chow diet.

Protective antibodies against Clostridium difficile are present in intravenous immunoglobulin and are retained in humans following its administration.

The prevalence of serum antibodies against Clostridium difficile (CD) toxins A and B in healthy populations have prompted interest in evaluating the therapeutic activity of intravenous immunoglobulin (IVIg) in individuals experiencing severe or recurrent C. difficile infection (CDI). Despite some promising case reports, a definitive clinical role for IVIg in CDI remains unclear. Contradictory results may be attributed to a lack of consensus regarding optimal dose, timing of administration and patient selection as well as variability in specific antibody content between commercial preparations. The purpose of this study was to investigate retrospectively the efficacy of three commercial preparations of IVIg for treating severe or recurrent CDI. In subsequent mechanistic studies using protein microarray and toxin neutralization assays, all IVIg preparations were analysed for specific binding and neutralizing antibodies (NAb) to CD antigens in vitro and the presence of anti-toxin NAbs in vivo following IVIg infusion. A therapeutic response to IVIg was observed in 41% (10 of 17) of the CDI patients. Significant variability in multi-isotype specific antibodies to a 7-plex panel of CD antigens and toxin neutralization efficacies were observed between IVIg preparations and also in patient sera before and after IVIg administration. These results extend our current understanding of population immunity to CD and support the inclusion of surface layer proteins and binary toxin antigens in CD vaccines. Future strategies could enhance IVIg treatment response rates by using protein microarray to preselect donor plasma/serum with the highest levels of anti-CD antibodies and/or anti-toxin neutralizing capacities prior to fractionation.

Dicarboxylic Acid Emissions from Aftertreatment Equipped Diesel Engines.

Dicarboxylic acids play a key role in atmospheric particle nucleation. Though long assumed to originate from primary sources, little experimental evidence exists directly linking combustion to their emissions. In this work, we sought definitive proof that dicarboxylic acids are produced in diesel engines and that they can slip through a modern aftertreatment system (ATS) at low exhaust temperatures. One difficulty in measuring dicarboxylic acid emissions is that they cannot be identified using conventional mass spectroscopy techniques. In this work, we refined a derivatization gas chromatography-mass spectroscopy technique to measure 11 mono- and dicarboxylic acids from plain and KOH impregnated quartz filters. Filters were loaded with exhaust from a modern passenger car diesel engine on a dynamometer sampled before and after an ATS consisting of an oxidation catalyst and diesel particulate filter. Our findings confirm that dicarboxylic acids are produced in diesel engine combustion, especially during low temperature combustion modes that emit significant concentrations of partially combusted hydrocarbons. Exhaust acids were largely removed by a fully warmed-up ATS, mitigating their environmental impact. Our results also suggest that dicarboxylic acids do not participate in primary particle formation in dilute engine exhaust as low quantities were collected on unimpregnated filters.

Obstetrical Providers' Management of Chronic Pain in Pregnancy: A Vignette Study.

Objective: Describe obstetrical providers' management of a hypothetical case on chronic pain in pregnancy and determine whether practices differ based on patient race. Design and Setting: This was a self-administered survey at a clinical conference. Subjects: Seventy-six obstetrician-gynecologists and one nurse practitioner were surveyed. Methods: A case-vignette described a pregnant patient presenting with worsening chronic lower back pain, requesting an opioid refill and increased dosage. We varied patient race (black/white) across two randomly assigned identical vignettes. Providers indicated their likelihood of prescribing opioids, drug testing, and referring on a 0 (definitely would not) to 10 (definitely would) scale; rated their suspicions/concerns about the patient on a 0-10 VAS scale; and ranked those concerns in order of importance. We calculated correlation coefficients, stratifying analyses by patient race. Results: Providers were not inclined to refill the opioid prescription (median = 3.0) or increase the dose (median = 1.0). They were more likely to conduct urine drug tests on white than black patients ( P = 0.008) and more likely to suspect that white patients would divert the medication ( P =0.021). For white patients, providers' highest-ranked concern was the patient's risk of abuse/addiction, whereas, for black patients, it was harm to the fetus. Suspicion about symptom exaggeration was more closely related to decisions about refilling the opioid prescriptions and increasing the dose for black patients (r = -0.357, -0.439, respectively), whereas these decisions were more closely correlated with concerns about overdose for white patients (r = -0.406, -0.494, respectively). Conclusions: Provider suspicion and concerns may differ by patient race, which may relate to differences in pain treatment and testing. Further study is warranted to better understand how chronic pain is managed in pregnancy.

Moving beyond descriptions of diversity: clinical and research implications of bacterial imbalance in chronic rhinosinusitis.

Chronic rhinosinusitis (CRS) is a debilitating disease which affects 5-16% of the general population and involves long-term inflammation of the sinonasal cavity. While microbial involvement in the pathogenesis of CRS has long been suspected, the exact role of microbes remains unclear. Recent application of cultivation-independent, molecular methods has provided much new information, taking advantage of developments in both laboratory- and bioinformatics-based analyses. The aim of this mini-review is to present a variety of available bioinformatics approaches, such as data classification techniques and network analyses, with proven applications in other aspects of human microbiome health and disease research. The uses of molecular techniques in the clinical setting are still in its infancy, but these tools can further our understanding of microbial imbalance during chronic disease and help guide effective patient treatment. The mini-review emphasises ways in which CRS bacterial gene-targeted sequencing data can progress beyond descriptive summaries and toward unlocking the mechanisms by which bacterial communities can be markers for sinus health.

Similarities Between Large Animal-Related and Motor Vehicle Crash-Related Injuries.

OBJECTIVE: Many Americans sustain large animal-related injuries (LARIs) from blunt trauma. We compare the injuries and management of LARI in our region of the United States with those of motor vehicle crashes (MVCs). METHODS: A 15-year retrospective study of trauma patients with LARI matched to MVC controls by Injury Severity Score (ISS), age, and sex was conducted. Values were statistically compared, and differences were considered statistically significant at P < .05. RESULTS: There were 156 LARI cases, of which 87% were related to horses, 8% to bulls, and the remainder to deer, mules, bison, cows, and rams. In the LARI group, the age was 42±18 years (mean±SD), ISS was 7±4, and 61% were females. The MVC group had a significantly longer length of hospital stay (5±5 vs 4±3 days) and blood alcohol concentration (35±84 vs 3±20 g/L). There were no significant differences in injury patterns between LARI and MVC; however, additional radiological studies (RS) were performed on MVC (9±6 vs 7±5). LARI patients were more often transferred from rural locations (39% vs 25%) and traveled further to our trauma center (40±32 vs 24±29 km) than did MVC patients. CONCLUSIONS: LARI has a similar pattern of injury to MVC, but fewer RS. LARI typically occurred further away, requiring transfer from rural areas to our institution. We recommend a similar approach for the evaluation of LARI and MVC.

Trade-offs between three forest ecosystem services across the state of New Hampshire, USA: timber, carbon, and albedo.

Forests are more frequently being managed to store and sequester carbon for the purposes of climate change mitigation. Generally, this practice involves long-term conservation of intact mature forests and/or reductions in the frequency and intensity of timber harvests. However, incorporating the influence of forest surface albedo often suggests that long rotation lengths may not always be optimal in mitigating climate change in forests characterized by frequent snowfall. To address this, we investigated trade-offs between three ecosystem services: carbon storage, albedo-related radiative forcing, and timber provisioning. We calculated optimal rotation length at 498 diverse Forest Inventory and Analysis forest sites in the state of New Hampshire, USA. We found that the mean optimal rotation lengths across all sites was 94 yr (standard deviation of sample means = 44 yr), with a large cluster of short optimal rotation lengths that were calculated at high elevations in the White Mountain National Forest. Using a regression tree approach, we found that timber growth, annual storage of carbon, and the difference between annual albedo in mature forest vs. a post-harvest landscape were the most important variables that influenced optimal rotation. Additionally, we found that the choice of a baseline albedo value for each site significantly altered the optimal rotation lengths across all sites, lowering the mean rotation to 59 yr with a high albedo baseline, and increasing the mean rotation to 112 yr given a low albedo baseline. Given these results, we suggest that utilizing temperate forests in New Hampshire for climate mitigation purposes through carbon storage and the cessation of harvest is appropriate at a site-dependent level that varies significantly across the state.

Boat noise impedes vocalizations of wild plainfin midshipman fish.

Marine noise is recognised as a growing threat that can induce maladaptive behavioural changes in many aquatic animals, including fishes. The plainfin midshipman is a soniferous fish with a prolonged breeding period, during which males produce tonal hums that attract females, and grunts and growls during agonistic interactions. In this study, we used acoustic recordings to assess the effects of boat noise on the presence, peak frequencies, and durations of plainfin midshipman calls in the wild. We found that all three call types were less likely to occur, and the peak frequencies of hums and grunts increased in the presence of boat noise. We also show that loud and quiet boat noise affected plainfin midshipman vocalizations similarly. As anthropogenic noise is likely to increase in the ocean, it will be important to understand how such noise can affect communication systems, and consequently population health and resiliency.

Lay Health Worker Interventions in Rheumatology: A Scoping Review.

OBJECTIVE: To determine next steps for lay health worker (LHW) intervention research, specifically in patients with rheumatic musculoskeletal diseases (RMDs), there is a need to establish what strategies have been effective for chronic disease management thus far. The goal of this scoping review is to collate the literature of LHW interventions for adults with RMDs to inform next steps for LHW research. METHODS: A comprehensive literature search was performed in the following databases from inception to September 2021: Ovid Medline, Ovid Embase, CINAHL, PsycINFO, and The Cochrane Library. Studies retrieved were then screened for eligibility against predefined inclusion and exclusion criteria. RESULTS: Twenty-two articles were eligible and included in this review. The most common RMDs studied, not mutually exclusive, were osteoarthritis (n = 13), rheumatoid arthritis (n = 9), and unspecified or other RMD (n = 14). Most studies had a homogenous patient population, enrolling White, non-Hispanic, or Latina women over the age of 60 (n = 13). Eight studies observed statistically significant results in the intervention arm compared with the control. Only one of these studies exhibited sustained treatment effects past one year. CONCLUSION: There are not enough data to conclude if LHW interventions have a positive, null, or negative effect on patients with RMDs. Future LHW interventions should specify a priori hypotheses, be powered to detect statistical significance for primary outcomes, employ a theoretical framework, include an active control, describe training protocols for LHWs, and increase minority representation to establish the effectiveness of LHWs for patients with RMDs.

Examining the Relationship Between Value and Patient Satisfaction With Treatment in Total Joint Arthroplasty.

Background: A shift toward performance, cost, outcomes, and patient satisfaction has occurred with healthcare reform promoting value-based programs. The purpose of this study was to evaluate the relationship between patient satisfaction and value with treatment in a cohort of patients undergoing total knee arthroplasty (TKA) and total hip arthroplasty (THA). Methods: Value was determined by the relationship of treatment outcome with episodic cost. Measurements included both clinical outcomes and patient-reported outcomes. Participating surgeons took part in the modified Delphi method resulting in an algorithm measuring patient value. Treatment outcome, cost, and resultant value (outcome/cost) of both TKA and THA were evaluated using binomial logistic regression by adjusting for age, gender, body mass index, Charlson comorbidity index, tobacco, education, and income with patient-reported satisfaction as the outcome. Results: This study had a total of 909 patients (TKA n = 438; THA n = 471), with an average age of 67 (TKA) and 65 (THA) years. Patient satisfaction shared a significant positive relationship with treatment outcome for TKA (odds ratio [OR] = 1.53, confidence interval [CI] = 1.35-1.73, P < .001) and THA (OR = 1.93, CI = 1.62-2.29, P < .001). Higher value was associated with a significantly higher odds of patient satisfaction for both TKA (OR = 1.39, CI = 1.25-1.54, P < .001) and THA (OR = 1.70, CI = 1.47-1.97, P < .001). Conclusions: This study showed a positive relationship between treatment outcome but not cost with subsequent value and patient satisfaction. This method provides a promising approach to comprehensively evaluate outcomes, cost, and value of total joint arthroplasty procedures. This approach can help predict the probability of value-driven patient satisfaction.

Attitudes toward Help-Seeking for Sexual Problems among College Women.

Prior research has indicated that 65-70% of college age women have experienced at least one sexual problem. Sexual problems are associated with higher rates of depression, anxiety, and relationship distress; however, few college age women seek professional help for sexual difficulties. The present study used the Theory of Planned Behavior to explore how adherence to traditional feminine norms and self-stigma are associated with intention to seek professional psychological help for a sexual difficulty. Results of structural equation modeling analysis demonstrated approximate fit in a sample of 259 college women. The structural model indicated that self-stigma and stronger endorsement of specific feminine gender norms (i.e., romantic relationship, sexual fidelity, sweet and nice) were found to be associated with less intention to seek help for a sexual problem. We discuss future research directions and implications for clinical services.

Acute dorsal genital nerve stimulation increases subjective arousal in women with and without spinal cord injury: a preliminary investigation.

Introduction: Female sexual dysfunction (FSD) impacts an estimated 40% of women. Unfortunately, female sexual function is understudied, leading to limited treatment options for FSD. Neuromodulation has demonstrated some success in improving FSD symptoms. We developed a pilot study to investigate the short-term effect of electrical stimulation of the dorsal genital nerve and tibial nerve on sexual arousal in healthy women, women with FSD, and women with spinal cord injury (SCI) and FSD. Methods: This study consists of a randomized crossover design in three groups: women with SCI, women with non-neurogenic FSD, and women without FSD or SCI. The primary outcome measure was change in vaginal pulse amplitude (VPA) from baseline. Secondary outcome measures were changes in subjective arousal, heart rate, and mean arterial pressure from baseline. Participants attended one or two study sessions where they received either transcutaneous dorsal genital nerve stimulation (DGNS) or tibial nerve stimulation (TNS). At each session, a vaginal photoplethysmography sensor was used to measure VPA. Participants also rated their level of subjective arousal and were asked to report any pelvic sensations. Results: We found that subjective arousal increased significantly from before to after stimulation in DGNS study sessions across all women. TNS had no effect on subjective arousal. There were significant differences in VPA between baseline and stimulation, baseline and recovery, and stimulation and recovery periods among participants, but there were no trends across groups or stimulation type. Two participants with complete SCIs experienced genital sensations. Discussion: This is the first study to measure sexual arousal in response to acute neuromodulation in women. This study demonstrates that acute DGNS, but not TNS, can increase subjective arousal, but the effect of stimulation on genital arousal is inconclusive. This study provides further support for DGNS as a treatment for female sexual dysfunction.

Return to Sport Using Corticosteroid Injections for Knee Pain in Triathletes.

Introduction Despite the prevalence of corticosteroid injections in athletes, little is known about their efficacy in triathletes. We aim to assess attitudes, use, subjective effectiveness, and time to return to sport with corticosteroid injections compared to alternative methods in triathletes with knee pain. Methods This is an observational study during the COVID-19 pandemic. Triathletes answered a 13-question survey posted to three triathlon-specific websites. Results Sixty-one triathletes responded, 97% of whom experienced knee pain at some point in their triathlete career; 63% with knee pain received a corticosteroid injection as treatment (average age 51 years old). The most popular attitude (44.3%) regarding corticosteroid injections was "tried them, with good improvement". Most found the cortisone injection helpful for two to three months (28.6%), or more than one year (28.6%); of individuals who found the injections useful for more than one year, four-eight (50%) had received multiple injections during that same period. After injection, 80.6% returned to sport within one month. The average age of people using alternative treatment methods was 39 years old; most returned to sport within one month (73.7%). Compared to alternative methods, there was an ~80% higher odds of returning to sport within one month using corticosteroid injections; however, this relationship was not significant (OR=1.786, p=0.480, 95% CI:0.448-7.09). Conclusion This is the first study to examine corticosteroid use in triathletes. Corticosteroid use is more common in older triathletes and results in subjective pain improvement. A strong association does not exist for a quicker return to sport using corticosteroid injections compared to alternative methods. Triathletes should be counseled on the timing of injections, duration of side effects, and be aware of potential risks.

Fluorescence in situ hybridisation in Carnoy's fixed tonsil tissue.

Fluorescence in situ hybridisation (FISH) is a powerful molecular technique that enables direct visualisation of specific bacterial species. Few studies have established FISH protocols for tonsil tissue in Carnoy's fixative, accordingly limiting its application to investigate the pathogenesis of tonsillar hyperplasia. Tonsil tissue from 24 children undergoing tonsillectomy for either recurrent tonsillitis or sleep-disordered breathing were obtained during a previous study. The specificity of each of the five FISH probes (Fusobacterium spp., Bacteroides spp., Streptococcus spp., Haemophilus influenzae and Pseudomonas spp.) were successfully optimised using pure and mixed bacterial isolates, and in Carnoy's fixed tonsil tissue. Bacteroides spp. were present in 100% of patients with microcolonies. In comparison, the prevalence of Fusobacterium spp. was 93.8%, Streptococcus spp. 85.7%, H. influenzae 82.35% and Pseudomonas spp. 76.5%. Notable differences in the organisation of bacterial taxa within a single microcolony were also observed. This is the first study to establish a robust FISH protocol identifying multiple aerobic and anaerobic bacteria in Carnoy's fixed tonsil tissue. This protocol provides a strong foundation for combining histological and microbiological analyses of Carnoy's fixed tonsil samples. It may also have important implications on the analysis of microorganisms in other human tissues prepared using the same techniques.