Melanoma secretion of transforming growth factor-ß2 leads to loss of epidermal AMBRA1 threatening epidermal integrity and facilitating tumour ulceration.

BACKGROUND: For patients with early American Joint Committee on Cancer (AJCC)-stage melanoma the combined loss of the autophagy regulatory protein AMBRA1 and the terminal differentiation marker loricrin in the peritumoral epidermis is associated with a significantly increased risk of metastasis. OBJECTIVES: The aim of the present study was to evaluate the potential contribution of melanoma paracrine transforming growth factor (TGF)-ß signalling to the loss of AMBRA1 in the epidermis overlying the primary tumour and disruption of epidermal integrity. METHODS: Immunohistochemistry was used to analyse AMBRA1 and TGF-ß2 in a cohort of 109 AJCC all-stage melanomas, and TGF-ß2 and claudin-1 in a cohort of 30 or 42 AJCC stage I melanomas, respectively, with known AMBRA1 and loricrin (AMLo) expression. Evidence of pre-ulceration was analysed in a cohort of 42 melanomas, with TGF-ß2 signalling evaluated in primary keratinocytes. RESULTS: Increased tumoral TGF-ß2 was significantly associated with loss of peritumoral AMBRA1 (P < 0·05), ulceration (P < 0·001), AMLo high-risk status (P < 0·05) and metastasis (P < 0·01). TGF-ß2 treatment of keratinocytes resulted in downregulation of AMBRA1, loricrin and claudin-1, while knockdown of AMBRA1 was associated with decreased expression of claudin-1 and increased proliferation of keratinocytes (P < 0·05). Importantly, we show loss of AMBRA1 in the peritumoral epidermis was associated with decreased claudin-1 expression (P < 0·05), parakeratosis (P < 0·01) and cleft formation in the dermoepidermal junction (P < 0·05). CONCLUSIONS: Collectively, these data suggest a paracrine mechanism whereby TGF-ß2 causes loss of AMBRA1 overlying high-risk AJCC early-stage melanomas and reduced epidermal integrity, thereby facilitating erosion of the epidermis and tumour ulceration.

Epidermal autophagy and beclin 1 regulator 1 and loricrin: a paradigm shift in the prognostication and stratification of the American Joint Committee on Cancer stage I melanomas.

BACKGROUND: The updated American Joint Committee on Cancer (AJCC) staging criteria for melanoma remain unable to identify high-risk stage I tumour subsets. OBJECTIVES: To determine the utility of epidermal autophagy and beclin 1 regulator 1 (AMBRA1)/loricrin (AMLo) expression as a prognostic biomarker for AJCC stage I cutaneous melanoma. METHODS: Peritumoral AMBRA1 expression was evaluated in a retrospective discovery cohort of 76 AJCC stage I melanomas. AMLo expression was correlated with clinical outcomes up to 12 years in two independent powered, retrospective validation and qualification cohorts comprising 379 AJCC stage I melanomas. RESULTS: Decreased AMBRA1 expression in the epidermis overlying primary melanomas in a discovery cohort of 76 AJCC stage I tumours was associated with a 7-year disease-free survival (DFS) rate of 81·5% vs. 100% survival with maintained AMBRA1 (P < 0·081). Following an immunohistochemistry protocol for semi-quantitative analysis of AMLo, analysis was undertaken in validation (n = 218) and qualification cohorts (n = 161) of AJCC stage I melanomas. Combined cohort analysis revealed a DFS rate of 98·3% in the AMLo low-risk group (n = 239) vs. 85·4% in the AMLo high-risk cohort (n = 140; P < 0·001). Subcohort multivariate analysis revealed that an AMLo hazard ratio (HR) of 4·04 [95% confidence interval (CI) 1·69-9·66; P = 0·002] is a stronger predictor of DFS than Breslow depth (HR 2·97, 95% CI 0·93-9·56; P = 0·068) in stage IB patients. CONCLUSIONS: Loss of AMLo expression in the epidermis overlying primary AJCC stage I melanomas identifies high-risk tumour subsets independently of Breslow depth. What's already known about this topic? There is an unmet clinical need for biomarkers of early-stage melanoma. Autophagy and beclin 1 regulator 1 (AMBRA1) is a proautophagy regulatory protein with known roles in cell proliferation and differentiation, and is a known tumour suppressor. Loricrin is a marker of epidermal terminal differentiation. What does this study add? AMBRA1 has a functional role in keratinocyte/epidermal proliferation and differentiation. The combined decrease/loss of peritumoral AMBRA1 and loricrin is associated with a significantly increased risk of metastatic spread in American Joint Committee on Cancer (AJCC) stage I tumours vs. melanomas, in which peritumoral AMBRA1 and loricrin are maintained, independently of Breslow depth. What is the translational message? The integration of peritumoral epidermal AMBRA1/loricrin biomarker expression into melanoma care guidelines will facilitate more accurate, personalized risk stratification for patients with AJCC stage I melanomas, thereby facilitating stratification for appropriate follow-up and informing postdiagnostic investigations, including sentinel lymph node biopsy, ultimately resulting in improved disease outcomes and rationalization of healthcare costs.

Musculoskeletal injuries in real tennis.

Introduction: Real tennis is a growing, unique, and well-established sport. To date, there has been no epidemiological data on real tennis injuries. The primary aim of this retrospective study is to record the incidence and document any trends in real tennis musculoskeletal injuries, so as to improve injury awareness of common and possibly preventable injuries. Methods: A surveillance questionnaire e-mailed to 2,036 Tennis & Rackets Association members to retrospectively capture injuries sustained by amateur and professional real tennis players over their playing careers. Results: A total of 485 (438 males and 47 females) questionnaires were fully completed over 4 weeks. A total of 662 musculoskeletal injuries were recorded with a mean of 1.4 injuries per player (range 0-7). The incidence of sustaining an acute real tennis musculoskeletal injury is 0.4/1000 hrs. The three main anatomical locations reported injured were elbow 15.6% (103/662), knee 11.6% (77/662), and face 10.0% (66/662). The most common structures reported injured were muscle 24% (161/661), tendon 23.4% (155/661), ligament 7.0% (46/661), soft tissue bruising 6.5% (43/661), and eye 6.2% (41/661). The majority of the upper limb injuries were gradual onset (64.7%, 143/221), and the lower limb injuries were sudden onset (72.0%, 188/261). Conclusion: This study uniquely provides valuable preliminary data on the incidence and patterns of musculoskeletal injuries in real tennis players. In addition, it highlights a number of reported eye injuries. The study is also a benchmark for future prospective studies on academy and professional real tennis players.

Ultrasonic preparation, stability and thermal conductivity of a capped copper-methanol nanofluid.

This paper describes a two-step method to prepare novel copper-methanol nanofluids capped with a short chain molecule, (3-Aminopropyl)trimethoxysilane (APTMS). Two commercial nanopowders were dispersed at various powers using a 20 kHz ultrasonic probe into solutions of methanol and the capping agent. Ultrasonic energy input was measured by calorimetry with z-average diameters, intensity and number size distributions recorded by a dynamic light scattering technique. The stability of the dispersion was monitored visually, and quantified by recording the zeta potential. Dispersions of the bare powder were used as a control. Absorption spectroscopy was used to confirm the presence of the capping agent. The thermal conductivities of 0 to 10% wt./vol. (1.1% vol.) dispersions of the capped copper-methanol nanofluid were determined using a C-Therm analyzer. Optimum ultrasonic de-agglomeration conditions gave dispersions with a z-average particle size of <200 nm and a PdI of <0.2. The capped particles showed good stability; up to six months in some instances, and an average zeta potential of +38 mV was recorded. The thermal conductivity of the nanofluid increased with concentration, and an enhancement of 9% over the base fluid was found at 10% wt./vol. (1.1% vol.). This innovative work has demonstrated the ultrasonic preparation and stability of copper nanoparticles protected with APTMS; a short chain molecule which binds to copper and prevents oxidation. The protected particles can enhance the thermal conductivity of methanol with no interference from the capping ligand.

Plains zebra (Equus quagga) adrenocortical activity increases during times of large aggregations in the Serengeti ecosystem.

Adverse environmental stimuli (stressors) activate the hypothalamic-pituitary-adrenal axis and contribute to allostatic load. This study investigates the contribution of environmental stressors and life history stage to allostatic load in a migratory population of plains zebras (Equus quagga) in the Serengeti ecosystem, in Tanzania, which experiences large local variations in aggregation. We expected higher fGCM response to the environmental stressors of feeding competition, predation pressure and unpredictable social relationships in larger than in smaller aggregations, and in animals at energetically costly life history stages. As the study was conducted during the 2016 El Niño, we did not expect food quality of forage or a lack of water to strongly affect fGCM responses in the dry season. We measured fecal glucocorticoid metabolite (fGCM) concentrations using an enzyme immunoassay (EIA) targeting 11ß-hydroxyetiocholanolone and validated its reliability in captive plains zebras. Our results revealed significantly higher fGCM concentrations 1) in large aggregations than in smaller groupings, and 2) in band stallions than in bachelor males. Concentrations of fGCM were not significantly higher in females at the energetically costly life stage of late pregnancy/lactation. The higher allostatic load of stallions associated with females, than bachelor males is likely caused by social stressors. In conclusion, migratory zebras have elevated allostatic loads in large aggregations that probably result from their combined responses to increased feeding competition, predation pressure and various social stressors. Further research is required to disentangle the contribution of these stressors to allostatic load in migratory populations.

Prevalence of pelvic floor symptoms in female patients attending the two-week wait clinic with suspected colorectal cancer.

Introduction The aim of this study was to establish the prevalence of pelvic floor symptoms in women referred to a colorectal two-week wait (2WW) clinic with suspected colorectal cancer. Methods A questionnaire assessing faecal incontinence (FI) (Wexner score) and obstructed defecation syndrome (ODS) (Renzi score) was offered to 98 consecutive female patients attending a colorectal 2WW clinic at a single trust. Results Overall, 56 (57%) of the 98 patients had significant ODS and/or FI (scores >9/20), 33 (34%) had ODS and 40 (41%) had FI. Seventeen patients (17%) had both ODS and FI. Analysis of the 63 patients referred with a change in bowel habit (CIBH) showed 40 (63%) to be Renzi and/or Wexner positive compared with 16 (46%) of the 35 patients who presented without CIBH (p=0.095, Fisher's exact test). Further analysis showed that 31 (78%) of the 40 patients with FI presented with CIBH compared with 32 (55%) of the 58 without FI (p=0.032). In terms of ODS, 23 (70%) of the 33 patients with ODS presented with CIBH compared with 40 (62%) of the 65 without ODS (p=0.506). Conclusions Over half of the female patients attending our colorectal 2WW clinic had significant pelvic floor dysfunction (FI/ODS), which may account for their symptoms (especially in the CIBH referral category). While it is important for malignancy to be excluded, many patients may benefit from investigation and management of their pelvic floor dysfunction as the cause for their presenting symptoms.

Morning vs evening dosing of the cathepsin K inhibitor ONO-5334: effects on bone resorption in postmenopausal women in a randomized, phase 1 trial.

UNLABELLED: The cathepsin K inhibitor, ONO-5334, improves bone mineral density in postmenopausal women with osteoporosis. The effects of morning versus evening administration of ONO-5334 were investigated by measuring bone turnover marker levels in healthy postmenopausal women. Morning administration of ONO-5334 showed a more consistent suppressive effect on bone resorption than evening administration. INTRODUCTION: Bone turnover is thought to be subject to circadian variation, and the efficacy of osteoporosis treatments may be optimized by regulating the time of dosing. This study assessed whether evening administration of the cathepsin K inhibitor, ONO-5334, had a differential effect on the bone turnover marker, C-terminal telopeptide of type I collagen (CTX-I), compared with morning administration. METHODS: This was a single-center, single blind crossover study. Fourteen healthy postmenopausal women were assigned to receive ONO-5334 150 mg once daily for 5 days in each period; they were randomized to receive either evening doses in the first period and morning doses in the second or vice versa. Serum and urinary levels of CTX-I were measured throughout the study. RESULTS: Both regimens showed similar patterns of reduction in serum and urinary CTX-I; however, CTX-I suppression was more consistently >60% over 24 h following morning administration. Morning administration led to 6% greater suppression of 24-h serum CTX-I area under the effect curve (AUE; 69 vs 63%; P < .05) and 7% greater suppression of urinary CTX-I/creatinine AUE (93 vs 86%; P < .01) than evening administration. Higher plasma ONO-5334 concentrations were observed between 12 and 24 h postdose following morning administration, with mean trough concentrations for the morning and evening regimens at 9.4 and 4.0 ng/mL, respectively. There were no safety findings of concern. CONCLUSION: Morning dosing of ONO-5334 is more efficacious at reducing markers of bone turnover in healthy postmenopausal women than evening dosing. TRIAL REGISTRATION: ClinicalTrials.gov: NCT01384188 , registered on June 27, 2011 EudraCT: 2008-006284-37.

Anti-NMDA Receptor Encephalitis in the Polar Bear (Ursus maritimus) Knut.

Knut the polar bear of the Berlin Zoological Garden drowned in 2011 following seizures and was diagnosed as having suffered encephalitis of unknown etiology after exhaustive pathogen screening. Using the diagnostic criteria applied to human patients, we demonstrate that Knut's encephalitis is almost identical to anti-NMDA receptor encephalitis which is a severe autoimmune disease representing the most common non-infectious encephalitis in humans. High concentrations of antibodies specific against the NR1 subunit of the NMDA receptor were detected in Knut's cerebrospinal fluid. Histological examination demonstrated very similar patterns of plasma cell infiltration and minimal neuronal loss in affected brain areas. We conclude that Knut suffered anti-NMDA receptor encephalitis making his the first reported non-human case of this treatable disease. The results suggest that anti-NMDA receptor encephalitis may be a disease of broad relevance to mammals that until now has remained undiagnosed.

First observations of nonhydrodynamic mix at the fuel-shell interface in shock-driven inertial confinement implosions.

A strong nonhydrodynamic mechanism generating atomic fuel-shell mix has been observed in strongly shocked inertial confinement fusion implosions of thin deuterated-plastic shells filled with 3He gas. These implosions were found to produce D3He-proton shock yields comparable to implosions of identical shells filled with a hydroequivalent 50∶50 D3He gas mixture. Standard hydrodynamic mixing cannot explain this observation, as hydrodynamic modeling including mix predicts a yield an order of magnitude lower than was observed. Instead, these results can be attributed to ion diffusive mix at the fuel-shell interface.

Polar bear encephalitis: establishment of a comprehensive next-generation pathogen analysis pipeline for captive and free-living wildlife.

This report describes three possibly related incidences of encephalitis, two of them lethal, in captive polar bears (Ursus maritimus). Standard diagnostic methods failed to identify pathogens in any of these cases. A comprehensive, three-stage diagnostic 'pipeline' employing both standard serological methods and new DNA microarray and next generation sequencing-based diagnostics was developed, in part as a consequence of this initial failure. This pipeline approach illustrates the strengths, weaknesses and limitations of these tools in determining pathogen caused deaths in non-model organisms such as wildlife species and why the use of a limited number of diagnostic tools may fail to uncover important wildlife pathogens.

Paediatric hand fractures.

This retrospective study reviews the demographics, morphology and management of paediatric hand fractures. Notes of all children with hand injuries attending a plastic surgery paediatric trauma clinic over a one-year period were reviewed. Non-bony injuries were excluded. A total of 303 fractures in 283 patients were included. Fracture incidence rose after the age of seven, peaking at 14 years of age; 76% of fractures occurred in males. Sporting injuries accounted for 47% of fractures. Physeal fractures were present in 39% of cases. Open fractures accounted for only 4% of all fractures. Management was primarily conservative. Only 5% of cases required surgical fixation; 6% of patients experienced complications.

Echinococcus and Taenia spp. from captive mammals in the United Kingdom.

Taeniid tapeworms which include Echinococcus and Taenia spp. are obligatory parasites of mammals with pathogenicity usually related to the larval stages of the life cycle. Two species (or genotypes) of Echinococcus, E. granulosus sensu stricto and E. equinus, as well as several Taenia spp. are endemic in the UK. Here we report on the occurrence of larval cystic stages of Echinococcus and Taenia spp. in captive mammals in the UK. Using molecular techniques we have identified E. granulosus (G1 genotype) in a guenon monkey and a Philippine spotted deer; E. equinus in a zebra and a lemur; E. ortleppi in a Philippine spotted deer; E. multilocularis in a macaque monkey and Taenia polyacantha in jumping rats. To the best of our knowledge this is the first report of E. multilocularis in a captive primate translocated to the UK. As far as we know these are the first reports of E. equinus in a primate (lemur) and in a zebra; as well as E. granulosus (G1 genotype) and E. ortleppi in a cervid translocated to the UK. These infections and implications of the potential establishment of exotic species of cestodes are discussed.

The genetic basis of divergent pigment patterns in juvenile threespine sticklebacks.

Animal pigment patterns are important for a range of functions, including camouflage and communication. Repeating pigment patterns, such as stripes, bars and spots have been of particular interest to developmental and theoretical biologists, but the genetic basis of natural variation in such patterns is largely unexplored. In this study, we identify a difference in a periodic pigment pattern among juvenile threespine sticklebacks (Gasterosteus aculeatus) from different environments. Freshwater sticklebacks exhibit prominent vertical bars that visually break up the body shape, but sticklebacks from marine populations do not. We hypothesize that these distinct pigment patterns are tuned to provide crypsis in different habitats. This phenotypic difference is widespread and appears in most of the freshwater populations that we sampled. We used quantitative trait locus (QTL) mapping in freshwater-marine F2 hybrids to elucidate the genetic architecture underlying divergence in this pigmentation pattern. We identified two QTL that were significantly associated with variation in barring. Interestingly, these QTL were associated with two distinct aspects of the pigment pattern: melanophore number and overall pigment level. We compared the QTL locations with positions of known pigment candidate genes in the stickleback genome. We also identified two major QTL for juvenile body size, providing new insights into the genetic basis of juvenile growth rates in natural populations. In summary, although there is a growing literature describing simple genetic bases for adaptive coloration differences, this study emphasizes that pigment patterns can also possess a more complex genetic architecture.

Distinct startle responses are associated with neuroanatomical differences in pufferfishes.

Despite the key function of the Mauthner cells (M-cells) in initiating escape responses and thereby promoting survival, there are multiple examples of M-cell loss across the teleost phylogeny. Only a few studies have directly considered the behavioral consequences of naturally occurring M-cell variation across species. We chose to examine this issue in pufferfishes, as previous research suggested that there might be variability in M-cell anatomy in this group of fish. We characterized the M-cell anatomy and fast-start responses of two pufferfish species, Tetraodon nigroviridis and Diodon holocanthus. T. nigroviridis showed robust fast-starts to both tactile and acoustic startling stimuli. These fast-starts occurred with a latency typical of M-cell initiation in other fish, and retrograde labeling of spinal-projection neurons revealed that T. nigroviridis does have M-cells. By contrast, D. holocanthus only rarely exhibited fast-start-like behavior, and these responses were at a substantially longer latency and were much less extensive than those of T. nigroviridis. Using three complementary anatomical techniques we were unable to identify obvious M-cell candidates in D. holocanthus. These results provide a clear correlation between M-cell presence or absence and dramatic differences in fast-start behavior. The rich diversity within the pufferfish clade should allow future studies investigating the factors that contribute to this correlated anatomical and behavioral variation.

Trichomonas gallinae in Mauritian columbids: implications for an endangered endemic.

Although well known as a widespread parasitic disease of columbids and birds of prey, there have been few studies of trichomonosis in populations of wild birds. In Mauritius, trichomonosis has been highlighted as a major threat to an endangered endemic, the Pink Pigeon (Neosoenas [Columba] mayeri). In this study, we examined the role that populations of other columbids in Mauritius might be playing as infectious reservoirs of the causal flagellate protozoan, Trichomonas gallinae. We screened 296 wild individuals of three columbid species (Madagascan Turtle Dove [Streptopelia picturata], Spotted Dove [Streptopelia chinensis], and Zebra Dove [Geopelia striata]) between September 2002 and April 2004. Prevalence varied significantly among species (ranging from 19% in S. chinensis to 59% in G. striata) and between S. picturata sampled from upland and coastal sites; S. picturata from upland sites (>500 m) were significantly less likely to be infected with T. gallinae than those from lowland sites (<50 m; 62% and 27% prevalence, respectively). There was no significant difference in the prevalence of T. gallinae at sites where Pink Pigeons were also present compared to those sampled at sites without Pink Pigeons. We show that T. gallinae infection prevalence is higher at sites and times of warmer temperatures and lower rainfall.

Avian blood parasites in an endangered columbid: Leucocytozoon marchouxi in the Mauritian Pink Pigeon Columba mayeri.

There is increasing evidence that pathogens can play a significant role in species decline. This study of a complete free-living species reveals a cost of blood parasitism to an endangered host, the Pink Pigeon Columba mayeri, endemic to Mauritius. We investigated the prevalence and effect of infection of the blood parasite, Leucocytozoon marchouxi, in the free-living Pink Pigeon population. Overall, L. marchouxi infection prevalence detected was 18.3%. Juveniles were more likely to be infected than older birds and there was geographical variation in infection prevalence. Survival of birds infected with L. marchouxi was lower than that of uninfected birds to 90 days post-sampling. This study suggests that while common haematozoa are well tolerated in healthy adults, these parasites may have greater pathogenic potential in susceptible juveniles. The study is unusual given its completeness of species sampling (96%) within a short time-period, the accurate host age data, and its focus on blood parasites in a threatened bird species. Species for which long-term life-history data are available for every individual serve as valuable models for dissecting the contribution of particular pathogens to species decline.

Solid-supported chemiluminescence and electrogenerated chemiluminescence based on a tris(2,2'-bipyridyl)ruthenium(II) derivative.

We report a simple and efficient technique for the covalent immobilisation of a tris(2,2'-bipyridyl)ruthenium(II) derivative suitable for both chemiluminescence and electrochemiluminescence detection.

Abyss1: a novel L2-like non-LTR retroelement of the snakelocks anemone (Anemonia sulcata).

Non-LTR retrotransposons are a diverse and taxonomically widely dispersed group of retroelements that can be divided into at least 14 distinguishable clades. Basal metazoans have not been examined in great detail for their retrotransposon content. In order to screen for the presence of reverse transcriptase (RT) related sequences in Cnidaria and Ctenophora, basal phyla of metazoans, PCR with highly degenerate oligonucleotides was performed and an RT-like sequence was identified from the sea anemone species Anemonia sulcata. Further screening identified a related element in another anemone species Actinia equina. Significant homology to non-LTR retrotransposon RTs was observed, particularly to L2-like elements of fish such as Maui. The sequence was not detected among other cnidarians and we have designated the A. sulcata and A. equina elements Abyss1 and Abyss2 respectively. Phylogenetic analysis of Abyss1 compared with members of 14 known non-LTR retroelement clades suggests that the sequence represents a novel L2 element.