Post-exercise dietary macronutrient composition modulates components of energy balance in young, physically active adults.

The effectiveness of exercise to reduce body mass is typically modest, partially due to energy compensation responses which may be linked to energy substrate availability around exercise. The present study aimed to investigate the effect of manipulating post-exercise energy substrate availability (high carbohydrate/low fat [HCLF] or low carbohydrate/high fat [LCHF] energy replacement) on energy balance components in the short-term (i.e., appetite, energy intake (EI) and energy expenditure (EE)). METHODS: Appetite, EI, activity- and total- EE were measured in twelve healthy, young (21.0 ± 2.3 years) physically active participants (10 men, 2 women) on two occasions across 4 days after a 75-min run and an isocaloric energy replacement drink (HCLF and LCHF). Appetite was measured daily by visual analogue scales, EI was calculated by subtracting the energy content of food leftovers from a provided food package, activity- and total- EE determined by heart-rate accelerometery. RESULTS: Composite appetite ratings between days were lower in HCLF (62.4 ± 12) compared to LCHF (68.3 ± 8.9 mm; p = 0.048). No differences between conditions were detected for EI. Cumulative activity-EE (HCLF=  20.9 ± 3.7, LCHF=  16.9 ± 3.1 MJ; p = 0.037), but not total-EE (HCLF=  44.6 ± 7.7, LCHF=  39.9 ± 4.7 MJ; p = 0.060), was higher for the HCLF condition than the LCHF across the measurement period. CONCLUSION: Compared with low carbohydrate/high fat, immediate post-exercise energy replacement with a high carbohydrate/low fat drink resulted in higher short-term activity energy expenditure and lower appetite ratings.

Validation of a combined ELISA to detect IgG, IgA and IgM antibody responses to SARS-CoV-2 in mild or moderate non-hospitalised patients.

BACKGROUND: Frequently SARS-CoV-2 results in mild or moderate disease with potentially lower concentrations of antibodies compared to those that are hospitalised. Here, we validated an ELISA using SARS-CoV-2 trimeric spike glycoprotein, with targeted detection of IgG, IgA and IgM (IgGAM) using serum and dried blood spots (DBS) from adults with mild or moderate disease. METHODS: Targeting the SARS-CoV-2 trimeric spike, a combined anti-IgG, IgA and IgM serology ELISA assay was developed using 62 PCR-confirmed non-hospitalised, mild or moderate COVID-19 samples, ≥14 days post symptom onset and 624 COVID-19 negative samples. The assay was validated using 73 PCR-confirmed non-hospitalised, mild or moderate COVID-19 samples, ≥14 days post symptom onset and 359 COVID-19 negative serum samples with an additional 81 DBSs. The assay was further validated in 226 PCR-confirmed non-hospitalised, mild or moderate COVID-19 samples, ≥14 days post symptom onset and 426 COVID-19 negative clinical samples. RESULTS: A sensitivity and specificity of 98.6% (95% CI, 92.6-100.0), 98.3% (95% CI, 96.4-99.4), respectively, was observed following validation of the SARS-CoV-2 ELISA. No cross-reactivities with endemic coronaviruses or other human viruses were observed, and no change in results were recorded for interfering substances. The assay was stable at temperature extremes and components were stable for 15 days once opened. A matrix comparison showed DBS to correlate with serum results. Clinical validation of the assay reported a sensitivity of 94.7% (95% CI, 90.9-97.2%) and a specificity of 98.4% (95% CI, 96.6-99.3%). CONCLUSIONS: The human anti-IgGAM SARS-CoV-2 ELISA provides accurate and sensitive detection of SARS-CoV-2 antibodies in non-hospitalised adults with mild or moderate disease. The use of dried blood spots makes the assay accessible to the wider community.

Measurement of Typhi Vi antibodies can be used to assess adaptive immunity in patients with immunodeficiency.

Vaccine-specific antibody responses are essential in the diagnosis of antibody deficiencies. Responses to Pneumovax II are used to assess the response to polysaccharide antigens, but interpretation may be complicated. Typhim Vi® , a polysaccharide vaccine for Salmonella typhoid fever, may be an additional option for assessing humoral responses in patients suspected of having an immunodeficiency. Here we report a UK multi-centre study describing the analytical and clinical performance of a Typhi Vi immunoglobulin (Ig)G enzyme-linked immunosorbent assay (ELISA) calibrated to an affinity-purified Typhi Vi IgG preparation. Intra- and interassay imprecision was low and the assay was linear, between 7·4 and 574 U/ml (slope = 0·99-1·00; R2  > 0·99); 71% of blood donors had undetectable Typhi Vi IgG antibody concentrations. Of those with antibody concentrations  > 7·4 U/ml, the concentration range was 7·7-167 U/ml. In antibody-deficient patients receiving antibody replacement therapy the median Typhi Vi IgG antibody concentrations were  < 25 U/ml. In vaccinated normal healthy volunteers, the median concentration post-vaccination was 107 U/ml (range 31-542 U/ml). Eight of eight patients (100%) had post-vaccination concentration increases of at least threefold and six of eight (75%) of at least 10-fold. In an antibody-deficient population (n = 23), only 30% had post-vaccination concentration increases of at least threefold and 10% of at least 10-fold. The antibody responses to Pneumovax II and Typhim Vi® correlated. We conclude that IgG responses to Typhim Vi® vaccination can be measured using the VaccZyme Salmonella typhi Vi IgG ELISA, and that measurement of these antibodies maybe a useful additional test to accompany Pneumovax II responses for the assessment of antibody deficiencies.

Lipolytic Markers, Insulin and Resting Fat Oxidation are Associated with Maximal Fat Oxidation.

The maximal capacity to oxidize fat during exercise (MFO) is associated with 24-h fat balance and insulin sensitivity. Understanding factors that influence MFO could have implications for metabolic health. We investigated relationships between selected plasma metabolites, hormones and overnight-fasted resting fat oxidation rates (Resting), with MFO. Resting fat oxidation and MFO was measured in 57 men with blood collected at rest and during exercise. Plasma glycerol (R=0.39, P=0.033), non-esterified fatty acids (NEFA: R=0.27, P=0.030) and insulin (R=- 0.36, P=0.007) measured at MFO correlated with MFO; only glycerol remained correlated when controlled for resting concentrations (R=0.36, P=0.008). The change in glycerol from rest to MFO correlated with exercise-induced fat oxidation (R=0.32, P=0.012). V˙O 2max correlated with resting fat oxidation (R=0.44, P=0.001) and MFO (R=0.52, P<0.001). Resting fat oxidation correlated with MFO (R=0.55, P<0.001); this remained when controlled for V˙O 2max (R=0.41, P=0.001). This study reports weak-to-moderate, albeit significant, relationships between plasma lipolytic markers, insulin and resting overnight-fasted fat oxidation with MFO and shows the plasma glycerol response to uniquely reflect exercise-induced fat oxidation. V˙O 2max correlates with fat oxidation but the relationship can be dissociated. Interventions to increase fat oxidation for optimal metabolic health would benefit from, but are not reliant on, increases in V˙O 2max.

Increasing shape modelling accuracy by adjusting for subject positioning: an application to the analysis of radiographic proximal femur symmetry using data from the Osteoarthritis Initiative.

In total hip arthroplasty, the shape of the contra-lateral femur frequently serves as a template for preoperative planning. Previous research on contra-lateral femoral symmetry has been based on conventional hip geometric measurements (which reduce shape to a series of linear measurements) and did not take the effect of subject positioning on radiographic femur shape into account. The aim of this study was to analyse proximal femur symmetry based on statistical shape models (SSMs) which quantify global femoral shape while also adjusting for differences in subject positioning during image acquisition. We applied our recently developed fully automatic shape model matching (FASMM) system to automatically segment the proximal femur from AP pelvic radiographs to generate SSMs of the proximal femurs of 1258 Caucasian females (mean age: 61.3 SD=9.0). We used a combined SSM (capturing the left and right femurs) to identify and adjust for shape variation attributable to subject positioning as well as a single SSM (including all femurs as left femurs) to analyse proximal femur symmetry. We also calculated conventional hip geometric measurements (head diameter, neck width, shaft width and neck-shaft angle) using the output of the FASMM system. The combined SSM revealed two modes that were clearly attributable to subject positioning. The average difference (mean point-to-curve distance) between left and right femur shape was 1.0mm before and 0.8mm after adjusting for these two modes. The automatic calculation of conventional hip geometric measurements after adjustment gave an average absolute percent asymmetry of within 3.1% and an average absolute difference of within 1.1mm or 2.9° for all measurements. We conclude that (i) for Caucasian females the global shape of the right and left proximal femurs is symmetric without isolated locations of asymmetry; (ii) a combined left-right SSM can be used to adjust for radiographic shape variation due to subject positioning; and (iii) adjusting for subject positioning increases the accuracy of predicting the shape of the contra-lateral hip.

Development of a fully automatic shape model matching (FASMM) system to derive statistical shape models from radiographs: application to the accurate capture and global representation of proximal femur shape.

OBJECTIVE: To evaluate the accuracy and sensitivity of a fully automatic shape model matching (FASMM) system to derive statistical shape models (SSMs) of the proximal femur from non-standardised anteroposterior (AP) pelvic radiographs. DESIGN: AP pelvic radiographs obtained with informed consent and appropriate ethical approval were available for 1105 subjects with unilateral hip osteoarthritis (OA) who had been recruited previously for The arcOGEN Study. The FASMM system was applied to capture the shape of the unaffected (i.e., without signs of radiographic OA) proximal femur from these radiographs. The accuracy and sensitivity of the FASMM system in calculating geometric measurements of the proximal femur and in shape representation were evaluated relative to validated manual methods. RESULTS: De novo application of the FASMM system had a mean point-to-curve error of less than 0.9 mm in 99% of images (n = 266). Geometric measurements generated by the FASMM system were as accurate as those obtained manually. The analysis of the SSMs generated by the FASMM system for male and female subject groups identified more significant differences (in five of 17 SSM modes after Bonferroni adjustment) in their global proximal femur shape than those obtained from the analysis of conventional geometric measurements. Multivariate gender-classification accuracy was higher when using SSM mode values (76.3%) than when using conventional hip geometric measurements (71.8%). CONCLUSIONS: The FASMM system rapidly and accurately generates a global SSM of the proximal femur from radiographs of varying quality and resolution. This system will facilitate complex morphometric analysis of global shape variation across large datasets. The FASMM system could be adapted to generate SSMs from the radiographs of other skeletal structures such as the hand, knee or pelvis.

Fully automatic segmentation of the proximal femur using random forest regression voting.

Extraction of bone contours from radiographs plays an important role in disease diagnosis, preoperative planning, and treatment analysis. We present a fully automatic method to accurately segment the proximal femur in anteroposterior pelvic radiographs. A number of candidate positions are produced by a global search with a detector. Each is then refined using a statistical shape model together with local detectors for each model point. Both global and local models use Random Forest regression to vote for the optimal positions, leading to robust and accurate results. The performance of the system is evaluated using a set of 839 images of mixed quality. We show that the local search significantly outperforms a range of alternative matching techniques, and that the fully automated system is able to achieve a mean point-to-curve error of less than 0.9 mm for 99% of all 839 images. To the best of our knowledge, this is the most accurate automatic method for segmenting the proximal femur in radiographs yet reported.

Accurate fully automatic femur segmentation in pelvic radiographs using regression voting.

Extraction of bone contours from radiographs plays an important role in disease diagnosis, pre-operative planning, and treatment analysis. We present a fully automatic method to accurately segment the proximal femur in anteroposterior pelvic radiographs. A number of candidate positions are produced by a global search with a detector. Each is then refined using a statistical shape model together with local detectors for each model point. Both global and local models use Random Forest regression to vote for the optimal positions, leading to robust and accurate results. The performance of the system is evaluated using a set of 519 images. We show that the fully automated system is able to achieve a mean point-to-curve error of less than 1 mm for 98% of all 519 images. To the best of our knowledge, this is the most accurate automatic method for segmenting the proximal femur in radiographs yet reported.

Pre-race dietary carbohydrate intake can independently influence sub-elite marathon running performance.

We examined whether selected anthropometric and nutritional factors influenced field-based marathon running performance. An internet-based data collection tool allowed competitors in the 2009 London Marathon (n=257, mean ± SD age: 39 ± 8 years, finish time: 273.8 ± 59.5 min) to record a range of anthropometric, training and nutritional predictors. Multivariate statistical methods were used to quantify the change in running speed mediated by a unit change in each predictor via the 95% confidence interval for each covariate-controlled regression slope ( B). Gender ( B=1.22 to 1.95 km/h), body mass index ( B=-0.14 to -0.27 km/h), training distance ( B=0.01 to 0.04 km/h) and the amount of carbohydrate consumed the day before the race ( B=0.08 to 0.26 km/h) were significant predictors, collectively accounting for 56% of the inter-individual variability in running speed (P<0.0005). Further covariate-adjusted analysis revealed that those competitors who consumed carbohydrate the day before the race at a quantity of >7 g/kg body mass had significantly faster overall race speeds (P=0.01) and maintained their running speed during the race to a greater extent than with those who consumed <7 g/kg body mass (P=0.02). We conclude that, in addition to gender, body size and training, pre-race day carbohydrate intake can significantly and independently influence marathon running performance.

Insights into the genetic architecture of osteoarthritis from stage 1 of the arcOGEN study.

OBJECTIVES: The genetic aetiology of osteoarthritis has not yet been elucidated. To enable a well-powered genome-wide association study (GWAS) for osteoarthritis, the authors have formed the arcOGEN Consortium, a UK-wide collaborative effort aiming to scan genome-wide over 7500 osteoarthritis cases in a two-stage genome-wide association scan. Here the authors report the findings of the stage 1 interim analysis. METHODS: The authors have performed a genome-wide association scan for knee and hip osteoarthritis in 3177 cases and 4894 population-based controls from the UK. Replication of promising signals was carried out in silico in five further scans (44,449 individuals), and de novo in 14 534 independent samples, all of European descent. RESULTS: None of the association signals the authors identified reach genome-wide levels of statistical significance, therefore stressing the need for corroboration in sample sets of a larger size. Application of analytical approaches to examine the allelic architecture of disease to the stage 1 genome-wide association scan data suggests that osteoarthritis is a highly polygenic disease with multiple risk variants conferring small effects. CONCLUSIONS: Identifying loci conferring susceptibility to osteoarthritis will require large-scale sample sizes and well-defined phenotypes to minimise heterogeneity.

Directional control-response relationships for mining equipment.

A variety of directional control-response relationships are currently found in mining equipment. Two experiments were conducted in a virtual environment to determine optimal direction control-response relationships in a wide variety of circumstances. Direction errors were measured as a function of control orientation (horizontal or vertical), location (left, front, right) and directional control-response relationships. The results confirm that the principles of consistent direction and visual field compatibility are applicable to the majority of situations. An exception is that fewer direction errors were observed when an upward movement of a horizontal lever or movement of a vertical lever away from the participants caused extension (lengthening) of the controlled device, regardless of whether the direction of movement of the control is consistent with the direction in which the extension occurs. Further, both the control of slew by horizontally oriented controls and the control of device movements in a frontal plane by the perpendicular movements of vertical levers were associated with relatively high rates of directional errors, regardless of the directional control-response relationship, and these situations should be avoided. STATEMENT OF RELEVANCE: The results are particularly applicable to the design of mining equipment such as drilling and bolting machines, and have been incorporated into MDG35.1 Guideline for bolting & drilling plant in mines (Industry & Investment NSW, 2010). The results are also relevant to the design of any equipment where vertical or horizontal levers are used to control the movement of equipment appendages, e.g. cranes mounted to mobile equipment and the like.

Shape learning and discrimination in reef fish.

Coral reef fish live in a complex world of colour and patterns. If they are to survive they need to be able to correctly identify the things they see (e.g. predators, prey) and act accordingly (e.g. flee, feed). This paper investigates whether discrimination is limited to ecologically relevant stimuli or is in fact more adaptable. Our work focuses on the reef damselfish Pomacentrus amboinensis. Within a day or two of capture the fish demonstrated an ability to associate an arbitrary stimulus with a food reward and to discriminate the reward stimulus from a distractor matched along various physical dimensions. In our initial experiments the reward was directly associated with the target. In the final experiment, however, the reward was separated from the target in both space and time, thereby eliminating a weakness applicable to the majority of food reward experiments involving fish; namely, the presence of olfactory cues emanating from the feeding tubes. All fish were not only able to solve this task but also showed anticipatory behaviour (also referred to as goal tracking). We conclude that freshly caught reef fish not only are able to quickly learn and discriminate between novel stimuli on the basis of shape but are also able to interpret stimuli as a predictor for the availability of food at a different time and place (anticipatory behaviour).

Colour vision in coral reef fish.

Over many millions of years, sea creatures have developed a range of light reflectance properties. One example is the large variation in the patterns and colours of fish inhabiting the world's coral reefs. Attempts to understand the significance of the colouration have been made, but all too often from the perspective of a human observer. A more ecological approach requires us to consider the visual system of those for whom the colours were intended, namely other sea life. A first step is to understand the sensitivity of reef fish themselves to colour. Physiological data has revealed wavelength-tuned photoreceptors in reef fish, and this study provides behavioural evidence for their application in colour discrimination. Using classical conditioning, freshly caught damselfish were trained to discriminate coloured patterns for a food reward. Within 3-4 days of capture the fish selected a target colour on over 75% of trials. Brightness of the distracter and target were systematically varied to confirm that the fish could discriminate stimuli on the basis of chromaticity alone. The study demonstrates that reef fish can learn to perform two-alternative discrimination tasks, and provides the first behavioural evidence that reef fish have colour vision.

Historic and contemporary levels of genetic variation in two New Zealand passerines with different histories of decline.

We compared historic and contemporary genetic variation in two threatened New Zealand birds (saddlebacks and robins) with disparate bottleneck histories. Saddlebacks showed massive loss of genetic variation when extirpated from the mainland, but no significant loss of variation following a severe bottleneck in the 1960s when the last population was reduced from approximately 1000 to 36 birds. Low genetic variation was probably characteristic of this isolated island population: considerably more genetic variation would exist in saddlebacks today if a mainland population had survived. In contrast to saddlebacks, contemporary robin populations showed only a small decrease in genetic variation compared with historical populations. Genetic variation in robins was probably maintained because of their superior ability to disperse and coexist with introduced predators. These results demonstrate that contemporary genetic variation may depend more greatly on the nature of the source population and its genetic past than it does on recent bottlenecks.

Mitochondrial phylogeography of New Zealand freshwater crayfishes, Paranephrops spp.

Tectonic movement at the boundary of the Indo-Australian and Pacific Plates during the Miocene and Pliocene is recognized as a driving force for invertebrate speciation in New Zealand. Two endemic freshwater crayfish (koura) species, Paranephrops planifrons White 1842 and Paranephrops zealandicus White 1842, represent good model taxa to test geological hypotheses because, due to their low dispersal capacity and life history, geographical restriction of populations may be caused by vicariant processes. Analysis of a mitochondrial DNA marker (cytochrome oxidase subunit I) reveals not two, but three major koura lineages. Contrary to expectation, the cryptic West Coast group appears to be more closely related to P. zealandicus than to P. planifrons and has diverged earlier than the final development (Late Pleistocene) of Cook Strait. Our date estimates suggest that koura lineage diversification probably coincided with early to mid-Alpine orogeny in the mid-Pliocene. Estimates of node ages and the phylogenies are inconsistent with both ancient Oligocene and recent postglacial Pleistocene range expansion, but suggest central to north colonization of North Island and west to east movement in South Island during mid- to late Pliocene. Crypsis and paraphyly of the West Coast group suggest that morphological characters presently used to classify koura species could be misleading.

Perceptions of orthopaedic surgeons regarding hepatitis C viral transmission: a questionnaire survey.

INTRODUCTION: Occupationally acquired hepatitis C viral infection is an important issue in surgery since there are no known vaccines or effective prophylaxis. MATERIALS AND METHODS: An anonymous questionnaire survey was performed to determine the attitudes and perception of risks of occupational acquired hepatitis C viral transmission in orthopaedic surgeons. RESULTS: A total of 763 questionnaires were posted to orthopaedic surgeons with various subspecialty interests and 261 surgeons responded (34.2%). Of respondents, 117 (47%) had sustained sharps injuries in the previous 12 months. Only 82 surgeons (33%) always reported such injuries, although 208 (84%) expressed concerns of occupationally acquired hepatitis C viral transmission. Orthopaedic surgeons were mostly unaware of the true prevalence of hepatitis C in high-risk groups, such as intravenous drug abusers. CONCLUSIONS: Greater awareness of all aspects of hepatitis C infection and its risks to the practice of surgery is required. Further debate is necessary on the role of routine testing of surgeons and patients.

Linkage to nodal osteoarthritis: quantitative and qualitative analyses of data from a whole-genome screen identify trait-dependent susceptibility loci.

OBJECTIVE: To identify susceptibility loci for nodal osteoarthritis. METHODS: A genome screen at an average marker spacing of 9.29 cM was carried out on 558 people from 202 families, of whom 491 had nodal osteoarthritis. All genotyped people were graded for the incidence and severity of distal interphalangeal (DIP) nodes, and radiographs from 354 people were graded for joint-space narrowing (JSN) and osteophytes (OSTs). Age-regressed indices for DIP nodes, JSN and OSTs were calculated using these phenotypic data. Affected sibling pair (ASP) and quantitative trait analyses were carried out using MERLIN. RESULTS: The data analysis identified suggestive linkage to loci on chromosomes 3 (for JSN and OST), 4 (for JSN), 8 (for DIP), 11 (for radiographic osteoarthritis) and 16 (for JSN). Both the ASP and quantitative analyses identified the loci on chromosomes 4 and 11. The loci on chromosomes 3 and 16 overlap with those previously identified for large-joint osteoarthritis. Of the loci identified by the quantitative analyses with the logarithm of the odds of linkage >1.5, two were linked to more than one trait, whereas nine were linked to single traits: one for DIP, six for JSN and two for OST. CONCLUSION: The ASP and quantitative analyses of the cohort with nodal osteoarthritis suggest that multiple susceptibility loci for osteoarthritis influence the traits, which combine to form the osteoarthritis phenotype, and that these loci may not act exclusively on the joints of the hand.

Transcription factor XBP-1 is expressed during osteoblast differentiation and is transcriptionally regulated by parathyroid hormone (PTH).

X-box-binding protein 1 (XBP-1) is a basic-region leucine zipper protein in the cyclic AMP response element binding protein/activating transcription factor (CREB/ATF) family of transcription factors involved in different cell-differentiation processes. We have investigated the expression of XBP-1 in differentiating MC3T3-E1 osteoblastic cells. Cultures were treated with ascorbic acid (AA) and beta-glycerophosphate (BGP) to induce differentiation. Under these conditions, the basal transcription of xbp-1 increases at day 2 following induction, peaks at day 5 and decreases thereafter. This result showed that xbp-1 gene is differentially expressed during MC3T3-E1 cell differentiation. Detection of XBP-1 by immunofluorescence at days 0 (control culture without AA and BGP), 8 and 21 showed that the protein has a major cytoplasmic perinuclear location. In addition, xbp-1 is transcriptionally upregulated by parathyroid hormone within 2.5 h of treatment and decreases thereafter.