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This article shares insights from the experiences of three high school practitioners and two university faculty who participated in a school-university-based action research program as a voluntary part of the teachers’ professional development. The three high school practitioners conducted action research projects around questions that stemmed from and were relevant to their own teaching practice. As part of the action research program, the practitioners were paired with university faculty to support the research. Building on practitioner inquiry traditions and critical case study methodologies, this study used qualitative methods to explore the experiences of practitioner action research processes. Drawing on in-person meeting notes and reflective memos, four key ideas emerged: Infrastructure, We are all Partners in Education, Engaging Pathway for Experienced Teachers, and Challenges. Insights gained from this inquiry will have implications for professional practices in the areas of school-university partnership, professional development, and action research process.

Montmorency cherry concentrate (MCC) supplementation enhances functional recovery from exercise, potentially due to antioxidant and anti-inflammatory effects. However, to date, supporting empirical evidence for these mechanistic hypotheses is reliant on indirect blood biomarkers. This study is the first to investigate functional recovery from exercise alongside molecular changes within the exercised muscle after MCC supplementation.Ten participants completed two maximal unilateral eccentric knee extension trials after MCC or placebo (PLA) supplementation for 7 d before and 48 h after exercise. Knee extension maximum voluntary contractions, maximal isokinetic contractions, single leg jumps, and soreness measures were assessed before, immediately, 24 h, and 48 h after exercise. Venous blood and vastus lateralis muscle samples were collected at each time point. Plasma concentrations of interleukin-6, tumor necrosis factor alpha, C-reactive protein, creatine kinase, and phenolic acids were quantified. Intramuscular mRNA expressions of superoxide dismutase 1 (SOD1), SOD3, glutathione peroxidase 1 (GPX1), GPX3, GPX4, GPX7, catalase, and nuclear factor erythroid 2-related factor 2 and relative intramuscular protein expressions of SOD1, catalase, and GPX3 were quantified.MCC supplementation enhanced the recovery of normalized maximum voluntary contraction 1-s average compared with PLA (postexercise PLA, 59.5% ± 18.0%, vs MCC, 76.5% ± 13.9%; 24 h PLA, 69.8% ± 15.9%, vs MCC, 80.5% ± 15.3%; supplementation effect P = 0.024). MCC supplementation increased plasma hydroxybenzoic, hippuric, and vanillic acid concentrations (supplementation effect P = 0.028, P = 0.002, P = 0.003); SOD3, GPX3, GPX4, GPX7 (supplement effect P0.05), and GPX1 (interaction effect P = 0.017) gene expression; and GPX3 protein expression (supplementation effect P = 0.004) versus PLA. There were no significant differences between conditions for other outcome measures.MCC supplementation conserved isometric muscle strength and upregulated antioxidant gene and protein expression in parallel with increased phenolic acid concentrations.

Shatavari has long been used as an Ayurvedic herb for women’s health, but empirical evidence for its effectiveness has been lacking. Shatavari contains phytoestrogenic compounds that bind to the estradiol receptor. Postmenopausal estradiol deficiency contributes to sarcopenia and osteoporosis. In a randomised double-blind trial, 20 postmenopausal women (68.5 ± 6 years) ingested either placebo (N = 10) or shatavari (N = 10; 1000 mg/d, equivalent to 26,500 mg/d fresh weight shatavari) for 6 weeks. Handgrip and knee extensor strength were measured at baseline and at 6 weeks. Vastus lateralis (VL) biopsy samples were obtained. Data are presented as difference scores (Week 6—baseline, median ± interquartile range). Handgrip (but not knee extensor) strength was improved by shatavari supplementation (shatavari +0.7 ± 1.1 kg, placebo −0.4 ± 1.3 kg; p = 0.04). Myosin regulatory light chain phosphorylation, a known marker of improved myosin contractile function, was increased in VL following shatavari supplementation (immunoblotting; placebo −0.08 ± 0.5 a.u., shatavari +0.3 ± 1 arbitrary units (a.u.); p = 0.03). Shatavari increased the phosphorylation of Aktser473 (Aktser473 (placebo −0.6 ± 0.6 a.u., shatavari +0.2 ± 1.3 a.u.; p = 0.03) in VL. Shatavari supplementation did not alter plasma markers of bone turnover (P1NP, β-CTX) and stimulation of human osteoblasts with pooled sera (N = 8 per condition) from placebo and shatavari supplementation conditions did not alter cytokine or metabolic markers of osteoblast activity. Shatavari may improve muscle function and contractility via myosin conformational change and further investigation of its utility in conserving and enhancing musculoskeletal function, in larger and more diverse cohorts, is warranted.

BackgroundShatavari has long been used as an Ayurvedic herb for women’s health, but empirical evidence for its effectiveness has been lacking. Shatavari contains phytoestrogenic compounds that bind to the estradiol receptor, and may therefore benefit postmenopausal women since postmenopausal estradiol deficiency contributes to sarcopenia and osteoporosis.MethodsIn a randomised double-blind trial, 20 postmenopausal women (68.5 ± 6 y) ingested either placebo (N=10) or shatavari (N=10; 1000 mg/d, equivalent to 26,500 mg/d fresh weight shatavari) for 6 weeks. Handgrip and knee extensor strength were measured at baseline and at 6 weeks. Vastus lateralis (VL) biopsy samples were obtained. Data are presented and analysed (t test/Mann Whitney U) as difference scores (Week 6 – baseline, median ± interquartile range).ResultsHandgrip, (but not knee extensor) strength was improved by shatavari supplementation (shatavari +0.7 ± 1.1 kg, placebo -0.4 ± 1.3 kg; p=0.04). Myosin regulatory light chain phosphorylation, a known marker of improved myosin contractile function, was increased in VL following shatavari supplementation (immunoblotting; placebo -0.08 ± 0.5 a.u. shatavari +0.3 ± 1 arbitrary units (a.u.); p=0.03). Shatavari increased phosphorylation of Aktser473 (Aktser473 (placebo -0.6 ± 0.6 a.u. shatavari +0.2 ± 1.3 a.u; p=0.03) in VL. Shatavari supplementation did not alter plasma markers of bone turnover (P1NP, β-CTX) and stimulation of human osteoblasts with pooled sera (N=8 per condition) from placebo and shatavari supplementation conditions did not alter cytokine or metabolic markers of osteoblast activity.ConclusionsShatavari may improve muscle function and contractility via myosin conformational change and warrants further investigation of its utility in conserving musculoskeletal function in postmenopausal women.Trial RegistrationRetrospectively registered at clinicaltrials.gov as NCT05025917 on 30/08/21.

Background: Mediterranean diets limit red meat consumption and increase intakes of high-phytate foods, a combination that could reduce iron status. Conversely, higher intakes of fish, a good source of selenium, could increase selenium status. Objectives: A 1-y randomized controlled trial [New Dietary Strategies Addressing the Specific Needs of the Elderly Population for Healthy Aging in Europe (NU-AGE)] was carried out in older Europeans to investigate the effects of consuming a Mediterraneanstyle diet on indices of inflammation and changes in nutritional status. Methods: Selenium and iron intakes and status biomarkers were measured at baseline and after 1 y in 1294 people aged 65–79 y from 5 European countries (France, Italy, the Netherlands, Poland, and the United Kingdom) who had been randomly allocated either to a Mediterranean-style diet or to remain on their habitual, Western diet. Results: Estimated selenium intakes increased significantly with the intervention group (P < 0.01), but were not accompanied by changes in serum selenium concentrations. Iron intakes also increased (P < 0.001), but there was no change in iron status. However, when stratified by study center, there were positive effects of the intervention on iron status for serum ferritin for participants in Italy (P = 0.04) and France (P = 0.04) and on soluble transferrin receptor (sTfR) for participants in Poland (P < 0.01). Meat intake decreased and fish intake increased to a greater degree in the intervention group, relative to the controls (P < 0.01 for both), but the overall effects of the intervention on meat and fish intakes were mainly driven by data from Poland and France. Changes in serum selenium in the intervention group were associated with greater changes in serum ferritin (P = 0.01) and body iron (P = 0.01), but not sTfR (P = 0.73); there were no study center × selenium status interactions for the iron biomarkers. Conclusions: Consuming a Mediterranean-style diet for 1 y had no overall effect on iron or selenium status, although there were positive effects on biomarkers of iron status in some countries. The NU-AGE trial was registered at clinicaltrials.gov as NCT01754012. Am J Clin Nutr 2019;00:1–12.

Introduction Low- and middle-income countries (LMICs) account for 70% of all global burns. Due to this significantly disproportionate burden, it’s critical we identify barriers to burn care and prevention in LMICs. As a result, this study aimed to elucidate trends in LMIC-related burn research to create focused strategies for burn care training, research, and innovation. Accomplishing meaningful change from the study’s findings will be guided by the first 4 steps of Dr. John Kotter’s “8-Step Process for Leading Change” – 1) create urgency for change, 2) build a guiding team, 3) develop a vision and plan, 4) communicate with key stakeholders to obtain buy-in. Methods Web of Science’s 7 citation databases were searched through March 2, 2021 using synonyms of “burns” and “low- and middle-income countries.” After screening articles, metadata were uploaded to VOSviewer (Leiden, Netherlands) where citation and network metrics were generated. The Kruskal-Wallis test and linear regression were used for bivariable and multivariable analysis of factors influencing publications, citations, and total link strength (TLS) – the strength of association between a given research article, other articles, and additional institutions. Results Bibliometric analysis identified 2,027 articles by 8,602 authors in 692 journals. Two-thirds of journals published a single article (n=453, 65.5%) whereas only 3.6% published ≥10 articles. One-quarter of LMIC burn research was published in ISBI’s Burns (n=417 articles, 20.6%) and ABA’s Journal of Burn Care & Research (n=89 articles, 4.4%). Most authors published < 5 articles (n=8521, 99.1%) but 19 (0.2%) had published ≥10. Authors were affiliated with 2,519 organizations in 132 countries. There was a strong positive correlation between total publications and citations (R=0.87, P< 0.001). In addition, there was a significant difference in the number of publications (P=0.003, 0.07), citations (P=0.005, 0.03), and TLS (P=0.009, 0.008) by geographic and economic categories - North America had the highest while Latin American and the Caribbean had the lowest. The USA (n = 563), India (n = 161), and China (n = 154) published the most articles. Conclusions Given the disproportionate representation of high-income countries and authors in the current LMIC burn research landscape, there must be a sense of urgency to develop pathways for facilitating change. Local and regional candidates for mentors and leaders were identified using bibliometric findings. Assembling teams with these individuals and prolific authors using a well-defined vision for change will facilitate sustainable communication and collaboration within LMIC research.

Simple Summary Animals provide food and other critical resources to much of the global population. Transboundary animal diseases are highly contagious or transmissible, epidemic diseases, with the potential to spread rapidly. They have the potential to cause negative socioeconomic and public health consequences. A greater understanding of the factors contributing to disease pathogenesis and spread is needed. Further work is also needed to improve the efficacy and cost of diagnostics and prevention measures for these diseases. This review aims to give a broad overview of 17 transboundary diseases, providing researchers and veterinarians with a current, succinct resource of salient details regarding these significant diseases. For each disease, we provide a synopsis of the disease and its status, species and geographic areas affected, a summary of research models, and when available, information regarding prevention or treatment. Abstract Animals provide food and other critical resources to most of the global population. As such, diseases of animals can cause dire consequences, especially disease with high rates of morbidity or mortality. Transboundary animal diseases (TADs) are highly contagious or transmissible, epidemic diseases, with the potential to spread rapidly across the globe and the potential to cause substantial socioeconomic and public health consequences. Transboundary animal diseases can threaten the global food supply, reduce the availability of non-food animal products, or cause the loss of human productivity or life. Further, TADs result in socioeconomic consequences from costs of control or preventative measures, and from trade restrictions. A greater understanding of the transmission, spread, and pathogenesis of these diseases is required. Further work is also needed to improve the efficacy and cost of both diagnostics and vaccines. This review aims to give a broad overview of 17 TADs, providing researchers and veterinarians with a current, succinct resource of salient details regarding these significant diseases. For each disease, we provide a synopsis of the disease and its status, species and geographic areas affected, a summary of in vitro or in vivo research models, and when available, information regarding prevention or treatment.

Incidence of Bordetella pertussis, the causative agent of whooping cough, is rising in some global human populations despite high vaccination rates, and significant research is underway to address the issue. Baboons are an established model for pertussis research, but like many mammals, they can be naturally infected with Bordetella bronchiseptica. Because B. bronchiseptica interferes with B. pertussis research, it must be excluded from baboons under consideration for enrollment in pertussis studies. In addition to research-related concerns, B. bronchiseptica can sometimes cause clinical disease in baboons and other nonhuman primates. This study examined the use of antibiotics to clear B. bronchiseptica in naturally infected baboons. Thirty-five juvenile baboons were divided into five treatment groups: oral sulfamethoxazole/trimethoprim (TMS), nebulized gentamicin (gentamicin), combination (TMS + gentamicin) in positive animals, combination (TMS + gentamicin) as a prophylactic in exposed animals and no treatment (control). Combination of oral TMS and nebulized gentamicin given to positive animals was most effective, producing long-term clearance in 11 out of 12 treated animals. To avoid unnecessary use of antibiotics, our primary management strategy is screening and separating to allow natural clearance and limiting exposure to non-infected animals, but this study investigates an antibiotic regimen that could be used in special circumstances.

Non-human primate models will expedite therapeutics and vaccines for coronavirus disease 2019 (COVID-19) to clinical trials. Here, we compare acute severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection in young and old rhesus macaques, baboons and old marmosets. Macaques had clinical signs of viral infection, mild to moderate pneumonitis and extra-pulmonary pathologies, and both age groups recovered in two weeks. Baboons had prolonged viral RNA shedding and substantially more lung inflammation compared with macaques. Inflammation in bronchoalveolar lavage was increased in old versus young baboons. Using techniques including computed tomography imaging, immunophenotyping, and alveolar/peripheral cytokine response and immunohistochemical analyses, we delineated cellular immune responses to SARS-CoV-2 infection in macaque and baboon lungs, including innate and adaptive immune cells and a prominent type-I interferon response. Macaques developed T-cell memory phenotypes/responses and bystander cytokine production. Old macaques had lower titres of SARS-CoV-2-specific IgG antibody levels compared with young macaques. Acute respiratory distress in macaques and baboons recapitulates the progression of COVID-19 in humans, making them suitable as models to test vaccines and therapies.

An urgent global quest for effective therapies to prevent and treat COVID-19 disease is ongoing. We previously described REGN-COV2, a cocktail of two potent neutralizing antibodies (REGN10987+REGN10933) targeting non-overlapping epitopes on the SARS-CoV-2 spike protein. In this report, we evaluate the in vivo efficacy of this antibody cocktail in both rhesus macaques and golden hamsters and demonstrate that REGN-COV-2 can greatly reduce virus load in lower and upper airway and decrease virus induced pathological sequalae when administered prophylactically or therapeutically. Our results provide evidence of the therapeutic potential of this antibody cocktail.

SummaryThere are no known cures or vaccines for COVID-19, the defining pandemic of this era. Animal models are essential to fast track new interventions and nonhuman primate (NHP) models of other infectious diseases have proven extremely valuable. Here we compare SARS-CoV-2 infection in three species of experimentally infected NHPs (rhesus macaques, baboons, and marmosets). During the first 3 days, macaques developed clinical signatures of viral infection and systemic inflammation, coupled with early evidence of viral replication and mild-to-moderate interstitial and alveolar pneumonitis, as well as extra-pulmonary pathologies. Cone-beam CT scans showed evidence of moderate pneumonia, which progressed over 3 days. Longitudinal studies showed that while both young and old macaques developed early signs of COVID-19, both groups recovered within a two-week period. Recovery was characterized by low-levels of viral persistence in the lung, suggesting mechanisms by which individuals with compromised immune systems may be susceptible to prolonged and progressive COVID-19. The lung compartment contained a complex early inflammatory milieu with an influx of innate and adaptive immune cells, particularly interstitial macrophages, neutrophils and plasmacytoid dendritic cells, and a prominent Type I-interferon response. While macaques developed moderate disease, baboons exhibited prolonged shedding of virus and extensive pathology following infection; and marmosets demonstrated a milder form of infection. These results showcase in critical detail, the robust early cellular immune responses to SARS-CoV-2 infection, which are not sterilizing and likely impact development of antibody responses. Thus, various NHP genera recapitulate heterogeneous progression of COVID-19. Rhesus macaques and baboons develop different, quantifiable disease attributes making them immediately available essential models to test new vaccines and therapies.

Mechanisms mediating postexercise cold-induced increases in PGC-1α gene expression in human skeletal muscle are yet to be fully elucidated but may involve local cooling effects on AMPK and p38 MAPK-related signaling and/or increased systemic β-adrenergic stimulation. Therefore, we aimed to examine whether postexercise cold water immersion enhancement of PGC-1α mRNA is mediated through local or systemic mechanisms. Ten subjects completed acute cycling (8 × 5 min at ~80% peak power output) followed by seated-rest (CON) or single-leg cold water immersion (CWI; 10 min, 8°C). Muscle biopsies were obtained preexercise, postexercise, and 3 h postexercise from a single limb in the CON condition but from both limbs in CWI [thereby providing tissue from a CWI and nonimmersed limb (NOT)]. Muscle temperature decreased up to 2 h postexercise following CWI (−5°C) in the immersed limb, with lesser changes observed in CON and NOT (−3°C, P < 0.05). No differences between limbs were observed in p38 MAPK phosphorylation at any time point ( P < 0.05), whereas a significant interaction effect was present for AMPK phosphorylation ( P = 0.031). Exercise (CON) increased gene expression of PGC-1α 3 h postexercise (~5-fold, P < 0.001). CWI augmented PGC-1α expression above CON in both the immersed (CWI; ~9-fold, P = 0.003) and NOT limbs (~12-fold, P = 0.001). Plasma normetanephrine concentration was higher in CWI vs. CON immediately postimmersion (860 vs. 665 pmol/l, P = 0.034). We report for the first time that local cooling of the immersed limb evokes transcriptional control of PGC-1α in the nonimmersed limb, suggesting increased systemic β-adrenergic activation of AMPK may mediate, in part, postexercise cold induction of PGC-1α mRNA. NEW & NOTEWORTHY We report for the first time that postexercise cold water immersion of one limb also enhances PGC-1α expression in a contralateral, nonimmersed limb. We suggest that increased systemic β-adrenergic stimulation, and not localized cooling per se, exerts regulatory effects on local signaling cascades, thereby modulating PGC-1α expression. Therefore, these data have important implications for research designs that adopt contralateral, nonimmersed limbs as a control condition while also increasing our understanding of the potential mechanisms underpinning cold-mediated PGC-1α responses.

The demand for measurement of vitamin D metabolites for clinical diagnosis and to advance our understanding of the role of vitamin D in human health has significantly increased in the last decade. New developments in technologies employed have enabled the separation and quantification of additional metabolites and interferences. Also, developments of immunoassays have changed the landscape. Programmes and materials for assay standardisation, harmonisation and the expansion of the vitamin D external quality assurance scheme (DEQAS) with the provision of target values as measured by a reference measurement procedure have improved standardisation, quality assurance and comparability of measurements. In this article, we describe developments in the measurement of the commonly analysed vitamin D metabolites in clinical and research practice. We describe current analytical approaches, discuss differences between assays, their origin, and how these may be influenced by physiological and experimental conditions. The value of measuring metabolites beyond 25 hydroxyvitamin D (25(OH)D), the marker of vitamin D status, in routine clinical practice is not yet confirmed. Here we provide an overview of the value and application of the measurement of 1,25 dihydroxyvitamin D, 24,25 dihydroxyvitamin D and free 25OHD in the diagnosis of patients with abnormalities in vitamin D metabolism and for research purposes.

Gingival lesions as the sole manifestation of African histoplasmosis (Histoplasma capsulatum var. duboisii) have never been reported in baboons. Grossly, lesions can be indistinguishable from bacterial ulcerative gingivitis or gingival hyperplasia. Clinical outcomes of primary gingival histoplasmosis in baboons are unknown, and, may complicate colony management decisions.

We assessed the effects of post‐exercise cold‐water immersion (CWI) in modulating PGC‐1α mRNA expression in response to exercise commenced with low muscle glycogen availability. In a randomized repeated‐measures design, nine recreationally active males completed an acute two‐legged high‐intensity cycling protocol (8 × 5 min at 82.5% peak power output) followed by 10 min of two‐legged post‐exercise CWI (8°C) or control conditions (CON). During each trial, one limb commenced exercise with low (LOW

Background: Nutritional status can be difficult to assess. Bioelectrical impedance analysis (BIA)-derived phase angle (PA), and the plasma markers citrulline and transthyretin (pre-albumin) have the potential to assist, but the protocol of fasting and resting for BIA renders the investigation impractical for routine use, especially so in populations at high risk of malnutrition. Aims: 1- To clarify whether starving and resting are necessary for reliable measurement of PA. 2- To identify whether PA, citrulline and transthyretin correlate with nutritional status. Methods: Eighty consenting adult in-patients were recruited. Nutritional status was determined by subjective global assessment (SGA) used as gold standard. The Malnutrition Universal Screening Tool (MUST) was used and anthropometric measurements were performed. Serum was analysed for citrulline and transthyretin. PA was measured using Bodystat 4000. The PA was considered to define malnutrition when lower than reference ranges for sex and age, and severe malnutrition if more than 2 integers below the lower limit. Anthropometric measurements were categorised according to WHO reference centiles. Ordinal logistic regression estimated the strength of association of PA, citrulline and transthyretin with SGA. PA values in the different metabolic states were compared using paired t tests. Results: All 80 subjects completed the BIA and the nutritional assessments in the 3 different states; 14 declined to provide blood samples for the biochemical assays. Malnutrition was identified in 32 cases, severe malnutrition in 14 cases, the remaining 34 cases were deemed not to be malnourished. PA was strongly inversely associated with SGA (Odds Ratio [OR] per unit increase = 0.21, CI 0.12-0.37, p < 0.001). PA was not influenced by exercise (p=0.134) or food intake (p=0.184). Transthyretin was inversely associated with malnourished/severely malnourished states (OR = 0.98, 95% CI 0.97 – 0.99, p = 0.001), but had poorer predictive values than PA. There was no significant association between citrulline concentration and SGA (OR = 1.01, 95% CI 0.99-1.04, p = 0.348). Conclusions: The BIA-derived PA reliably identifies malnutrition. It is strongly associated with SGA but requires less skill and experience, and out-performs circulating transthyretin, rendering it a promising and less operator-dependent tool for assessing nutritional status in hospital patients. Our novel demonstration that fasting and bed-rest are unnecessary consolidates that position.

Background Currently, implantation of patient-specific cranial prostheses requires reoperation after a period for design and formulation by a third-party manufacturer. Recently, 3-dimensional (3D) printing via fused deposition modeling has demonstrated increased ease of use, rapid production time, and significantly reduced costs, enabling expanded potential for surgical application. Three-dimensional printing may allow neurosurgeons to remove bone, perform a rapid intraoperative scan of the opening, and 3D print custom cranioplastic prostheses during the remainder of the procedure. Objective To evaluate the feasibility of using a commercially available 3D printer to develop and produce on-demand intraoperative patient-specific cranioplastic prostheses in real time and assess the associated costs, fabrication time, and technical difficulty. Methods Five different craniectomies were each fashioned on 3 cadaveric specimens (6 sides) to sample regions with varying topography, size, thickness, curvature, and complexity. Computed tomography-based cranioplastic implants were designed, formulated, and implanted. Accuracy of development and fabrication, as well as implantation ability and fit, integration with exiting fixation devices, and incorporation of integrated seamless fixation plates were qualitatively evaluated. Results All cranioprostheses were successfully designed and printed. Average time for design, from importation of scan data to initiation of printing, was 14.6 min and average print time for all cranioprostheses was 108.6 min. Conclusion On-demand 3D printing of cranial prostheses is a simple, feasible, inexpensive, and rapid solution that may help improve cosmetic outcomes; significantly reduce production time and cost-expanding availability; eliminate the need for reoperation in select cases, reducing morbidity; and has the potential to decrease perioperative complications including infection and resorption.

AB Purpose: Strenuous endurance exercise increases biochemical markers of bone resorption but not formation, although the effect of recovery duration between consecutive bouts of exercise is unknown. We examined the effect of recovery duration on the bone metabolic response to two bouts of running. Methods: Ten physically active men completed two 9-d trials. On days 4 and 5 (D4 and D5), participants completed two 60-min bouts of running at 65% V[spacing dot above]O2max separated by either a 23-h (LONG) or a 3-h (SHORT) recovery period. Osteoprotegerin (OPG), parathyroid hormone (PTH), albumin-adjusted calcium (ACa), and phosphate (PO4) were measured from blood samples obtained before and for 3 h after exercise and on four follow-up days (D6-D9). Markers of bone resorption (C-terminal telopeptide region of collagen type 1) and bone formation (N-terminal propeptides of procollagen type 1 and bone alkaline phosphatase) were measured in early morning fasted samples on D4-D9. Results: There were no significant changes in C-terminal telopeptide region of collagen type 1, N-terminal propeptides of procollagen type 1, or bone alkaline phosphatase with either protocol. OPG, PTH, ACa, and PO4 concentrations increased with all exercise bouts, but the response to the second bout was not altered by recovery duration. Conclusions: Two 60-min bouts of running at 65% V[spacing dot above]O2max separated by either 23 or 3 h had no effect on the markers of bone resorption or formation from 1 to 4 d after exercise. Reducing recovery duration from 23 to 3 h between two bouts of running did not alter the increase in OPG, PTH, ACa, and PO4 to the second bout.

Background: Pyridinium cross-links Pyridinoline (PYD) and Deoxypyridinoline (DPD) are established markers of collagen degradation. Measurement of PYD and DPD can be used to evaluate changes in bone turnover in patients with metabolic bone disease and to monitor response to anti-resorptive treatment. Objective: To develop a method to extract and measure urine free PYD (fPYD) and free DPD (fDPD) by Liquid Chromatography Tandem Mass Spectrometry (LC-MS/MS). The method was used to quantify urine samples from 172 healthy individuals and 63 patients diagnosed with metabolic bone disease. Method: Acidified urine samples were extracted using solid phase extraction with cellulose slurry. PYD and DPD were separated by reversed-phase, ion-paired chromatography prior to MS/MS detection. Results: The fully validated method showed good agreement with other laboratories in the UK National External Proficiency Scheme (UK NEQAS). The method was compared against two commercial immunoassays for fDPD and pyridinium cross-links, r2 were 0.906 and 0.816 respectively. Urine concentrations of fDPD/Cr and fPYD/Cr were significantly higher in the patients than healthy individuals (p

Background: Population-based research on vitamin D has increased dramatically in recent years. Such studies are typically reliant on assay procedures to measure reliable and comparable levels of 25-hydroxyvitamin D [25(OH)D] concentrations. Methods: Concentrations of 25(OH)D3 and 25(OH)D2 were measured using LC-MS/MS in 5,915 participants (aged 31 years) of Northern Finland Birth Cohort 1966. Blood samples were assayed in batches over a course of 18 months. As anomalies were present in the measurements, 200 samples were reassayed using Diasorin RIA. Agreement between measurements was assessed by Passing-Bablok regression and limits of agreement (LoA). To harmonize LC-MS/MS with Diasorin RIA measurements, formulae were derived from the LoA. Results: Concentrations measured by LC-MS/MS were much higher than those measured by Diasorin RIA, with a mean difference of 12.9 ng/ml. Constant variation was evident between batch measurements after log transformation. Statistical formula was applied separately for each batch of LC-MS/MS measurements, enabling us to remove both the constant and proportional bias that was evident prior to the transformation. Conclusion: Despite the introduction of schemes/programs to improve accuracy of assays to measure 25(OH)D, significant differences can still happen. In these instances, methods to harmonize measurements based on a relatively small number of replicates can be successfully applied to establish confidence and to enable between-study comparisons. Refereed/Peer-reviewed

Studying the early stages of cancer can provide important insight into the molecular basis of the disease. We identified a preneoplastic stage in the patched (ptc) mutant mouse, a model for the brain tumor medulloblastoma. Preneoplastic cells (PNCs) are found in most ptc mutants during early adulthood, but only 15% of these animals develop tumors. Although PNCs are found in mice that develop tumors, the ability of PNCs to give rise to tumors has never been demonstrated directly, and the fate of cells that do not form tumors remains unknown. Using genetic fate mapping and orthotopic transplantation, we provide definitive evidence that PNCs give rise to tumors, and show that the predominant fate of PNCs that do not form tumors is differentiation. Moreover, we show that N-myc, a gene commonly amplified in medulloblastoma, can dramatically alter the fate of PNCs, preventing differentiation and driving progression to tumors. Importantly, N-myc allows PNCs to grow independently of hedgehog signaling, making the resulting tumors resistant to hedgehog antagonists. These studies provide the first direct evidence that PNCs can give rise to tumors, and demonstrate that identification of genetic changes that promote tumor progression is critical for designing effective therapies for cancer.

Providing adequate analgesia while minimizing handling and stress post-surgery can be challenging. Recently, there have been commercial products made available for providing long acting analgesia in rodents. However, we find there are limitations for use in mice due to the viscosity of the product and the small dosing volumes needed. This project evaluated an in-house compounded formulation of buprenorphine easily made in the laboratory using pharmaceutical grade products. The release of buprenorphine was evaluated when compounded with two types of hydrogels (Pluronic(®) F-127 and F-68). Mice given buprenorphine in hydrogel (BP) demonstrated higher serum levels of buprenorphine for a longer period of time compared to mice given standard buprenorphine (Bup). However, the rate of decline in serum levels between the groups was similar; thus, it is more likely that the higher buprenorphine concentration seen in the BP group is due to the higher dose of buprenorphine given, rather than a slower release of product. Feed consumption was decreased in both groups one day after dosing; however, there was no difference in body weights. Increased activity in the open field was observed with both buprenorphine formulations, and lipemia was observed in mice given BP which persisted to at least 96 h. Based on our results, we conclude that this formulation did not sustain the release of buprenorphine or eliminate the increased activity commonly seen in mice given buprenorphine. In addition, the lipemia may confound research parameters, especially in cardiac studies and lipid metabolism studies. Therefore, we cannot recommend this formulation for use.

The purpose of this study was to compare the sympathoadrenergic and metabolic responses following 30 s of maximal high intensity cycle ergometry exercise when cradle resistive forces were derived from total-body mass (TBM) or fat-free mass (FFM). Increases in peak power output (PPO) and pedal velocity were recorded when resistive forces reflected FFM (953 +/- 114 W vs 1020 +/- 134 W; 134 +/- 8 rpm vs 141 +/- 7 rpm ; P < 0.05). No differences were observed between mean power output (MPO), fatigue index (FI%), work done (WD) or heart rate (HR) when the TBM and FFM protocols were compared. There were no differences between the TBM and FFM protocols for adrenaline (A), noradrenaline (NA) or blood lactate concentrations ([La-]B) recorded at rest, immediately post or 24 h post exercise. However, increases in blood concentrations of A and NA (P < 0.05) were recorded for both the TBM and FFM protocol immediately post exercise. Significant correlations (P < 0.05) were recorded between PPOs, immediate post- exercise NA and [La-]B for both the TBM and FFM protocols. [La-]B levels were also significantly elevated (P < 0.01) immediately post exercise for both the TBM and FFM protocols. The results from this study suggest that greater peak power outputs are obtainable with no subsequent differences in neurophysiological or metabolic stress as determined by plasma A, NA and [La-]B concentrations when resistive forces reflect FFM and not TBM during loading procedures. The findings also indicate that immediate post exercise concentrations return to resting levels 24 h post exercise.

Background Alkaptonuria is a rare debilitating autosomal recessive disorder of tyrosine metabolism, where deficiency of homogentisate 1,2-dioxygenase results in increased homogentisic acid. Homogentisic acid is deposited as an ochronotic pigment in connective tissues, especially cartilage, leading to a severe early onset form of osteoarthritis, increased renal and prostatic stone formation and hardening of heart vessels. Treatment with the orphan drug, nitisinone, an inhibitor of 4-hydroxyphenylpyruvate dioxygenase has been shown to reduce urinary excretion of homogentisic acid. Method A reverse phase liquid chromatography tandem mass spectrometry method has been developed to simultaneously analyse serum homogentisic acid, tyrosine and nitisinone. Using matrix-matched calibration standards, two product ion transitions were identified for each compound (homogentisic acid, tyrosine, nitisinone) and their respective isotopically labelled internal standards (13C6-homogentisic acid, d2-tyrosine, 13C6-nitisinone). Results Intrabatch accuracy was 94–108% for homogentisic acid, 95–109% for tyrosine and 89–106% for nitisinone; interbatch accuracy (n = 20) was 88–108% for homogentisic acid, 91–104% for tyrosine and 88–103% for nitisinone. Precision, both intra- and interbatch were Conclusions The method developed and validated shows good precision, accuracy and linearity appropriate for the monitoring of alkaptonuria patients, pre- and post-nitisinone therapy.

Background Alkaptonuria (AKU) is a serious genetic disease characterised by premature spondyloarthropathy. Homogentisate-lowering therapy is being investigated for AKU. Nitisinone decreases homogentisic acid (HGA) in AKU but the dose-response relationship has not been previously studied. Methods Suitability Of Nitisinone In Alkaptonuria 1 (SONIA 1) was an international, multicentre, randomised, open-label, no-treatment controlled, parallel-group, dose-response study. The primary objective was to investigate the effect of different doses of nitisinone once daily on 24-h urinary HGA excretion (u-HGA 24 ) in patients with AKU after 4 weeks of treatment. Forty patients were randomised into five groups of eight patients each, with groups receiving no treatment or 1 mg, 2 mg, 4 mg and 8 mg of nitisinone. Findings A clear dose-response relationship was observed between nitisinone and the urinary excretion of HGA. At 4 weeks, the adjusted geometric mean u-HGA 24 was 31.53 mmol, 3.26 mmol, 1.44 mmol, 0.57 mmol and 0.15 mmol for the no treatment or 1 mg, 2 mg, 4 mg and 8 mg doses, respectively. For the most efficacious dose, 8 mg daily, this corresponds to a mean reduction of u-HGA 24 of 98.8% compared with baseline. An increase in tyrosine levels was seen at all doses but the dose-response relationship was less clear than the effect on HGA. Despite tyrosinaemia, there were no safety concerns and no serious adverse events were reported over the 4 weeks of nitisinone therapy. Conclusions In this study in patients with AKU, nitisinone therapy decreased urinary HGA excretion to low levels in a dose-dependent manner and was well tolerated within the studied dose range. Trial registration number EudraCT number: 2012-005340-24. Registered at ClinicalTrials.gov: NCTO1828463.

Alkaptonuria (AKU) is a rare debilitating autosomal recessive disorder of tyrosine metabolism. Deficiency of homogentisate 1,2-dioxygenase results in increased homogentisic acid (HGA) which although excreted in gram quantities in the urine, is deposited as an ochronotic pigment in connective tissues, especially cartilage. Ochronosis leads to a severe, early-onset form of osteoarthritis, increased renal and prostatic stone formation and hardening of heart vessels. Treatment with the orphan drug, Nitisinone, an inhibitor of the enzyme 4-hydroxyphenylpyruvate dioxygenase has been shown to reduce urinary excretion of HGA, resulting in accumulation of the upstream pre-cursor, tyrosine. Using reverse phase LC-MS/MS, a method has been developed to simultaneously quantify urinary HGA and tyrosine. Using matrix-matched calibration standards, two product ion transitions were identified for each compound and their appropriate isotopically labelled internal standards. Validation was performed across the AKU and post-treatment concentrations expected. Intrabatch accuracy for acidified urine was 96-109% for tyrosine and 94-107% for HGA; interbatch accuracy (n=20 across ten assays) was 95-110% for tyrosine and 91-109% for HGA. Precision, both intra- and interbatch was

This paper reports research on whether online delivery performs as well as traditional lecture delivery for a computer science course at North Carolina State University. The comparisons made are for two large sections of the course for which almost the only difference was that one section attended on-campus lectures and the other did not. Where significant differences in outcomes appear for students who completed the course, they favor the online students. However, online students who started the course were less likely to complete it.

Background: Analysis of urinary free catecholamines was automated recently, but analysis of plasma samples posed special difficulties. The present study was undertaken to evaluate a new method for the automated analysis of plasma catecholamines. Methods: The procedure is based on an improved sample handling system that includes dialysis and sample clean-up on a strong cation trace-enrichment cartridge. The catecholamines norepinephrine, epinephrine, and dopamine are then separated by reversed-phase ion-pair chromatography and quantified by electrochemical detection. Results: Use of a 740-μL sample is required to give the catecholamine detection limit of 0.05 nmol/L and analytical imprecision (CV) between 1.1% and 9.3%. The assay can be run unattended, although >12 h of analysis time is not recommended without cooling of the autosampler rack. Comparison (n = 68) of the automated cation-exchange clean-up with the well-established manual alumina procedure gave excellent agreement (mean, 3.78 ± 2.76 and 3.8 ± 2.89 nmol/L for norepinephrine and 0.99 ± 1.72 and 1.08 ± 1.78 nmol/L for epinephrine). Hemodialysis had no clear effect on plasma norepinephrine. Epinephrine concentrations were similar (0.05 < P < 0.1) in chronic renal failure patients (0.24 ± 0.3 nmol/L; n = 15) and healthy controls (0.5 ± 0.24 nmol/L; n = 31). Dopamine was not quantified, being usually Conclusion: The availability of such a fully automated procedure should encourage the more widespread use of plasma catecholamine estimation, e.g., after dialysis, exercise, or trauma/surgery and in the investigation of catecholamine-secreting tumors, particularly in the anuric patient.

The São Tomé white-toothed shrew Crocidura thomensis is endemic to the island of São Tomé in the Gulf of Guinea. This species was discovered in 1886 and since then has been recorded from fewer than 10 locations. This paper summarizes all published and unpublished information on C. thomensis. The species is currently listed by the IUCN as ‘Insufficiently Known’, but using the new IUCN Red List Categories is re-classified as ‘Vulnerable B1 + 2c’. Threats to the survival of C. thomensis include deforestation, plantation redevelopment and predation by introduced mammals.

There are few data on urinary markers of collagen breakdown in children. We have determined a normal range for urinary pyridinoline and deoxypyridinoline in children, assessed the variability in excretion in individual children and examined the effect of GH treatment on the excretion of these collagen cross-links.A cross-sectional study of a group of healthy children and sequential samples from children receiving GH treatment.One hundred and nine healthy children aged 2-15 years, 8 healthy children aged 4-11 years and 4 children receiving GH treatment.Total pyridinoline and deoxypyridinoline excretion were measured by high performance liquid chromatography after initial acid hydrolysis and cellulose extraction steps. Serum parathyroid hormone was measured using a two-site immunoradiometric assay and urinary hydroxyproline by Ehrlich's reaction using a colorimetric assay. Pyridinoline and deoxypyridinoline excretion were expressed as a ratio against urine creatinine.High excretion of pyridinoline (Pyr) and deoxypyridinoline (DPyr) was seen at all ages with no apparent relation to age (mean Pyr/Cr 115 nmol/mmol and DPyr/Cr 31 nmol/mmol). No correlation was found with serum parathyroid hormone or urinary hydroxyproline excretion. Marked day to day variation was seen in individual children. A progressive rise in excretion was seen in children receiving GH treatment with no significant correlation to height velocity.There is a high excretion of the pyridinium cross-linking amino acids in children of all ages compared to adults. However, a high variability exists in single morning urine samples which will limit the usefulness of these markers in growing children.

To compare the cytokine response to exhaustive running in recreationally-active (RA) and endurance-trained (ET) men. Eleven RA men (VO2max 55 ± 7 mL·min(-1)·kg(-1)) and 10 ET men (VO₂max 68 ± 7 mL·min(-1)·kg(-1)) followed a controlled diet and refrained from volitional exercise for 8 days. On the fourth day, participants completed 60 min of treadmill running (65 % VO₂max), followed by intermittent running to exhaustion (70 % VO₂max). Fasting blood was obtained at baseline, after 20, 40 and 60 min of exercise, at the end of intermittent exercise, during 2 h of recovery and on four follow-up days (FU1-FU4). Tumour necrosis factor-alpha (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6), interleukin-1 receptor antagonist (IL-1ra) and creatine kinase (CK) were measured. Exercise increased the concentrations of all cytokines and CK, but there were no significant differences between groups. IL-1β increased (2.2-2.5-fold, P < 0.001) during exercise, while TNF-α was increased (1.6-2.0-fold, P < 0.001) during exercise and for 2 h post-exercise. IL-6 (71-84-fold, P < 0.001) and IL-1ra (52-64-fold, P < 0.001) were increased throughout exercise and up to FU1, peaking immediately after exercise and at 1.5-2 h post-exercise, respectively. CK concentrations were increased (P < 0.001) throughout exercise and up to FU4, peaking at FU1, but were not associated with changes in any cytokines. Exhaustive running resulted in modest and transient increases in TNF-α and IL-1β, and more marked and prolonged increases in IL-6 and IL-1ra, but improved training status did not affect this response. Increased CK might indicate either exercise-induced muscle cell disruption or increased cell permeability, although neither appears to have contributed to the increased cytokine concentrations.

Cerebellar granule neurons are the most abundant neurons in the brain, and a critical element of the circuitry that controls motor coordination and learning. In addition, granule neuron precursors (GNPs) are thought to represent cells of origin for medulloblastoma, the most common malignant brain tumor in children. Thus, understanding the signals that control the growth and differentiation of these cells has important implications for neurobiology and neurooncology. Our previous studies have shown that proliferation of GNPs is regulated by Sonic hedgehog (Shh), and that aberrant activation of the Shh pathway can lead to medulloblastoma. Moreover, we have demonstrated that Shh-dependent proliferation of GNPs and medulloblastoma cells can be blocked by basic fibroblast growth factor (bFGF). But while the mitogenic effects of Shh signaling have been confirmed in vivo, the inhibitory effects of bFGF have primarily been studied in culture. Here, we demonstrate that mice lacking FGF signaling in GNPs exhibit no discernable changes in GNP proliferation or differentiation. In contrast, activation of FGF signaling has a potent effect on tumor growth: treatment of medulloblastoma cells with bFGF prevents them from forming tumors following transplantation, and inoculation of tumor-bearing mice with bFGF markedly inhibits tumor growth in vivo. These results suggest that activators of FGF signaling may be useful for targeting medulloblastoma and other Shh-dependent tumors.

Background Teicoplanin is an antibiotic used for the treatment of endocarditis, osteomyelitis, septic arthritis and methicillin-resistant Staphylococcus aureus. Teicoplanin is emerging as a suitable alternative antibiotic to vancomycin, where their trough serum levels are monitored by immunoassay routinely. This is the first report detailing the development of a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for measuring teicoplanin in patients' serum. Methods An Acquity™ UPLC (ultra-pressure liquid chromatography) tandem mass spectrometer was used to measure teicoplanin concentrations in samples from patients, quality assurance schemes and quality control preparations. Ristocetin was successfully implemented as a suitable internal standard. Ion suppression, linearity, stability, matrix effects, recovery, imprecision, lower limits of quantification and detection, interference and method comparison against immunoassay were all assessed. Results Teicoplanin and ristocetin had elution times of 1.39 and 1.24 min, respectively. Ion suppression was shown to be negligible, and linear calibration curves (0–200 μg/mL) were consistently reproduced to have r2 values >0.99. Postextraction stability was achieved up to 20 h, while matrix effects were minimal coupled with sample recovery of >93%. The lower limit of quantification was 1 μg/mL, and 0.2 μg/mL was the lower limit of detection. Interference with other antibiotics was dependent on the combination of drugs present in patients’ serum. A method comparison between immunoassay and LC-MS/MS suggested a negative bias for tandem mass spectrometry. Conclusions This novel method of teicoplanin determination by LC-MS/MS is proven to be a robust protocol that is consistent and reproducible. Clinicians searching for alternatives in therapeutic drug monitoring may have an additional option that is potentially more accurate and specific.