Amoebic gill disease increases energy requirements and decreases hypoxia tolerance in Atlantic salmon (Salmo salar) smolts.

Globally, Atlantic salmon (Salmo salar Linnaeus) aquaculture is now routinely affected by amoebic gill disease (AGD; Neoparamoeba perurans). The disease proliferates throughout the summer and is implicated in decreasing tolerance of salmon to environmental perturbations, yet little empirical evidence exists to support these observations. Using salmon acclimated to 15 or 19 °C, our aim was to determine the effects of clinically light-moderate (industry-relevant) AGD on metabolism (MO2rest and MO2max), aerobic scope (MO2max - MO2rest), excess post-exercise oxygen consumption (EPOC), and hypoxia tolerance. An increase in MO2rest (~8% and ~ 13% increase within the 15 and 19 °C acclimation groups, respectively) with increasing disease signs demonstrated an increase in baseline energy requirements as the disease progressed. Conversely, MO2max remained stable at both temperatures (~364 mg O2 kg-1 h-1), resulting in a decline in aerobic scope by 13 and 19% in the 15 and 19 °C groups, respectively. There was evidence of a decrease in hypoxia tolerance as the dissolved oxygen concentrations at loss of equilibrium increased by ~8% with more severe lesion coverage of the gills. These results suggest an increase in basal energy requirements and reduction in hypoxia tolerance as AGD proliferates, lending support to the idea that AGD reduces environmental tolerance. However, the lack of an effect of acclimation temperature indicates that the temperature-disease interaction may be more complicated than currently thought.

An Economic Evaluation of the Costs and Benefits of Providing Comprehensive Supports to Students in Elementary School.

There is growing evidence that out-of-school factors, such as physical and mental health, family support, and social and emotional development, significantly affect student learning (Berliner 2009). To address challenges related to poverty, schools are being charged with serving as a focal point in providing and coordinating support services for students and their families (Adelman and Taylor 2002; Dryfoos 2002). In many schools these support services are provided in fragmented ways that do not address the needs of all students or engage teachers in connecting these services to the academic mission of the school (Walsh and DePaul 2008). An emerging school-based model, broadly termed "comprehensive student support" (Walsh et al. 2016), is designed to overcome such fragmentation. In this paper, we build upon previous effectiveness work with an economic evaluation of a successful support model, City Connects. We find that the benefits of the program exceed the costs, indicating that the program is a sound investment and should be considered an option to address the needs of students and to prevent future crises from disrupting their learning.

Clinical implications of germline mutations in breast cancer genes: RECQL.

BACKGROUND: The identification of new hereditary breast cancer genes is an area of highly active research. In 2015, two independent studies provided initial evidence for a novel breast cancer susceptibility gene, RECQL, a DNA helicase which plays an important role in the DNA damage response. Several subsequent studies in independent patient cohorts have provided further data on RECQL variant frequency in additional populations, some of which have brought in to question the increased breast cancer risk associated with RECQL mutations. RESULTS: The initial reports present findings from whole exome sequencing of high-risk familial breast cancer cases in the French-Canadian, Polish and Han Chinese populations and estimate the carrier frequency of pathogenic RECQL mutations in high-risk breast cancer patients who have previously tested negative for BRCA1 and BRCA2 mutations to be approximately 1-2%. Proposed founder mutations were identified in French-Canadian and Polish populations. Functional studies support loss of function of the helicase activity of RECQL for some of the reported pathogenic mutations. An additional study in a cohort of Southern Chinese high-risk breast cancer patients estimated the frequency of pathogenic RECQL mutations to be 0.54%. A possible Chinese founder mutation was identified, but only a small number of controls were sequenced. Subsequent case-control studies screening for the Polish founder mutation in patients from Germany and Belarus did not find any evidence for increased breast cancer risk for this variant. An Australian case-control study also failed to identify an increased risk of breast cancer associated with RECQL loss of function variants. CONCLUSIONS: RECQL plays an important role in DNA repair, and is a plausible candidate breast cancer susceptibility gene. Initial studies showed evidence of an association between variants in this gene and an increased breast cancer risk in three separate populations, and identified founder mutations with significantly increased odds ratios. However, several subsequent studies have failed to support the association. With the limited and conflicting evidence available, there remains debate as to whether there is an increased breast cancer risk in individuals carrying RECQL loss of function variants. Further studies are required to better quantify the risks associated with RECQL variants and the current evidence base is not sufficient to justify routine inclusion of RECQL on breast cancer gene panels in clinical use. Management of patients in whom RECQL variants have been identified should be based on clinician assessment, in the context of the family history. Further studies are required to better quantify the risks to RECQL mutation carriers and may also guide management and potential therapeutic targeting for patients.

UX007 for the treatment of long chain-fatty acid oxidation disorders: Safety and efficacy in children and adults following 24weeks of treatment.

BACKGROUND: Long-chain fatty acid oxidation disorders (LC-FAOD) lead to accumulation of high concentrations of potentially toxic fatty acid intermediates. Newborn screening and early intervention have reduced mortality, but most patients continue to experience frequent hospitalizations and significant morbidity despite treatment. The deficient energy state can cause serious liver, muscle, and heart disease, and may be associated with an increased risk of sudden death. Triheptanoin is a medium odd-chain fatty acid. Anaplerotic metabolites of triheptanoin have the potential to replace deficient tricarboxylic acid (TCA) cycle intermediates, resulting in net glucose production as a novel energy source for the treatment of LC-FAOD. STUDY DESIGN: A single-arm, open-label, multicenter Phase 2 safety and efficacy study evaluated patients with severe LC-FAOD evidenced by ongoing related musculoskeletal, cardiac, and/or hepatic events despite treatment. After a four-week run-in on current regimen, investigational triheptanoin (UX007) was titrated to a target dose of 25-35% of total daily caloric intake. Patients were evaluated on several age/condition-eligible endpoints, including submaximal exercise tests to assess muscle function/endurance (12-minute walk test; 12MWT) and exercise tolerance (cycle ergometry), and health related quality of life (HR-QoL). Results through 24weeks of treatment are presented; total study duration is 78weeks. RESULTS: Twenty-nine patients (0.8 to 58years) were enrolled; most qualified based on severe musculoskeletal disease. Twenty-five patients (86%) completed the 24-week treatment period. At Week 18, eligible patients (n=8) demonstrated a 28% increase (LS mean=+181.9 meters; p=0.087) from baseline (673.4meters) in 12MWT distance. At Week 24, eligible patients (n=7) showed a 60% increase in watts generated (LS mean=+409.3W; p=0.149) over baseline (744.6W) for the exercise tolerance test. Improvements in exercise tests were supported by significant improvements from baseline in the adult (n=5) self-reported SF-12v2 physical component summary score (LS mean=+8.9; p<0.001). No difference from baseline was seen in pediatric parent-reported (n=5) scores (SF-10) at Week 24. Eighteen patients (62%) had treatment-related adverse events, predominantly gastrointestinal (55%), mild-to-moderate in severity, similar to that seen with prior treatment with medium chain triglyceride (MCT) oil. One patient experienced a treatment-related serious adverse event of gastroenteritis. One patient discontinued from study due to diarrhea of moderate severity; the majority of patients (25/29; 86%) elected to continue treatment in the extension period. CONCLUSIONS: In patients with severe LC-FAOD, UX007 interim study results demonstrated improved exercise endurance and tolerance, and were associated with positive changes in self-reported HR-QoL.

Alterations in gill structure in tropical reef fishes as a result of elevated temperatures.

Tropical regions are expected to be some of the most affected by rising sea surface temperatures (SSTs) because seasonal temperature variations are minimal. As temperatures rise, less oxygen dissolves in water, but metabolic requirements of fish and thus, the demand for effective oxygen uptake, increase. Gill remodelling is an acclimation strategy well documented in freshwater cyprinids experiencing large seasonal variations in temperature and oxygen as well as an amphibious killifish upon air exposure. However, no study has investigated whether tropical reef fishes remodel their gills to allow for increased oxygen demands at elevated temperatures. We tested for gill remodelling in five coral reef species (Acanthochromis polyacanthus, Chromis atripectoralis, Pomacentrus moluccensis, Dascyllus melanurus and Cheilodipterus quinquelineatus) from populations in northern Papua New Guinea (2° 35.765' S; 150° 46.193' E). Fishes were acclimated for 12-14 days to 29 and 31°C (representing their seasonal range) and 33 and 34°C to account for end-of-century predicted temperatures. We measured lamellar perimeter, cross-sectional area, base thickness, and length for five filaments on the 2nd gill arches and qualitatively assessed 3rd gill arches via scanning electron microscopy (SEM). All species exhibited significant differences in the quantitative measurements made on the lamellae, but no consistent trends with temperature were observed. SEM only revealed alterations in gill morphology in P. moluccensis. The overall lack of changes in gill morphology with increasing temperature suggests that these near-equatorial reef fishes may fail to maintain adequate O2 uptake under future climate scenarios unless other adaptive mechanisms are employed.

The National Neonatal Transport Programme (NNTP) 2004-2009.

A retrospective analysis of all National Neonatal Transport Programme (NNTP) transport data from 2004-2009 was performed. 1621 transports were conducted during this period with a yearly average of 271. The majority (96%) were ground transports. 1118 (69%) were forward transfers. Of the 446 (27.5%) retrotransfers, 411 (91%.) were to tertiary centres. When transported, 592 infants (36.5%) were <48 hours old and 770 (47.5%) were <1 week old. 902 transports (55%) involved infants <32 wks birth-gestation and 720 (44%) involved infants weighing <1500gms. Transport for management of patent ductus arteriosus accounted for 357 (22%). The average mobilisation time was 34 minutes. 54% of transports were completed after scheduled service hours. The NNTP currently transports similar numbers of critically ill infants during its 8hr service compared with neonatal transport services that operate 24hr services. Performing PDA ligations in a tertiary neonatal unit would significantly reduce the number of neonatal transports required annually.

Inhaled vs subcutaneous effects of a dual IL-4/IL-13 antagonist in a monkey model of asthma.

BACKGROUND: Pitrakinra is a recombinant protein derived from human interleukin-4 (IL-4) that binds to IL-4Ralpha and acts as a competitive antagonist of IL-4 and IL-13. The studies reported here compare the dose-ranging effects of pitrakinra on allergen-induced airway hyperresponsiveness (AHR) and airway eosinophilia when administered subcutaneously (s.c.) or by inhalation to the Ascaris suum-sensitive cynomolgus monkey for the purpose of elucidating the primary site of pitrakinra's anti-asthmatic action. METHODS: Airway responsiveness to inhaled methacholine and bronchoalveolar lavage cell composition was determined before and after three allergen exposures with a 1-week course of twice-daily (b.i.d.) s.c. or inhaled pitrakinra or placebo treatment. RESULTS: Treatment with s.c. pitrakinra significantly reduced allergen-induced AHR, with a maximum effect of a 2.8- to 3.8-fold increase in methacholine PC(100) relative to control (P < 0.05) observed at b.i.d. s.c. doses of 0.05-0.5 mg/kg. Inhaled pitrakinra also significantly reduced AHR with a similar maximum effect of a 2.8- to 3.2-fold increase in methacholine PC(100) relative to control (P < 0.05) at nominal b.i.d. doses of 3-100 mg. The maximal effect on AHR following inhalation was observed at a plasma concentration which exhibited no efficacy via the subcutaneous route. The effect of pitrakinra on lung eosinophilia was not statistically significant following either route of administration, although lung eosinophil count was reduced in all studies relative to control. CONCLUSION: Local administration of pitrakinra to the lung is sufficient to inhibit AHR, one of the cardinal features of asthma, indicating the therapeutic potential of inhaled pitrakinra in the treatment of atopic asthma.

Genetics of Degenerative Cervical Myelopathy: A Systematic Review and Meta-Analysis of Candidate Gene Studies.

Degenerative cervical myelopathy (DCM) is estimated to be the most common cause of adult spinal cord impairment. Evidence that is suggestive of a genetic basis to DCM has been increasing over the last decade. A systematic search was conducted in MEDLINE, EMBASE, Cochrane, and HuGENet databases from their origin up to 14th December 2019 to evaluate the role of single genes in DCM in its onset, clinical phenotype, and response to surgical intervention. The initial search yielded 914 articles, with 39 articles being identified as eligible after screening. We distinguish between those contributing to spinal column deterioration and those contributing to spinal cord deterioration in assessing the evidence of genetic contributions to DCM. Evidence regarding a total of 28 candidate genes was identified. Of these, 22 were found to have an effect on the radiological onset of spinal column disease, while 12 genes had an effect on clinical onset of spinal cord disease. Polymorphisms of eight genes were found to have an effect on the radiological severity of DCM, while three genes had an effect on clinical severity. Polymorphisms of six genes were found to have an effect on clinical response to surgery in spinal cord disease. There are clear genetic effects on the development of spinal pathology, the central nervous system (CNS) response to bony pathology, the severity of both bony and cord pathology, and the subsequent response to surgical intervention. Work to disentangle the mechanisms by which the genes that are reviewed here exert their effects, as well as improved quality of evidence across diverse populations is required for further investigating the genetic contribution to DCM.

Horse owners' knowledge, and opinions on recognising colic in the horse.

BACKGROUND: Colic is the most common emergency problem in the horse. An owner's ability to recognise colic and seek assistance is a critical first step in determining case outcome. OBJECTIVES: The aim of this study was to assess horse owners' knowledge and opinions on recognising colic. STUDY DESIGN: Cross-sectional study. METHODS: An online questionnaire was distributed to horse owners with open and closed questions on their knowledge of normal clinical parameters in the horse, confidence and approach to recognising colic (including assessment through case scenarios), and their demographics. Descriptive and chi squared statistical analyses were performed. RESULTS: There were 1564 participants. Many respondents either did not know or provided incorrect estimates for their horse's normal clinical parameters: only 45% (n = 693/1540) gave correct normal values for heart rate, 45% (n = 694/1541) for respiratory rate and 67% (n = 1028/1534) for rectal temperature. Knowledge of normal values was statistically associated with participants' educational qualifications (P<0.01). Owners said if they suspected their horse had colic they would assess faecal output (76%; n = 1131/1486), gastrointestinal sounds (75%; n = 1113/1486), respiratory rate (65%; n = 967/1486) and heart rate (54%; n = 797/1486). There was a lack of consensus on whether to call a vet for behavioural signs of colic, unless the signs were severe or persistent. The majority of participants (61%) were confident that they could recognise most types of colic. In the case scenarios, 49% were confident deciding that a surgical case had colic, but 9% were confident deciding an impaction case had colic. MAIN LIMITATIONS: Most respondents were UK based; risk of self-selection bias for owners with previous experience of colic. CONCLUSIONS: There was marked variation in horse owners' recognition and responses to colic, and significant gaps in knowledge. This highlights the need for the development of accessible educational resources to support owners' decision-making.

Allergen dose dependency of the early- and late-phase cutaneous response in the cynomolgus monkey.

BACKGROUND: Cutaneous administration of allergen provides a means to confirm an allergic status, investigate the pathogenesis of allergic diseases, and/or provide a mechanism to evaluate the benefit of new potential therapeutics. OBJECTIVE: Studies were performed to characterize the allergen-induced cutaneous early- and late-phase response (EPR and LPR) in the cynomolgus monkey. METHODS: Following intradermal injections of Ascaris suum allergen, the cutaneous weal and flare EPR was measured 15 min post-injection, and skin biopsies were collected at 8-24 h to determine the optimal time of LPR occurrence. Biopsies were analysed for epidermal and dermal inflammatory changes. RESULTS: The EPR was dose related with a reproducible, measurable response at 1 : 10 000 and maximal at a 1 : 100 allergen dilution. In contrast, the threshold dose required for a reproducible LPR was much greater requiring a dilution of 6 : 100, suggesting independent mechanisms for the EPR and LPR. The LPR 20 h post-allergen injection induced an inflammatory response in the upper and deep dermis. The response was characterized by a moderate perivascular to diffuse inflammation consisting of mononuclear cells, neutrophils and eosinophils. Dexamethasone, while having no effect on the EPR, reduced dermal inflammation (upper dermis, P=0.004; deep dermis, P=0.03). Similarly, dermal eosinophilia was also reduced (upper dermis, P<0.001; deep dermis, P=0.02). CONCLUSION: Collectively, the results indicate the dose dependency of the EPR and LPR. Furthermore, our observations indicate the value of the LPR response in the cynomolgus monkey to evaluate new therapeutics for the treatment of allergic diseases such as atopic dermatitis.

The pH dependence of the apparent equilibrium constant, K', of a biochemical reaction.

Biochemical reactions can be discussed in terms of chemical equations or biochemical equations. Both are right and both are needed. Therefore, it is important to be able to distinguish between them at a glance. Sometimes the relations between the apparent equilibrium constant 'K" for a biochemical reaction and the equilibrium constant, K, for a reference reaction written as a chemical equation is simple, and sometimes it is complicated. This article will discuss several examples.

Hexokinase and 'glucokinase' in liver metabolism.

Rat liver contains four hexokinase isoenzymes, one of which, despite often being called 'glucokinase', is no more specific for glucose than the others. However, it does differ from them in displaying a sigmoid kinetic response to glucose, requiring much higher glucose concentrations for activity, and being insensitive to physiological concentrations of glucose 6-phosphate.

Principles for the safety evaluation of cosmetic powders.

Consumer exposure to cosmetic (personal care) products is mostly by dermal contact, however additional considerations with regards to potential inhalation exposure from some cosmetics, such as sprays and powders, may be needed for a robust and reliable safety assessment. To get a deeper understanding of the exposure to airborne particles and droplets during product application, a team of international experts was founded under the umbrella of the European Association of the Cosmetic Industry "Cosmetics Europe" (CE) in Brussels. This expert team has worked out a pragmatic strategy how small and medium sized enterprises (SMEs), but also relevant authorities, could handle the safety evaluation of cosmetic powder products. Sufficient information on the aerodynamic diameter of sprayed droplets and here specifically of airborne particles is essential in addition to knowing the exposure after typical product application. The current article is focused on the determination of inhalation exposure to solids, and the derivation of safe exposure levels for cosmetic powder products found in the market. The principles described herein are very similar to spray products as published earlier and should be applied in a similar way (Steiling et al., 2014). Prediction models for the best estimate of inhalation exposure, developed with data from computer simulation programs, individual real-time measurements or finally by experience from the market were introduced and applied. Safety assessment approaches for exposure from powder spray products were developed and have been already considered in regulatory guidelines like the EC Cosmetics Regulation.

Estimation of Michaelis constant and maximum velocity from the direct linear plot.

When estimates of Michaelis-Menten parameters are obtained from kinetic observations taken in pairs, as in the direct linear plot, bias can arise in the final estimates if any pairs lead to negative values of the maximum velocity V. This bias can be removed by treating such negative values as if they were large and positive, and by treating the corresponding values of Km in the same way.

Validation of a micro-CT technique for measuring volumetric wear in retrieved acetabular liners.

In this study, a novel micro-CT-based technique for evaluating wear in retrieved acetabular liners was introduced and validated. Six UHMWPE acetabular components ranging in implantation time from 2.7 to 14.4 years were collected and evaluated with the use of a high-resolution micro-CT scanner. The components were scanned with a uniform volumetric resolution of 74 microns (16-bit precision) with the use of a 1,024 x 1,024 in-plane image matrix. Manual rigid 3D image registration of the interior hemispherical portion of the acetabular cup with geometric primitives by trained observers allowed for isolation, visualization, and measurement of the wear volume. Results for these six components indicated an average wear rate of 65 mm(3)/year. Overall scanner error was quantified gravimetrically and associated with a maximum uncertainty of 0.6%. Intra-- and interobserver uncertainty analysis showed the method to be both accurate and repeatable.

Clinical Research Abstracts of the British Equine Veterinary Association Congress 2015.

REASONS FOR PERFORMING STUDY: Colic is the most common equine emergency problem, and one of the main causes of equid deaths. The horse owners' ability to recognise colic is a critical first step in determining case outcome. OBJECTIVES: To assess equine owners' knowledge and recognition of colic. STUDY DESIGN: Online questionnaire of horse owners. METHODS: An online survey was designed to evaluate owners' approach to colic in the horse. The survey included questions on owner demographics, their recognition of colic (including owner's opinions of their ability to recognise colic, their approach, and their recognition of colic using case vignettes), and their knowledge of normal ranges for clinical parameters. Descriptive and chi squared statistical analysis was performed. RESULTS: The survey was completed by 1061 UK respondents. Six per cent of owners thought they could recognise all types of colic, 61% said they could recognise most cases and 30% said they could recognise some but not all cases. Owners said they would assess faecal output (73% of respondents), gastrointestinal sounds (69%), respiratory rate (62%) and heart rate (50%) in horses with suspected colic. One fifth (22%) of owners would call a vet immediately without assessing any parameters. Many respondents either did not know, or provided incorrect estimates of normal values for clinical parameters: 30.4% were 'unsure' of the normal heart rate and 35.5% gave heart rate values which were outside reference ranges; only 24.5% gave appropriate values for normal respiratory rates and only 31% gave normal temperature values. There was no statistical significance between participants' age, educational qualifications, or their experience with horses and their knowledge of normal clinical parameters. CONCLUSIONS: Owners varied in their approach and ability to recognise colic, and many had significant gaps in their knowledge of normal parameters. Educational materials and/or training to assist owners could help address these issues. Ethical animal research: The study did not involve animal research. The study was reviewed and approved by the Ethics Committee, School of Veterinary Medicine and Science, University of Nottingham. The questionnaire was conducted in accordance with the 1998 Data Protection Act, and the British Educational Research Association's Revised Ethical Guidelines for Educational Research (2004). SOURCE OF FUNDING: Adelle Bowden's studentship is funded by the School of Veterinary Medicine and Science, University of Nottingham. Competing interests: None declared.

Why is uncompetitive inhibition so rare? A possible explanation, with implications for the design of drugs and pesticides.

Uncompetitive inhibition is much less common in nature than consideration of enzyme structure and mechanism might lead one to expect. A possible explanation may be that uncompetitive inhibition of an enzyme in a metabolic pathway can have enormously larger effects on the concentrations of metabolic intermediates than competitive inhibition, under circumstances where their effects on the kinetics of the isolated enzyme are very similar. The severely toxic effects that an uncompetitive inhibitor might be expected to have may have caused enzymes to have evolved in such a way that there has been selection against structures that might favour uncompetitive inhibition.

Regulating the cellular economy of supply and demand.

Cellular metabolism is a molecular economy that is functionally organised into supply and demand blocks linked by metabolic products and cofactor cycles. Supply-demand analysis allows the behaviour, control and regulation of metabolism as a whole to be understood quantitatively in terms of the elasticities of supply and demand, which are experimentally measurable properties of the individual blocks. The kinetic and thermodynamic aspects of regulation are clearly distinguished. One important result is the demonstration that when flux is controlled by one block, the other block determines to which degree the concentration of the linking metabolite is homeostatically maintained.

Evidence for requirement of tyrosine phosphorylation in endothelial P2Y- and P2U- purinoceptor stimulation of prostacyclin release.

1. The release of prostacyclin (PGI2) from vascular endothelial cells is stimulated by ATP acting at G protein-coupled P2-purinoceptors. Here we investigate the hypothesis that tyrosine protein phosphorylations are involved in this response. 2. The use of Western blots with anti-phosphotyrosine antibodies showed that 30 microM 2MeSATP (selective for P2Y-purinoceptors), 300 microM UTP (selective for P2U-purinoceptors) and 300 microM ATP (effective at both these purinoceptors), each stimulate the tyrosine phosphorylation of proteins in bovine cultured aortic endothelial cells. Each of these agonists also stimulates 6-keto PGF1 alpha accumulation in the medium (an index of PGI2 release) in these cells in the same period. 3. The tyrosine kinase inhibitor, genistein, inhibits the 6-keto PGF1 alpha response with the same concentration-dependency (1-100 microM) as the tyrosine phosphorylation response. 4. Tyrphostin, a structurally and functionally distinct tyrosine kinase inhibitor, is also a potent inhibitor (0.1-10 microM) of the 6-keto PGF1 alpha response. 5. Neither tyrphostin nor genistein inhibit the phospholipase C response to P2-purinoceptor stimulation. Furthermore, these inhibitors do not affect the 6-keto PGF1 alpha response to ionomycin. 6. These results show that the regulation of vascular endothelial cells by ATP acting at both P2Y- and P2U-purinoceptors involves the stimulation of tyrosine phosphorylation, and suggest that this is a necessary event for the purinoceptor-mediated stimulation of PGI2 production.