bluuebunny/crossref_metadata_2025 huggingface dataset:
Modification of the Metabolomic Profile Induced by Chronic Growth Hormone Administration in a Doping Model
Authors: Barrey E , Triba M , Messier F , Le Moyec L
Published: June 2014

Introduction

Horses performance can be enhanced via growth hormone (GH) doping. The detection time for GH in blood is too short to allow detection during competition. GH not only affects the proteosynthesis in cells and tissues but can also cause long term metabolic variations. This study aimed to show the metabolomic modifications occurring after three months of GH treatment.

Methods

The study was performed on 20 Welsh ponies. 10 were injected with REPORCIN TM, an approved recombinant GH (30 mg.kg−1 per day) for 3 months and the other 10 comprised the control group. Blood samples were collected before GH treatment (T0), then 3 months later (T3). The NMR spectra acquired on the 40 serum samples were classified according to the factors (season, GH treatment) using the supervised multivariate orthogonal projection on latent structure (OPLS) method. A first OPLS model was computed with the control ponies (T0‐T3). The predictive component of this model was subtracted from the whole data (treated, controls at T0 and T3) to only retain the GH‐dependent variations.

Results

Before filtering seasonal effects, glucose, lipids and phospholipids were metabolites discriminating between T0 and T3 samples. After removing the seasonal effect, the OPLS model revealed the effect of GH treatment on protein metabolism through the modification of aromatic amino acids and branched chain amino acids. The variations of other metabolites (glycoproteins and lactate) were enhanced.

Conclusions

This study showed that chronic GH injection induced systemic metabolism modifications that can be detected using the NMR metabolomic method without neglecting the misleading effect of season.

Ethical Animal Research

Studies approved by “Comité regional d′éthique pour l′expérimentation animale Centre Limousin” Saisine N° CL2008_015. Sources of funding: French Agency against Doping (AFLD). Competing interests: none.
Heart Rate Responses in Show‐Jumpers Over A Three‐Day Training Session
Authors: Harris P , Roberts C , Armstrong S , Murray R , Handel I
Published: June 2014

Introduction

The objective of this study was to provide more information regarding the relationship between heart rate (HR) and performance in show‐jumpers.

Methods

Ten mixed breed horses (9.1 ± 1.9 yrs) were assessed at a British Equestrian Federation (BEF) World Class Performance three‐day training session. Riders received coaching during warm‐up (WU) and after jumping a 15 fence (1.35–1.45 m) course once on day‐1, twice on days 2 & 3; on day‐3 the second round was an 8‐fence jump‐off. Clinical assessments were performed, and plasma lactate concentrations determined, pre‐ and post‐exercise. Take‐off distance (TDF) was determined using high‐speed motion capture. The coach graded horse jumping technique (validated methodology). HR was recorded telemetrically from WU until 10 mins post‐exercise. Variables were compared using regression procedures/Spearman correlation and paired Students t‐test (P<0.05).

Results

HR was not significantly different between days or consecutive jumping rounds. Mean HR was lower in WU than during the course (127 ± 11 vs 136 ± 16: P = 0.027), likely related to increased speed during the course, although HRpeak tended to be higher during WU day‐ 1 possibly reflecting excitement. Mean HR increased for each quarter during the course. Higher mean course and third‐quarter HR were associated with higher blood lactate concentrations (P = 0.047), greater gluteal and longissimus dorsi muscle spasm on palpation (P<0.01), more faults (P = 0.044) and lower technique scores (P = 0.019). TDF was negatively correlated with mean course and third‐quarter HR, indicating that closer take‐off may be harder work (P<0.01).

Conclusions

HR increases throughout a jumping course. Higher HRs may be associated with jumping inefficiency and muscle spasm, with potential performance implications for multi‐day competitions.

Ethical Animal Research

Explicit owner informed consent for participation in this study is not stated. Sources of funding: The World Class Development Programme is funded by the Department for Culture, Media and Sport, through Exchequer and National Lottery funds, via UK Sport. Competing interests: none.
Blood Biochemistry and Hemogasometry of Horses Performing Incremental Jumping and Velocity Tests
Authors: Rodrigues TN , Miranda AC , Ramos MT , Godoi FN , Almeida FQ
Published: June 2014

Introduction

Biochemical and hemogasometric parameters have been used to evaluate equine performance. This study evaluated blood biochemical and hemogasometry variables of horses during incremental jumping and speed tests.

Methods

Eighteen horses performed a jumping test with four steps over a vertical obstacle of 1.10 m height (10, 15, 20 and 25 jumps, respectively) totaling 70 jumps, and a speed test with six steps of one minute each with increasing speed from 2 to 8 m/s. Blood samples were collected before and immediately after each test to determine plasma lactate, hematocrit, total plasma protein, glucose, uric acid and hemogasometry variables. Results were analysed by ANOVA and compared by 5% Tukey test.

Results

Immediately after tests the hematocrit and total plasma protein decreased from 52.4% to 38.9%, and from 7.2 to 6.7 g/dl respectively, and the glucose increased from 98.3 to 109.8 mg/dl (P<0.05). The lactate and uric acid concentrations were higher with speed testing (7.97 ± 4.8 mmol/l and 1.33 ± 0.8 mg/dl respectively) than jumping testing (3.92 ± 3.0 mmol/l and 1.24 ± 0.9 mg/dl respectively). The blood pH was lower after speed testing (7.36 ± 0.1) than the jumping test (7.40 ± 0.04) (P<0.05). Blood pCO2, Na and K concentrations increased more after the speed test (53.38 ± 7.3 mmHg/l; 139 ± 3.0 mEq/l; 4.7 ± 0.8 mEq/l respectively) than after the jumping test (49.34 ± 3.2 mmHg/l; 138.3 ± 2.3 mEq/l; 4.51 ± 0.6 mEq/l respectively) (P<0.05). Blood pO2 was not different among tests (P>0.05).

Conclusions

Incremental jumping and velocity tests are useful tools for assessing performance of jumping horses. Differences among tests could evidence a smaller anaerobic contribution during jumping test.

Ethical Animal Research

The study was approved by UFRRJ Ethics Committee on Animal Research: number 263/2012. Sources of funding: CNPq, CAPES and FAPERJ. Competing interests: none.
Electrolyte Supplementation Prior to Prolonged Submaximal Exercise Results in Decreased PCO2 and tCO2 During Exercise and Recovery
Authors: Waller AP , Ecker G , Lindinger MI
Published: June 2014

Introduction

Electrolyte supplementation prior to exercise is known to benefit both performance and recovery; however its effects on acid‐base status remain not well characterized. The purpose of this study was to use the physicochemical approach to provide a comprehensive time‐course of the acid‐base alterations that occur following pre‐exercise electrolyte supplementation in horses. It was hypothesized that electrolyte supplementation will attenuate the alkalosis occurring with prolonged moderate intensity exercise.

Methods

In a randomized crossover experiment four conditioned mares were given, via nasogastric tube, 3 L of a commercial electrolyte solution (ES; designed to fully replace sweat electrolyte and fluid losses), or water alone (Control), 1 hour before they exercised on a treadmill at 30% VO2max until voluntary fatigue (inability to keep pace with belt speed). Blood was sampled from jugular venous catheters at frequent intervals before, during, and until 20 h of exercise recovery, and analyzed for all dependent and independent plasma acid‐base variables. Differences (P<0.05) over time and between groups were assessed by 1‐ or 2‐way repeated measures ANOVA.

Results

Horses exercised for 57.5 ± 10.3 and 72.5 ± 17.0 min and lost 2.73 ± 0.82 and 4.63 ± 1.35 L in Control and ES trials, respectively. ES horses had lower pCO2 and total carbon dioxide (TCO2) concentrations during exercise and early recovery. Control horses exhibited decreased [H+]. pCO2 was the primary contributor to changes in [H+] and [TCO2], while there were no differences in strong ion difference ([SID]) or total weak acid concentration ([Atot]).

Conclusions

Electrolyte supplementation prior to prolonged exercise leads to sustained effects on equine acid‐base variables, even in the absence of significant changes in plasma electrolytes or hydration.

Ethical Animal Research

The experiments and use of animals were approved by the University of Guelph Animal Care Committee, in accordance with the guidelines of the Canadian Council on Animal Care. Sources of funding: Ontario Horse Racing Industry Association and the Natural Sciences and Engineering Research Council of Canada. Competing interests: none.
Differentiating Pathological from Physiological Left Ventricular Hypertrophy by Quantifying Myocardial Function
Authors: Decloedt A , Ven S , Van Der Vekens N , De Clercq D , van Loon G
Published: June 2014

Introduction

Differentiating left ventricular (LV) dilatation caused by aortic regurgitation (AR) from athletic eccentric LV hypertrophy can be challenging. In human medicine, tissue Doppler imaging (TDI) and 2D speckle tracking (2DST) are used to distinguish ‘athlete's heart’ from pathological conditions.

Methods

40 healthy horses (group 1) were compared to 13 horses with moderate AR (group 2) and 11 horses with severe AR (group 3), all with an end diastolic LV internal diameter (LVIDd) within maximal reference limits (≤14.2 cm). Fractional shortening (FS) was measured from a short‐axis M‐mode at chordal level. Peak TDI velocities during systole (S), early (E) and late (A) diastole were measured in the LV free wall (LVFW) and interventricular septum (IVS). Global longitudinal strain was measured by 2DST. Groups were compared by a one‐way ANOVA with Bonferroni correction.

Results

The study included Warmbloods (n = 52), trotters (n = 9) and Thoroughbreds (n = 3), used for jumping, dressage, racing and eventing. Group 2 (15 ± 5 years; P<0.001) and 3 (20 ± 4 years; P<0.001) were older than group 1 (8 ± 4 years). LVIDd in group 3 (13.1 ± 0.7 cm) was higher compared to group 1 (11.5 ± 0.8 cm; P<0.001) and 2 (12.0 ± 0.8 cm; P = 0.003) but FS was not different. Peak S in IVS was higher in group 2 (6.6 ± 1.5 cm/s; P = 0.001) and 3 (9.0 ± 1.7 cm/s; P<0.001) compared to group 1 (4.4 ± 1.8 cm/s), indicating hyperkinetic septal motion. Peak E in LVFW was lower in group 2 (18.6 ± 5.2 cm/s;P = 0.001) and 3 (19.3 ± 3.9 cm/s; P = 0.006) compared to group 1 (24.4 ± 4.6 cm/s), indicating decreased early filling velocity. Longitudinal strain was not different between groups.

Conclusions

TDI shows potential for differentiating AR‐induced LV remodeling from athletic eccentric hypertrophy in horses.

Ethical Animal Research

Horses with aortic regurgitation were examined in a prospective clinical study with owner informed consent. Sources of funding: The corresponding author is a postdoctoral research fellow of the Research Foundation Flanders. Competing interests: none.
Longitudinal Follow‐Up of Bronchoalveolar Lavage Cytology in Standardbred Racehorses
Authors: Depecker M , Leleu C , Richard E , Couroucé‐Malblanc A
Published: June 2014

Introduction

Bronchoalveolar lavage (BAL) fluid cytology is a major diagnostic method for detecting exercise‐induced pulmonary haemorrhage (EIPH) and inflammatory airway disease (IAD) in racehorses. Our hypothesis was that training and racing would increase the incidence of IAD and EIPH.

Methods

On three different occasions (day‐1, −30 and −60), BAL fluid was obtained from both lungs of 14 Standardbred trotters with similar age and training level. Cytological inclusion criteria were neutrophils >10%, and/or mast cells ≥5%, and/or eosinophils ≥5% for IAD, and haemosiderophage/macrophage (H/M) ratio >20% for EIPH. Since previously found not to be equivalent, both samples from each horse were independently investigated. Horse status (‘IAD’,‘EIPH’, ‘mixed’, or ‘control’) at each time point was based on cytological profiles from both lungs. Cell proportions were compared by repeated measures ANOVA.

Results

Haemosiderophage proportions and H/M were significantly higher on day‐60 compared to day‐1. No significant difference was observed for other cell proportions. On day‐1, 5 horses were ‘control’, 2 ‘IAD’, 2 ‘EIPH, and 5 were ‘mixed’. On day‐30, 3 horses were ‘control’, 4 ‘IAD’, 2 ‘EIPH’, and 5 were ‘mixed’. On day‐60, 3 horses were ‘control’, 0 ‘IAD’, 3 ‘EIPH’, and 8 ‘mixed’. Despite the different horses′ distribution within each category, an overall increase in EIPH was observed. Five of the 7 horses classified ‘control’ or ‘IAD’ on day‐1 presented EIPH on day‐30 and/or −60. EIPH was detected in 11/14 horses (78.6%) on day‐60, versus 7/14 horses (50%) on day‐1.

Conclusions

The incidence of EIPH, but not IAD, significantly increased during a racing season. This evolution should be considered when assessing BAL fluid cytology in racehorses.

Ethical Animal Research

The study was approved by the regional Animal Ethics Committee (n°CEEA.2012.179) and all owners signed a consent form. Sources of funding: IFCE (National Studs) and SECF (French Trotters Racing Association). Competing interests: none.
Hemodynamic and Endocrine Changes Associated with Splenic Reserve Mobilization
Authors: McKeever K , Lehnhard R , Hinchcliff K
Published: June 2014

Introduction

In theory, splenic reserve mobilization should elicit a cardiopulmonary baroreceptor mediated neuroendocrine response (the Gauer‐Henry reflex).

Six intact (IN) and 5 (SP) splenectomized Standardbred mares (421–491 kg, ∼9 yr) performed 10 min of treadmill exercise (80% VO2max) to test the hypothesis that hemodynamic and endocrine responses to exercise would differ between IN and SP horses

Methods

Blood samples were obtained before and at 1, 2, 4, 6, 8 and 10 min of exertion. Right ventricular pressure (RVP), right atrial pressure (RAP), arterial pressure (MAP), heart rate (HR), and cardiac output (CO) were measured and total peripheral resistance (TPR) calculated. Plasma atrial natriuretic peptide (ANP) and vasopressin (AVP) concentrations were measured using RIA. Data analysis used ANOVA for repeated measures and SNK. Significance was set at P<0.05.

There were no differences (P>0.05) between IN and SP for any variable at rest. All variables except TPR and RAP increased (P<0.05) during exertion in both IN and SP. Means (±SE) for RAP (18 ± 3 vs. 1 ± 2 mmHg), CO (255 ± 21 vs.177 ± 19 L/min), MAP (147 ± 3 vs. 116 ± 8 mmHg), and ANP (66 ± 3 vs. 50 ± 8 pg/mL) were greater for IN vs. SP horses at 10 min of exertion. Mean plasma AVP (95 ± 19 vs. 23 ± 3 pg/mL) was greater (P<0.05) for SP than for IN horses at 10 min of exertion.

Ethical Animal Research

The study was approved by the Institutional Animal Care and Use Committee. Sources of funding: Ohio Thoroughbred and Standardbred Research Fund and the New Jersey State Initiative on Equine. Competing interests: none.
The Effect of Land and Underwater Treadmill Training on Metabolic Responses to Exercise and Properties of Superficial Digital Flexor and Gluteal Muscle
Authors: Firshman A , Borgia L , Valberg S
Published: June 2014

Introduction

Resistance training provided by underwater (UWT) treadmills may enhance strength and fatigue resistance of superficial digital flexor muscle thereby reducing tendon injury. This study examined effects of UWT and land (LT) treadmill training on SDF and gluteal muscle.

Methods

Six unfit horses (3 × 2 crossover design, 60d detraining period) walked on UWT and LT for 5d/week for 8 weeks (up to 40 min/day). A standardized exercise test (SET) was performed 2 weeks before, and after L or UW training, and V180 bpm and La10.5 m/s calculated. Resting and post‐SET gluteal and SDF muscle biopsies were obtained. Muscle [glycogen], [lactate] and [ATP], fiber type composition and min/max fiber diameters were analysed. Comparisons were made using GLM or ANOVA, mean ± SD and significance of P<0.05.

Results

Post‐exercise SDF muscle lactate increased 3–4 fold to 30 ± 12 (UWT) and 41 ± 15 (LT) mmol/kg dw post‐exercise. Post‐exercise gluteal muscle lactate increased 2 fold to 55 ± 18 (UWT) and 62 ± 18 (LT) mmol/kg. No significant differences were found between UW and L post‐exercise muscle [glycogen], [lactate] and [ATP], V180 or LA10.5 or fiber type composition. Gluteal fiber diameters decreased significantly with UW and L training. Fiber diameters in SDF muscle decreased with training to a significantly lesser extent after UW vs. L training.

Conclusions

At maximal speed, both SDF and gluteal muscle have a significant anaerobic response which was not impacted by UWT or LT training at a walk. UWT may be of benefit by attenuating the training induced decrease in fiber sizes in SDF muscle thereby enhancing SDF muscle strength.

Ethical Animal Research

All procedures were approved by the Institutional Animal Care and Use Committee of the University of Minnesota. Sources of funding: Supported by University of Minnesota Equine Center with funds provided by the Minnesota Racing Commission, University of Minnesota Agricultural Experiment Station, and private donors. Competing interests: none.
Can Stalled Horses Provided Light Exercise and “Weekend” Turnout Maintain Bone Mineral Content?
Authors: Spooner H , Hoffman R , Haffner J
Published: June 2014

Introduction

Previous research suggests stalling may have a negative influence on bone mineral content (BMC). Yet slow, long‐distance exercise has not been successful in improving BMC compared to free‐choice exercise. Still, it is a common practice to stall horses with light exercise and minimal turnout. We hypothesized stalled horses, despite “weekend” turnout and light exercise, would have lower estimated BMC compared to pasture turnout.

Methods

Thirteen mature horses were split into two groups, Control (C, n = 6) and Stalled (S, n = 7). Digital radiographs were taken of the third metacarpal for estimation of BMC by radiographic bone aluminum equivalence (RBAE). After a 42‐d period (P1) additional radiographs were taken and S‐horses stalled for 4‐d (receiving light under‐saddle exercise), followed by 72‐h pasture turnout. This pattern repeated for an additional 42‐d (P2) followed by radiographs. Radiographs were analyzed for optical density of bone cortices and total BMC compared to an aluminum step‐wedge on each cassette. Changes in BMC were analyzed using PROC mixed in SAS with repeated measures.

Results

No treatment differences were observed in the medial (P = 0.72) or lateral (P = 0.46) cortices. Dorsal and palmar cortices exhibited a treatment by period interaction (P<0.01, P<0.02), remaining unchanged in C but S decreased over P1 and returned to baseline in P2. Total RBAE was not different by treatment or period.

Conclusions

Contrary to our hypothesis, horses stalled with light exercise and “weekend” turnout maintained BMC compared to themselves and controls at pasture. Such management may be considered for maintaining BMC when horses must be stalled for weekday riding or training. Further research is necessary to determine minimum turnout and exercise needs to maintain BMC when stalling.

Ethical Animal Research

The study was approved by the Middle Tennessee State University Institutional Animal Care and Use Committee Sources of funding: The Horse Science Program and Office of Research at Middle Tennessee State University. Competing interests: none.
Does Phenylbutazone Administration Contribute to Gastric Permeability and Ulceration in Horses?
Authors: Kenny J , Nielsen B , Robison C , Larson C , Schott II H
Published: June 2014

Introduction

Gastric ulcers are commonly found in equine athletes and there is evidence suggesting phenylbutazone may increase the incidence. In human athletes, gastric permeability has been shown to increase with non‐steroidal anti‐inflammatory use with potential to negatively affect performance and recovery. The aim of this study was to examine the effects of phenylbutazone on gastric mucosa and permeability.

Methods

18 Arabian horses were examined via gastroscopy on day 0, 42, 49, 56, and 63, with ulceration in both the squamous and glandular regions scored from 0 to 4. For seven days commencing after the day 42 evaluation, horses received 4.4 mg/kg BW phenylbutazone twice daily PO. On day 49, sucrose (1 g/kg bwt) was administered via nasogastric tube, with blood and urine samples collected for 90 min and 4 h, respectively, to assess gastric permeability. Data were analyzed using SAS 9.2 with Proc MIXED.

Results

Ulcer scores increased from day 42 to day 49 (P<0.0001) with prevalence of grade 2 or greater ulceration being 33% on day 42 compared with 94% on day 49. Scores decreased from day 49 to 56 and, though continuing to decrease, had not returned to baseline scores by day 63. Squamous scores increased from day 0 to day 49 and remained elevated through day 63, while glandular scores were highest on day 49 but had decreased by day 56 (P<0.05). Sucrose concentration increased (P<0.0001) in both plasma and urine after phenylbutazone administration.

Conclusions

Besides exacerbating gastric ulcers, phenylbutazone contributes to gastric permeability and, particularly when combined with strenuous exercise, may impair recovery and increase illness susceptibility.

Ethical Animal Research

All methods were approved by the Michigan State University Institutional Animal Care and Use Committee (approval number 10/11‐216‐00). Sources of funding: Zinpro Corporation. Competing interests: none.
A Comparison of Racing Results Between Standardbred Horses Fed a Forage‐Only Diet and Their Siblings
Authors: Jansson A , Ringmark S
Published: June 2014

Introduction

High concentrate diets increase the risk of several health problems and high energy forage diets could therefore be an alternative. The aim of this study was to compare the number of horses and the number of races performed at the age of three by a group of 16 Standardbred horses fed a forage only‐diet from the age of 1.5 years (F) with the results of their siblings (S).

Methods

To F‐horses, a forage diet was offered ad libitum (10.2–11.7 MJ metabolisable energy, 10–16% crude protein/kg dry matter) and supplemented daily with minerals and vitamins to meet requirements. F‐horses were trained by pupils at the National Trotting School during supervision of professional trainers. For each individual in the F‐group, the mean racing results from a maximum of five older siblings (same mother) were used for the S‐group (5 siblings, n = 9; 4‐3 siblings, n = 4 and 1–2 siblings, n = 3). The siblings selected were born closest in time in relation to the F individual. S‐horses were trained by different trainers and it was assumed that they had been fed a conventional high concentrate diet. Racing results were obtained from the Swedish Trotting Association. ANOVA and a Tukey test were used to analyse differences between groups.

Results

There was no difference in the number of horses that competed (F: 56% vs S: 54%, SE 10) and in the number of races performed (F: 1.6 vs S: 2.9, SE 0.5).

Conclusions

The study indicates that a forage‐only diet may not be a limitation to achieve conventional training results and could therefore be an alternative to conventional diets.

Ethical Animal Research

The study was approved by the local ethics committee in Umeå, Sweden. Sources of funding: Swedish University of Agricultural Sciences, the Swedish National Trotting School at Wången, the Swedish Trotting Association, The Swedish Horse Council Foundation, the Swedish Horse Racing Totalisator Board (ATG), Trioplast AB and Dow Chemicals. Competing interests: none.
Oil Supplementation Produces an Increase in Antioxidant Biomarkers in Four‐Beat Gaited Horses
Authors: Melo S , Wanderley E , Diniz I , Manso H , Manso Filho H
Published: June 2014

Introduction

Marcha is typically a low intensity/medium duration exercise that has the potential to induce oxidative stress. The objective of the present study was to test the hypothesis that supplementation with oil would alter biomarkers of oxidative stress in four‐beat gaited horses during marcha training.

Methods

Eleven gaited horses underwent training for 60 min (∼60% consisting of marcha at ∼3.5 m/s) 3 d/wk for eight weeks. Subjects were fed elephant grass (∼15 kg/day) and concentrate (∼5 kg/day). They were also supplemented with 300 mL of oil per os (vitamin E: 300 IU/kg, omega‐3: 300 g/kg; omega‐6: 80 g/kg) and had free access to water and salt. Blood samples were collected before and after four and eight weeks of supplementation for the measurement of GPx, SOD, RBC, hemoglobin, hematocrit, WBC, TPP, urea, creatinine and uric acid. ANOVA and Tukey tests were used for statistical analysis (P<0.05).

Results

Increases in GPx (∼12%), SOD (∼51%) and uric acid (∼19%) were observed after 8 week of supplementation (P<0.05). There were changes (P<0.05) in [urea] and [creatinine]; however, there were no changes (P>0.05) in RBC, hemoglobin, hematocrit or WBC. Published research has demonstrated that administration of supplements with antioxidants enhances the antioxidant system and protects against oxidative stress during exercise. Supplementation with oil, rich in vitamin E, omega‐3 and ‐6, produced a beneficial increase in antioxidant capacity in gaited horses during training.

Conclusions

The elevation in [GPx], [SOD] and [uric acid] in the present study offers a strategy to counterbalance the physiologic processes that overwhelm endogenous antioxidants during training.

Ethical Animal Research

Methods were approved by UFRPE Animal Care Unit (#026/2013). Sources of funding: INTEGRALMIX Animal Nutrition (Fortaleza‐CE), Haras Cascatinha (Camaragibe‐PE) and CAPES. Competing interests: none.
Enzymatic properties and substrate specificity of a bacterial phosphatidylcholine synthase
Authors: Aktas Meriyem , Köster Stefan , Kizilirmak Sarah , Casanova Javier C. , Betz Heidi , Fritz Christiane , Moser Roman , Yildiz Özkan , Narberhaus Franz
Published: June 2014

Phosphatidylcholine (PC) is a rare membrane lipid in bacteria, but is crucial for virulence of the plant pathogen Agrobacterium tumefaciens and various other pathogens. Agrobacterium tumefaciens uses two independent PC biosynthesis pathways. One is dependent on the integral membrane protein PC synthase (Pcs), which catalyzes the conversion of cytidine diphosphate–diacylglycerol (CDP–DAG) and choline to PC, thereby releasing a cytidine monophosphate (CMP). Here, we show that Pcs consists of eight transmembrane segments with its N‐ and C‐termini located in the cytoplasm. A cytoplasmic loop between the second and third membrane helix contains the majority of the conserved amino acids of a CDP‐alcohol phosphotransferase motif (DGX2ARX12GX3DX3D). Using point mutagenesis, we provide evidence for a crucial role of this motif in choline binding and enzyme activity. To study the catalytic features of the enzyme, we established a purification protocol for recombinant Pcs. The enzyme forms stable oligomers and exhibits broad substrate specificity towards choline derivatives. The presence of CDP–DAG and manganese is a prerequisite for cooperative binding of choline. PC formation by Pcs is reversible and proceeds via two successive reactions. In a first choline‐ and manganese‐independent reaction, CDP–DAG is hydrolyzed releasing a CMP molecule. The resulting phosphatidyl intermediate reacts with choline in a second manganese‐dependent step to form PC.

Structured digital abstract

Pcs and Pcs bind by molecular sieving (1, 2, 3)
Evaluating pharmacist prescribing for minor ailments
Authors: Mansell Kerry , Bootsman Nicole , Kuntz Arlene , Taylor Jeff
Published: June 2014

Objectives

Saskatchewan is the second Canadian province to allow pharmacists to prescribe medications for minor ailments and the only province that remunerates for this activity. The aim of this project was to determine whether patients prescribed such treatment by a pharmacist symptomatically improve within a set time frame.

Methods

Pharmacists were asked to hand a study-invitation card to anyone for whom they prescribed a medication for a minor ailment during the 1-year study period. Consenting participants contacted the study researchers directly and were subsequently instructed to complete an online questionnaire at the appropriate follow-up time.

Key findings

Ninety pharmacies in Saskatchewan participated, accruing 125 participants. Cold sores were the most common minor ailment (34.4%), followed by insect bites (20%) and seasonal allergies (19.2%). Trust in pharmacists and convenience were the most common reasons for choosing a pharmacist over a physician, and 27.2% would have chosen a physician or emergency department if the minor ailment service were not available. The condition significantly/completely improved in 80.8%; only 4% experienced bothersome side effects. Satisfaction with the pharmacist and service was strong; only 5.6% felt a physician would have been more thorough.

Conclusions

Participants were very satisfied with their symptomatic improvement and with the service in general, albeit for a small number of conditions. Participants reported getting better, and side effects were not a concern. These results are encouraging for pharmacists; however, a comparison of physician care with pharmacist care and unsupported self-care is required to truly know the benefit of pharmacist prescribing.
Efficacy of traditional and alternative sunitinib treatment schedules in Japanese patients with metastatic renal cell carcinoma
Authors: Ohzeki Takayuki , Fukasawa Satoshi , Komaru Atsushi , Namekawa Takeshi , Sato Yosuke , Takagi Kimiaki , Kobayashi Masayuki , Uemura Hirotsugu , Ichikawa Tomohiko , Ueda Takeshi
Published: June 2014

We report the adverse events and efficacy of traditional (4 weeks on 2 weeks off) and alternative sunitinib treatment schedules for Japanese patients with metastatic renal cell carcinoma. We retrospectively investigated 54 patients who received sunitinib for metastatic renal cell carcinoma between May 2006 and June 2012: 32 received a traditional treatment schedule and 22 received an alternative schedule. According to the Memorial Sloan‐Kettering Cancer Center risk classification, five patients had favorable prognoses, 42 had intermediate prognoses and seven had poor prognoses. The mean observation periods were 16.3 and 20 months for the traditional and alternative schedule groups, respectively. Adverse events were significantly less common in the alternative schedule group, including most high‐grade events. In the traditional and alternative schedule groups, median times to failure were 4.1 and 11.6 months (P = 0.040), median progression‐free survival times were 4.1 and 11.3 months (P = 0.031), and median overall survival times were 12.0 and 32.1 months (P = 0.018), respectively. Each of these measures was better in the group of patients who received an alternative treatment schedule, suggesting that individualized changes to the sunitinib administration schedule can be effective.
Trends in Causes of Death in Low‐Mortality Countries: Implications for Mortality Projections
Authors: Bongaarts John
Published: June 2014

This study examines the potential role that information about trends in causes of death could have in improving projections of mortality in low‐mortality countries. The article first summarizes overall trends in mortality by cause since the middle of the twentieth century. Special attention is given to the crucial impact of the smoking epidemic on mortality and on cause‐of‐death patterns. The article then discusses the implications for projections and reaches two conclusions. First, mortality projections can be improved by taking into account the distorting effects of smoking. Mortality attributable to smoking has risen in the past but has now leveled off or declined, thus boosting improvements in life expectancy. Second, making cause‐specific projections is not likely to be helpful. Trends in specific medical causes of death have exhibited discontinuities in the past, and future trends are therefore difficult to predict.
Nontraditional Family‐Related Attitudes in Japan: Macro and Micro Determinants
Authors: Choe Minja Kim , Bumpass Larry L. , Tsuya Noriko O. , Rindfuss Ronald R.
Published: June 2014

Attitudinal data are commonly used to measure values, which in turn represent moral doctrines that are resistant to change and are the foundation for societal norms. This article examines changes in the attitudinal climate in Japan. Three national surveys (1994, 2000, and 2009) are used to examine a range of attitudes that measure a) the centrality of marriage and childbearing, b) nontraditional family behaviors, and c) gender roles in the work and family spheres. There is strong evidence of movement toward less‐traditional attitudes during 1994–2000, followed by limited change in the 2000s. Period factors were paramount in the 1990s. Across the board, women hold less‐traditional attitudes than men, and this difference has increased over time. Both engaging in nontraditional family behaviors (being married but remaining childless) and knowing someone who has engaged in nontraditional family behaviors (cohabitation) causally lead to holding nontraditional attitudes, suggesting mechanisms whereby changes in individual behavior can lead to changes in societal values.
Effects of Parents' Migration on the Education of Children Left Behind in Rural China
Authors: Zhou Minhui , Murphy Rachel , Tao Ran
Published: June 2014

This essay draws on an original cross‐sectional survey of 1,010 children and their guardians in highly migratory regions of Anhui and Jiangxi provinces located in China's interior. It uses propensity score matching, a technique that mitigates endogenity, to examine the impact of parental migration and post‐migration guardianship arrangements on the children's educational performance as measured by test scores for Chinese and mathematics. One core finding is that the educational performance of children is adversely affected by parental migration only when both parents migrate or when a non‐parent guardian is the principal carer. Additionally, longer durations of parental absence are associated with poorer educational performance. The migration of two parents only significantly adversely affects the educational performance of boys. There is no significant effect on the educational performance of girls. On the basis of our findings we argue that rather than support left‐behind children within the countryside, the long‐term policy response should be to remove the institutional obstacles that prevent family resettlement in the cities.
A Transaction Cost Approach to Outsourcing by Households
Authors: Raz‐Yurovich Liat
Published: June 2014

Below‐replacement fertility and late marriage reflect, in part, the incompatibility of women's family and paid work roles. The outsourcing of childcare and housework to market and state service providers offers a strategy for reconciling work–family conflicts. By referring to the household as an organizational unit, I use the transaction cost approach (TCA) of organizational economics to discuss the factors that facilitate or impede outsourcing by households. In my analysis the frequency, specificity, and uncertainty level of the transaction, as well as normative and social beliefs, can facilitate or impede the household's decision to outsource. Monetary considerations, preferences, and government policies might moderate the effect of the transaction cost on this decision. The analysis further demonstrates that gender is an important factor, because transaction costs are often not distributed equally within households.
Reasons for Contraceptive Nonuse among Women Having Unmet Need for Contraception in Developing Countries
Authors: Sedgh Gilda , Hussain Rubina
Published: June 2014

The level of unmet need for contraception—an important motivator of international family planning programs and policies—has declined only slightly in recent decades. This study draws upon data from 51 surveys conducted between 2006 and 2013 in Africa, Asia, and Latin America and the Caribbean to provide an updated review of the reasons why many married women having unmet need are not practicing contraception. We examine the reasons for contraceptive nonuse and how these reasons vary across countries and according to national levels of unmet need and contraceptive use. We present specific findings regarding the most widespread reasons for nonuse, particularly infrequent sex and concerns regarding side effects or health risks. Our findings suggest that access to services that provide a range of methods from which to choose, and information and counseling to help women select and effectively use an appropriate method, can be critical in helping women having unmet need overcome obstacles to contraceptive use.
Are testicular mast cells involved in the regulation of germ cells in man?
Authors: Windschüttl S. , Nettersheim D. , Schlatt S. , Huber A. , Welter H. , Schwarzer J. U. , Köhn F. M. , Schorle H. , Mayerhofer A.
Published: June 2014

Protease activated receptor‐2 (PAR‐2) is the receptor for the prototype mast cell product tryptase. PAR‐2 expression by cells of the human germinal epithelium was reported, but the exact cellular sites of testicular expression remained unknown. That became of interest, because mast cells, expressing tryptase, were found in the walls of seminiferous tubules of patients suffering from sub‐ and infertility. This location suggested that mast cells via tryptase might be able to influence PAR‐2‐expressing cells in the germinal epithelium. To explore these points, we used testicular paraffin‐embedded sections for immunohistochemistry. PAR‐2‐positive cells were mostly basally located cells of the seminiferous epithelium, namely spermatogonia. Some stained for the receptor for GDNF (GFRalpha‐1), and possibly represent spermatogonial stem cells (SSCs). As true human SSCs could not be examined, we turned to TCam‐2 seminoma cells, expressing PAR‐2 and stem cell markers, including GFRalpha‐1. TCam‐2 cells robustly responded to stimulation with a specific PAR‐2 agonist (SLIGKV) by increased intracellular Ca2+ levels. Recombinant tryptase and trypsin, but not a control peptide (VKGILS) evoked this response, implying functional PAR‐2. Video imaging and caspase 3/7 assays showed that SLIGKV and tryptase prevented spontaneous apoptosis and increased proliferation of TCam‐2 cells. The expression of the marker of pluripotency OCT3/4 was unchanged upon activation of PAR‐2, suggesting that the stem cell‐like character is not changed. Furthermore, human germ cell cancers were examined. A subset of seminoma and carcinoma in situ samples expressed PAR‐2, indicating that yet unknown subgroups exist. Collectively, the descriptive data obtained in human testicular sections, in germ cell cancers and the functional results in TCam‐2 cells imply a trophic role of mast cell‐derived tryptase for human germ cells. This may be relevant for subtypes of human germ cell cancers, and possibly SSCs. It also raises the possibility that PAR‐2 agonists might be useful for the in vitro propagation of human SSCs.
Expression of testicular androgen receptor in non‐obstructive azoospermia and its change after hormonal therapy
Authors: Kato Y. , Shiraishi K. , Matsuyama H.
Published: June 2014

Several trials aimed at improving the sperm retrieval from men with non‐obstructive azoospermia (NOA) by optimizing intratesticular testosterone (ITT) have reported partial responses, however, an appropriate level of ITT has not been identified. In this study, we examined the expression of the testicular androgen receptor (AR) in NOA and investigated its correlation with clinical and pathological parameters. Expression of the testicular AR was investigated in 52 men with NOA and 22 men with obstructive azoospermia (OA). Twenty‐two patients for whom sperm retrieval failed during microdissection testicular sperm extraction (micro‐TESE) were enrolled in hormonal therapy using hCG with or without recombinant human follicle stimulating hormone (rhFSH) prior to a second micro‐TESE. Sertoli cells were identified by vimentin immunostaining, and positivity in Sertoli cells was used as the AR index. AR immunostaining was robust in the nuclei of Sertoli cells [Sertoli cell androgen receptor (SCAR)] in both OA and NOA. The mean AR index in NOA was significantly higher than that in OA (p < 0.05). In NOA patients, there was no correlation between the AR index and the clinical parameters, whereas the AR index of early maturation arrest (MA) was significantly lower than that of Sertoli cell only, late MA and hypospermatogenesis (p < 0.05). A significant increase in the AR index after salvage hormonal therapy was shown, particularly when using rhFSH. The AR index in patients from whom spermatozoa could be retrieved at the second micro‐TESE increased significantly after hormonal therapy. In human testes, the expression of AR is dominant in Sertoli cells, and the expression of SCAR is upregulated by FSH. Germ cell maturation, especially during spermatogonia to spermatocyte stage, has been shown to be SCAR‐dependent. Taken together, the results indicate that SCAR elevation is closely associated with sperm retrieval after hormonal therapy and that FSH‐based hormonal therapy is potentially effective in NOA men with MA.
Out of thin air: Why banks must be allowed to create money
Authors: Pettifor Ann
Published: June 2014

Framing money creation as a ‘bottom‐up, even democratic’ process, Ann Pettifor challenges the argument that private banks should be restricted to making loans from existing deposits only, a plan she says promises to ‘return society to the dark ages’.
Planktonic or non‐planktonic food in young‐of‐the‐year European perch Perca fluviatilis in ponds
Authors: Bláha M. , Šetlíková I. , Peterka J. , Musil J. , Policar T.
Published: June 2014

Higher biomass especially of some aquatic macrophyte species offered a higher density of phytophilous zoobenthos, but did not increase the proportion of non‐planktonic to planktonic prey in young‐of‐the‐year perch Perca fluviatilis. Both abundance and biomass of non‐planktonic prey dominated over planktonic prey in the pond with lower biomass of aquatic macrophytes and lower food. Survival of P. fluviatilis was lower (20%) in the pond with lower food than in the other pond (34%), however, specific growth rate (1·3% day−1) and final Fulton's condition factor of P. fluviatilis were similar in both ponds.
Association between fluid balance and survival in critically ill patients
Authors: Lee J. , de Louw E. , Niemi M. , Nelson R. , Mark R. G. , Celi L. A. , Mukamal K. J. , Danziger J.
Published: June 2014

Objective

Although the consequences of chronic fluid retention are well known, those of iatrogenic fluid retention that occurs during critical illness have not been fully determined. Therefore, we investigated the association between fluid balance and survival in a cohort of almost 16 000 individuals who survived an intensive care unit (ICU) stay in a large, urban, tertiary medical centre.

Design

Longitudinal analysis of fluid balance at ICU discharge and 90‐day post‐ICU survival.

Measurements

Associations between fluid balance during the ICU stay, determined from the electronic bedside record, and survival were tested using Cox proportional hazard models adjusted for severity of critical illness.

Results

There were 1827 deaths in the first 90 days after ICU discharge. Compared with the lowest quartile of discharge fluid balance [median (interquartile range) −1.5 (−3.1, −0.7) L], the highest quartile [7.6 (5.7, 10.8) L] was associated with a 35% [95% confidence interval (CI) 1.13–1.61)] higher adjusted risk of death. Fluid balance was not associated with outcome amongst individuals without congestive heart failure or renal dysfunction. Amongst patients with either comorbidity, however, fluid balance was strongly associated with outcome, with the highest quartile having a 55% (95% CI 1.24–1.95) higher adjusted risk of death than the lowest quartile. Isotonic fluid balance, defined as the difference between intravenous isotonic fluid administration and urine output, was similarly associated with 90‐day outcomes.

Conclusion

Positive fluid balance at the time of ICU discharge is associated with increased risk of death, after adjusting for markers of illness severity and chronic medical conditions, particularly in patients with underlying heart or kidney disease. Restoration of euvolaemia prior to discharge may improve survival after acute illness.
Mindfulness as a cognitive–emotional segmentation strategy: An intervention promoting work–life balance
Authors: Michel Alexandra , Bosch Christine , Rexroth Miriam
Published: June 2014

Employees struggling with work‐related cognitions, emotions, and associated energy levels during non‐work time can find their private roles impaired and work–life balance derogated. To reduce unwanted psychological preoccupation with work concerns, boundary theory suggests employees find their ideal way to integrate or segment both life domains. In this study, the authors design and evaluate an intervention teaching mindfulness as a cognitive–emotional segmentation strategy to promote work–life balance. They use a randomized waitlist control group design to evaluate effects of a 3‐week online self‐training intervention, with 246 employees participating at pre‐ and post‐test, and 191 participating at a 2‐week follow‐up. As expected, experimental group participants, compared with control group participants, experienced significantly less strain‐based work–family conflict and significantly more psychological detachment and satisfaction with work–life balance.

Practitioner points

Mindfulness, a cognitive–emotional segmentation strategy, enables employees to balance between work and private life.

Voluntary organizational health and work–life balance programmes should include low‐cost but effective brief mindfulness interventions.
Transcriptional profiles of SHH pathway genes in keratocystic odontogenic tumor and ameloblastoma
Authors: Gurgel Clarissa Araújo Silva , Buim Marcilei Eliza Cavichiolli , Carvalho Kátia Cândido , Sales Caroline Brandi Schlaepfer , Reis Mitermayer Galvão , de Souza Renata Oliveira , de Faro Valverde Ludmila , de Azevedo Roberto Almeida , dos Santos Jean Nunes , Soares Fernando Augusto , Ramos Eduardo Antônio Gonçalves
Published: June 2014

Background

Sonic hedgehog (SHH) pathway activation has been identified as a key factor in the development of many types of tumors, including odontogenic tumors. Our study examined the expression of genes in the SHH pathway to characterize their roles in the pathogenesis of keratocystic odontogenic tumors (KOT) and ameloblastomas (AB).

Methods

We quantified the expression of SHH, SMO, PTCH1, SUFU, GLI1, CCND1, and BCL2 genes by qPCR in a total of 23 KOT, 11 AB, and three non‐neoplastic oral mucosa (NNM). We also measured the expression of proteins related to this pathway (CCND1 and BCL2) by immunohistochemistry.

Results

We observed overexpression of SMO, PTCH1, GLI1, and CCND1 genes in both KOT (23/23) and AB (11/11). However, we did not detect expression of the SHH gene in 21/23 KOT and 10/11 AB tumors. Low levels of the SUFU gene were expressed in KOT (P = 0.0199) and AB (P = 0.0127) relative to the NNM. Recurrent KOT exhibited high levels of SMO (P = 0.035), PTCH1 (P = 0.048), CCND1 (P = 0.048), and BCL2 (P = 0.045) transcripts. Using immunolabeling of CCND1, we observed no statistical difference between primary and recurrent KOT (P = 0.8815), sporadic and NBCCS‐KOT (P = 0.7688), and unicystic and solid AB (P = 0.7521).

Conclusions

Overexpression of upstream (PTCH1 and SMO) and downstream (GLI1, CCND1 and BCL2) genes in the SHH pathway leads to the constitutive activation of this pathway in KOT and AB and may suggest a mechanism for the development of these types of tumors.