Quantitative analysis of short- and long-distance racing performance in young and adult horses and association analysis with functional candidate genes in Spanish Trotter horses.

The association of five candidate genes with sporting performance in young and adult Spanish Trotter horses (STHs) was performed according to a previous selection based on quantitative analysis of the trait time per kilometre (TPK). A total of 334 516 records of TPK from 5958 STHs were used to estimate the estimated breeding values (EBVs) at different age groups (young and adults horses) throughout the range of distances (1600-2700 m) using a bicharacter random regression model. The heritability estimated by distance ranged from 0.16 to 0.40, with a different range for the two age groups. Considering the animals with the best and the worst deregressed EBV, 321 STHs were selected for SNP genotyping in MSTN, COX4I2, PDK4, DMRT3 and CKM genes. An association analysis based on ridge and logistic regression revealed that the young trotters with genotype GG in PDK4 (p < 0.05) and AA of DMRT3 (p < 0.001) SNPs show the best potential in short-distance races, while those carrying the genotype AA in DMRT3 (p < 0.001) and CC in CKM (p < 0.05) genes seem to be the best in long-distance races. Adult trotters with genotype AA in DMRT3 also display greater speed (p < 0.05) and endurance (p < 0.001).

Implementation of Optimum Contributions selection in endangered local breeds: the case of the Menorca Horse population.

Limiting the inbreeding rate (∆F) while maximizing genetic gain for any trait of economic interest is especially important in small populations of local breeds, like the Menorca Horse. In this breed, dressage performance is important for the profitability of the breed and should be accounted in the selection criterion. The aim of this study was to assess if a breeding programme aiming at improved dressage performance is feasible in such a small breed. To perform the analysis, animals that were currently available for breeding (between 3 and 20 years) were used. Selection was based on the estimated breeding values for dressage obtained by BLUP. The pedigree and molecular coancestry between potential breeding horses was used (separately or in combination) to account for the restriction on ∆F. Results show that it is possible to avoid large increases in inbreeding while obtaining acceptable levels of genetic gain (i.e. a ∆F of 1% would imply a maximum loss in genetic gain of 2%). Thus, the Menorca Horse population is suitable for a management procedure which jointly optimizes the response to selection and the levels of variability and inbreeding (Optimal Contribution selection). Regarding the source of information used to calculate the relationships, molecular information would provide a greater range of solutions to increase genetic gain than using pedigree coancestry (gain was 1-4% higher for the same levels of restriction on the increase in inbreeding).

Anti-endotoxin antibodies consumption in cardiovascular collapse: Pathophysiological concerns.

OBJECTIVE: Abnormal endotoxin activity in critically ill patients has been described in the absence of Gram-negative bacterial (GNB) infection. As disease severity seems to be crucial in the detection of this phenomenon, we decided to assess and compare endotoxin exposure in those patients representing the critical situation: septic shock and cardiogenic shock. DESIGN: Prospective, observational non intervention study. SETTING: Critical Care Department of a University tertiary hospital. PATIENTS: Cardiogenic shock (CS) and septic shock (SS) patients. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Follow-up was performed for the first three days. Inflammatory biomarkers (C-reactive protein, procalcitonin and interleuquin-6) and IgM antiendotoxin-core antibodies titter (IgM EndoCAb) were daily analyzed. Sixty-two patients were included; twenty-five patients with SS and thirty-seven with CS. Microbial etiology was established in 23 SS patients (92%) and GNB were present in 13 cases (52%). Although infection was suspected and even treated in 30 CS patients (81%), any episode could be finally confirmed. EndoCAb consumption was more intense in SS patients, although twenty-two CS patients (59.5%) had IgM anti-endotoxin value below 10th percentile range for healthy people. No statistically significant difference in endotoxin exposure was detected between Gram-positive and Gram-negative infections in the SS group. Endotoxin exposure ability to distinguish between SS and CS was moderate (AUC 0.7892, 95% IC: 0.6564-0.9218). CONCLUSIONS: In the severely ill patient some mechanisms take place allowing endotoxin incursion and therefore blurring the limits of diseases pathophysiology. Our work representatively shows how exposure to endotoxin was not fully capable of distinguishing between CS and SS.

Prevalence of Trypanosoma cruzi infection in pregnant Latin American women and congenital transmission rate in a non-endemic area: the experience of the Valencian Health Programme (Spain).

This study describes the results of the health programme implemented in the Valencian Community (Spain) to achieve an early diagnosis of Chagas disease in pregnant Latin American women and their newborns. During 2009 and 2010, 1975 women living in the health districts of three university hospitals were enrolled via midwives or at the time of delivery. Diagnosis of disease was performed using two serological tests with different antigens. Congenital infection was diagnosed by parasitological, molecular or serological methods from blood samples obtained at birth or in subsequent controls. The overall seroprevalence of Chagas infection in pregnant women from 16 different endemic countries was 11·4%. Infection was higher in those from countries in the Gran Chaco Region (Bolivia, 34·1%; Paraguay, 7·4%; Argentina, 5·3%). Eight newborn infants from Bolivian mothers had congenital Chagas which represents a vertical transmission rate of 3·7%. In conclusion, this work supports the benefits of offering an early diagnosis to pregnant women and newborns during routine prenatal healthcare.

Estimation of genetic parameters for racing speed at different distances in young and adult Spanish Trotter horses using the random regression model.

A total of 71 522 records (from 3154 horses) with the times per kilometre (TPK), recorded in Spanish Trotter horses (individual races) from racing performances held from 1991 to 2007, were available for this study. The TPK values for the different age groups (young and adult horses) and different distances (1600-2700 m) were considered as different traits, and a bi character random regression model (RRM) was applied to estimate the (co)variance components throughout the trajectory of age groups and distances. The following effects were considered as fixed: the combination of hippodrome-date of race (404 levels); sex of the animals (3 levels); type of start (2 levels) and a fixed regression of Legendre polynomials (order 2). Those considered as random effects were the random regression Legendre polynomial (order 1) for animals (9201 animals in the pedigree); the individual environment permanent (3154 animals with data) and the driver (n = 957 levels). The residual variance was considered as heterogeneous with two classes (ages). The heritability estimated by distance ranged from 0.12 to 0.34, with a different trajectory for the two age groups. Within each age group, the genetic correlations between adjacent distances were high (>0.90), but decreased when the differences between them were over 400 metres for both age groups. The genetic correlations for the same distance across the age groups ranged from 0.47 to 0.78. Accordingly, the analysed trait (TPK) can be considered as positive genetic correlated but as different traits along the trajectory of distance and age. Therefore, some re-ranking should be expected in the breeding value of the horses at different characteristics of the racing. The use of RRM is recommended because it allows us to estimate the breeding value along the whole trajectory of race competition.

[Impact of a short stay unit in a tertiary hospital]. / Impacto de una unidad de corta estancia en un hospital de tercer nivel.

OBJECTIVE: To analyze the health care activity indicators of a short-stay unit (SSU) and compare them with those of other services in a Conventional Hospital. MATERIAL AND METHODS: A descriptive, retrospective study was conducted of the patients admitted to SSU during 2003-2007. Number of admissions, mean stay (MS), mortality and destination were analyzed. Mean stay and number of admissions of the main diagnosis-related groups (DRGs) in the SSU were compared with other services of conventional hospitalization. RESULTS: A total of 15.3% of the patients who came to the Emergency Service were admitted to the SSU. Mean stay was significantly lower in the SSU than in the rest of the hospital (year 2003, SSU 3.2 days, rest of the hospital 10.2 days; year 2007, SSU 3.3 days, rest of the hospital 8.6 days). The 4 most common DRGs (71.3% of all those admitted to the SSU) were COPD, respiratory infection/bronchitis, simple pneumonia/pleuritis and heart failure/shock. Mean stay for the 4 DRGs in the SSU was significantly lower (p<0.01) than in Internal Medicine, Pneumology and Cardiology (heart failure/shock). CONCLUSIONS: The SSU handles its most prevalent diseases with greater agility compared to other conventional hospitalization services.

Phenotypic and genetic analysis of reproductive traits in horse populations with different breeding purposes.

Reproductive traits have a major influence on the economic effectiveness of horse breeding. However, there is little information available. We evaluated the use of reproductive traits as selection criteria in official breeding programs to increase the reproductive efficiency of breeding studs, analysing 696 690 records from the pedigree data of eight Spanish horse populations, with different breeding purposes. The reproductive parameters studied in both sexes were age at first foaling (AFF), age at last foaling, average reproductive life and generational interval. In the females, the average interval between foaling (AIF) and interval between first and second foaling were also studied. There were clear differences between sexes and breeds, which may be due to management practices, breeding purposes and the status of the populations, rather than to differences in actual physiological conditions. Riding mares were the most precocious (AFF, 1937.64 to 2255.69 days) and had a more intensive reproductive use (AIF, 625.83 to 760.07 days), whereas sires used for meat production were the most precocious males (AFF, 1789.93 to 1999.75 days), although they had a shorter reproductive life (1564.34 to 1797.32 days). Heritabilities (0.02 to 0.42 in females and 0.04 to 0.28 in males) evidenced the genetic component of the reproductive traits, with Sport Horses having the higher average values. These results support the selection by AFF to improve reproductive aspects because of its medium-high heritability and its positive correlations with other important reproductive traits. The inclusion of the AIF is also recommended in sport populations, because this determines the length of the breaks between foaling and conditions the reproductive performance of the dams, as well as their selective intensity, genetic gain and genetic improvement. It is therefore an important economic parameter in breeding studs.

Breed effect on carcass and meat quality of foals slaughtered at 24months of age.

In some areas, horsemeat may be considered as an alternative to beef. And recent studies have shown that it may be positive from a nutritional point of view. However, little research has been done on the effects that influence horse carcass traits and meat quality. Breed effect has been studied by comparing two Spanish local breeds (Burguete and Hispano-Bretón) reared following the same traditional production system (24months old). Some differences between breeds were observed for carcass quality measurements and colour parameters. Proximate composition was generally not affected by breed. Burguete foals had lower SFA and MUFA and higher PUFA than Hispano-Bretón ones. PUFA/SFA ratio from both breeds was higher than 0.4 and CLA levels were similar to those found in ruminants.

[Final size attained in type 1 diabetes children]. / Talla final en diabéticos tipo 1 diagnosticados en la edad pediátrica.

OBJECTIVE: To describe the final height and height-gain in relation to target height, in children with type 1 diabetes mellitus, and analyse their relationship to different variables. PATIENTS AND METHODS: Retrospective analysis of the growth data of 52 children (27 girls) diagnosed with type 1 diabetes mellitus before 14 years old, and followed up until their final height was attained. MAIN VARIABLES: final height, target height, illness duration, glycated haemoglobin (HbA1c), insulin dose, BMI, and other autoimmune diseases. RESULTS: The height SDS (standard deviation scale) at diagnosis was slightly higher (0.734 in boys and 0.563 in girls). During the development of the disease, a growth reduction was seen, which was significantly higher in boys of prepubertal age (p = 0.016). The mean final height attained was 173.14 +/- 5.28 cm in boys and 161.9 +/- 6.97 cm in girls. Height gain was 1.56 +/- 3.66 in boys (SDS = -0.034) and 2.26 +/- 6.13 in girls (SDS = 0.385). The only variable significantly related to height gain was mean glycated-haemoglobin (growth reduction of 2 cm for every increment of 1% in mean glycated-haemoglobin). CONCLUSIONS: At onset, diabetic children were slightly taller than the general population. A growth reduction was shown as the disease developed, significantly higher in boys of prepubertal age. The final height in boys was slightly lower than the mean, but in girls was similar to the general population. Both sexes attained their target height, although the height gain was less in boys. Poorer metabolic control was associated with reduced height gain.

Genetic parameters of biokinematic variables of the trot in Spanish Purebred horses under experimental treadmill conditions.

The purpose of this study was to estimate the genetic parameters of biokinematic variables in Spanish Purebred (SPB) horses in order to select those of sufficient interest to be measured in the pre-selection of the animals for possible inclusion in the breeding programme. Kinematic analysis of 130 SPB horses 4.6+/-1.5 years old were recorded at the trot (4m/s) on a treadmill. Genetic parameters were estimated using VCE software and a bivariate mixed animal model including age and stud as fixed effects and animal additive genetic effect and residual error as random effects. In general, heritabilities were high (0.33-0.88). The angular variables presented the lowest heritabilities, whereas the maximum height of the fore-hoof and the duration of swing phase in the hindlimb gave the highest scores. Genetic correlations were also very high, so it was possible to reduce the number of breeding programme characteristics to stride duration, hindlimb swing phase duration, range of stifle and elbow angles, minimal angle of carpus, and minimal retraction-protraction angle of the hindlimb.

Heritability and factors associated with number of harness race starts in the Spanish Trotter horse population.

BACKGROUND: Longevity/durability is a relevant trait in racehorses. Genetic analysis and knowledge of factors that influence number of harness race starts would be advantageous for both horse welfare and the equine industry. OBJECTIVES: To perform a genetic analysis on harness racing using number of races as a measure of longevity/durability and to identify factors associated with career length in Spanish Trotter Horses (STH). STUDY DESIGN: Longitudinal study. METHODS: Performance data (n = 331,970) on the STH population for harness racing at national level between 1990 and 2014 were used. A grouped data model was fitted to assess factors influencing the risk of ending harness racing career and to estimate the heritability and breeding values for total number of harness races starts as an indicator of horses' longevity and durability. The model included sex, age at first race and first start earnings as time-independent effects, and the calendar year, driver, trainer, racetrack category and season of competition as time-dependent effects. RESULTS: Across the whole dataset, the average number of harness races horses achieved in Spain was 54.7 races, and this was associated with the horses' sex, age at first race and first start earnings, calendar year, driver, racetrack category, and season. The heritability estimated (0.17 ± 0.01) for number of harness race starts indicates that a beneficial response to direct genetic selection can be expected. MAIN LIMITATIONS: Data on horses' health status were not available. CONCLUSIONS: Horses' total number of harness race starts is a promising tool for genetic analysis and the evaluation of racing longevity and durability. The estimated heritability provides evidence to support the application of genetic selection of total career number of races to improve longevity/durability of STH.

Blockage of the light-sensitive conductance in hyperpolarizing photoreceptors of the scallop. Effects of tetraethylammonium and 4-aminopyridine.

The tight-seal whole-cell recording technique was used to examine the effect of tetraethylammonium (TEA) and 4-aminopyridine (4-AP) on the photocurrent of hyperpolarizing ciliary photoreceptors isolated from the distal retina of the bay scallop (Pecten irradians). In these cells, light causes an increase in a conductance that is highly selective to potassium ions. Extracellular application of TEA at a concentration of 50 mM produced a modest, reversible block (approximately 35% at -20 mV holding potential). The blockage was weakly voltage dependent, increasing by approximately 20% for a 20-mV hyperpolarization, suggestive of a site of interaction superficially located within the electric field of the membrane. Treatment with TEA produced no significant changes either in the light sensitivity of the photocurrent or in its kinetics. The effects of superfusion with 4-AP were more dramatic: the light-evoked current was nearly abolished (> 95%) at submillimolar concentrations, with a half-maximal dose of approximately 0.6 microns. The blockage had a rapid onset and was slowly reversible. No significant use or voltage dependency were observed. A number of control experiments indicated that the phototransduction cascade remained functional during treatment with 4-AP: the early receptor current, the prolonged after current and its suppression, the photoresponse kinetics and the light sensitivity of the cell were little affected by 4-AP, suggesting that the suppression of the photocurrent is due to blockage of the light-sensitive channels, rather than impairment of some of the activation steps. The results are discussed in the light of a possible kinship between the light-activated potassium channels of invertebrate hyperpolarizing photoreceptors and the family of rapidly-inactivating voltage-dependent potassium channels, which typically exhibit high susceptibility to blockage by this drug.

Ion permeation through light-activated channels in rhabdomeric photoreceptors. Role of divalent cations.

The receptor potential of rhabdomeric photoreceptors is mediated primarily by a Na influx, but other ions must also permeate through light-dependent channels to account for some properties of the photoresponse. We examined ion conduction in macroscopic and single-channel light-induced currents of slug and scallop photoreceptors. In the absence of Na, a fivefold change in extracellular K shifted the reversal voltage of the photocurrent (Vrev) by approximately 27 mV. Because the dependency of Vrev on [K]o was sub-Nernstian, and Vrev in each condition was more positive than Ek, some other ion(s) with a positive equilibrium potential must be implicated, in addition to K. We assessed the participation of calcium, an important candidate because of its involvement in light adaptation. Three strategies were adopted to minimize the impairments to cytosolic Ca homeostasis and loss of responsiveness that normally result from the required ionic manipulations: (a) Internal dialysis with Na-free solutions, to prevent reverse operation of the Na/Ca exchanger. (b) Rapid solution changes, temporally limiting exposure to potentially detrimental ionic conditions. (c) Single-channel recording, exposing only the cell-attached patch of membrane to the test solutions. An inward whole-cell photocurrent could be measured with Ca as the only extracellular charge carrier. Decreasing the [Ca]o to 0.5 mM reduced the response by 43% and displaced the reversal potential by -4.3 mV; the shift was larger (delta Vrev = -44 mV) when intracellular permeant cations were also removed. In all cases, however, the current carried by Ca was < 5% of that measured with normal [Na]o. Unitary light-activated currents were reduced in a similar way when the pipette contained only divalent cations, indicating a substantial selectivity for Na over Ca. The fall kinetics of the photoresponse was slower when external Ca was replaced by Ba, or when the membrane was depolarized; however, dialysis with 10 mM BAPTA failed to antagonize this effect, suggesting that mechanisms other than the Ca influx participate in the modulation of the time course of the photocurrent.

The placental blood program of the Barcelona Cord Blood Bank.

Cord blood has recently become an alternative to bone marrow transplantation, generating the need for cord blood banks where large numbers of frozen cord blood units can be stored. The Barcelona Cord Blood Bank (bcB) was created in October 1995. Initially, several methods for volume reduction were tested, including Ficoll, Percoll, Gelatin and HES sedimentation. Of these, HES sedimentation (88% +/- 11 CD34+ cells recovery) was the one chosen for routine banking. Up to November 1997, the bank has processed 754 units with a median of 1.05 x 10(9) nucleated cells and 2.5 x 10(6) CD34+ cells stored per unit. Nine of these units have been shipped for transplantation.

Physiology and molecular biology of multidrug resistance in Entamoeba histolytica.

In this paper, we present the most relevant facts on multidrug resistance (MDR) in the protozoan parasite Entamoeba histolytica. MDR in E. histolytica presents characteristics similar to transformed mammalian cells. E. histolytica drug resistant mutants show cross-resistance to several drugs, and as in mammalian cells the resistance is reverted by verapamil. Six P-glycoprotein-like genes (EhPgp) have been cloned and characterized. Apparently, four of these genes are transcribed in drug-resistant mutants (EhPgp1, EhPgp2, EhPgp5 and EhPgp6), although only EhPgp1, EhPgp5 and EhPgp6 transcripts were clearly detected. The open reading frame (ORF) of the four completely full length genes is about 1300 amino acids long. EhPgp1, EhPgp2 and EhPgp5 have between 64 and 67% of positional identity among them, while EhPgp6 shows 38 to 46% positional identity to the other ameba genes. Interestingly, the phylogenetic tree suggested that Entamoeba P-glycoproteins are more related to the human and mouse P-glycoproteins than to the Plasmodium and Leishmania P-glycoproteins. Differential gene expression in drug-resistant mutants was detected when specific probes for each Ehpgp gene were used. To understand the differential expression of EhPgp genes we initiated the characterization of the upstream flanking regions of EhPgp1 and EhPgp5 genes. Upstream sequences showed between 53 and 66% of positional identity to Dictyostelium discoideum promoters.

First- and second-derivative spectrophotometric determination of imipenem and cilastatin in injections.

First- and second-derivative spectrophotometry has been used for the quantitation of mixtures of imipenem and cilastatin sodium, compounds that have closely overlapping spectral bands. Beer's law was obeyed at concentrations up to 100 micrograms ml-1 of imipenem in both the first- and second-derivative modes and up to 75 micrograms ml-1 of cilastatin in the first-derivative mode. Detection limits at the P = 0.05 level of significance were calculated to be 0.40 and 0.52 micrograms ml-1 of imipenem and cilastatin sodium, respectively, in the first-derivative mode, and in a range from 0.45 to 0.68 micrograms ml-1 for imipenem in the second-derivative mode. The method, which is rapid, simple and does not require a separation step, has been successfully applied to the assay of commercial injections.

First and second derivative spectrophotometric determination of cefoperazone and sulbactam in injections.

A simple, spectrophotometric assay to measure the concentrations of cefoperazone and sulbactam in injectable formulations is described. Since zero-order spectra are subject to interference, derivative spectrophotometry was used to enhance the spectral details. A linear relationship between derivative amplitudes and the concentrations of the compounds was found. Beer's law is obeyed up to 75 and 80 micrograms ml-1 of cefoperazone in the first and second derivative modes, respectively, and up to 75 micrograms ml-1 of sulbactam in the second derivative mode. Detection limits were 0.64 and 0.88 microgram ml-1, respectively for cefoperazone in the first and second derivative modes and 0.30 micrograms ml-1 for sulbactam in the second derivative mode. The method is rapid, simple, does not require a separation step and has successfully been applied to the assay of commercial injections containing cefoperazone and sulbactam.

Simultaneous determination of cefepime and L-arginine in injections by second-derivative spectrophotometry.

A simple spectrophotometric assay for the determination of cefepime and L-arginine in injections is described. Since zero-order spectra showed considerable overlap, second-derivative spectrophotometry was used to enhance the spectral details. A linear relationship between second-derivative amplitude and concentration of each compound was found. Beer's law was obeyed up to 50 and 22 micrograms ml-1 of cefepime and arginine, respectively, in the second-derivative mode. Detection limits were 0.31 and 0.58 micrograms ml-1 for cefepime and arginine, respectively. The method, which is rapid, simple and does not require any separation step, has been successfully applied to the assay of commercial injections containing cefepime and arginine.

Evaluation of conformation against traits associated with dressage ability in unridden Iberian horses at the trot.

The work describes and compares the conformation traits and gait characteristics in the main native Iberian dressage Purebreds (Lusitano, Spanish and Menorca horses). In total, 4 strides belonging to each of 66 males (4-14 years old) were recorded at handled trot, using a 3D semi-automatic movement analysis system. Principal component analysis (PCA) and intraclass Correlations (iC) were computed. The Menorca Purebred showed proportionally higher limbs (23.8 and 15.1cm for the forelimb cannon and pastern lengths), the Lusitano Purebred had larger inclinations (66.1°, 37.7° and 47.9° for the scapula and pastern angles), and the Spanish Purebred presented a longer scapula (41.5 cm) and a larger hip angle (132.1°), apparently showing a better conformation for dressage gaits. However, the three breeds presented undesirable tarsus joint conformation (<155.5°) for dressage performance. In general, all of them showed good features in the kinematics of the forelimbs. Nevertheless, the functional capacity of their hindlimbs and linear parameters probably need to be improved for their competitiveness.