Application of embryo biopsy and sex determination via polymerase chain reaction in a commercial equine embryo transfer program in Argentina.

Embryo biopsy for fetal sexing has clinical application, but few reports are available of its use within an active embryo transfer program. We evaluated results on biopsy of 459 embryos over one breeding season. There were no significant differences in pregnancy rate between biopsied and non-biopsied embryos (72% vs 73%) or for biopsied embryos recovered at the centre (73%) compared with those shipped overnight (72%). However, the pregnancy rate decreased significantly in shipped embryos biopsied ≥20h after collection. Overall, 86% of biopsies provided a sex diagnosis. The likelihood of a positive genomic (g) DNA result was significantly higher for biopsies from large blastocysts (96%) than from smaller embryos (70-85%). In total, 38% of biopsies were positive for Y chromosome DNA (Y-DNA) and were diagnosed as male. Subsequently, 95% of Y-DNA-positive embryos were confirmed as male and 78% of Y-DNA-negative embryos were confirmed as female. The accuracy of prediction of female (Y-DNA negative) was significantly higher when the biopsy sample was probed for Y-DNA only compared with probing for both gDNA and Y-DNA. We estimate that by transferring only Y-DNA-negative embryos, 3% of potential female pregnancies may have been lost, and production of male pregnancies was reduced by 72%.

Characterization of third-generation cephalosporin-resistant Escherichia coli from slaughter calves and fattening pigs: A pilot study for monitoring antimicrobial resistance by whole genome sequencing in Switzerland.

INTRODUCTION: Whole genome sequencing (WGS) was introduced into Swiss antimicrobial resistance monitoring in 2022 as an additional method to phenotypic antimicrobial susceptibility testing by broth microdilution to characterize presumptive third-generation cephalosporin-resistant (3GC-R) Escherichia coli. Caecal samples from Swiss slaughter calves and fattening pigs, as well as beef and pork meat from Swiss retail taken in 2021, were analyzed for the presence of 3GC-R E. coli according to European harmonized protocols. In 2021, 3GC-R E. coli was detected in 23,8 % of slaughter calves, 5,9 % of fattening pigs, and 0 % of meat. Comparative analysis of the antimicrobial resistance results obtained by phenotypic measurement and those obtained by the detection of corresponding underlying molecular mechanisms by WGS showed very high agreement (99 %). Resistance to third-generation cephalosporins (3GCs) was mainly associated with the presence of blaCTX-M-15 in E. coli isolates from calves and blaCTX-M-1 in E. coli isolates from pigs and mutations in the ampC-promoter (g.-42 C>T) in E. coli isolates from both animal species. Moreover, WGS data were used for phylogenetic analysis based on multi locus sequence types (MLST) and core genome MLST(cgMLST) revealing that 3GC-R E. coli isolated from Swiss slaughter calves and fattening pigs were genetically diverse. In this study, it was shown that using WGS alone to monitor antimicrobial resistance could detect trends in known molecular antimicrobial resistance mechanisms while also providing other valuable information about the isolates, such as genetic relatedness. However, only by combining phenotypic susceptibility testing and WGS early detection of previously unknown resistance mechanisms will be possible.

INTRODUCTION: Le séquençage du génome entier (Whole Genome Sequencing, WGS) a été introduit dans la surveillance suisse de la résistance aux antibiotiques en 2022 en tant que méthode supplémentaire aux tests phénotypiques de sensibilité aux antibiotiques pour caractériser les Escherichia coli résistants aux céphalosporines de troisième génération (3GC-R). Des échantillons de cæcum pris en 2021 à l'abattoir de veaux et de porcs suisses, ainsi que de viande de bœuf et de porc provenant de détaillants suisses ont été analysés pour détecter la présence d'E. coli 3GC-R conformément aux protocoles européens harmonisés. En 2021, les E. coli 3GC-R ont été détectés dans 23,8 % des veaux d'abattage, 5,9 % des porcs d'engraissement et 0 % dans la viande. Les résultats de résistance aux antibiotiques obtenus par mesure phénotypique et ceux obtenus par la détection des mécanismes moléculaires sous-jacents concordaient à 99 %. La résistance aux céphalosporines de troisième génération était principalement associée à la pré-sence de blaCTX-M-15 dans les isolats d'E. coli provenant de veaux et de blaCTX-M-1 dans les isolats d'E. coli provenant de porcs et à des mutations dans le promoteur ampC (g.-42 C>T) dans les isolats d'E. coli provenant des deux espèces animales. Les données WGS ont également été utilisées pour une analyse phylogénétique basée sur les types de séquences multilocus (MLST) et MLST du génome de base (cgMLST) révélant que les E. coli 3GC-R isolés des veaux et des porcs suisses étaient génétiquement divers. Dans cette étude, il a été démontré que l'utilisation du WGS seul pour surveiller la résistance aux antibiotiques pouvait détecter des tendances dans les mécanismes moléculaires connus de la résistance aux antibiotiques tout en fournissant d'autres informations précieuses sur les isolats, comme la parenté génétique. Cependant, ce n'est qu'en combinant les tests de sensibilité phénotypique avec le WGS que la détection pré-coce de mécanismes de résistance inconnus sera possible.

Novel DNA methylation signatures of tobacco smoking with trans-ethnic effects.

BACKGROUND: Smoking remains one of the leading preventable causes of death. Smoking leaves a strong signature on the blood methylome as shown in multiple studies using the Infinium HumanMethylation450 BeadChip. Here, we explore novel blood methylation smoking signals on the Illumina MethylationEPIC BeadChip (EPIC) array, which also targets novel CpG-sites in enhancers. METHOD: A smoking-methylation meta-analysis was carried out using EPIC DNA methylation profiles in 1407 blood samples from four UK population-based cohorts, including the MRC National Survey for Health and Development (NSHD) or 1946 British birth cohort, the National Child Development Study (NCDS) or 1958 birth cohort, the 1970 British Cohort Study (BCS70), and the TwinsUK cohort (TwinsUK). The overall discovery sample included 269 current, 497 former, and 643 never smokers. Replication was pursued in 3425 trans-ethnic samples, including 2325 American Indian individuals participating in the Strong Heart Study (SHS) in 1989-1991 and 1100 African-American participants in the Genetic Epidemiology Network of Arteriopathy Study (GENOA). RESULTS: Altogether 952 CpG-sites in 500 genes were differentially methylated between smokers and never smokers after Bonferroni correction. There were 526 novel smoking-associated CpG-sites only profiled by the EPIC array, of which 486 (92%) replicated in a meta-analysis of the American Indian and African-American samples. Novel CpG sites mapped both to genes containing previously identified smoking-methylation signals and to 80 novel genes not previously linked to smoking, with the strongest novel signal in SLAMF7. Comparison of former versus never smokers identified that 37 of these sites were persistently differentially methylated after cessation, where 16 represented novel signals only profiled by the EPIC array. We observed a depletion of smoking-associated signals in CpG islands and an enrichment in enhancer regions, consistent with previous results. CONCLUSION: This study identified novel smoking-associated signals as possible biomarkers of exposure to smoking and may help improve our understanding of smoking-related disease risk.

Effect of 3basic life support training programs in future primary school teachers. A quasi-experimental design. / Efecto de 3métodos de enseñanza en soporte vital básico en futuros maestros de Educación Primaria. Un diseño cuasiexperimental.

AIM: To evaluate the learning of basic life support (BLS) measures on the part of laypersons after 3different teaching programs. DESIGN: A quasi-experimental before-after study involving a non-probabilistic sample without a control group was carried out. SCOPE: Primary school teacher students from the University of Santiago (Spain). PARTICIPANTS: A total of 124 students (68.8% women and 31.2% men) aged 20-39 years (M=22.23; SD=3.79), with no previous knowledge of BLS, were studied. INTERVENTIONS: Three teaching programs were used: a traditional course, an audio-visual approach and feedback devices. MAIN VARIABLES OF INTEREST: Chest compressions as sole cardiopulmonary resuscitation skill evaluation: average compression depth, compression rate, chest recoil percentage and percentage of correct compressions. Automated external defibrillator: time needed to apply a shock before and after the course. RESULTS: There were significant differences in the results obtained after 2minutes of chest compressions, depending on the training program received, with feedback devices having a clear advantage referred to average compression depth (p<0.001), compression rate (p<0.001), chest recoil percentage (p<0.001) and percentage of correct compressions (p<0.001). Regarding automated external defibrillator, statistically significant differences were found in Tafter (p=0.025). CONCLUSIONS: The teaching course using feedback devices obtained the best results in terms of the quality of chest compressions, followed by the traditional course and audio-visual approach. These favorable results were present in both men and women. All 3teaching methods reached the goal of reducing defibrillation time.

Fast-track recovery technique applied to primary total hip and knee replacement surgery. Analysis of costs and complications. / Técnica de recuperación acelerada (fast-track) aplicada a cirugía protésica primaria de rodilla y cadera. Análisis de costos y complicaciones.

PURPOSE: To determine the cost reduction and complication rates of using an enhanced recovery pathway (Fast-track) when compared to traditional recovery in primary total hip replacement (THR) and total knee replacement (TKR), as well as to determine if there were significant differences in complication rates. MATERIAL AND METHODS: Retrospective review of 100 primary total arthroplasties using the Fast-track recovery system and another 100 using conventional recovery. Gender, Charlston comorbidity index, ASA score, length of stay and early complications were measured, as well in-hospital complications and those in the first six months, re-admissions and transfusion rates. The total and daily cost of stay was determined and the cost reduction was calculated based on the reduction in the length of stay found between the groups. RESULTS: Both groups where comparable as regards age, gender, ASA score, and Charlston index. The mean reduction in length of stay was 4.5 days for TKR and 2.1 days for THR. The calculated cost reduction was 1266 euros for TKR and 583 euros for THR. There were no statistically significant differences between groups regarding in-hospital complications, transfusion requirements, re-admissions and complication rates in the first six months. DISCUSSION: There are few publications in the literature reviewed that analyse the cost implications of using fast-track recovery protocols in arthroplasty. Several published series comparing recovery protocols found no significant differences in complication rates either. The use of a fast-track recovery protocol resulted in a significant cost reduction of 1266 euros for the TKR group and 583 for the THR group, without affecting complication rates.

Modeling photosynthesis in olive leaves under drought conditions.

We quantified parameters for a model of leaf-level photosynthesis for olive, and tested the model against an independent dataset. Specific temperature-dependence parameters of the model for olive leaves were measured, as well as the relationship of the model parameters with area-based leaf nitrogen (N) content. The effect of soil water deficit on leaf photosynthesis was examined by applying two irrigation treatments to 29-year-old trees growing in a plantation: drip irrigation sufficient to meet the crop water requirements (I) and dry-farming (D). In both treatments, leaves had a higher photosynthetic capacity in April than in August. In August, photosynthetic capacity was lower in D trees than in I trees. Leaf photosynthetic capacity was linearly and positively related to leaf N content on an area basis (N(a)) and to leaf mass per unit area (LMA), and the regression slope varied with irrigation treatment. The seasonal reduction in N(a) was used in the model to predict photosynthesis under drought conditions. Olive leaves showed a clear limitation of photosynthesis by triose phosphate utilization (TPU) even at 40 degrees C, and the data suggest that olive invests fewer resources in TPU than other species. The seasonal decrease in photosynthetic capacity moderated the stomatal limitation to carbon dioxide (CO(2)) fixation as soil water deficit increased. Further, it enabled leaves to operate close to the transition point between photosynthetic limitation due to RuBP carboxylation capacity and that due to RuBP regeneration capacity, and resulted in a near constant value of internal CO(2) concentration from April to August. Under well watered conditions, N-use efficiency of the olive leaves was enhanced at the expense of reduced water-use efficiency.

Calibration of sap flow estimated by the compensation heat pulse method in olive, plum and orange trees: relationships with xylem anatomy.

The compensation heat pulse method is widely used to estimate sap flow in conducting organs of woody plants. Being an invasive technique, calibration is crucial to derive correction factors for accurately estimating the sap flow value from the measured heat pulse velocity. We compared the results of excision and perfusion calibration experiments made with mature olive (Olea europaea L. 'Manzanilla de Sevilla'), plum (Prunus domestica L. 'Songal') and orange (Citrus sinensis (L.) Osbeck. 'Cadenero') trees. The calibration experiments were designed according to current knowledge on the application of the technique and the analysis of measured heat pulse velocities. Data on xylem characteristics were obtained from the experimental trees and related to the results of the calibration experiments. The most accurate sap flow values were obtained by assuming a wound width of 2.0 mm for olive and 2.4 mm for plum and orange. Although the three possible methods of integrating the sap velocity profiles produced similar results for all three species, the best results were obtained by calculating sap flow as the weighted sum of the product of sap velocity and the associated sapwood area across the four sensors of the heat-pulse-velocity probes. Anatomical observations showed that the xylem of the studied species can be considered thermally homogeneous. Vessel lumen diameter in orange trees was about twice that in the olive and plum, but vessel density was less than half. Total vessel lumen area per transverse section of xylem tissue was greater in plum than in the other species. These and other anatomical and hydraulic differences may account for the different calibration results obtained for each species.

Induction of drug resistance in embryonal rhabdomyosarcoma treated with conventional chemotherapy is associated with HLA class I increase.

Effectiveness of conventional cytotoxic treatment of rhabdomyosarcoma (RMS) may be limited by the development of multidrug resistance (MDR) mediated by mdr1 gene. This gene codes for P-glycoprotein (P-gp) which has been related to a immunoregulatory function. Modulation of HLA expression by P-gp has been described in different types of tumor cells including RMS. However, very little is known about biological implications of the P-gp expression in RMS patients treated with conventional chemotherapy. In order to study the problem, we used embryonal RMS tissue samples from treated patients. Our results indicated that positive RMS samples to mdr1 show higher HLA class I expression than those which were negative to mdr1 PCR, what indicates a significant correlation between the expression of both molecules. In addition, we developed two resistant RMS cell lines (A-204-1 and 2) using similar concentrations of actinomycin D as are plasma levels in clinical situation. Both resistant cell lines showed mdr1 expression and an increase of HLA class I expression which was dose-dependent. These results demonstrated that conventional chemotherapy of embryonal RMS is able to induce resistance which can modulate HLA class I expression and suggest that immunological studies of these tumors may be necessary to the design new specific therapeutic strategies.

HLA class I and II expression in rhabdomyosarcomas.

The expression of class I and II histocompatibility antigens was studied in 43 specimens of skeletal and in 36 specimens of cardiac muscle. Expression was also analyzed in the tumoral counterpart of striated muscle tissue, in 21 rhabdomyosarcomas (13 embryonic, 2 alveolar and 6 pleomorphic). Normal striated muscle showed very weak HLA class I antigen expression and no class II expression. The rhabdomyosarcomas showed increased class I expression, possibly related to malignant transformation of this type of tissue and with the degree of cellular differentiation. In our series of rhabdomyosarcomas, we observed class II neoexpression only in some HLA class I positive specimens, which was unrelated to the degree of cellular differentiation in these tumors.

Production and characterization of a new monoclonal antibody, GR-ICOR-2, recognizing sarcomeric actin: analysis of the expression in the developing chick heart.

We produced and characterized a specific monoclonal antibody (mAB) designated GR-ICOR-2. This mAb recognizes sarcomeric actin molecules (43 kDa) and was used in an immunohistochemical analysis of staining patterns in Hamburger and Hamilton's stages 18, 22 and 25 (HH 18, 22 and 25) embryonic chick hearts. Staining showed a mainly cytoplasmic distrubition in three regions: the atrioventricular (AV) canal cushion tissue, the primitive ventricle, and conal crests. In addition, this mAb-cross-reacted with rabbit and human cardiac and skeletal muscle tissue; but not with smooth muscle tissue.

Influence of fibric acid derivatives on intermediate filament proteins in myocardiocyte cultures.

We analyzed desmin and vimentin accumulation in chick myocardiocyte cultures treated with the fibric acid derivatives bezafibrate, fenofibrate and gemfibrozil. The most noteworthy finding was the 50% decrease in the cytoplasmic desmin fraction in cells treated with gemfibrozil in comparison to control cultures, and the 19% increase in the cytoskeletal fraction in cultures treated with gemfibrozil and with bezafibrate. Vimentin accumulation by cells treated with bezafibrate was similar to that in control cultures, however the cytoskeletal vimentin fraction rose by 26% after treatment with gemfibrozil, and fell 13% after treatment with fenofibrate. No vimentin was found in the cytoplasmic fraction of cell treated with bezafibrate. Given the role of intermediate filaments in heart muscle contraction, fibric acid derivative- induced changes in the cytoplasmic and cytoskeletal concentrations of intermediate filament proteins may be related with the secondary effects of these drugs on heart rate.

Photon backscattering tissue characterization by energy dispersive spectroscopy evaluations.

Techniques for in vivo tissue characterization based on scattered photons have usually been confined to evaluating coherent and Compton peaks. However, information can also be obtained from the energy analysis of the Compton scattered distribution. This paper looks at the extension of a technique validated by the authors for characterizing tissues composed of low-atomic-number elements. To this end, an EDXRS (energy dispersive x-ray spectrometry) computer simulation procedure was performed and applied to test the validity of a figure of merit able to characterize binary compounds. This figure of merit is based on the photon fluence values in a restricted energy interval of the measured distribution of incoherently scattered photons. After careful experimental tests with 59.54 keV incident photons at scattering angles down to 60degrees, the simulation procedure was applied to quasi-monochromatic and polychromatic high-radiance sources. The results show that the characterization by the figure of merit, which operates satisfactorily with monochromatic sources, is unsatisfactory in the latter cases, which seem to favour a different parameter for compound characterization.

Expression of epidermal growth factor receptor in chick embryo myocardiocytes: relation with desmin expression during cardiac development.

The epidermal growth factor receptor is related with processes of cell replication and differentiation. We used the intermediate filament protein desmin as a marker to study the relation between muscle cell differentiation and modifications in the expression of this receptor during heart development in the chick embryo. Epidermal growth factor receptor was expressed as early as Hamburger and Hamilton's stage 17, when myocardiocytes are still poorly differentiated and desmin-negative. Expression became steadily weaker as the heart matured, and decreased after Hamburger and Hamilton's stage 25, a key stage in heart maturation characterized by a sharp increase in desmin expression. Our findings suggest that in the chick embryo, the expression of epidermal growth factor receptor becomes steadily weaker as myocardiocyte differentiation progresses.

Effects of fibric acid derivatives on accumulation of actin in myocardiocytes.

We used sodium dodecylsulphate polyacrylamide gel electrophoresis and immunoblotting to analyze the effects of the fibric acid derivatives bezafibrate, fenofibrate and gemfibrozil on the accumulation of actin in the cytoplasmic and cytoskeletal fraction of cultured myocardiocytes. All three drugs tested modified cellular and subcellular actin in different ways, and the findings are thought to be related with the secondary effect of arrhythmia known to be caused by these drugs. Bezafibrate and gemfibrozil more markedly affected accumulation of actin by myocytes, while fenofibrate interfered less notably with the accumulation of this protein.

Analysis of the distribution of human CD38 and of its ligand CD31 in normal tissues.

Human CD38 is a 45 kD ectoenzyme endowed with ADP-ribosyl cyclase and hydrolase activities. The molecule plays a central role in lymphocyte activation, proliferation and selectin-type adhesion with endothelial cells (HEC). A HEC surface molecule displaying all the features of a CD38L has been identified by means of a mAb (Moon-1), able to block CD38-mediated adhesion processes. The 130 kD molecule recognized by Moon-1 is CD31, a member of the Ig superfamily. This paper reports on the analysis of the surface expression of CD38 and CD38L in various human tissues of adult origin and compared in some instances to the fetal (9-14 weeks) counterparts. This was achieved by means of immunohistochemical techniques and analysis of purified cell populations. Among the specimens analyzed, CD38 is expressed by a vast array of cells of lymphatic origin as well as by the skeletal and cardiac muscle fibers, the bronchi (epithelial cells), the parotid gland (ductal epithelial cells) and hepatic sinusoids. On the contrary, CD31 proved constantly expressed by HEC at high levels, independent of the organ or tissue analyzed or of the kind of vessel. Other cells expressing CD38L were found in the lymphoid compartment (follicle mantle B cells and plasma cells), in the lungs (alveolar ducts, alveoli and lymphatic vessels) and in the kidney (glomerular cells). Interestingly, no fetal organ or tissue ever expressed CD38 and its ligand. The above results were enriched by the analysis of the expression of the two molecules on purified populations including mononuclear cells from the lamina propria of the gastro-intestinal tract and broncho-alveolar lavage lymphocytes.

Creatine kinase isoenzymes as markers of differentiation in rhabdomyosarcoma cells in culture.

Rhabdomyosarcomas have been classified on the basis of their degree of differentiation, a feature closely related with their response to chemotherapy. We recently reported the role of creatine kinase isoenzymes as tumoral markers in the diagnosis of rhabdomyosarcoma. The present study was designed to determine whether these isoenzymes are also good markers of the degree of differentiation of these neoplasm. Dimethyl sulfoxide, a well-known differentiating agent, was used to induce myogenic differentiation. This agent increased CK-MM and/or CK-MB fractions, and decreased CK-BB isoenzyme in rhabdomyosarcoma cell lines. The variable behavior of MCK-1 macromolecule was unrelated to the degree of differentiation. Our findings suggest that CK-MM, CK-MB and CK-BB isoenzymes are good markers of the degree of differentiation in rhabdomyosarcomas and could be used as specific markers of prognostic and diagnostic value.

Design-oriented functional capacity evaluation: the available motions inventory--a review.

The accommodation of a task to the unique abilities of an individual with a disability is becoming increasingly the responsibility of the employer. Functional capacity evaluation (FCE) provides the information necessary for rational design decisions when modifying work stations. The currently used FCE systems have been designed to assess an individual's performance for a range of reasons other than industrial task accommodation. The Available Motions Inventory (AMI) is a system that has been developed specifically to address the design issues raised when accommodating an industrial task to a person with a neuromuscular impairment. This system has a unique scoring system that allows intra-individual ability comparisons that support task allocation decisions. Some general principles of task design are developed for persons with cerebral palsy, but the strength of assessment lies in its application to a specific individual performing a specific task.

[Modulation of cardiac contractility in myogenesis using antidesmin monoclonal antibody]. / Modulación de la contractilidad cardíaca en la miogénesis mediante el anticuerpo monoclonal antidesmina.

The cytoskeleton of the myocardiocyte is a complex network of protein filaments. The intermediate filaments, consisting fundamentally of desmin, are considered markers of cellular differentiation. We used indirect immunofluorescence with specific monoclonal antibody to characterize the expression of desmin during the different phases of heart maturation, and demonstrated a direct relationship between the expression of this protein and myocardiocyte development. Our results show that in auricular myocardiocytes, desmin is present in early stages of development, whereas in ventricular myocardiocytes, this protein, which is intimately involved in contractile function, first appears in HH stage 25.

Molecular coherent scattering data for tissue in photon transport Monte Carlo codes.

The suitability of introducing molecular interference effects into coherent scattering phenomenology is considered here in the case of X and gamma photon transport in biological tissues. In order to make its implementation practicable in most of the existing open user Monte Carlo codes, a self-consistent basic set of tabulations for the linear differential scattering coefficient was proposed. The tabulations were critically reviewed by testing their validity over the entire range of the momentum transfer variable from 0 to 10(10) nm(-1). Implementation in three different Monte Carlo codes was performed, and they proved to be reliable.

Kienböck's disease. II. Risk factors, diagnosis, and ergonomic interventions.

The term CTD has been used to refer to those musculoskeletal impairments that appear to be work-related and tend to be chronic. Upper extremity CTD are categorized in three major groups: tendon disorders, neurovascular disorders, and nerve. Kienböck's disease, on the other hand, has been documented in the workplace and has a similar etiology, but due to current anatomic classifications cannot be considered a CTD. With this in mind, the present article reviews risk factors, diagnosis, and treatment, with the purpose of providing physicians, engineers, and designers with information to reduce the risk of workers developing this potentially debilitating disease. Furthermore, additional support will be accumulated and combined with Part I of this article with the intent of introducing into the literature a new category of upper extremity CTD: Bone Disorders.