Baseline concussion assessment performance by sex in military service academy rugby players: findings from the CARE Consortium.

BACKGROUND: Normative student-athlete concussion assessment data may not be appropriate for service academy members (SAMs), particularly rugby players, because of the uniqueness of their academic/military training environment. Having accurate baseline data for this population is important because of their high risk for concussion and frequent lack of assigned sports medicine professional. The primary purpose of this study was to characterise baseline performance on a concussion assessment battery, with secondary purpose to determine effect of sex and concussion history on these measures among SAM rugby players. METHODS: 601 rugby-playing SAMs (19.3±1.5 years, 37.9% female) completed baseline concussion assessments: the Sport Concussion Assessment Tool (SCAT) Symptom and Symptom Severity Checklist, Standard Assessment of Concussion (SAC) and a neuropsychological test (either ImPACT (Immediate Post Concussion Assessment and Cognitive Testing) or ANAM (Automated Neuropsychological Assessment Metrics)). Groups were compared using an independent samples t-test or Mann-Whitney U test. A 2 (sex) × 2 (concussion history) ANOVA was conducted to determine the effects of sex and concussion history on outcomes. RESULTS: Women reported greater SCAT total symptoms (3.3 vs 2.8, p<0.001, r=0.143) and symptom severities (5.7 vs 4.3, p<0.001, r=0.139), and performed worse on ImPACT Visual Memory (79.3 vs 82.6, p=0.002, r=0.144) than men. Women performed better than men on SAC (28.0 vs 27.7, p=0.03, r=0.088), ImPACT Reaction Time Composite (0.59 vs 0.61, p=0.04, r=0.092) and ANAM Code Substitution Delayed (64.3 vs 61.5, p=0.04, d=0.433). Individuals with a history of concussion reported lower ImPACT Symptom Severity (2.6 vs 4.2, p=0.02, r=0.110). There was no interaction between concussion history and sex on outcomes. CONCLUSIONS: These findings provide reference data for SAM rugby players on baseline assessments and to help in clinical decision-making when managing sports-related concussion in absence of baseline data.

Use of internal-external diversion stent in open pyeloplasty in patients under one year of age. / Uso de catéter de derivación interno-externo en la pieloplastia abierta en menores de un año.

INTRODUCTION: There are various alternatives available for renal pelvis drainage following pyeloplasty. One of them is the use of an internal-external diversion stent, which according to our protocol, is knotted 48 hours following surgery, prior to discharge, and removed 7 days later on an outpatient consultation basis, with no sedation or analgesia required. OBJECTIVE: To analyze the results of patients under one year of age who underwent open pyeloplasty associated with an outpatient internal-external diversion stent. MATERIALS AND METHODS: A retrospective, descriptive analysis of 28 patients (31 renal units) undergoing surgery from 2011 to 2021 was carried out. Diagnostic methods, indications, surgical approach, and postoperative progression were assessed. RESULTS: 28 patients (23 male) prenatally diagnosed with hydronephrosis confirmed by ultrasonography and/or renogram underwent pyeloplasty at a median age of 3 months (15 days-11 months). Pyeloplasty was conducted according to the Anderson-Hynes technique or dismembered pyeloplasty in 28 renal units, and according to the Culp-DeWeerd technique or spiral flap in 3. In all cases, an internal-external diversion stent was used according to our protocol. Mean hospital stay was 3.5 days (2-7 days), with a good postoperative progression. 2 patients had complications (urinary infection requiring intravenous antibiotics, and pyonephrosis requiring re-pyeloplasty). CONCLUSIONS: Using an internal-external diversion stent following pyeloplasty in patients under 1 year of age with ureteropelvic junction obstruction is a simple and safe option that allows for early discharge with outpatient management. It also avoids a second general anesthesia for drainage catheter removal purposes.

INTRODUCCION: Existen diversas alternativas para el drenaje de la pelvis renal tras una pieloplastia. Una de ellas es la utilización de un catéter de derivación interno-externo que, según nuestro protocolo, se anuda a las 48 horas posoperatorias previas al alta y se retira a los siete días de forma ambulatoria en consulta, sin necesidad de sedoanalgesia. OBJETIVO: Analizar los resultados de los pacientes menores de un año intervenidos mediante pieloplastia abierta, asociando un catéter de derivación interno-externo de manejo ambulatorio. MATERIAL Y METODOS: Análisis descriptivo retrospectivo de 28 pacientes (31 unidades renales) intervenidos entre los años 2011 y 2021. Se evaluaron métodos diagnósticos, indicaciones, abordaje quirúrgico y evolución posoperatoria. RESULTADOS: Veintiocho pacientes (23 varones) con diagnóstico prenatal de hidronefrosis confirmado con ecografía y/o renograma, fueron intervenidos mediante pieloplastia a una mediana de edad de tres meses (15 días-11 meses). Se realizó pieloplastia según técnica de Anderson-Hynes o pieloplastia desmembrada en 28 unidades renales y según técnica de Culp-DeWeerd o colgajo en espiral en 3. En todos los casos se utilizó un catéter de derivación interno-externo según protocolo. El tiempo medio de ingreso fue 3,5 días (2-7 días) con buena evolución posoperatoria. Dos pacientes presentaron complicaciones (infección urinaria que requirió antibioterapia intravenosa y pionefrosis que requirió repieloplastia). CONCLUSIONES: Asociar un catéter de derivación interno-externo a la pieloplastia en pacientes menores de un año con estenosis de la unión pieloureteral es una opción sencilla y segura que permite un alta precoz con manejo ambulatorio y evita una segunda anestesia general para la retirada del catéter de drenaje.

Uso de catéter de derivación interno-externo en la pieloplastia abierta en menores de un año / Use of internal-external diversion stent in open pyeloplasty in patients under one year of age

Introducción: Existen diversas alternativas para el drenaje de la pelvis renal tras una pieloplastia. Una de ellas es la utilización de un catéter de derivación interno-externo que, según nuestro protocolo, se anuda a las 48 horas posoperatorias previas al alta y se retira a los siete días de forma ambulatoria en consulta, sin necesidad de sedoanalgesia. Objetivos: Analizar los resultados de los pacientes menores de un año intervenidos mediante pieloplastia abierta, asociando un catéter de derivación interno-externo de manejo ambulatorio. Material y métodos: Análisis descriptivo retrospectivo de 28 pacientes (31 unidades renales) intervenidos entre los años 2011 y 2021. Se evaluaron métodos diagnósticos, indicaciones, abordaje quirúrgico y evolución posoperatoria. Resultados: Veintiocho pacientes (23 varones) con diagnóstico prenatal de hidronefrosis confirmado con ecografía y/o renograma, fueron intervenidos mediante pieloplastia a una mediana de edad de tres meses (15 días-11 meses). Se realizó pieloplastia según técnica de Anderson-Hynes o pieloplastia desmembrada en 28 unidades renales y según técnica de Culp-DeWeerd o colgajo en espiral en 3. En todos los casos se utilizó un catéter de derivación interno-externo según protocolo. El tiempo medio de ingreso fue 3,5 días (2-7 días) con buena evolución posoperatoria. Dos pacientes presentaron complicaciones (infección urinaria que requirió antibioterapia intravenosa y pionefrosis que requirió repieloplastia). Conclusiones: Asociar un catéter de derivación interno-externo a la pieloplastia en pacientes menores de un año con estenosis de la unión pieloureteral es una opción sencilla y segura que permite un alta precoz con manejo ambulatorio y evita una segunda anestesia general para la retirada del catéter de drenaje.(AU)

Introduction: There are various alternatives available for renal pelvis drainage following pyeloplasty. One of them is the use of an internal-external diversion stent, which according to our protocol, is knotted 48 hours following surgery, prior to discharge, and removed 7 days later on an outpatient consultation basis, with no sedation or analgesia required. Objective: To analyze the results of patients under one year of age who underwent open pyeloplasty associated with an outpatient internalexternal diversion stent. Materials and methods: A retrospective, descriptive analysis of 28 patients (31 renal units) undergoing surgery from 2011 to 2021 was carried out. Diagnostic methods, indications, surgical approach, and postoperative progression were assessed. Results: 28 patients (23 male) prenatally diagnosed with hydronephrosis confirmed by ultrasonography and/or renogram underwent pyeloplasty at a median age of 3 months (15 days-11 months). Pyeloplasty was conducted according to the Anderson-Hynes technique or dismembered pyeloplasty in 28 renal units, and according to the Culp-DeWeerd technique or spiral flap in 3. In all cases, an internal-external diversion stent was used according to our protocol. Mean hospital stay was 3.5 days (2-7 days), with a good postoperative progression. 2 patients had complications (urinary infection requiring intravenous antibiotics, and pyonephrosis requiring re-pyeloplasty). Conclusions: Using an internal-external diversion stent following pyeloplasty in patients under 1 year of age with ureteropelvic junction obstruction is a simple and safe option that allows for early discharge with outpatient management. It also avoids a second general anesthesia for drainage catheter removal purposes.(AU)

Common surgical training program: standardization of learning quality. / Programa de formación quirúrgica común: uniformidad en la calidad del aprendizaje.

INTRODUCTION: The various surgical specialties in our center have used the simulation and experimental surgery resources available for their training tasks in minimally invasive surgery (MIS) in an individualized manner. With this learning model, a great dispersion of effort and expense was observed, so it was decided to create a unified program based on the following: shared learning, synergy among specialties, moderation of the economic cost, and rational use of the facilities. OBJECTIVE: To describe and assess our consensually designed training program in order to consolidate a shared learning strategy that will enable our residents to acquire and perfect surgical skills in MIS. MATERIALS AND METHODS: The program consists of various increasingly complex phases implemented on a continuous basis throughout the period of specialized training in the virtual laboratory and experimental operating room. The assessment methods were based on quantifiable criteria: percentage of efficiency and completion time of the "McGill Inanimate System for Training and Evaluation of Laparoscopic Skills" (MISTELS) exercises at the beginning and end of the program. An economic study was also conducted. RESULTS: 20 residents have completed the program. Mean times show a significant reduction in each of the exercises. The efficiency percentages at the end of the program were higher than at the beginning (p < 0.001). The cost of the program represented a saving of 67.89%. CONCLUSION: The new MIS training program improved the quality of learning in a safe environment, establishing common criteria among the different specialties and an improved use of resources.

INTRODUCCION: Las diferentes especialidades quirúrgicas de nuestro centro han usado los recursos de simulación y cirugía experimental para sus tareas de formación en cirugía mínimamente invasiva (CMI) de manera individualizada. Con este modelo de aprendizaje se detectó una gran dispersión de esfuerzos y gasto, por lo que se decidió crear un programa unificado basado en: aprendizaje compartido, sinergia entre especialidades, moderación del coste económico y uso racional de las instalaciones. OBJETIVO: Describir y evaluar nuestro programa de entrenamiento diseñado por consenso de cara a la consolidación de una estrategia de aprendizaje compartido que permita a nuestros residentes adquirir y perfeccionar habilidades quirúrgicas en CMI. MATERIAL Y METODOS: El programa consta de diferentes fases con complejidad creciente desarrolladas durante todo el periodo de formación especializada de forma continuada en laboratorio virtual y quirófano experimental. Los criterios de evaluación se basaron en criterios cuantificables: porcentaje de eficiencia y tiempo de realización de los ejercicios de McGill Inanimate System for Training and Evaluation of Laparoscopic Skills (MISTELS) al inicio y final del programa. Se realizó también el estudio económico. RESULTADOS: Han completado el programa 20 residentes. Los tiempos medios demuestran una reducción significativa en cada uno de los ejercicios. Los porcentajes de eficiencia al final fueron mayores que al inicio del programa (p < 0,001). El coste del programa supuso un ahorro del 67,89%. CONCLUSION: El nuevo programa de entrenamiento en CMI mejoró la calidad de aprendizaje en un entorno seguro, estableciendo criterios comunes entre las diferentes especialidades y un mayor aprovechamiento de los recursos.

Programa de formación quirúrgica común: uniformidad en la calidad del aprendizaje / Common surgical training program: standardization of learning quality

Introducción: Las diferentes especialidades quirúrgicas de nuestrocentro han usado los recursos de simulación y cirugía experimentalpara sus tareas de formación en cirugía mínimamente invasiva (CMI)de manera individualizada. Con este modelo de aprendizaje se detectóuna gran dispersión de esfuerzos y gasto, por lo que se decidió crear unprograma unificado basado en: aprendizaje compartido, sinergia entreespecialidades, moderación del coste económico y uso racional de lasinstalaciones. Objetivo: Describir y evaluar nuestro programa de entrenamientodiseñado por consenso de cara a la consolidación de una estrategia deaprendizaje compartido que permita a nuestros residentes adquirir yperfeccionar habilidades quirúrgicas en CMI. Material y métodos: El programa consta de diferentes fases concomplejidad creciente desarrolladas durante todo el periodo de forma-ción especializada de forma continuada en laboratorio virtual y quiró-fano experimental. Los criterios de evaluación se basaron en criterioscuantificables: porcentaje de eficiencia y tiempo de realización de losejercicios de McGill Inanimate System for Training and Evaluation ofLaparoscopic Skills (MISTELS) al inicio y final del programa. Se realizótambién el estudio económico. Resultados: Han completado el programa 20 residentes. Los tiem-pos medios demuestran una reducción significativa en cada uno de losejercicios. Los porcentajes de eficiencia al final fueron mayores queal inicio del programa (p < 0,001). El coste del programa supuso unahorro del 67,89%. Conclusión: El nuevo programa de entrenamiento en CMI mejoróla calidad de aprendizaje en un entorno seguro, estableciendo criterioscomunes entre las diferentes especialidades y un mayor aprovechamientode los recursos.(AU)

Introduction: The various surgical specialties in our center haveused the simulation and experimental surgery resources availablefor their training tasks in minimally invasive surgery (MIS) in anindividualized manner. With this learning model, a great dispersionof effort and expense was observed, so it was decided to create aunified program based on the following: shared learning, synergyamong specialties, moderation of the economic cost, and rationaluse of the facilities. Objective: To describe and assess our consensually designed train-ing program in order to consolidate a shared learning strategy that willenable our residents to acquire and perfect surgical skills in MIS. Materials and methods: The program consists of various increas-ingly complex phases implemented on a continuous basis throughout theperiod of specialized training in the virtual laboratory and experimentaloperating room. The assessment methods were based on quantifiablecriteria: percentage of efficiency and completion time of the “McGillInanimate System for Training and Evaluation of Laparoscopic Skills”(MISTELS) exercises at the beginning and end of the program. Aneconomic study was also conducted. Results: 20 residents have completed the program. Mean timesshow a significant reduction in each of the exercises. The efficiencypercentages at the end of the program were higher than at the begin-ning (p < 0.001). The cost of the program represented a saving of67.89%.(AU)

[Teaching experience of the anesthesiology training unit at Hospital Universitario Nuestra Señora de Candelaria]. / Experiencia de la unidad docente de Anestesiología y Reanimación del Hospital Universitario Nuestra Señora de Candelaria.

BACKGROUND: Health care in Spain has improved progressively and professionals are now required to meet competency levels that safeguard the citizen's right to health protection. To achieve this, instructors in residency training programs and resident physicians themselves are calling for a common framework for training to ensure quality and consistency. Given the scarcity of articles related to training in our journal and following the First Meeting of Residency Program Instructors of the Sociedad Española de Anestesiologia y Reanimación (SEDAR), there has arisen a need to explain how SEDAR's training unit is organized. METHODS: In order to facilitate the sharing of experiences of those involved in training anesthesiology medical residents, we undertook a descriptive analysis of our hospital's curriculum. RESULTS: The structure and operation of the department are described in this report. The results of anonymous surveys completed annually show the satisfaction of residents (9.4 out of 10) and physicians (8.7 out of 10). An audit by the Ministry of Health showed that the curriculum met 100% of the required criteria.

ANALYSIS OF TWO POPULATIONS OF THE URUGUAYAN CANINE BREED CIMARRON (Canis familiaris) USING RAPD MARKERS / ANÁLISIS DE DOS POBLACIONES DE CANINOS (Canis familiaris) DE LA RAZA CIMARRÓN URUGUAYO UTILIZANDO RAPDs COMO MARCADORES MOLECULARES

Objective. To analyze the genetic variability in two populations of the native canine breed "Uruguayan Cimarron" from different areas of the country. Materials and methods. DNA was isolated from 40 animals of the Cimarron breed. Using PCR, 11 RAPD (random amplified polymorphism of DNA) molecular markers were analyzed. Results. 14.9% of loci were polymorphic. We found an index of shared bands of 0.91, a genetic distance of 0.16 and a Nei's average diversity index of 0.08. Conclusions. Both populations present a high homogeneity and genetic identity for these markers suggesting that they belong to a common genetic nucleus.

Objetivo. Analizar la variabilidad genética en dos poblaciones de caninos de la raza autóctona "Cimarrón uruguayo" provenientes de distintas zonas del País. Materiales y métodos. Se aisló ADN de 40 animales de la raza Cimarrón. Mediante la técnica de PCR se analizaron 11 marcadores moleculares RAPD (amplificación al azar de ADN polimórfico). Resultados. Se detectó un 14.9% de loci polimórficos. Se obtuvo un índice de bandas compartidas de 0.91, una distancia genética de 0.16 e índice promedio de diversidad de Nei's de 0.08. Conclusiones. Ambas poblaciones presentan una alta homogeneidad e identidad genética para estos marcadores y pertenecerían a un núcleo genético común.

Avian adeno-associated virus-based expression of Newcastle disease virus hemagglutinin-neuraminidase protein for poultry vaccination.

The avian adeno-associated virus (AAAV) is a replication-defective nonpathogenic virus member of the family Parvoviridae that has been proved to be useful as a viral vector for gene delivery. The use of AAAV for transgenic expression of Newcastle disease virus (NDV) hemagglutinin-neuraminidase (HN) protein and its ability to induce immunity in chickens were assessed. Proposed advantages of this system include no interference with maternal antibodies, diminished immune response against the vector, and the ability to accommodate large fragments of genetic information. In this work the generation of recombinant AAAV virions expressing the HN protein (rAAAV-HN) was demonstrated by electron microscopy, immunocytochemistry, and western blot analysis. Serological evidence of HN protein expression after in ovo or intramuscular inoculation of the recombinant virus in specific-pathogen-free chickens was obtained. Serum from rAAAV-HN-vaccinated birds showed a systemic immune response evidenced by NDV-specific enzyme-linked immunosorbent assay and hemagglutination inhibition testing. Positive virus neutralization in embryonated chicken eggs and indirect immunofluorescence detection of NDV infected cells by serum from rAAAV-HN vaccinated birds is also reported. A vaccine-challenge experiment in commercial broiler chickens using a Venezuelan virulent viscerotropic strain of NDV was performed. All unvaccinated controls died within 5 days postchallenge. Protection up to 80% was observed in birds vaccinated in ovo and revaccinated at 7 days of age with the rAAAV-HN. The results demonstrate the feasibility of developing and using an AAAV-based gene delivery system for poultry vaccination.

Protection against infectious bursal disease virulent challenge conferred by a recombinant avian adeno-associated virus vaccine.

The development and use of recombinant vaccine vectors for the expression of poultry pathogens proteins is an active research field. The adeno-associated virus (AAV) is a replication-defective virus member of the family Parvoviridae that has been successfully used for gene delivery in humans and other species. In this experiment, an avian adeno-associated virus (AAAV) expressing the infectious bursal disease virus (IBDV) VP2 protein (rAAAV-VP2) was evaluated for protection against IBDV-virulent challenge. Specific pathogen free (SPF) birds were inoculated with rAAAV-VP2 or with a commercial intermediate IBDV vaccine and then challenged with the Edgar strain. IBDV-specific antibody levels were observed in all vaccinated groups; titers were higher for the commercial vaccine group. The live, commercial vaccine induced adequate protection against morbidity and mortality; nevertheless, initial lymphoid depletion and follicular atrophy related to active viral replication was observed as early as day 14 and persisted up to day 28, when birds were challenged. No bursal tissue damage due to rAAAV-VP2 vaccination was observed. Eight-out-of-ten rAAAV-VP2-vaccinated birds survived the challenge and showed no clinical signs. The bursa:body weight ratio and bursa lesion scores in the rAAAV-VP2 group indicated protection against challenge. Therefore, transgenic expression of the VP2 protein after rAAAV-VP2 vaccination induced protective immunity against IBDV challenge in 80% of the birds, without compromising the bursa of Fabricius. The use of rAAAV virions for gene delivery represents a novel approach to poultry vaccination.

Primary chicken tracheal cell culture system for the study of infection with avian respiratory viruses.

A major route of infection of avian influenza virus (AIV) and Newcastle disease virus (NDV) in chickens is through cells of the respiratory epithelium. Here we describe the development of a method for culture of tracheal epithelial cells from chicken embryos as well as their use in studies of infection with avian respiratory viruses such as low-pathogenicity AIV and lentogenic NDV. Positive immunostaining for cytokeratin, the presence of cilia and microvilli, and microarray analysis of transcribed RNA demonstrated that the isolated cells were epithelial in nature. Infection of the epithelial cell cultures with AIV and NDV was demonstrated using immunofluorescence or green fluorescence protein fluorescence microscopy, respectively. Growth curves of AIV and NDV in tracheal epithelial cells revealed that tracheal epithelial cells can fully support AIV and NDV growth and reinfection. This system, which mimics that of the natural infection, will be useful to study the mechanisms of early viral infection and cellular host transcriptional responses.

High frequency of beta-catenin mutations in borderline endometrioid tumours of the ovary.

Some low-grade endometrioid carcinomas arise from a background of endometrioid tumours of borderline malignancy. To determine the molecular mechanisms involved in the initiation of endometrioid carcinoma, the present study investigated whether the genetic alterations reported in these tumours (mutations in PTEN, KRAS, and beta-catenin genes, and microsatellite instability) are already present in endometrioid tumours of borderline malignancy. Eight endometrioid tumours of borderline malignancy were studied. By immunohistochemistry, beta-catenin was expressed in the nuclei of all tumours, suggesting the presence of stabilizing beta-catenin mutations. By mutational analysis, five different beta-catenin mutations were found in seven of eight cases (90%), affecting codons 32, 33, and 37. In contrast, only one tumour harboured a PTEN mutation, which affected codon 130. Neither KRAS mutations nor microsatellite instability was detected. A review of the literature indicated that beta-catenin mutations are characteristic of well-differentiated endometrioid carcinomas, since they were present in nearly 60% of grade I but in less of 3% of grade III tumours. In conclusion, the present study identifies beta-catenin mutation as a nearly constant molecular alteration in borderline endometrioid tumours, whereas PTEN and KRAS mutations and microsatellite instability are very infrequent. The findings in the present study, and previously reported data, strongly suggest that beta-catenin mutation is an early event in endometrioid ovarian carcinogenesis, and that it is involved in the development of low-grade endometrioid tumours.

On the conformational memory in the photodissociation of formic acid.

The photodissociation of formic acid at 248 and 193 nm was investigated by classical trajectory and RRKM calculations using an interpolated potential energy surface, iteratively constructed using the B3LYP/aug-cc-pVDZ level of calculation. Several sampling schemes in the ground electronic state were employed to explore the possibility of conformational memory in formic acid. The CO/CO2 branching ratios obtained from trajectories initiated at the cis and at the trans conformers are almost identical to each other and in very good accordance with the RRKM results. In addition, when a specific initial excitation that simulates more rigorously the internal conversion process is used, the calculated branching ratio does not vary with respect to those obtained from cis and trans initializations. This result is at odds with the idea of conformational memory in the ground state proposed recently for the interpretation of the experimental results. It was also found that the calculated CO vibrational distributions after dissociation of the parent molecule at 248 nm are in agreement with the experimental available data.

Sequence analysis, viral rescue from infectious clones and generation of recombinant virions of the avian adeno-associated virus.

Aiming at the generation of a viral-vectored system for gene delivery and vaccination in poultry, the entire genomes of the VR-865 and DA-1 strains of the avian adeno-associated virus have been cloned and sequenced. Sequence analysis of the clones showed that the genomic distribution of the structural and non-structural protein-coding genes of these viruses is conserved and in agreement with what has been previously described for the primate adeno-associated viruses. Amino acid differences between the avian adeno-associated viruses and the primate adeno-associated viruses are more evident in the genes that code for the non-structural (Rep) proteins of the virus, while the Cap region amino acid sequence was found to be more conserved. Since all the regulatory and coding sequences of the virus were present in the plasmids obtained, complete infectious viral particles were rescued from these clones, and these rescued viral populations were amplified by co-infecting primary embryo liver cells with the rescued virus and the CELO strain of the avian adenovirus type 1. As a proof of concept of the validity of this system for the purpose of gene delivery, recombinant viruses encoding for the LacZ gene as a reporter system were also generated. These recombinant viruses were used to express beta galactosidase activity in primary chicken embryo cell cultures.

Transgenic planarian lines obtained by electroporation using transposon-derived vectors and an eye-specific GFP marker.

To generate transgenic planarians we used a set of versatile vectors for animal transgenesis based on the promiscuous transposons, mariner, Hermes and piggyBac, and a universal enhanced GFP (EGFP) marker system with three Pax6 dimeric binding sites, the 3xP3-EGFP developed by Berghammer et al. [Berghammer, A. J., Klinger, M. & Wimmer, E. A. (1999) Nature 402, 370-371]. This marker is expressed specifically in the eyes of various arthropod taxa. Upon microinjection into the parenchyma of adult planarians and subsequent electroporation, these vectors transpose efficiently into the planarian genome. One of the cell types transformed are the totipotent "neoblast" stem cells present in the adults, representing 30% of total cells. The neoblast represents a unique cell type with the capacity to proliferate and to differentiate into all somatic cell types as well as into germ cells. All three transposon vectors have high transformation efficiency, but only Hermes and piggyBac show stable integration. The mariner vector is frequently lost presumably because of the presence of active mariner-type transposons in the genome of the Girardia tigrina. Transformed animals are mosaics containing both transformed and untransformed neoblasts. These differentiate to form EGFP-positive and -negative photoreceptor cells. Such mosaicism is maintained through several cycles of regeneration induced by decapitation or asexual reproduction. Transformed neoblasts also contribute to the germ line, and can give rise to pure transgenic planarian lines in which EGFP is expressed in all photoreceptor cells after sexual reproduction. The presence of the transgenes was confirmed by PCR, plasmid rescue assay, inverse PCR, and Southern blotting. Our results with the 3xP3-EGFP marker confirm the presence of Pax6 activity in the differentiated photoreceptor cells of planarian eyes. Transgenesis will be an important tool to dissect developmental molecular mechanisms in planarian regeneration, development and stem cell biology, and may also be an entry point to analyze the biology of parasitic Platyhelminthes.

Molecular characterization of seven field isolates of infectious bursal disease virus obtained from commercial broiler chickens.

Specific-pathogen-free sentinel birds were used as an initial biological system to isolate infectious bursal disease virus (IBDV) field isolates from commercial broiler farms exhibiting recurrent respiratory problems and poor performance. Reverse transcription (RT)-polymerase chain reaction (PCR) was used to amplify a 248-bp product encompassing the hypervariable region of the IBDV VP2 gene. Restriction fragment length polymorphism (RFLP) analysis of the RT-PCR products was performed with the restriction endonucleases DraI, SadI, TaqI, StyI, BstNI, and SspI. Two isolates (619 and 850) exhibited a RFLP pattern characteristic of Delaware variant E IBDV. Restriction enzyme digestion for four isolates (625, 849, 853, and 11,153) revealed unmatched RFLP patterns when compared with reference IBDV strains. Nucleotide and deduced amino acid sequence analyses of the VP2 hypervariable region for these six isolates revealed identity (96.3% up to 98%) with Delaware E variant IBDV strain. However, serine at position 254, which is characteristic of Delaware variant strains, was substituted by asparagine in these six isolates. The seventh IBDV isolate (9109) also exhibited a unique RFLP pattern, which included the SspI restriction site, which is characteristic of very virulent (vv) IBDV strains. Nucleotide and amino acid sequence analyses of the hypervariable region for this isolate revealed identity (90%) with the standard challenge strain. However, the leucine residue at position 294 was substituted by isoleucine. This substitution corresponds to one of the amino acids that are conserved in the vvIBDV strains. Antigenic index studies of the predicted amino acid sequence of the hypervariable region of VP2 from isolates 619, 625, 849, 850, 853, and 11,153 exhibited a profile almost identical to variant E, whereas the isolate 9109 exhibited a profile characteristic of standard IBDV strains.

Reactivity of alkyl versus silyl peroxides. The consequences of 1, 2-silicon bridging on the epoxidation of alkenes with silyl hydroperoxides and bis(trialkylsilyl)peroxides.

The bond dissociation energies for a series of silyl peroxides have been calculated at the G2 and CBS-Q levels of theory. A comparison is made with the O-O BDE of the corresponding dialkyl peroxides, and the effect of the O-O bond strength on the activation barrier for oxygen atom transfer is discussed. The O-O bond dissociation enthalpies (DeltaH(298)) for bis (trimethylsilyl) peroxide (1) and trimethylsilyl hydroperoxide (2) are 54.8 and 53.1 kcal/mol, respectively at the G2 (MP2) and CBS-Q levels of theory. The O-O bond dissociation energies computed at G2 and G2(MP2) levels for bis(tert-butyl) peroxide and tert-butyl hydroperoxide are 45.2 and 48.3 kcal/mol, respectively. The barrier height for 1,2-methyl migration from silicon to oxygen in trimethylsilyl hydroperoxide is 47.9 kcal/mol (MP4//MP2/6-31G). The activation energy for the oxidation of trimethylamine to its N-oxide by bis(trimethylsilyl) peroxide is 28.2 kcal/mol (B3LYP/6-311+G(3df,2p)// B3LYP/6-31G(d)). 1,2-Silicon bridging in the transition state for oxygen atom transfer to a nucleophilic amine results in a significant reduction in the barrier height. The barrier for the epoxidation of E-2-butene with bis(dimethyl(trifluoromethyl))silyl peroxide is 25.8 kcal/mol; a reduction of 7.5 kcal/mol relative to epoxidation with 1. The activation energy calculated for the epoxidation of E-2-butene with F(3)SiOOSiF(3) is reduced to only 2.2 kcal/mol reflecting the inductive effect of the electronegative fluorine atoms.

[Types of microbial contaminants in pharmaceutical raw materials]. / Tipos de contaminantes microbianos de materias primas farmacéuticas.

In order to analyze the significance of the microbial content of pharmaceutical raw materials contributed to the finished pharmaceutical products, we have carried out a study of contamination taking into account aerobic bacteria, anaerobic bacteria and fungi. None or only low numbers of pathogenic microorganisms was found in most analyzed products but in some materials, specially those of natural origin, we have detected high bacterial and fungal contamination. Microorganisms of the genus Bacillus have been the aerobic bacteria most frequently isolated; Bifidobacterium and Clostridium were the most common anaerobic bacteria and with respect to the fungi, Penicillium and Aspergillus have been found with the highest frequency. These microorganisms can produce problems in pharmaceutical finished products, due to their enzymatic or toxigenic activities.

[Minor malformations and variations from normality at birth. II: Minor body anomalies excluding craniofacial anomalies]. / Malformaciones menores y variantes de la normalidad al nacimiento. II: Anomalías menores corporales con exclusión de las craneofaciales.

Authors studied in a total of 189 newborns, incidence of minor and variant malformation normalities, excluding craniofacial ones (already analyzed in a previous paper). After grouping the results in different corporal zones where these could be found, they find a greater incidence, in the cutaneous system and less in other corporeal areas. Since results differ mainly from those which already exist, therefore, they think that new studies are necessary in distinct geographic areas due to variation o their incidence in them.

[Minor malformations and variants of normality at birth. I: Minor craniofacial anomalies]. / Malformaciones menores y variantes de la normalidad al nacimiento. I: Anomalías menores craneofaciales.

Authors have studied the presence of minor variant malformations of craniofacial normalities in 189 newborns. Results were grouped according to different anatomical areas where they settled. They have found a greater expressiveness of minor anomalies and, above all, in the auricular external car and, in minor proportion, in the nose and ocular orbs. Results are similar in some cases with the scarce preexistent data, but they differ meaningly with others; maybe that this is produced by the own distinct geographic areas variations.