Isolation of subtelomeric sequences of porcine chromosomes for translocation screening reveals errors in the pig genome assembly.

Balanced chromosomal aberrations have been shown to affect fertility in most species studied, often leading to hypoprolificacy (reduced litter size) in domestic animals such as pigs. With an increasing emphasis in modern food production on the use of a small population of high quality males for artificial insemination, the potential economic and environmental costs of hypoprolific boars, bulls, rams etc. are considerable. There is therefore a need for novel tools to facilitate rapid, cost-effective chromosome translocation screening. This has previously been achieved by standard karyotype analysis; however, this approach relies on a significant level of expertise and is limited in its ability to identify subtle, cryptic translocations. To address this problem, we developed a novel device and protocol for translocation screening using subtelomeric probes and fluorescence in situ hybridisation. Probes were designed using BACs (bacterial artificial chromosomes) from the subtelomeric region of the short (p-arm) and long (q-arm) of each porcine chromosome. They were directly labelled with FITC or Texas Red (p-arm and q-arm respectively) prior to application of a 'Multiprobe' device, thereby enabling simultaneous detection of each individual porcine chromosome on a single slide. Initial experiments designed to isolate BACs in subtelomeric regions led to the discovery of a series of incorrectly mapped regions in the porcine genome assembly (from a total of 82 BACs, only 45 BACs mapped correctly). Our work therefore highlights the importance of accurate physical mapping of newly sequenced genomes. The system herein described allows for robust and comprehensive analysis of the porcine karyotype, an adjunct to classical cytogenetics that provides a valuable tool to expedite efficient, cost effective food production.

Aneuploidy in dizygotic twin sheep detected using genome-wide single nucleotide polymorphism data from two commonly used commercial vendors.

Early detection of karyotype abnormalities, including aneuploidy, could aid producers in identifying animals which, for example, would not be suitable candidate parents. Genome-wide genetic marker data in the form of single nucleotide polymorphisms (SNPs) are now being routinely generated on animals. The objective of the present study was to describe the statistics that could be generated from the allele intensity values from such SNP data to diagnose karyotype abnormalities; of particular interest was whether detection of aneuploidy was possible with both commonly used genotyping platforms in agricultural species, namely the Applied BiosystemsTM AxiomTM and the Illumina platform. The hypothesis was tested using a case study of a set of dizygotic X-chromosome monosomy 53,X sheep twins. Genome-wide SNP data were available from the Illumina platform (11 082 autosomal and 191 X-chromosome SNPs) on 1848 male and 8954 female sheep and available from the AxiomTM platform (11 128 autosomal and 68 X-chromosome SNPs) on 383 female sheep. Genotype allele intensity values, either as their original raw values or transformed to logarithm intensity ratio (LRR), were used to accurately diagnose two dizygotic (i.e. fraternal) twin 53,X sheep, both of which received their single X chromosome from their sire. This is the first reported case of 53,X dizygotic twins in any species. Relative to the X-chromosome SNP genotype mean allele intensity values of normal females, the mean allele intensity value of SNP genotypes on the X chromosome of the two females monosomic for the X chromosome was 7.45 to 12.4 standard deviations less, and were easily detectable using either the AxiomTM or Illumina genotype platform; the next lowest mean allele intensity value of a female was 4.71 or 3.3 standard deviations less than the population mean depending on the platform used. Both 53,X females could also be detected based on the genotype LRR although this was more easily detectable when comparing the mean LRR of the X chromosome of each female to the mean LRR of their respective autosomes. On autopsy, the ovaries of the two sheep were small for their age and evidence of prior ovulation was not appreciated. In both sheep, the density of primordial follicles in the ovarian cortex was lower than normally found in ovine ovaries and primary follicle development was not observed. Mammary gland development was very limited. Results substantiate previous studies in other species that aneuploidy can be readily detected using SNP genotype allele intensity values generally already available, and the approach proposed in the present study was agnostic to genotype platform.

Characterization of an X-chromosomal non-mosaic monosomy (59, X0) dairy heifer detected using routinely available single nucleotide polymorphism genotype data.

Evidence exists from a range of species on the impact of karyotype abnormalities on reproductive performance. Despite this, cytogenetic analyses of cattle, especially females, are not routinely undertaken. Genome-wide single nucleotide polymorphism (SNP) genotype data are now, however, routinely being generated in many species globally at a relatively low cost. The objective of the present study was to evaluate the potential of routinely available SNP genotype data to identify sex-chromosome aberrations using X chromosome monosomy 59,X0 as a case study for illustration. A single 2.5-yr old Holstein-Friesian heifer was detected with a mean allelic intensity of SNP on the X chromosome almost 17 standard deviations less than the mean of other genotyped females ( = 103,326). Following cytogenetic analysis (10 replicates by karyotyping and a further 140 by FISH), the female was deduced to be a non-mosaic 59,X0. The female had never produced a calf and, although gross examination revealed no physical abnormalities, she was smaller in size than expected based on her breed and age. Given the age of the animal at slaughter, the uterus and uterine tubes appeared immature and inactive. The oviduct appeared normal while the single ovary present contained a markedly reduced number of follicles. There was, however, some evidence of prior ovulation and formation of corpora lutea. The approach proposed in the present study to identify allosome aneuploidy from routinely available genotype data can be used to screen for such abnormalities at no additional cost to the breeder or producer.

Effect of irradiation on microorganisms in strawberries.

Seventeen samples of strawberries from seven different growers were analysed for total counts, Enterobacteriaceae, fluorescent pseudomonads and yeast and mould counts before and after irradiation at 1.2 or 2 kGy. Enterobacteriaceae were absent (less than 5 cfu/g) from all irradiated strawberries but were always detected at counts of greater than 30 cfu/g in untreated samples. This criterion was true for both fresh and stored (5 days at 8 degrees C) strawberries. Assuming no other sanitizing treatment, the Enterobacteriaceae count appears to be suitable for differentiating between irradiated and non-irradiated strawberries. The other counts used in this study were not suitable for this purpose. Isolates from irradiated strawberries could be classified into four types based on colony morphology. Three types consisted of aerobic spore-forming bacteria whilst the fourth group consisted of yeasts.

The rate of clinically significant plain radiograph misinterpretation by faculty in an emergency medicine residency program.

OBJECTIVE: To estimate the rate of clinically significant discrepancies between radiograph interpretations by attending radiologists and emergency medicine (EM) faculty in 2 academic EDs, using a unique scoring system. METHODS: A retrospective comparison of radiographic agreement between EM and radiology faculty members was performed. All plain films initially interpreted by EM faculty or by EM residents with immediate reinterpretation by EM faculty were subsequently reviewed by attending radiologists. All discrepancies between these readings were reported to the ED on the following day for review by an EM faculty member (usually different from the initial EM faculty reader) who determined the need for treatment or follow-up changes. A secondary chart review by a quality assurance faculty member determined whether radiographic findings not noted on the x-ray log were present on the ED record All discrepancies from February to June 1994 were reviewed. A severity score was assigned based on the following criteria. Q-0: There was no change in treatment or follow-up; or the initial interpretation by EM faculty was validated by repeat or additional views. Q-1: Discrepancy is minor. Q-2: Discrepancy is significant, with potential for injury or bad outcome. Q-3: Discrepancy is significant, with actual injury or bad outcome. RESULTS: Of 14,046 radiographic studies eligible for enrollment, there were 134 discrepancies (0.95%). Only 28 cases (0.2%) were found to be clinically significant. Of these, 25 were scored Q-1, 3 were scored Q-2, and 0 were scored Q-3. These clinically significant discrepancy rates were highest for the finger, skull, elbow, hand, and lumbar spine. CONCLUSION: Emergency medicine faculty provide highly accurate rates of plain radiograph interpretation, particularly when adjusted for clinical significance and actual impact on patient care.

Reducing the rate of paramedic needlesticks in emergency medical services: the role of self-capping intravenous catheters.

OBJECTIVE: To determine whether the frequency of unintentional needlesticks can be reduced by replacing conventional i.v. catheters with self-capping ones. METHODS: Retrospective cohort, historically controlled study, conducted in an emergency medical services advanced life support (ALS) service. The ALS service annually transports 12,000 patients, for whom i.v. therapy is attempted in about 65% of cases. The needlestick rate per 1,000 patients receiving attempts at i.v. access was examined during the 2 10-month periods, before and after introduction of a self-capping i.v. catheter. RESULTS: For the 2 periods, the percentage of patients for whom i.v. access was attempted remained constant at 65%. The success rate for i.v. access was statistically unchanged from 88% to 90% (p > 0.5, power = 0.995). During the period prior to use of the new catheter, 44 injuries were reported overall. Of these, 15 were due to unintentional needlesticks, 11 associated with contaminated needles. Following the system-wide introduction of the new catheter, only 1 of 31 reported injuries was due to needlestick (uncontaminated). The extrapolated annual incidence of contaminated needlesticks decreased from 169 (95% CI; 85, 253) to 0 (95% CI; 0, 46) per 100,000 i.v.attempts. The extrapolated incidence for all needlesticks decreased from 231 (95% CI; 132, 330) to 15 (95% CI; 0, 40) per 100,000 i.v. attempts. The absolute number of needlesticks and the proportion of injuries due to needlesticks decreased significantly (p < 0.005). CONCLUSION: The use of i.v. catheters with self-capping needles was associated with a significant reduction in the absolute number of inadvertent needlesticks as well as the proportion of injuries due to needlesticks among ALS providers. The use of self-capping i.v. catheters was feasible and did not appear to be a deterrent to initiating i.v. therapy in the out-of-hospital environment.

EMS systems: foundations for the future.

Emergency medical services (EMS) occupy a unique position in the continuum of emergency health care delivery. The role of EMS personnel is expanding beyond their traditional identity as out-of-hospital care providers, to include participation and active leadership in EMS administration, education, and research. With these roles come new challenges, as well as new responsibilities. This paper was developed by the SAEM EMS Task Force and provides a discussion of these new concepts as well as recommendations for the specialty of emergency medicine to foster the continued development of all of the potentials of EMS.

"Ping-pong" gaze in severe monoamine oxidase inhibitor toxicity.

Clinical features of monoamine oxidase inhibitor (MAOI) toxicity include hypertension, hyperthermia, tachycardia, and muscular agitation. We report two cases in which some of these signs of severe toxicity were seen in association with a unique periodic alternating gaze disturbance known as "ping-pong" gaze.

Thoracentesis in the emergency department.

Presented is a review of thoracentesis, a procedure with which the emergency physician should be familiar. The pathophysiology of pleural effusions is described and is followed by a review of the clinical presentation and diagnosis. Special attention is given to technique and interpretation of results.

The effect of a quality improvement feedback loop on paramedic skills, charting, and behavior.

OBJECTIVE: A mechanism was initiated for conveying quality improvement (QI) results to paramedics as a means of improving chart documentation in difficult-to-correct areas. This study examines the impact of this QI feedback loop on charting, resuscitation rates from cardiac arrest, endotracheal intubation (ETI) success rates, and trauma scene times. DESIGN: Paramedic trip sheets were reviewed before and after the institution of the QI feedback loop in this interrupted time series design. SETTING: The New Castle County, Delaware, Paramedic Program. PARTICIPANTS: All New Castle County paramedics participated in the study. INTERVENTIONS: In January 1990, the medical director began to circulate a QI summary among the paramedics in an effort to improve performance and chart documentation. The summary focused on the management of respiratory distress or arrest, cardiac arrest, and major trauma. The success rate for ETI was compared with the rate of field resuscitation from cardiac arrest, the percentage of unjustified prolonged trauma scene times (longer than 10 minutes), and the percent compliance with minimum endotracheal intubation documentation (ETID) requirements from a six-month period before institution of the QI feedback mechanism with data obtained from a six-month period after the program had been operational for one year. RESULTS: Comparing results from before with after the initiation of the QI program, the ETI success rate was 273 of 295 (92.5%) before and 300 of 340 (88.2%) after (chi 2 = 3.04, p < .1, ns); field resuscitations totaled 26 of 187 (13.9%) before and 44 of 237 (18.6) after (chi 2 = 1.40, p < .25, ns); ETID rate was 249 of 295 (84.4%) before and 336 of 340 (98.8%) after (chi 2 = 44.24, p < .001), and unjustified prolonged trauma scene times were 69 of 278 (24.8%) before and seven of 501 (1.4%) after (chi 2 = 320.5, p < .001). CONCLUSION: The use of QI feedback had little effect on psychomotor skills as the ETI success rate or resuscitation rate, but had a dramatic effect on chart documentation, as evidenced by ETID rate, and behavior, as evidenced by the reduction in prolonged trauma scene times. The use of QI feedback is recommended as a means of correcting charting deficiencies or modifying behavior.

Two approaches to reading instruction with children with disabilities: does program design make a difference?

This study examined the effects on reading achievement of variation in program design and tested the hypothesis this Distar Reading Mastery's (Engelmann & Bruner, 1988b) demonstrated effects with disadvantaged children in Project Follow Through can be generalized to children with disabilities. We compared the effects of two synthetic phonics reading programs, Direct Instruction (DI) Reading Mastery I and II and Addison Wesley's Meet the Superkids and The Superkids' Club, (Rowland, 1982a, 1982b) for 81 children in transitional kindergarten special education classes. No significant achievement differences were evident for the instructional program either at the end of the treatment year, or on follow-up testing 1 year later. However, among children who made advanced progress, the DI group showed larger reading gains.

Teaching phonological awareness to young children with learning disabilities.

This study examined the feasibility of teaching phonological manipulation skills to preschool children with disabilities. Forty-seven children, 4-6 years old, enrolled in a special education preschool, were randomly assigned to receive training in one of three categories of phonological tasks (rhyming, blending, and segmenting) or a control group. Results indicated that children were able to make significant progress in each experimental category, but that they demonstrated little or no generalization either within a category (e.g., from one type of blending task to another type of blending task) or between categories (e.g., from blending to segmenting). Although the children's level of cognitive development significantly predicted some learning outcomes, it did not appear to limit the learning of phonological tasks.

Medicare HMOs: part of the solution or part of the problem in the struggle for cost containment?

The correct answer for salvage of the Medicare program while maintaining acceptable medical care is clearly not available today. Managed care programs for seniors have been an option in the last decade and plans are expanding rapidly into previously untapped areas. Some programs can indeed provide a multiplicity of services and thereby truly integrate and improve existing care plans. Others have less clear cut advantages. For any senior contemplating the options available for medical care under Medicare, HMOs and Medigap type instruments, the most salient advice is "buyer beware." A complete understanding of the options gained and lost by changing medical insurance coverage is essential.

Cesarean section in a severely traumatized patient.

Massive trauma in a near-term pregnant patient challenges the emergency physician with very difficult treatment choices. We present the case of a severely traumatized pregnant female, near term, in whom the decision to proceed with a perimortem cesarean section in the Emergency Department produced a viable infant. We discuss the guidelines that are used in making such a decision and their rationale. The physiologic changes of pregnancy that may complicate maternal assessment and the effect of shock on the fetus are also discussed.

The prehospital use of nebulized albuterol on patients with wheezing whose chief complaint is shortness of breath.

The use of prehospital nebulized beta-agonists has become widespread, and their safety and efficacy has been documented. Our purpose was to study their broadened use and determine their effectiveness in specific sub-sets of wheezing patients. We conducted a six and one-half month prospective study to determine the benefit of nebulized albuterol treatments on a variety of wheezing patients whose chief complaint to paramedics was shortness of breath. Sixty-two patients were enrolled in the study and were subdivided into four groups based on patient history; asthma, COPD, asthma & COPD (A/C), and non-Asthma/non-COPD (NANC). The effectiveness of the treatment was evaluated objectively by peak expiratory flow rates (PEFR) obtained before and immediately after treatment and subjectively by the patients' evaluation of their own dyspnea. Changes in PEFR were subjected to analysis by a paired T-test. Albuterol was effective in increasing the PEFR in patients with asthma, COPD and NANC. Patients with both asthma and COPD did not demonstrate increased PEFR after treatment. The majority of all patients were subjectively improved after nebulized albuterol treatments. We conclude that aerosolized albuterol is safe and effective in the prehospital treatment of patients complaining of dyspnea who are wheezing.

Delaware Emergency Medical Services System: cardiac resuscitation.

The Delaware EMS system has been in existence for 20 years. Initially begun as one paramedic unit serving New Castle County, it now comprises 15 units in a statewide system. The goal of this report is to detail the EMS system's impact on prehospital cardiac resuscitation and airway management. Emergency Medical Services (EMS) encompass all aspects of managing a sick or injured patient prior to arrival to the hospital. From the time a patient dials 911 until they arrive in the care of a doctor in an emergency room, the EMS system provides initial medical evaluation and care as well as transportation to the hospital. The components of Delaware's EMS system include: 1. Bystanders--the public is often called upon to perform CPR until trained rescuers arrive. 2. Medical Dispatchers--they receive incoming 911 calls and determine the personnel needed. 3. First responders--ambulance crews trained in basic life support (BLS). 4. Second responders--ambulance crews trained in advanced life support (ALS), paramedics. 5. Medical Control--doctors who are in radio contact with paramedics to provide medical advice.

Bolus administration of 5 mg epinephrine as a treatment of last resort for cardiac arrest.

There is some evidence that the dose of epinephrine currently recommended for resuscitation is low. This study examines the effect of bolus administration of 5 mg of epinephrine when given as an agent of last resort for cardiac arrest. The experimental design called for the administration of high-dose epinephrine at the point where resuscitative efforts would have ordinarily been stopped. Resuscitation was then continued for a minimum of five minutes so that any changes in rhythm or return of spontaneous circulation could be noted. Eighty-five consecutive patients undergoing resuscitation in our Emergency Department were enrolled in this study. Twenty-eight (33 percent) exhibited a change in cardiac rhythm. Of these 28 patients, seven (8 percent) developed pulses. Of these seven patients, four expired in the Emergency Department, one survived to admission but expired two days later, and two patients survived to discharge. We conclude that bolus administration of 5 mg of epinephrine may be useful in treating cardiac arrest refractory to conventional therapy.