Single closed-tube quantitative real-time PCR assay with dual-labelled probes for improved sex determination of equine embryos.

In addition to fulfilling many breeders' curiosity, equine embryonic sex determination can have a profound commercial impact. However, the application of currently described assays for equine embryonic sexing has rendered variable diagnosis and validation rates, with sensitivity being the main problem. In addition, while pregnancy results of in vivo-flushed equine embryos following a needle aspiration biopsy equal those of non-biopsied embryos, the effect on in vitro-produced embryos is unknown. Here, we aimed to develop a highly sensitive and specific assay for equine sex determination that can be directly performed on few embryonic cells, and to test the effect of a needle aspiration biopsy on the viability of the in vitro-produced embryo. To this end, a multiplex quantitative real-time PCR (qPCR) assay with dual-labelled probes was designed to allow the simultaneous generation of both male-specific and control fragments in a single closed-tube reaction, avoiding potential sample loss or contamination. To improve sensitivity, multicopy and polymeric genes were chosen to be specifically amplified, i.e., eight copies of Y-chromosomal ETSTY5 as male-specific and four autosomal UBC monomers as control fragment. Specificity was enhanced by the equine-specific character of ETSTY5 and by using dual-labelled probes. The assay was optimised with equine male and female genomic DNA and demonstrated a 100% accuracy and a >95% qPCR efficiency down to 10 pg of DNA. The assay was subsequently applied to determine the sex of 44 in vitro-produced embryos, collecting trophectoderm biopsies by means of a needle aspiration biopsy and herniating cells. Of all trophectoderm biopsies and herniating cell samples (n = 54), 87% could be diagnosed. Assay results were validated on a second sample obtained from the biopsied embryo (n = 18) or, by ultrasound-based sex determination of the foetus (n = 7) following the transfer of the biopsied embryo to a recipient mare, with about half of the embryos being fillies and colts. The needle aspiration biopsy procedure did not impair initial pregnancy rate or early pregnancy losses as compared to non-biopsied embryos. In conclusion, we report a safe, reliable, fast, and cost-effective assay for equine sex determination which was validated for the sex determination of in vitro-produced embryos based on few embryonic cells, and needle aspiration biopsy did not impair the embryo's viability. The assay and safe biopsy strategy hold potential for other applications.

Malnourished adults' receipt of hospital discharge nutrition care instructions: a pilot study.

BACKGROUND: Malnutrition remains an important yet under-recognised problem among hospitalised adults. Although interventions exist aiming to improve nutritional status beyond hospitalisation, few studies examine how often and what type of nutrition care instructions are given at discharge. The present study sought to review nutrition-focused discharge care provided to malnourished adults. METHODS: We reviewed the electronic medical record for discharge nutrition care instructions provided to adult patients identified by dietitians as malnourished over a 4-month period. RESULTS: Seventy-six eligible patients were identified during the study period. More than half of malnutrition cases (64.5%) were attributed to chronic illness. According to electronic medical record documentation, 6.6% received discharge instructions to consume oral nutrition supplements and 30.3% received new or changed prescriptions for vitamins/noncaloric supplements. Almost half of patients (47.4%) received general diet instructions that did not address malnutrition and 44.8% received inappropriate instructions to limit caloric intake. CONCLUSIONS: A majority of malnourished adult patients receive inappropriate or inadequate nutrition care instructions at the time of discharge. Clinician education and redesign of nutrition care options in the electronic medical record may improve the provision of post-discharge nutrition care instructions.

Early rapid weight gain among formula-fed infants: Impact of formula type and maternal feeding styles.

BACKGROUND: What and how infants are fed are considered important determinants for the risk factor of early rapid gain weight. OBJECTIVES: We conducted secondary analyses on data from a randomized clinical trial, wherein infants randomized to feed cow milk formula had double the incidence of early rapid weight gain than those fed extensively hydrolyzed protein formula, to determine whether maternal feeding styles had independent effects or interactive effects with infant formula type on early rapid weight gain. METHODS: Anthropometry and feeding patterning (number of daily formula feeds) were measured monthly, and maternal feeding styles were measured at 0.5, 3.5, and 4.5 months. Longitudinal models were fitted using generalized estimating equations and separate logistic models conducted. RESULTS: The treatment groups did not differ in formula feeding patterning or in maternal feeding styles, which were stable across the first 4.5 months. Feeding styles had no significant effects on early rapid weight gain and did not interact with formula group. However, type of infant formula had a direct and independent impact on early rapid weight gain (P = 0.003). CONCLUSIONS: The type of infant formula had a differential impact on early rapid weight gain independent of maternal feeding style, highlighting the self-regulatory capabilities of infants.

Total Energy Expenditure of Infants with Congenital Heart Disease Who Have Undergone Surgical Intervention.

Growth failure is often observed in infants with congenital heart disease (CHD); it is unclear, however, whether growth failure is due to increased total energy expenditure (TEE). An observational study of infants with CHD and surgical intervention within the first 30 days of life and healthy infants of similar age was undertaken. TEE was measured using the doubly labeled water method in 3-month-old infants (n = 15 CHD, 12 healthy) and 12-month-old infants (n = 11 CHD, 12 healthy). Multiple linear regression models were fit to examine the association between health status (CHD vs. healthy) and TEE. The accuracy of equations for calculating TEE was also determined. TEE for CHD infants was not significantly different from healthy infants at 3 and 12 months; TEE in CHD infants was 36.4 kcal/day higher (95 % CI -46.3, 119.2; p = 0.37) and 31.7 kcal/day higher, (95 % CI -71.5, 134.8; p = 0.53) at 3 and 12 months, respectively, compared to healthy infants. The 2002 Dietary Reference Intake (DRI) equation and the 1989 Recommended Dietary Allowance equation over-estimated measured TEE to a lesser extent than CHD specific equations; the 2002 DRI yielded the smallest mean difference between calculated versus measured TEE (difference 79 kcal/day). During the first year of life, TEE of infants with CHD and interventional surgery within the first month of life was not different than age-matched healthy infants. When calculating TEE of ≤12-month-old infants with CHD who have undergone surgical intervention, the 2002 DRI equation may be used as a starting point for estimating initial clinical energy intake goals.

[Comparative study of the treatment of pertrochanteric fractures--trochanteric gamma nail vs. proximal femoral nail]. / Vergleichsstudie der Versorgung pertrochantärer Femurfrakturen -- trochanteric Gamma-Nagel vs. PFN.

AIM: We have performed a retrospective comparative study between the trochanteric gamma nail (TGN) and the proximal femoral nail (PFN). METHOD: During the period 1998-2003, 97 TGN and 83 PFN were used for the treatment of pertrochanteric fractures. Most of the fractures were of the A3 type according to the AO/ASIF classification. Clinical and radiological follow-ups were available for 87 TGN and 65 PFN. RESULTS: The mean operative time for the TGN was shorter than that for the PFN. Intraoperative complications were noted in 17.5 % and 28.8 % for the TGN and PFN groups, respectively. Late complications occurred in 18.4 % for TGN compared to 27.6 % of the PFN. Union was achieved in 94.2 % and 89.3 % of the patients treated with the TGN and PFN, respectively. The reoperation rates were 10.3 % and 24.6 % for the TGN and the PFN, respectively. Clinical outcomes were good for both groups (65 % in the TGN, 62 % in the PFN group). CONCLUSION: Treatment of pertrochanteric fractures using the TGN and PFN implants is quite reliable. The major complication was cut-out and occurred mostly in the PFN group, while varus deformity was more frequent in the TGN group. The PFN was associated with a higher rate of reoperation and longer operative time, probably due to a more demanding technique.

Evaluation of the offensive behavior of elite soccer teams.

The purpose of the present study was to evaluate the offensive behavior of the four elite teams (France, Brazil, Croatia, and Holland) using data from the semifinals of the 18th World Soccer Championship in France in 1998. 28 videotaped soccer games were observed, 7 for each team. The protocol contained the following parameters of evaluation: (a) successful pass in the defensive and middle area, (b) unsuccessful pass in the defensive and middle area, (c) attempt on goal in the offensive area, and (d) cross and follow-up action. A multivariate analysis of variance showed the teams' plan was significantly different only in playing the ball back to the goalkeeper. This last action, used more often by Holland than by the other teams, indicated its restrained offensive behavior, which may be one of the reasons for its defeat in some games.

Tribal-based cancer control activities among Alaska Natives: services and perceptions.

Cancer has become a significant health problem in American Indian and Alaskan Native (AI/AN) communities. Despite the precipitous rise in cancer rates, limited data are available concerning cancer control services operative in these communities. To address this issue, a cross-sectional survey of all federally recognized tribes was undertaken to ascertain the breadth of cancer control activities offered and Tribal Health Directors perceptions of and priorities ascribed to cancer. This article presents responses given by AN Health Directors juxtaposed to those proffered by AI Health Directors. Nearly three-quarters (71%) of respondents perceived cancer rates to be increasing. Cancer was found to rank third, fifth among AI Health Directors, among seven health conditions when Directors were asked to rank their Tribe's commitment to confronting each one. Awareness of cancer as a public health concern coupled with competing health problems relegates cancer control activities to a lower priority. Findings underscore the need to elevate the issue of cancer in Indian Country as well as to educate investigators to become more sensitive and responsive to other Tribal health issues.

An introduction to benesh movement notation and its relevance to physiotherapy.

During the last few years attention has been concentrated on the analysis of movement in patients with disorders of the central nervous system. The phenomenon of movement has been looked at not as the operation of a mechanism made up of individual parts, but as the result of the integration of motor patterns in the central nervous system. The traditional neurological examination is inadequate for the assessment of movement patterns and as a result a specific method called "motoscopic" examination has been developed for this purpose. This has been described elsewhere (Milani Comparetti and Gidoni, 1967) and consists of the systematic visual observation of: (a) spontaneous posture and motor behaviour, (b) the performance of a series of movements as requested by the examiner (c) patterns of movement under certain stimulus conditions.