Functional assessments for decision-making regarding return to sports following ACL reconstruction. Part I: development of a new test battery.

PURPOSE: Return to activity remains the most common concern following an injury. To facilitate the decision regarding a patient's return to sport, we developed a standardized and easy-to-use test battery to enable an objective evaluation of knee function. METHODS: The test battery consisted of seven functional tests: the two-leg stability test, one-leg stability test (OL-ST), two-leg countermovement jump (CMJ), one-leg CMJ (OL-CMJ), plyometric jumps, speedy test and quick feet test. For each test, the reliability was determined based on the intraclass correlation coefficient. For all one-leg tests, the limb symmetry index (LSI) was calculated. RESULTS: All tests showed a moderate-to-high reliability. Normative data from 434 participants were included in the analysis. The subjects were categorized according to age as follows: children (10-14 years), youth (15-19 years), young adults (20-29 years) and adults (30-50 years). The establishment of the functional test values allowed the classification into five normative categories. The LSI for the OL-ST (98 %) indicated a better performance of the non-dominant leg. In contrast, high LSI values were found for the OL-CMJ (124 %), indicating a better performance of the dominant leg. CONCLUSION: Each test was found to be reliable and simple to perform. The better performance of the non-dominant leg in stability tasks must be considered when interpreting side-to-side differences. The established norm data from healthy individuals of each test battery represents an important basis for a clinical setting. Test results from an ACL-reconstructed patient should be at least classified as a functionally average outcome to support a safe return to sports. LEVEL OF EVIDENCE: IV.

Behaviour of postnatally growth-impaired mice during malnutrition and after partial weight recovery.

OBJECTIVES: Early malnutrition is a highly prevalent condition in developing countries. Different rodent models of postnatal early malnutrition have been used to approach the subject experimentally, inducing early malnutrition by maternal malnutrition, temporal maternal separation, manipulation of litter size or the surgical nipple ligation to impair lactation. Studies on the behaviour of (previously) malnourished animals using animal models have produced sometimes contradictory results regarding the effects of early postnatal malnutrition and have been criticized for introducing potential confounding factors. The present paper is a first report on the behavioural effects of early malnutrition induced by an alternative approach: mice nursed by α-casein-deficient knockout dams showed a severe growth delay during early development and substantial catch-up growth after weaning when compared with animals nursed by wild-type females. METHODS: Established behavioural tests were used to study the consequences of early postnatal malnutrition on mouse pups at weaning and after partial weight recovery. RESULTS: Despite the impaired growth, the only behavioural difference between malnourished and normally growing animals was found in exploratory behaviour during acute malnutrition at the time of weaning. After partial catch-up in weight early protein malnourished animals showed no indication of lasting effects on general activity, emotionality and exploration, memory, and pain reactivity. DISCUSSION: These results suggest that the role of early nutrition on behavioural development after recovery in animal models may have been overestimated. Further careful examination of this animal model in terms of maternal care and offspring behaviour will be necessary to confirm if mice nursed by α-casein-deficient dams offer an alternative to existing models while eliminating potential confounding factors.

Welfare assessment in transgenic pigs expressing green fluorescent protein (GFP).

Since large animal transgenesis has been successfully attempted for the first time about 25 years ago, the technology has been applied in various lines of transgenic pigs. Nevertheless one of the concerns with the technology--animal welfare--has not been approached through systematic assessment and statements regarding the welfare of transgenic pigs have been based on anecdotal observations during early stages of transgenic programs. The main aim of the present study was therefore to perform an extensive welfare assessment comparing heterozygous transgenic animals expressing GFP with wildtype animals along various stages of post natal development. The protocol used covered reproductory performance and behaviour in GFP and wildtype sows and general health and development, social behaviour, exploratory behaviour and emotionality in GFP and wildtype littermates from birth until an age of roughly 4 months. The absence of significant differences between GFP and wildtype animals in the parameters observed suggests that the transgenic animals in question are unlikely to suffer from deleterious effects of transgene expression on their welfare and thus support existing anecdotal observations of pigs expressing GFP as healthy. Although the results are not surprising in the light of previous experience, they give a more solid fundament to the evaluation of GFP expression as being relatively non-invasive in pigs. The present study may furthermore serve as starting point for researchers aiming at a systematic characterization of welfare relevant effects in the line of transgenic pigs they are working with.

The relationship between ACL injuries and physical fitness in young competitive ski racers: a 10-year longitudinal study.

BACKGROUND: Epidemiological studies have shown a high incidence of anterior cruciate ligament (ACL) injuries among competitive alpine skiers. Little is known regarding modifiable risk factors in young skiers. There are still uncertainties in gender-related risk factors. OBJECTIVE: The purpose of this study was to determine the relationship between ACL injuries and internal risk factors. METHODS: Retrospective data analyses were performed based on a group of 175 female and 195 male alpine ski racers between the ages of 14 and 19 years. The athletes underwent physical testing annually from 1996 to 2006. Z score transformations normalised the age groups. Multivariate binary logistic regressions were calculated for men and women separately to detect significant predictors of ACL ruptures. t Tests were computed to reveal the differences in test scores between injured and non-injured athletes. RESULTS: A total of 57 (15%) ACL injuries occurred. The female-male risk ratio (RR) was higher in females (2.3, 95% CI 1.3 to 4.2). Z scores for relative leg force, ratio of absolute core flexion to extension force, relative core strength and reactive strength index were predictive variables for men. Z scores of all of these predictive variables except relative leg force were higher in the non-injured group. The ratios of absolute flexion to extension force and absolute core strength were predictive covariates for women. Z scores for absolute core strength were higher in the non-injured group. CONCLUSIONS: The risk of ACL injury was greater in female athletes. The findings suggest that core strength is a predominant critical factor for ACL injuries in young ski racers.

Chromosomal rearrangements in post-Chernobyl papillary thyroid carcinomas: evaluation by spectral karyotyping and automated interphase FISH.

Structural genomic rearrangements are frequent findings in human cancers. Therefore, papillary thyroid carcinomas (PTCs) were investigated for chromosomal aberrations and rearrangements of the RET proto-oncogene. For this purpose, primary cultures from 23 PTC have been established and metaphase preparations were analysed by spectral karyotyping (SKY). In addition, interphase cell preparations of the same cases were investigated by fluorescence in situ hybridisation (FISH) for the presence of RET/PTC rearrangements using RET-specific DNA probes. SKY analysis of PTC revealed structural aberrations of chromosome 11 and several numerical aberrations with frequent loss of chromosomes 20, 21, and 22. FISH analysis for RET/PTC rearrangements showed prevalence of this rearrangement in 72% (16 out of 22) of cases. However, only subpopulations of tumour cells exhibited this rearrangement indicating genetic heterogeneity. The comparison of visual and automated scoring of FISH signals revealed concordant results in 19 out of 22 cases (87%) indicating reliable scoring results using the optimised scoring parameter for RET/PTC with the automated Metafer4 system. It can be concluded from this study that genomic rearrangements are frequent in PTC and therefore important events in thyroid carcinogenesis.

Effects of Regular Long-Term Circuit Training (Once per Week) on Cardiorespiratory Fitness in Previously Sedentary Adults.

The purpose of the study was (1) to investigate the effects of regular long-term circuit training (once per week) on cardiorespiratory fitness (CRF) in sedentary adults and (2) to compare training progress with the effects of continued exercise participation by regularly active age-matched individuals. Ten sedentary, middle-aged (51 ± 6 years) individuals (sedentary group, SG) of both sexes performed 32 weeks (1 training session/week) of supervised circuit training and 10 weeks of self-managed training. Effects were compared to an age-matched group (51 ± 8 years; n = 10) of regularly active individuals (active group, AG). CRF (expressed as peak oxygen uptake: VO2peak; peak power output: PPO) and systemic blood pressure (BP) during the incremental test were measured at the start and after the training intervention. CRF decreased significantly within the AG (VO2peak: 43.1 ± 7.3 vs. 40.3 ± 6.5 mL/min/kg, p < 0.05; PPO: 3.3 ± 0.6 vs. 3.1 ± 0.6; p < 0.05) but was maintained in the SG. In addition, significant improvements in restoration of the oxygen level in leg muscles after exercise and reduced systolic BP (180 ± 14 vs. 170 ± 17 mmHg, p = 0.01) at submaximal exercise were found within the SG. However, differences in changes from pre to post did not reach significance between groups. In contrast to the regularly active individuals, circuit training once per week over 32 weeks prevented the aging-related decline of CRF in previously sedentary subjects and reduced systolic BP during submaximal exercise, indicating improved exercise tolerance.

Chromosomal changes characterize head and neck cancer with poor prognosis.

It is well established that genetic alterations may be associated to prognosis in tumor patients. This study investigates chromosomal changes that predict the clinical outcome of head and neck squamous cell carcinoma (HNSCC) and correlate to characteristic clinicopathological parameters. We applied comparative genomic hybridization (CGH) to tissue samples from 117 HNSCC patients scheduled for radiotherapy. Genomic aberrations occurring in more than five patients were studied for impact on locoregional progression (LRP)-free survival. p values were adjusted by the Hochberg-Benjamini procedure and significant aberrations and clinical variables subjected to a stepwise backwards Cox proportional model. Significant alterations were further analyzed by array-CGH and fluorescence in situ hybridization (FISH). In multivariate survival analysis gains on 1q and 16q predict reduced LRP-free survival independently from known prognostic factors. Cluster analysis separated the HNSCC cases into two groups (cluster 1 and 2) that are characterized by significant differences for imbalances in 13 chromosomal regions. Moreover, it became apparent that cluster 1 correlates to nonanemic patients, while cluster 2 represents predominantly anemic cases. Array-CGH pinpoints 16q24.3 to be the region of interest on chromosome 16 which was further verified by FISH analysis where an increased copy number of FANCA, a member of the Fanconi anemia/breast cancer pathway, could be identified. This study demonstrates that chromosomal gains on 1q and 16q as well as chromosomal loss on 18q represent prognostic markers in HNSCC and that these alterations may explain to some extent the dismal course of a subgroup of patients.

Intra-oesophageal pH profiles and pharmacokinetics of pantoprazole and esomeprazole: a crossover study in patients with gastro-oesophageal reflux disease.

AIM: To compare the effect of pantoprazole and esomeprazole on intra-oesophageal pH and investigate their pharmacokinetics in patients with symptomatic gastro-oesophageal reflux disease (GORD). METHODS: Double-blind, randomized, two-period crossover study. Caucasian men with symptomatic GORD (n=48) were selected on the basis of clinical records of typical GORD symptoms, confirmed by a pathological reflux time (oesophageal pH<4 for > or =6% of the time). They received oral pantoprazole 40 mg once daily (od) or esomeprazole 40 mg od for seven days. Continuous 24 h oesophageal pH-metry was performed at baseline and day 7. Evaluations included: pre- and post-treatment differences in the percentage of time with pH<4.0 and <3.0 between baseline and day 7; area under the curve (AUC), Cmax, and T(1/2); point estimates and 90% confidence intervals (CI) on days 1 and 7, calculated for ratios of the AUC and Cmax. RESULTS: Both drugs decreased the mean total number of reflux episodes and reduced the percentage of reflux time within 24 h to <3%. No pathological reflux was detectable after repeated administration of either drug. The 90% CI were within the predefined range at all time points; thus, equivalence of pantoprazole and esomeprazole was concluded. For pantoprazole, Cmax and AUC were unchanged on day 7 vs day 1, confirming its high and constant bioavailability. For esomeprazole, Cmax and AUC were increased on day 7 vs day 1 by 80% and 50%, respectively, indicating low initial bioavailability. No clinically relevant side effects were seen for either drug. CONCLUSION: Pantoprazole and esomeprazole have equivalent effect on oesophageal pH, since no pathological reflux was detected after treatment with either drug. For esomeprazole, the Cmax and AUC increased after multiple dosing; for pantoprazole the pharmacokinetics were predictable and independent of the number of administered doses.

The influence of electromyographic biofeedback therapy on knee extension following anterior cruciate ligament reconstruction: a randomized controlled trial.

BACKGROUND: Loss of knee extension and a deficit in quadriceps strength are frequently found following anterior cruciate ligament (ACL) reconstruction. The aim of this study was to investigate whether the addition of Eletromyographic Biofeedback (EMG BFB) therapy for the vastus medialis muscle to the in the early phase of the standard rehabilitation programme could improve the range of knee extension and strength after ACL reconstruction more than a standard rehabilitation programme. The correlation between EMG measurement and passive knee extension was also investigated. METHOD: Sixteen patients, all of whom underwent endoscopic ACL reconstruction using patellar tendon autograft, were randomly assigned to two groups:• Control group (8 patients): standard rehabilitation protocol; with full weight-bearing postoperative, knee brace (0° extension, 90° flexion), electrical stimulation, aquatics and proprioceptive training.• The EMG BFB group (8 patients): EMG BFB was added to the standard rehabilitation protocol within the first postoperative week and during each session for the next 6 weeks.Each patent attended a total of 16 outpatient physiotherapy sessions following surgery. High-Heel-Distance (HHD) Test, range of motion (ROM) and integrated EMG (iEMG) for vastus medialis were measured preoperatively, and at the 1, 2, 4 and 6-week follow ups. Additionally, knee function, swelling and pain were evaluated using standardized scoring scales. RESULTS: At 6 weeks, passive knee extension (p < 0.002) and the HHD Test were significantly (p < 0.01) better in the EMG BFB group compared to controls. Integrated EMG (vastus medialis) of the EMG BFB group also showed a significant increase after 2 (p < 0.01) and 6 (p < 0.01) weeks. At the 6-week follow up, no significant (p > 0.01) differences were found between the two groups for the assessment of knee function, swelling and pain. CONCLUSION: The results indicate that EMG BFB therapy, in the early phase of rehabilitation after ACL reconstruction, is useful in enhancing knee extension. Improved innervation of the vastus medialis can play a key role in the development of postoperative knee extension. EMG BFB therapy is a simple, inexpensive and valuable adjunct to conventional therapeutic modalities.

Chromosomal radiosensitivity and acute radiation side effects after radiotherapy in tumour patients--a follow-up study.

BACKGROUND: Radiotherapists are highly interested in optimizing doses especially for patients who tend to suffer from side effects of radiotherapy (RT). It seems to be helpful to identify radiosensitive individuals before RT. Thus we examined aberrations in FISH painted chromosomes in in vitro irradiated blood samples of a group of patients suffering from breast cancer. In parallel, a follow-up of side effects in these patients was registered and compared to detected chromosome aberrations. METHODS: Blood samples (taken before radiotherapy) were irradiated in vitro with 3 Gy X-rays and analysed by FISH-painting to obtain aberration frequencies of first cycle metaphases for each patient. Aberration frequencies were analysed statistically to identify individuals with an elevated or reduced radiation response. Clinical data of patients have been recorded in parallel to gain knowledge on acute side effects of radiotherapy. RESULTS: Eight patients with a significantly elevated or reduced aberration yield were identified by use of a t-test criterion. A comparison with clinical side effects revealed that among patients with elevated aberration yields one exhibited a higher degree of acute toxicity and two patients a premature onset of skin reaction already after a cumulative dose of only 10 Gy. A significant relationship existed between translocations in vitro and the time dependent occurrence of side effects of the skin during the therapy period. CONCLUSIONS: The results suggest that translocations can be used as a test to identify individuals with a potentially elevated radiosensitivity.

Milk lacking α-casein leads to permanent reduction in body size in mice.

The major physiological function of milk is the transport of amino acids, carbohydrates, lipids and minerals to mammalian offspring. Caseins, the major milk proteins, are secreted in the form of a micelle consisting of protein and calcium-phosphate.We have analysed the role of the milk protein α-casein by inactivating the corresponding gene in mice. Absence of α-casein protein significantly curtails secretion of other milk proteins and calcium-phosphate, suggesting a role for α-casein in the establishment of casein micelles. In contrast, secretion of albumin, which is not synthesized in the mammary epithelium, into milk is not reduced. The absence of α-casein also significantly inhibits transcription of the other casein genes. α-Casein deficiency severely delays pup growth during lactation and results in a life-long body size reduction compared to control animals, but has only transient effects on physical and behavioural development of the pups. The data support a critical role for α-casein in casein micelle assembly. The results also confirm lactation as a critical window of metabolic programming and suggest milk protein concentration as a decisive factor in determining adult body weight.

Establishment and Molecular Cytogenetic Characterization of a Cell Culture Model of Head and Neck Squamous Cell Carcinoma (HNSCC).

Cytogenetic analysis of head and neck squamous cell carcinoma (HNSCC) established several biomarkers that have been correlated to clinical parameters during the past years. Adequate cell culture model systems are required for functional studies investigating those potential prognostic markers in HNSCC. We have used a cell line, CAL 33, for the establishment of a cell culture model in order to perform functional analyses of interesting candidate genes and proteins. The cell line was cytogenetically characterized using array CGH, spectral karyotyping (SKY) and fluorescence in situ hybridization (FISH). As a starting point for the investigation of genetic markers predicting radiosensitivity in tumor cells, irradiation experiments were carried out and radiation responses of CAL 33 have been determined. Radiosensitivity of CAL 33 cells was intermediate when compared to published data on tumor cell lines.