Changes in the expression of calcitonin gene-related peptide after exposure to injurious stretch-shortening contractions.

UNLABELLED: One of the factors that can result in musculoskeletal injuries, and time off work, is exposure to repetitive motion. The goal of this study was to determine if skeletal muscle injury induced by exposure to injurious stretch-shortening cycles (iSSCs), resulted in hyperalgesia in the hind limb and changes in calcitonin-gene related peptide (CGRP) immunolabeling in the dorsal root ganglia (DRG) in young and old male rats. METHODS: Young (3months) and old (30months) male Fisher 344×BN F1 rats were anesthetized with isoflurane and the left hind limbs were exposed to 15 sets of 10 SSCs. Control animals were exposed to a single bout of SSCs of equal intensity. Sensitivity to mechanical stimulation was assessed using von Frey filaments prior to beginning the experiment, and on days 2 and 9 following exposure to iSSCs. Rats were euthanized one, 3 or 10days after the exposure. The ipsilateral DRG were dissected from the L4-5 region of the spine, along with the left tibialis anterior (LTA) muscle. RESULTS: Rats exposed to iSSCs were more sensitive to mechanical stimulation than control rats 2days after the exposure, and showed a reduction in peak force 3days after exposure. Changes in sensitivity to pressure were not associated with increases in CGRP labeling in the DRG at 3days. However, 9days after exposure to iSSCs, old rats still displayed an increased sensitivity to mechanical stimulation, and this hyperalgesia was associated with an increase in CGRP immunolabeling in the DRG. Young rats exposed to iSSC did not display a change in CGRP immunolabeling and sensitivity to mechanical stimulation returned to control levels at 10days. CONCLUSIONS: These findings suggest that hyperalgesia seen shortly after exposure to iSSC is not influenced by CGRP levels. However, in cases where recovery from injury may be slower, as it is in older rats, CGRP may contribute to the maintenance of hyperalgesia.

Detecting a thienopyridine effect by platelet reactivity assessment and its implications for risk stratification.

BACKGROUND: On-treatment platelet reactivity (OTR) is a predictor of clinical outcomes in patients receiving thienopyridine therapy. OBJECTIVE: To assess whether point-of-care platelet reactivity testing can discriminate between patients who have and have not received a thienopyridine. PATIENTS/METHODS: This was an analysis of a randomized, multicenter, pharmacodynamic trial. Subjects with coronary artery disease treated with aspirin were randomly assigned to clopidogrel 75 mg daily or prasugrel 10 mg daily for 7 days. Platelet reactivity assessment with the VerifyNow P2Y12 test was performed before study drug admistration and 24 h after the final dose. Optimal cut-offs for a detectable drug effect were identified by the use of receiver operating characteristic curve analysis. RESULTS: A total of 54 subjects were enrolled and completed the study. The c-statistic for the identification of a thienopyridine effect was highly significant (0.93, P < 0.001), including for the clopidogrel and prasugrel groups considered separately (P < 0.001 for both). The optimal cut-off was < 213 P2Y12 reaction units (PRU), which provided a sensitivity of 80% and a specificity of 98%. This cut-off provided a sensitivity of 58% and a specificity of 100% for a clopidogrel effect, and a sensitivity of 100% and specificity of 96% for a prasugrel effect. CONCLUSIONS: OTR of < 213 PRU is highly specific for exposure to either clopidogrel or prasugrel. This may be useful in the management of thienoypridine-treated patients who require surgery. Furthermore, this diagnostic cut-off is similar to levels of OTR that have been associated with ischemic events in thienopyridine-treated patients, supporting the contention that a lack of drug effect is the mechanistic basis for the prognostic relationship between OTR and clinical outcomes.

Characterization of isometric contractions of rat skeletal muscle in vivo: duty cycle effects.

Many work related injuries stem from the exertion of skeletal muscle forces over an extended period of time. Musculoskeletal injury can be caused by muscle's inability to maintain force during occupational exposure. The goal of the present study is to test how various rest times (duty cycles) between long isometric contractions will affect decrements in force, and develop a model that characterizes force decrements due to skeletal muscle fatigue. All tests were performed in vivo on the tibialis anterior muscle of anesthetized Sprague-Dawley rats. Animals were randomly assigned to either a 10 second (N=8), 1 minute (N=8), or 5 minute (N=8) duty cycle group. All animals were then subjected to 7 isometric contractions (duration of 2.8 seconds). A model was constructed to characterize forces changes over the duration of a contraction and over multiple contractions. The model consisted of a power law and an exponential component; these two components were combined by using an exponential weighting function. Overall, the combination of a power law and exponential model with a weighting function satisfactorily characterized the changes in isometric force for the 10 second duty cycle, but a simpler exponential model could be used where longer duty cycles are performed.

A randomized double-blind placebo-controlled clinical trial of a product containing ephedrine, caffeine, and other ingredients from herbal sources for treatment of overweight and obesity in the absence of lifestyle treatment.

OBJECTIVE: To evaluate the efficacy and side effects of an herbal formulation to promote weight loss, as compared to placebo. DESIGN: 12-week multicenter double-blind, placebo-controlled, randomized parallel groups design. Study conducted at three clinical sites in New York State. Subjects were randomized to receive either the 'active' product or a 'placebo' supplement for 12 weeks. Minimal steps were taken to influence lifestyle changes with regard to diet or exercise. SUBJECTS: 102 overweight/obese (30

Impact of muscle length during stretch-shortening contractions on real-time and temporal muscle performance measures in rats in vivo.

The objective of the present study was to investigate the impact of muscle length during stretch-shortening cycles on static and dynamic muscle performance. Animals were randomly assigned to an isometric (control, Con, n = 12), a short-muscle-length (S-Inj, 1.22-2.09 rad, n = 12), or a long-muscle-length (L-Inj, 1.57-2.44 rad, n = 12) group. The dorsiflexor muscles were exposed in vivo to 7 sets of 10 stretch-shortening contractions (conducted at 8.72 rad/s) or 7 sets of isometric contractions of the same stimulation duration by using a custom-designed dynamometer. Performance was characterized by multipositional isometric exertions and positive, negative, and net work before exposure, 6 h after exposure, and 48 h after exposure to contractions. Real-time muscle performance during the stretch-shortening cycles was characterized by stretch-shortening parameters and negative, positive, and net work. The S-Inj group recovery (force difference) was similar to the Con group force difference at 48 h, whereas the L-Inj group force difference was statistically greater at 1.39, 1.57, and 1.74 rad than the Con group force difference (P < 0.05). Negative work (P < 0.05) and net work (P < 0.05) were statistically lower in the S-Inj and L-Inj groups than in the Con group 48 h after exposure to contractions. Of the real-time parameters, there was a difference in cyclic force with treatment during the stretch-shortening cycles (P < 0.0001), with the L-Inj group being the most affected. Thus longer ranges of motion result in a more profound isometric force decrement 48 h after exposure to contractions and in real-time changes in eccentric forces.

Comparative genomic hybridization analysis of hepatoblastomas.

Prior cytogenetic analyses of hepatoblastomas have shown the most common recurring abnormalities to be trisomy for chromosomes 2 and 20, and a recurrent translocation involving chromosomes 1 and 4 identified in a minority of cases. Four cases have shown double minute chromosomes, which provide cytogenetic evidence for gene amplification, although no particular genes or genetic regions have been shown to be amplified. To further investigate the cytogenetic changes involved in the pathogenesis and progression of hepatoblastoma, this study analyzes 10 tumors by comparative genomic hybridization. Regions of relative gain or loss were found in nine tumors. The most common recurrent abnormalities were gain of the long arm of chromosome 1 (six tumors), gain of chromosomes 2 (seven tumors), 17 (four tumors), and 20 (three tumors), and loss of chromosomes 4 and 11 (two tumors each). Four cases showed restricted regions of high-level gain at 1q32 or 2q24, regions that have previously been reported to be amplified in other tumors, but not in hepatoblastomas. A specific amplified gene has yet to be identified at these loci, although candidate genes have been proposed and may offer targets for future studies. Genes Chromosomes Cancer 27:196-201, 2000.

DNA content of ovarian immature teratomas and malignant germ cell tumors.

OBJECTIVE: Ovarian germ cell tumors (GCT) show greater histologic and biologic heterogeneity than their testicular counterparts and remain poorly understood. Ploidy analysis was performed on ovarian GCT registered on Pediatric Oncology Group germ cell tumor protocols 9048 and 9049 to distinguish biologically distinct subsets of immature teratomas and malignant ovarian germ cell tumors. METHODS: Tumors from 22 patients (mean age 12 years) were analyzed and classified according to the submitting diagnosis; when pure samples of different histologic subtypes within a single tumor were possible, these were analyzed separately. Archival tissue was disaggregated and Feulgen stained; DNA index (DI) was determined by static image analysis utilizing internal normal cells as diploid controls. RESULTS: 26 histologic subtypes from 22 patients were analyzed. The tumors of 18 patients were composed of a single histologic subtype according to the submitting institution, including 6 dysgerminomas, 8 immature teratomas (IT), and 4 endodermal sinus tumors (EST). Two tumors contained both IT and EST components that were separately analyzed. Two tumors were classified as mixed germ cell tumors; 1 showed multiple intermingling subtypes unable to be separately analyzed and the second showed three histologic subtypes separately analyzed (IT, EST, embryonal carcinoma). From a total of 15 malignant histologic GCT subtypes in 14 patients, all but 2 demonstrated a DI of 1.4-2.4 (mean 1.85). Two diploid malignant GCT (1 EST, 1 dysgerminoma) were both associated with gonadoblastoma. Overall, 11 IT subtypes were analyzed and 9 were diploid (2 grade 1, 5 grade 2, and 2 grade 3). Two tumors originally submitted and classified as pure IT (grades 2 and 3) were aneuploid with a dominant diploid and a secondary aneuploid peak (both DI 1.7). On central review, both of these tumors demonstrated the presence of subtle patterns of EST that were unrecognized by the submitting institution and were much too small for separate analysis. Analysis of the 3 patients containing sufficient IT and EST to be separately analyzed all showed a diploid IT component and an aneuploid EST component. CONCLUSIONS: Analysis of ploidy data suggests that polyploidization is a consistent finding in malignant ovarian GCT arising in normal patients, similar to the data for adult testicular GCT. Immature teratomas in this pediatric population, however, are most commonly diploid, regardless of grade. The development of EST within an IT is associated with the development of an aneuploid clone. Therefore, the finding of such a clone in an IT may be of diagnostic utility, as EST may be difficult to recognize. Last, the development of a malignant GCT in patients with gonadal dysgenesis may be pathogenetically different from those arising in normal patients, in that polyploidization is not required.

Dietary fibre, exercise and serum lipids and lipoprotein cholesterols in 12 to 15 year olds.

Serum lipid and lipoprotein cholesterol levels track from childhood and are associated with risk of coronary heart disease. There is some evidence that these are influenced by dietary intake and exercise. Serum lipid and lipoprotein cholesterols were measured in a cohort of 119 British children aged 12-15 y who completed a dietary assessment and exercise questionnaire. The ratio of total- to high-density lipoprotein cholesterol fell with increasing fibre intake, but after adjustment for age, body mass index, sex and other dietary factors, this was not statistically significant. Children exercising at least once a day had significantly lower serum total cholesterol and low density lipoprotein cholesterol levels than those exercising less frequently, even after adjustment for the above factors and dietary fibre intake. No dietary factor was significantly associated with any lipid measure after adjustment for the above factors. The challenge is how to optimize exercise level in adolescent children.

Inability of the activated partial thromboplastin time to predict heparin levels. Time to reassess guidelines for heparin assays.

BACKGROUND: In treating venous thromboembolic disorders, patient outcomes appear to correlate with heparin levels. Due to pharmacokinetic and pharmacodynamic variations, a relationship between heparin dose and level cannot be reliably predicted in individual patients. Some patients have low heparin levels despite therapeutic activated partial thromboplastin times (aPTTs), which may increase their risk for recurrent thromboembolism. Patients with high heparin requirements appear to have fewer bleeding episodes with heparin level-guided therapy. The aPTT does not reliably correlate with heparin blood concentrations or antithrombotic effects. Consequently, heparin therapy monitored with heparin levels may be more effective and safer. OBJECTIVES: To prospectively determine whether (1) the aPTT therapeutic range adequately predicts heparin levels in 38 patients used to establish the therapeutic aPTT range as is currently recommended and (2) whether 3 paired sets of aPTT-antifactor Xa levels provide the basis for using aPTTs to predict subsequent heparin levels in individual patients (n = 27) receiving intravenous heparin for coronary artery disease or venous thromboembolic disease. RESULTS: In the therapeutic aPTT range established, the R2 value for the relationship was 0.4. Prediction intervals were wide. For an aPTT of 60 seconds, the 95% prediction interval estimates were heparin levels of 0.05 to 1.0 U/mL. In individual patients, the aPTT-antifactor Xa relationship had an average R2 value of 0.75. There was no consistent relationship between the aPTT and anti-factor Xa level in a significant number of patients. CONCLUSIONS: The aPTT does not appear to be a useful surrogate for heparin levels. These findings suggest that the current recommendations on the use of heparin levels should be expanded.

Nutrition in pregnant or lactating rats programs lipid metabolism in the offspring.

Epidemiological studies in human show that size in early life is related to blood cholesterol concentrations in adult life, raising the hypothesis that early nutrition programs later lipid metabolism, affecting risk for later vascular disease. Here, we tested the hypothesis that nutrition during pregnancy or lactation in the rat programs lipid metabolism in the offspring, studied in adult life (mean 6 months). Rats (n 35) from normally-fed dams (controls) were compared with (1) rats (n 22) from dams protein-restricted in pregnancy and lactation; (2) rats (n 9) born to normally-fed mother crossed to protein-restricted lactating dams and (3) those (n 9) born of protein-restricted dams and crossed to normally-fed lactating animals. In these latter three groups the offspring showed long-term reduction in plasma cholesterol, HDL-cholesterol and triacylglycerol concentrations compared with controls. The effects were predominantly in males. These findings suggest that in the rat the sensitive period for nutritional programming of cholesterol and triacylglycerol metabolism is both pre- and postnatal (pre-weaning) and that rats may be 'indirectly' programmed by altering the maternal nutritional milieu during gestation or lactation. Whilst it has been hypothesized that early human undernutrition programs risk for vascular disease, one aspect of undernutrition, low maternal protein intake, in this rat model programmed lower plasma cholesterol and triacylglycerol concentrations.

Randomized outcome trial of human milk fortification and developmental outcome in preterm infants.

Despite potential benefits, human milk may fail to meet preterm infants' nutrient requirements. We tested the hypothesis that fortified breast milk, fed alone or with preterm formula, would improve neurodevelopment and growth at 18-mo follow-up without adverse short-term clinical or biochemical consequences. Two hundred seventy-five preterm infants from two medical centers (birth weight < 1850 g; mean gestation 29.8 +/- 2.7 wk) whose mothers chose to provide breast milk were randomly assigned to receive for a mean of 39 d a multinutrient fortifier or control supplement containing phosphate and vitamins. Breast milk comprised 47.6% and 46.4% of enteral intake in fortified and control groups, respectively; preterm formula supplements were used when insufficient breast milk was available. Overall, there were no significant growth advantages with fortification; although, when breast milk exceeded 50% of intake, fortification promoted faster weight gain (an advantage of 1.6 g.kg-1.d-1; 95% CI: 0.1, 3.1; P < 0.05). Compared with control infants, the fortified group showed 1) higher plasma urea from week 2 (P = 0.04), 2) higher plasma calcium (mean 2.34 +/- 0.01 compared with 2.27 +/- 0.02 mmol/L; P = 0.003), 3) a greater rise in alkaline phosphatase by week 6 (P = 0.04), 4) more clinical infections (suspected plus proven; 43% compared with 31%, P = 0.04), 5) a nonsignificantly increased incidence of necrotizing enterocolitis (5.8% compared with 2.2%, P = 0.12), and 6) higher white cell and platelet counts. Developmental scores at 18 mo were slightly but not significantly higher in the fortified group. This study confirmed that breast milk fortifiers can improve short-term growth (when breast milk intakes are high); but beneficial effects on long-term development remained unproven. Future research is required to evaluate potential adverse consequences and explore more optimal fortification strategies.

Circulatory support of cardiac interventional procedures with the Hemopump cardiac assist system.

The Hemopump is a catheter-mounted mechanical circulatory assist device which can support the majority of the circulation and significantly reduce left-ventricular work. It can be introduced via a peripheral artery or the ascending aorta. The following is an overview of the operation of the Hemopump and the results of a clinical trial in which the Hemopump was used to treat cardiogenic shock. In addition, the results of pilot experiences using the Hemopump for circulatory support during aorto-coronary artery surgery and high-risk angioplasty will be discussed.

Hyperphenylalaninaemia and outcome in intravenously fed preterm neonates.

Hyperphenylalaninaemia is likely to have occurred in many infants fed the intravenous amino acid solution Vamin 9. In this study of 336 preterm infants plasma phenylalanine was measured weekly during their hospital stay. Reference data on plasma phenylalanine were prepared for 243 infants who did not receive Vamin. Only 1% of these infants had a peak plasma phenylalanine concentration greater than 150 mumol/l (maximum 202 mumol/l) compared with 23% in 93 infants fed Vamin 9, seven of whom had concentrations > 300 mumol/l (maximum 704 mumol/l). High concentrations only occurred when the total energy to protein energy ratio in the intravenous solutions decreased to less than 8.5:1 and always occurred with a ratio less than 6.5:1, implying that hyperphenylalaninaemia may be minimised with an intravenous energy intake of greater than 34 kcal (142 kJ)/g protein. Nevertheless, follow up at 18 months post-term showed that increased plasma phenylalanine in this instance was not associated with any impairment of the Bayley mental development index (or subscales including fine motor, cognitive, or language development), the psychomotor development index, or the social maturity quotient. Thus, despite theoretical concern, an adverse outcome after hyperphenylalaninaemia induced by intravenous feeding has not been observed.

Nutrient utilisation in muscle and in the whole body of patients receiving total parenteral nutrition.

Forearm metabolite exchange was assessed by the arterio-venous catheterization technique in 5 parenterally fed patients (weight 55.22 kg +/- 4.18 kg; height 1.71 m +/- 0.04 m), who received an 'all-in-one' nutrition regimen whilst in remission from Crohn's disease. All patients received 12.8 g N, 4725 kJ from carbohydrate and 4200 kJ from fat (10416 kJ total energy). The exchanges were related to nutrient oxidation and nutrient balances in the whole body as assessed by indirect calorimetry and nitrogen excretion. At rest, the subjects were found to be in positive balances for carbohydrate (+0.78 +/- 0.13 kJ/min), fat (+1.85 +/- 0.26 kJ/min) and protein (+0.240 +/- 0.04 kJ/min). Resting forearm muscle was also in positive amino acid balance and positive carbohydrate balance. Despite the large estimated uptake of glucose by forearm muscle (+1860 +/- 84 nmol/100 ml tissue/min) there was no net release of pyruvate and lactate. Glutamate and the branched chain amino acids (BCAA) were the dominant amino acids taken up by muscle (26% and 30% of total uptake respectively) and glutamine was the dominant amino acid carrying nitrogen out of muscle (78% of total amino acid nitrogen release). The energy taken up by muscle as non-esterified fatty acids, triacylglycerol and ketone bodies was small relative to that associated with glucose uptake. The results suggest that during the hypercaloric parenteral nutrition regimen, a) increased peripheral glucose uptake is not necessarily associated with increased release of glycolytic products, b) in the absence of glutamine intake for at least 10 days, muscle retains enough capacity to synthesise and release sufficient quantities of glutamine so that it remains the dominant amino acid carrying nitrogen out of muscle, c) despite the use of the intravenous route for administration of nutrients, and unusual amino acid composition of the regimen, the overall pattern of forearm metabolism bears many similarities to that which occurs after a mixed meal in normal subjects.

A pyrolysis product, anhydroecgonine methyl ester (methylecgonidine), is in the urine of cocaine smokers.

A method using combined gas chromatography/mass spectrometry (GC/MS) for determination of the cocaine pyrolysis product anhydroecgonine methyl ester (AEME) in urine is described. Using this method, we found that human subjects who smoked cocaine under laboratory conditions excreted substantial amounts of AEME in their urine. Little, if any, AEME was excreted in the urine when the same subjects were administered cocaine by intravenous and intranasal routes. AEME may be a useful marker for cocaine (crack) smoking. The pharmacology of AEME is unknown. The possibility that AEME may play a role in the effects associated with cocaine smoking needs to be examined.

Plasma prolactin and clinical outcome in preterm infants.

Plasma prolactin was measured weekly in 280 preterm infants. The complex gestational age dependent pattern of postnatal prolactin release has been defined and reference standards provided. Plasma prolactin was higher in girls, with increasing divergence between the sexes from the third week onwards, and higher after two weeks, in infants of mothers with pregnancy related hypertension. Diet, assigned randomly, exerted a major effect on plasma prolactin, with significantly higher values in infants fed donor breast milk or standard formula than in those fed a protein, energy, and mineral enriched preterm formula. After adjusting for confounding factors, infants with the lowest plasma prolactin concentrations (less than 1000 mU/l, 32.9 micrograms/l) occurring usually at a nadir between days 5 and 12, showed a 120% increase in the duration of ventilatory assistance required, a 20% increase in the number of days to attain full enteral feeds, and a 30% decrease in length gain. We suggest preterm birth disrupts the normal perinatal pattern of prolactin release and that those infants who develop relatively low plasma concentration have an adverse outcome. Our data add to the broader debate on whether preterm infants require multiple endocrine replacement treatment.

The pattern of childhood hepatitis B infection in two Gambian villages.

Serologic markers of hepatitis B virus (HBV) infection were measured in children from Manduar and Keneba, two adjacent villages in The Gambia, in 1980 and in 1984. The rate of HBV infection over the 4 years differed markedly: in Manduar 71% of children who were less than 5 years of age in 1980 became infected, whereas in Keneba only 37% became infected. Male children were more frequent carriers of either HBs or e antigen than were female children. Marked clustering of hepatitis B surface antigen (HBsAg) antigenemia within sibling relationships was shown in both villages. The chance of the youngest child in a household being a carrier of HBsAg was strongly related to the number of antigen-positive siblings. Four years later, 53% of children who were initially positive for HBsAg and 33% who were positive for hepatitis B e antigen still carried these antigens. Jaundice was not observed.