Life in the fat lane: seasonal regulation of insulin sensitivity, food intake, and adipose biology in brown bears.

Grizzly bears (Ursus arctos horribilis) have evolved remarkable metabolic adaptations including enormous fat accumulation during the active season followed by fasting during hibernation. However, these fluctuations in body mass do not cause the same harmful effects associated with obesity in humans. To better understand these seasonal transitions, we performed insulin and glucose tolerance tests in captive grizzly bears, characterized the annual profiles of circulating adipokines, and tested the anorectic effects of centrally administered leptin at different times of the year. We also used bear gluteal adipocyte cultures to test insulin and beta-adrenergic sensitivity in vitro. Bears were insulin resistant during hibernation but were sensitive during the spring and fall active periods. Hibernating bears remained euglycemic, possibly due to hyperinsulinemia and hyperglucagonemia. Adipokine concentrations were relatively low throughout the active season but peaked in mid-October prior to hibernation when fat content was greatest. Serum glycerol was highest during hibernation, indicating ongoing lipolysis. Centrally administered leptin reduced food intake in October, but not in August, revealing seasonal variation in the brain's sensitivity to its anorectic effects. This was supported by strong phosphorylated signal transducer and activator of transcription 3 labeling within the hypothalamus of hibernating bears; labeling virtually disappeared in active bears. Adipocytes collected during hibernation were insulin resistant when cultured with hibernation serum but became sensitive when cultured with active season serum. Heat treatment of active serum blocked much of this action. Clarifying the cellular mechanisms responsible for the physiology of hibernating bears may inform new treatments for metabolic disorders.

A protocol for the isolation and cultivation of brown bear (Ursus arctos) adipocytes.

Brown bears (Ursus arctos) exhibit hyperphagia each fall and can become obese in preparation for hibernation. They do this without displaying the physiological problems typically seen in obese humans, such as Type 2 diabetes and heart disease. The study of brown bear hibernation biology could therefore aid in the development of novel methods for combating metabolic diseases. To this end, we isolated mesenchymal stem cells from subcutaneous fat biopsies, and culture methods were developed to differentiate these into the adipogenic lineage. Biopsies were taken from 8 captive male (N = 6) and female (N = 2) brown bears, ages 2-12 years. Plastic adherent, fibroblast-like cells were proliferated and subsequently cryopreserved or differentiated. Differentiation conditions were optimized with respect to fetal bovine serum content and time spent in differentiation medium. Cultures were characterized through immunostaining, RT-qPCR, and Oil red O staining to quantify lipid accumulation. Adiponectin, leptin, and glycerol medium concentrations were also determined over the course of differentiation. The culturing protocol succeeded in generating hormone-sensitive lipase-expressing, lipid-producing white-type adipocytes (UCP1 negative). Serum concentration and time of exposure to differentiation medium were both positively related to lipid production. Cells cultured to low passage numbers retained similar lipid production and expression of lipid markers PLIN2 and FABP4. Ultimately, the protocols described here may be useful to biologists in the field investigating the health of wild bear populations and could potentially increase our understanding of metabolic disorders in humans.

Response characteristics for thermal and pressure devices commonly used for monitoring nasal and oral airflow during sleep studies.

We examined thermocouple and pressure cannulae responses to oral and nasal airflow using a polyester model of a human face, with patent nasal and oral orifices instrumented with a dual thermocouple (F-ONT2A, Grass) or a dual cannula (0588, Braebon) pressure transducer (± 10 cm H2O, Celesco) system. Tidal airflow was generated using a dual compartment facemask with pneumotachographs (Fleisch 2) connected to the model orifices. During nasal breathing: thermocouple amplitude = 0.38 Ln [pneumotachograph amplitude] + 1.31 and pressure cannula amplitude = 0.93 [pneumotachograph amplitude](2.15); during oral breathing: thermocouple amplitude = 0.44 Ln [pneumotachograph amplitude] + 1.07 and pressure cannula amplitude = 0.33 [pneumotachograph amplitude](1.72); (all range ∼ 0.1-∼ 4.0 L s(-1); r(2) > 0.7). For pneumotachograph amplitudes <1 L s(-1) (linear model) change in thermocouple amplitude/unit change in pneumotachograph amplitude was similar for nasal and oral airflow, whereas nasal pressure cannula amplitude/unit change in pneumotachograph amplitude was almost four times that for oral. Increasing oral orifice area from 0.33 cm(2) to 2.15 cm(2) increased oral thermocouple amplitude/unit change in pneumotachograph amplitude by ∼ 58% but decreased pressure cannula amplitude/unit change in pneumotachograph amplitude by 49%. For pneumotachograph amplitudes up to 1 L s(-1), alterations in inspiratory/expiratory ratios or total respiratory time did not affect the sensitivity of either nasal or oral pressure cannulae or the nasal thermocouple, but the oral thermocouple sensitivity was influenced by respiratory cycle time. Different nasal and oral responses influence the ability of these systems to quantitatively assess nasal and oral airflow and oro-nasal airflow partitioning.

TSSi--an R package for transcription start site identification from 5' mRNA tag data.

UNLABELLED: High-throughput sequencing has become an essential experimental approach for the investigation of transcriptional mechanisms. For some applications like ChIP-seq, several approaches for the prediction of peak locations exist. However, these methods are not designed for the identification of transcription start sites (TSSs) because such datasets contain qualitatively different noise. In this application note, the R package TSSi is presented which provides a heuristic framework for the identification of TSSs based on 5' mRNA tag data. Probabilistic assumptions for the distribution of the data, i.e. for the observed positions of the mapped reads, as well as for systematic errors, i.e. for reads which map closely but not exactly to a real TSS, are made and can be adapted by the user. The framework also comprises a regularization procedure which can be applied as a preprocessing step to decrease the noise and thereby reduce the number of false predictions. AVAILABILITY: The R package TSSi is available from the Bioconductor web site: www.bioconductor.org/packages/release/bioc/html/TSSi.html.

Public support for the use of newborn screening dried blood spots in health research.

OBJECTIVES: The level of support among Michigan adults for the use of residual newborn screening dried blood spots (DBS) was investigated. METHODS: We analyzed data from 4 questions on the 2008 Michigan Behavioral Risk Factor Surveillance System (n = 3,108). The questions asked about general support for the use of DBS for research and for research investigating childhood diseases, adult diseases and diseases related to environmental exposures. RESULTS: The majority of adults (72.3%) favored the use of DBS for research intended to benefit the health of residents. With more question specificity, a higher proportion of adults (84.2%-86.8%) were found to favor the use of DBS for research, and a lower proportion had no opinion. The odds of favoring use were higher among those who were younger, female, white, healthy, or with at least a high school degree. CONCLUSIONS: This is the first population survey of adult attitudes regarding use of DBS for different types of health research, with results showing considerable public support. The findings are being used in community outreach efforts and highlight the need to investigate opposition in vulnerable populations.

Long-term effects of levetiracetam and topiramate in clinical practice: A head-to-head comparison.

OBJECTIVE: Two of the most commonly prescribed new antiepileptic drugs as add-on therapy for patients with chronic refractory epilepsies are topiramate and levetiracetam. In regulatory trials, both drugs were characterized as very promising new antiepileptic drugs. However, results from these highly controlled short-term clinical trials cannot simply be extrapolated to everyday clinical practice, also because head-to-head comparisons are lacking. Therefore, results from long-term open label observational studies that compare two or more new AEDs are crucial to determine the long-term performance of competing new antiepileptic drugs in clinical practice. METHOD: We analyzed all patients referred to a tertiary epilepsy centre who had been treated with topiramate from the introduction of the drug in spring 1993 up to a final assessment point mid-2002 and all patients who had been treated with LEV in the same centre from the introduction of the drug in early 2001 up to a final assessment point end-2003 using a medical information system. RESULTS: Three hundred and one patients were included for levetiracetam and 429 patients for TPM. Retention rate after 1 year was 65.6% for LEV-treated patients and 51.7% for TPM-treated patients (p=0.0015). Similarly, retention rates for LEV were higher at the 24-month mark: 45.8% of LEV-treated patients and 38.3% of TPM-treated patients were still continuing treatment (p=0.0046). Adverse events led to drug discontinuation in 21.9% of TPM-treated patients compared to 6.0% of LEV-treated patients (p<0.001). The number of patients discontinuing treatment because of lack of efficacy was similar for both groups. Seizure freedom rates varied between 11.6 and 20.0% for TPM and between 11.1 and 14.3% for LEV per 6-months interval. Several important AED specific adverse events leading to drug discontinuation were identified, including neurocognitive side effects from TPM and mood disorders from LEV. CONCLUSION: The retention rate for LEV is significantly higher than for TPM. LEV had a more favourable side effect profile than TPM with comparable efficacy. Patients on TPM discontinued treatment mainly because of neurocognitive side effects. In the treatment with LEV, the effects on mood must not be underestimated.

Levetiracetam in clinical practice: long-term experience in patients with refractory epilepsy referred to a tertiary epilepsy center.

OBJECTIVE: For the treatment of patients with chronic refractory epilepsies, development of new antiepileptic drugs is crucial. Three regulatory trials have demonstrated that add-on levetiracetam is efficacious in patients with localization-related epilepsy. However, results from these highly controlled short-term clinical trials cannot simply be extrapolated to everyday clinical practice. Therefore, more information is needed about the long-term profile of a new antiepileptic drug in clinical practice. METHOD: We analyzed all patients who had been treated with levetiracetam in the Epilepsy Centre Kempenhaeghe from the introduction of the drug in early 2001 up to a final assessment point, at the end of 2003, using a medical information system. RESULTS: In total, 301 patients were included. One hundred thirty-eight patients (45.8%) discontinued LEV treatment during the 24-month follow-up period. Reasons for discontinuation were lack of efficacy in 15.9% of patients, adverse events in 6.0% of patients, and a combination of lack of efficacy and adverse events in 16.3% of patients. By Kaplan-Meier survival analysis, the continuation rate was 65.6% after 1 year and 45.8% after 2 years. About 15% of patients in this highly refractory group had a 3-month remission, whereas 10% of patients became seizure-free for longer periods. The most frequently reported side effects at the time of discontinuation were mood disorders, tiredness, and sleepiness. Variables predicting (dis)continuation of levetiracetam treatment could not be identified. CONCLUSION: Levetiracetam is a new antiepileptic drug that appears to be a useful add-on treatment in patients with refractory epilepsy. Its side effect profile is mild, with mood disorders being the most dominant adverse event.

Nasal resistance and flow resistive work of nasal breathing during exercise: effects of a nasal dilator strip.

Using posterior rhinomanometry, we measured nasal airflow resistance (Rn) and flow-resistive work of nasal breathing (WONB), with an external nasal dilator strip (ENDS) and without (control), in 15 healthy adults (6 men, 9 women) during exclusive nasal breathing and graded (50-230 W) exercise on a cycle ergometer. ENDS decreased resting inspiratory and/or expiratory Rn (at 0.4 l/s) by >0.5 cmH(2)O. l(-1). s in 11 subjects ("responders"). Inspired ventilation (VI) increased with external work rate, but tended to be greater with ENDS. Inspiratory and expiratory Rn (at 0.4 l/s) decreased as VI increased but, in responders, tended to remain lower with ENDS. Inspiratory (but not expiratory) Rn at peak nasal airflow (Vn) increased as VI increased but, again, was lower with ENDS. At a VI of approximately 35 l/min, ENDS decreased flow limitation and hysteresis of the inspiratory transnasal pressure-flow curve. In responders, ENDS reduced inspiratory WONB per breath and inspiratory nasal power values during exercise. We conclude that ENDS stiffens the lateral nasal vestibule walls and, in responders, may reduce the energy required for nasal ventilation during exercise.

Abdominal wall hernia: an uncommon complication of in utero vesicoamniotic shunt placement.

Three cases are reported in which placement of a vesicoamniotic shunt in utero for the treatment of obstructive uropathy led to the rare complication of abdominal wall hernia. All 3 patients underwent vesicoamniotic shunt placement for severe oligohydramnios and a markedly dilated bladder in an effort to preserve renal function and to prevent pulmonary hypoplasia. All three shunts were initially placed at or above the umbilicus. The abdominal wall hernias were closed postnatally at the time of temporary vesicostomy for urethral obstruction. All 3 patients had sufficient pulmonary development, but 2 of 3 had renal failure, requiring dialysis. Management and potential etiology of this rare complication are discussed.

Evaluation of signal-averaged cardiokymography for the detection of ischaemic left ventricular dysfunction.

UNLABELLED: Cardiokymography (CKG) is a non-invasive method for the detection of patients with coronary artery disease (CAD). Issues of the present study were to evaluate the feasibility, sensitivity and specificity of a recently developed signal-averaged CKG system for detecting patients with pharmacologically induced ischaemic left ventricular wall motion abnormalities (WMA) during pharmacologic stress echocardiography (SE). Precordial CKG curves were recorded in 100 consecutive patients who underwent dobutamine-SE for suspected CAD. For interpretation, CKG curves were classified into three different types, depending on the degree of systolic outward motion. CKG test results were regarded as positive (indicating myocardial ischaemia) if there was a change of the baseline CKG type at peak pharmacologic stress. The CKG test results were positive in 18 of 27 patients with a pathologic dobutamine-SE (sensitivity 67%), but did not show any change of the prior CKG type in 57 of 69 patients with a normal SE (specificity 83%). Patients with a true positive CKG test had significantly (P<0.05) more echocardiographic segments with WMA than patients with a false negative CKG test. CONCLUSIONS: Signal-averaged CKG can detect patients with ischaemic ventricular dysfunction. Sensitivity of CKG in detecting patients with WMA depends on the extent of left ventricular ischaemia. Further studies are needed to define the diagnostic value of signal-averaged CKG in the non-invasive detection of patients with suspected CAD.

Effect of metoprolol and amlodipine on myocardial total ischaemic burden in patients with stable angina pectoris.

OBJECTIVE: A randomized double-blind cross-over study to assess the effect on myocardial total ischaemic burden and the anti-anginal efficacy of the beta-1-blocker metoprolol given as metoprolol CR/Zok versus the calcium channel blocker amlodipine, both given in the recommended and commonly used doses of 100 mg o.d. and 5 mg o.d., respectively. METHOD: Fifty-two patients with a history of stable exercise-induced angina pectoris and at least six episodes of significant ST-segment depression during 24-h ambulatory electrocardiographic monitoring after 9 days of placebo were included in the study. The patients first completed a 9-day placebo run-in phase with additional administration of a long-acting nitrate during the first 7 days. They then received in a randomized sequence metoprolol CR/Zok and amlodipine each for 4 weeks. During placebo treatment and at the end of each phase of active treatment the patients' clinical progress was assessed and a 24-h ECG monitor performed. RESULTS: Forty-seven patients completed the two 4-week treatment periods. Five patients withdrew from the study. The number of ischaemic episodes during 24 h was 30.4 at baseline with placebo, which was reduced significantly by both treatments (P<0.005) to 6.8 episodes after metoprolol and 15.8 episodes after amlodipine treatment (P<0.001 for the difference between the treatment groups). Metoprolol and amlodipine reduced the total duration of ischaemic episodes from 86.0 min to 15-1 and 48.3 min with a mean episode duration of 1.1 and 2.6 min, respectively. Twenty patients on metoprolol (42.6%) and four on amlodipine (8.4%) showed no ST-segment depression at the end of treatment. Baseline heart rate of 80 b.p.m. decreased by 11.1 b.p.m. after metoprolol and increased by 4b.p.m. after amlodipine. Anginal attacks were reduced from 14.8 attacks per week at baseline to 2.4 attacks on metoprolol and 4.4 attacks on amlodipine treatment. For all variables the observed changes from baseline were significant after either treatment (P<0.005) with a significantly more pronounced effect in favour of metoprolol (P<0.0001). Ten patients reported nine different adverse events during metoprolol treatment. On amlodipine, 12 patients were affected by 11 different symptoms leading to three treatment withdrawals. On metoprolol one patient withdrew due to adverse events. CONCLUSION: Both drugs reduce total ischaemic burden by reducing ischaemic episodes and antianginal attacks, with a significantly greater effect from metoprolol 100 mg o.d. as compared to amlodipine 5 mg o.d.

Cardiovascular risk factors, ECG abnormalities and quality of life in subjects with atrial fibrillation.

In Central European regions epidemiologic findings for atrial fibrillation (AF) in a randomly selected population are not available. Therefore, information obtained by a standardized examination procedure including resting 12 lead ECG of 4003 participants (2014 men, 1989 women), aged 25 to 64 years, of the MONICA-Augsburg Survey 1984/85 were analysed. Reexamination of 3753 subjects took place three years later (Follow-up Study 1987/88). Persons with AF in the baseline survey (n = 13) were compared with an age-and sex-matched control group (n = 156) without AF, chosen from the same population sample, with regard to cardiovascular risk factors, associated disease and disturbances in the subjects' general well-being. In 1984/85 thirteen cases with AF (6 males, 7 females) were observed, giving an age-standardized prevalence of AF in males of 0.22% and in females of 0.34%. The age of men with AF ranged between 50 and 63 years and of women between 61 and 64 years. No significant differences were observed in persons with AF compared to the control group in risk factor levels and alcohol consumption; however, significant differences could be seen concerning disturbances in quality of life like self-reported health status (p < 0.001), sleep disturbances (p < 0.05), antihypertensive medication (p < 0.001). AF cases were found to have further ECG abnormalities significantly more often (left anterior hemiblock: p < 0.05; ventricular premature beats: p < 0.05). In all subjects with AF in the initial examination 1984/85 AF was found three years later (chronic AF). Overall 13 new cases (7 men, 6 women) were identified in the 1987/88 follow-up. The prevalence of AF in a South German population is comparable with AF prevalences reported from studies in other populations (e.g.) Framingham 1950, Reykjavik 1967/70). Associated ECG abnormalities were found more frequently in subjects with AF. Cases with AF have considerable disturbances in their general well-being.

Assessment of global and regional myocardial function using the Minnesota Q/QS codes. A comparison with clinical ECG interpretation.

The authors investigated 244 consecutive patients with suspected coronary artery disease by coronary angiography and quantitative left ventriculography to compare the Minnesota Q/QS code (MC) with clinical electrocardiographic (ECG) interpretation. Patients who were suspected to have wall motion abnormalities for reasons other than coronary artery disease for possible regional wall motion abnormalities were excluded. Out of 244 patients, 159 (65%) had wall motion abnormalities. The sensitivity for detecting wall motion abnormalities was 21% for MC 1.1 and 51% for MC 1.1-3, whereas clinical ECG interpretation showed a sensitivity of 73%. Specificity for MC 1.1 was 93% and for MC 1.1-3 it was 84%. Specificity of clinical ECG interpretation (84%) was comparable. Compared to the MC, clinical ECG interpretation showed a stronger association with left ventricular ejection fraction, number of segments with abnormal wall motion, and severity of wall motion abnormality. Anterior myocardial infarction presented more often with clinical ECG changes (71%) and with a Q/QS code (50%) than inferior myocardial infarction (61% and 41%, respectively). In summary, in contrast to clinical ECG criteria, the MC has high specificity at the expense of a low sensitivity.

The diagnostic performance of computer programs for the interpretation of electrocardiograms.

BACKGROUND: Computer programs for the interpretation of electrocardiograms (ECGs) are now widely used. However, a systematic assessment of various computer programs for the interpretation of ECGs has not been performed. METHODS: We undertook a large international study to compare the performance of nine electrocardiographic computer programs with that of eight cardiologists in interpreting ECGs in 1220 clinically validated cases of various cardiac disorders. ECGs from the following groups were included in the sample: control patients (n = 382); patients with left ventricular hypertrophy (n = 183), right ventricular hypertrophy (n = 55), or biventricular hypertrophy (n = 53); patients with anterior myocardial infarction (n = 170), inferior myocardial infarction (n = 273), or combined myocardial infarction (n = 73); and patients with combined infarction and hypertrophy (n = 31). The interpretations of the computer programs and the cardiologists were compared with the clinical diagnoses made independently of the ECGs, and the computer interpretations were compared with those of the cardiologists. RESULTS: The percentage of ECGs correctly classified by the computer programs (median, 91.3 percent) was lower than that of the cardiologists (median, 96.0 percent; P less than 0.01). The median sensitivity of the computer programs was also significantly lower than that of the cardiologists in diagnosing left ventricular hypertrophy (56.6 percent vs. 63.9 percent, P less than 0.02), right ventricular hypertrophy (31.8 percent vs. 46.6 percent, P less than 0.01), anterior myocardial infarction (77.1 percent vs. 84.9 percent, P less than 0.001), and inferior myocardial infarction (58.8 percent vs. 71.7 percent, P less than 0.0001). The median total accuracy level (the percentage of correct classifications) was 6.6 percent lower for the computer programs (69.7 percent) than for the cardiologists (76.3 percent; P less than 0.001). However, the performance of the best programs nearly matched that of the most accurate cardiologists. CONCLUSIONS: Our study shows that some but not all computer programs for the interpretation of ECGs perform almost as well as cardiologists in identifying seven major cardiac disorders.

[24-hour blood pressure profile: reproducibility of automatic ambulatory measurement]. / 24-Stunden-Blutdruckprofil: Reproduzierbarkeit bei ambulanter automatischer Messung.

Twelve patients (8 males, 4 females; mean age 53 [43-60] years) who were undergoing rehabilitation treatment after myocardial infarction were studied to ascertain the reproducibility of ambulatory automatic blood pressure measurements. All were in the last stage of a rehabilitation programme (tolerance to ordinary activity; symptom-free exercise at 75 W). Within two weeks 2, 3 or 4 blood-pressure profiles over 24 hours (total of 32 readings) were obtained by automatic measurement and the records and mean values were compared. Single mild stresses, such as gymnastic exercise or visit to the doctor, were identifiable on the records, but did not alter the overall profile or mean values. Normotensives, hypertensives and borderline hypertensives had 24-hour profiles which were nearly identical with regard to the curve "envelope", day-night profile and mean values in the individual patients. Mean values of diastolic and systolic pressures day by day in each patient showed no deviations greater than 5 mm Hg. Therapeutic measures were recognizable by parallel fall of the curve "envelope", as well as by a reduction in mean value.--At least in these selected patients a single 24-hour profile would in principle have sufficed to describe blood pressure behaviour.