Absence of the kinase S6k1 mimics the effect of chronic endurance exercise on glucose tolerance and muscle oxidative stress.

OBJECTIVE: Ribosomal protein S6 Kinase-1 (S6K1) has been linked to resistance exercise-mediated improvements in glycemia. We hypothesized that S6K1 may also play a role in regulating glycemic control in response to endurance exercise training. METHODS: S6k1-knockout (S6K1KO) and WT mice on a 60 cal% high-fat diet were trained for 4 weeks on treadmills, metabolically phenotyped, and compared to sedentary controls. RESULTS: WT mice showed improved glucose tolerance after training. In contrast, S6K1KO mice displayed equally high glucose tolerance already in the sedentary state with no further improvement after training. Similarly, training decreased mitochondrial ROS production in skeletal muscle of WT mice, whereas ROS levels were already low in the sedentary S6K1KO mice with no further decrease after training. Nevertheless, trained S6K1KO mice displayed an increased running capacity compared to trained WT mice, as well as substantially reduced triglyceride contents in liver and skeletal muscle. The improvements in glucose handling and running endurance in S6K1KO mice were associated with markedly increased ketogenesis and a higher respiratory exchange ratio. CONCLUSIONS: In high-fat fed mice, loss of S6K1 mimics endurance exercise training by reducing mitochondrial ROS production and upregulating oxidative utilization of ketone bodies. Pharmacological targeting of S6K1 may improve the outcome of exercise-based interventions in obesity and diabetes.

The inter-individual variability outperforms the intra-individual variability of differentially expressed proteins prior and post irradiation in lymphoblastoid cell lines.

CONTEXT: Radio-sensitivity in normal tissue is characterized by heterogeneity throughout the population and the absence of pre-diagnostic biomarkers. OBJECTIVE: We conducted a proteomic approach to search for radiation characteristic protein regulation. MATERIALS AND METHODS: Cell lines were 10 Gy irradiated and analysed by 2D-DIGE after 24 h. RESULTS were analysed intra- and inter-individually. The principal component analysis and hierarchical clustering was applied to all datasets. RESULTS: Differences in intra-individual spot abundance prior and post irradiation exactly show the separation of sample classes in two groups: sham-irradiated and irradiated. The inter-individual datasets clustered according to the cell line. The intra-individual differences on protein level after gamma-irradiation are very low, compared with the inter-individual differences among cell lines derived from the same tissue. CONCLUSION: The application of 2-D DIGE may offer a realistic chance for a better molecular characterization of radio-sensitivity and for the discovery of candidate biomarkers.

Knockdown of PRAS40 inhibits insulin action via proteasome-mediated degradation of IRS1 in primary human skeletal muscle cells.

AIMS/HYPOTHESIS: The proline-rich Akt substrate of 40 kDa (PRAS40) is a component of the mammalian target of rapamycin complex 1 (mTORC1) and among the most prominent Akt substrates in skeletal muscle. Yet the cellular functions of PRAS40 are incompletely defined. This study assessed the function of PRAS40 in insulin action in primary human skeletal muscle cells (hSkMC). METHODS: Insulin action was examined in hSkMC following small interfering RNA-mediated silencing of PRAS40 (also known as AKT1S1) under normal conditions and following chemokine-induced insulin resistance. RESULTS: PRAS40 knockdown (PRAS40-KD) in hSkMC decreased insulin-mediated phosphorylation of Akt by 50% (p < 0.05) as well as of the Akt substrates glycogen synthase kinase 3 (40%) and tuberous sclerosis complex 2 (32%) (both p < 0.05). Furthermore, insulin-stimulated glucose uptake was reduced by 20% in PRAS40-KD myotubes (p < 0.05). Exposing PRAS40-KD myotubes to chemokines caused no additional deterioration of insulin action. PRAS40-KD further reduced insulin-mediated phosphorylation of the mTORC1-regulated proteins p70S6 kinase (p70S6K) (47%), S6 (43%), and eukaryotic elongation 4E-binding protein 1 (100%), as well as protein levels of growth factor receptor bound protein 10 (35%) (all p < 0.05). The inhibition of insulin action in PRAS40-KD myotubes was associated with a reduction in IRS1 protein levels (60%) (p < 0.05), and was reversed by pharmacological proteasome inhibition. Accordingly, expression of the genes encoding E3-ligases F-box protein 32 (also known as atrogin-1) and muscle RING-finger protein-1 and activity of the proteasome was elevated in PRAS40-KD myotubes. CONCLUSIONS/INTERPRETATION: Inhibition of insulin action in PRAS40-KD myotubes was found to associate with IRS1 degradation promoted by increased proteasome activity rather than hyperactivation of the p70S6K-negative-feedback loop. These findings identify PRAS40 as a modulator of insulin action.

Pigment epithelium-derived factor (PEDF) is one of the most abundant proteins secreted by human adipocytes and induces insulin resistance and inflammatory signaling in muscle and fat cells.

OBJECTIVE: Pigment epithelium-derived factor (PEDF) is a multifunctional protein with neurotrophic and anti-angiogenic properties. More recently it became evident that PEDF is upregulated in patients with type 2 diabetes and also contributes to insulin resistance in mice. During characterization of the secretome of in vitro differentiated human adipocytes by two-dimensional polyacrylamide gel electrophoresis and matrix-assisted laser desorption/ionization-MS, we found that PEDF is one of the most abundant proteins released by adipocytes. The aim of this study was to investigate the regulation and autocrine function of PEDF in human adipocytes and to determine its paracrine effects on human skeletal muscle cells (hSkMC) and human smooth muscle cells (hSMC). METHODS AND RESULTS: Human primary adipocytes secrete 130 ng ml(-1) PEDF over 24 h from 1 million cells, which is extremely high as compared with adiponectin, interleukin-6 (IL-6) or IL-8. This release of PEDF is significantly higher than from other primary cells, such as adipose-tissue located macrophages (50-times), hSkMC and hSMC (5-times). PEDF protein expression significantly increases during adipogenesis, which is paralleled by increased PEDF secretion. Furthermore, tumor necrosis factor-α and hypoxia significantly downregulate PEDF protein levels. PEDF secretion was significantly reduced by troglitazone and hypoxia and significantly increased by insulin. Treatment of adipocytes and hSkMC with PEDF induced insulin resistance in adipocytes, skeletal and smooth muscle cells at the level of insulin-stimulated Akt phosphorylation, which was dose dependent and more prominent in adipocytes. Furthermore, inflammatory nuclear factor-κB (NF-κB) signaling was induced by PEDF. In hSMC, PEDF induced proliferation (1.7-fold) and acutely activated proliferative and inflammatory signaling pathways (NF-κB, p38 mitogen-activated protein kinase and mammalian target of rapamycin). CONCLUSION: PEDF is one of the most abundant adipokines and its secretion is inversely regulated by insulin and hypoxia. PEDF induces insulin resistance in adipocytes and hSkMC and leads to inflammatory signaling in hSMC. Because of these diverse actions, PEDF is a key adipokine, which could have an important role in diabetes and obesity-related disorders.

[Pari sinus inhalation versus metered pump spray in postoperative care after FESS]. / Topische nasale Therapie nach endonasaler NNH-Op: pulsierende Inhalation versus Pumpspray.

OBJECTIVE: To evaluate which way of topical drug application would result in a better deposition pattern after FESS. MATERIAL AND METHODS: We compared the deposition pattern of a nasal steroid after application with a metered pump spray or inhalation using the Pari sinus device. Visualization was achieved via colouring using 1% Sodium-fluorescein solution. All patients had a well healed sinus system with an endoscopically wide open access after FESS. We looked for the deposition with Blue-light-endoscopy directly after application and after washing out using a nasal douche. Analysis was performed blinded by two independent experienced sinus surgeons via representative single shots. RESULTS: Data of 11 patients revealed, that deposition after metered pump spray application was superior than after inhalation for all localisations. By far most part of the drug was deposited in the anterior nasal cavity, followed by the head of the middle turbinate, ethmoid. Small amounts reached the maxillary sinus, the anterior wall of the sphenoid sinus, the olfactory cleft and the entrance to the frontal sinus. The frontal sinus itself was not reached in any case. Washing out led to a decrease of deposition intensity, in some cases to a better distribution into the maxillary and sphenoid sinus and frontal recess. CONCLUSIONS: Postoperative deposition of topical nasal steroids after FESS is insufficient. We need better methods and devices to optimize efficacy of topical treatment.

Anti-apoptotic action of exendin-4 in INS-1 beta cells: comparative protein pattern analysis of isolated mitochondria.

Glucagon like peptide-1 (Glp-1) exhibits beneficial effects on beta cell mass by both enhancing proliferation and inhibiting apoptosis. The precise mechanism of the anti-apoptotic effect of Glp-1 and Glp-1 mimetics like exendin-4 has remained elusive. Here, we studied cytokine-induced apoptosis in the pancreatic beta cell line INS-1 and performed a comparative mitochondrial protein pattern analysis using two-dimensional difference gel electrophoresis (2D-DIGE). Cytokine incubation of INS-1 cells increased caspase-3 activity about 3-fold, which was reduced by 60% in the presence of exendin-4. Production of reactive oxygen species in response to cytokines was completely prevented after preincubation with exendin-4. Highly purified mitochondria were obtained and mitochondrial proteins were labeled with Cy-dyes and separated on overlapping zoom 2D gels spanning a pH-range of 4-9. Protein spots with significant changes after cytokine and exendin-4 treatment were identified by MALDI mass spectrometry. Comparing all treatment conditions, comparative mitochondrial proteome analysis allowed to identify 33 different proteins, which were significantly altered between comparison groups. Changes in protein patterns revealed involvement of cytokine-induced electron transport chain damage. Thus, cytochrome bc1 complex subunit I and ATP synthase subunit beta were downregulated by 30-40%. This was abrogated by the presence of exendin-4. In conclusion, this study provides further insights into the role of mitochondria in cytokine-induced apoptosis. We show here that exendin-4 significantly counter-regulates the reduced abundance of electron transport chain proteins, leading to a reduction of oxidative stress and most likely contributing to the anti-apoptotic action of this drug.

A key to canthopexy: the tarsal strap. A fresh cadaveric study.

BACKGROUND: This article describes a poorly understood yet clinically significant eyelid structure the authors have labeled the tarsal strap. The tarsal strap anchors the tarsus to the periosteum of the inferolateral orbit. Releasing this tethering band is the cornerstone of achieving lasting, superior surgical results during lateral canthopexy and periorbital restoration in aesthetic and reconstructive surgery. METHODS: An 87-year-old, male, fresh cadaveric head of Caucasian origin was studied. The dissection was repeated on 15 cadaver heads, including both male and female cadavers, from various ethnic backgrounds, to confirm the anatomy of the tarsal strap described in this article. RESULTS: The tarsal strap was identified in all 16 cadaver dissections and appeared in a consistently similar location. CONCLUSIONS: Until now, the tarsal strap has been a poorly understood entity, despite being an anatomically and clinically significant eyelid structure. Knowledge of its significance is essential to restoring the lateral canthus and returning periorbital structures to their youthful state. It should be routinely sought out and properly released.

Ambient odors of orange and lavender reduce anxiety and improve mood in a dental office.

The goal of this study was to investigate the impact of the essential oils of orange and lavender on anxiety, mood, alertness and calmness in dental patients. Two hundred patients between the ages of 18 and 77 years (half women, half men) were assigned to one of four independent groups. While waiting for dental procedures patients were either stimulated with ambient odor of orange or ambient odor of lavender. These conditions were compared to a music condition and a control condition (no odor, no music). Anxiety, mood, alertness and calmness were assessed while patients waited for dental treatment. Statistical analyses revealed that compared to control condition both ambient odors of orange and lavender reduced anxiety and improved mood in patients waiting for dental treatment. These findings support the previous opinion that odors are capable of altering emotional states and may indicate that the use of odors is helpful in reducing anxiety in dental patients.

The presence of IgG antibodies against beta2-glycoprotein I predicts the risk of thrombosis in patients with the lupus anticoagulant.

BACKGROUND: Lupus anticoagulant (LA) is a strong risk factor of thrombosis. However, a subgroup of patients positive for LA is unaffected by thrombosis and currently no predictive markers are available to identify patients positive for LA at increased risk for thrombosis. OBJECTIVE: The aim of the study was to investigate whether anti-beta-2-glycoprotein I (anti-beta2GPI) or anticardiolipin antibodies (ACA) are associated with an increased risk of thrombosis in patients persistently positive for LA. PATIENTS AND METHODS: A cohort of 87 consecutive patients persistently positive for LA was investigated, 55 with and 32 without a history of thrombosis. Immunoglobulin G (IgG) and M (IgM) antibodies against beta2GPI and cardiolipin were determined by enzyme-linked immunoassay. RESULTS: Patients positive for LA with thrombosis had significantly higher levels of anti-beta2GPI IgG (median 16.7 standard units, interquartile range 3.0-75.2, P = 0.002) and of ACA IgG (41.1 IgG phospholipid units per mL, 8.9-109.0, P = 0.002) than those without thrombosis (2.6, 1.4-7.9 and 9.7, 4.6-22.1, respectively). Levels of anti-beta2GPI IgM and ACA IgM did not differ significantly between LA patients with and without thrombosis (P = 0.25 and 0.12, respectively). Elevated anti-beta2GPI IgG was associated with an increased risk for thrombosis (OR = 4.0, 95% CI 1.2-13.1), especially for venous thromboembolism (OR = 5.2, 95% CI 1.5-18.0). CONCLUSIONS: Increased levels of anti-beta2GPI IgG were associated with thrombosis. We conclude that anti-beta2GPI IgG levels above normal predict an increased risk of thrombosis in patients persistently positive for LA.

Significant weight loss after laparoscopic Nissen fundoplication.

BACKGROUND: Laparoscopic Nissen fundoplication (LNF) has evolved as a gold standard in antireflux surgery. However, the association between body weight and gastroesophageal reflux disease (GERD) is still unclear, and no data are available concerning the effect of fundoplication on body weight. We present the first report elucidating the impact of LNF on body weight in GERD patients with special emphasis on patients' quality of life. METHODS: From July 2000 to March 2003, LNF was carried out in 213 patients (85 women and 128 men) after thorough preoperative examination including clinical interview with standardized assessment of symptoms and quality of life (QoL), endosocopy, barium swallow, 24-h pH-metry, and manometry. Follow-up investigations were performed 3 and 12 months after LNF obtainable from 209 patients (98.1%) and 154 patients (72.3%), respectively. RESULTS: The mean body mass index (BMI) decreased significantly after LNF (27.6 +/- 5.6 kg/m(2) before LNF vs 26.0 +/- 3.8 kg/m(2) after LNF, p < 0.001). Twelve months after LNF, neither a tendency toward a renewed increase nor a further decrease in BMI was observable. The average body weight loss was 3.9 kg. BMI reduction was higher in women than in men (p < 0.002), and obese patients lost more weight than lean patients (p < 0.001). There was no association between BMI reduction and dysphagia. Plasma cholesterol and triglyceride levels did not change after LNF. The mean general score of the Gastrointestinal Quality of Life Index markedly improved (90.1 +/- 21.3 before LNF vs 118.0 +/- 16.2 after LNF, p < 0.01), as did the GERD-Health Related Quality of Life Index (21.9 +/- 6.4 before LNF vs 3.5 +/- 2.7 after LNF, p < 0.001). However, there was no association between changes in BMI and QoL. CONCLUSION: LNF leads to significant and persistent body weight loss.

Changes in bone mineral density in the proximal femur after cementless total hip arthroplasty. A five-year longitudinal study.

We investigated prospectively the bone mineral density (BMD) of the proximal femur after implantation of a tapered rectangular cementless stem in 100 patients with a mean age of 60 years (16 to 87). It was determined using dual energy x-ray absorptiometry, performed one week after surgery and then every six months until the end-point of five years. The BMD increased significantly in Gruen zones 2, 4 and 5 by 11%, 3% and 11% respectively, and decreased significantly in Gruen zones 1, 6 and 7 by 3%, 6% and 14% respectively, over the five-year period. The net mean BMD did not change over this time period. The changes in the BMD were not confined to the first 12 months after surgery. This investigation revealed no change in the overall periprosthetic BMD, but demonstrated a regional redistribution of bone mass from the proximal to distal zones.

A randomized, double-blind, placebo controlled study on analgesic effects of botulinum toxin A.

OBJECTIVE: Botulinum toxin type A (BTXA) is used to treat neurologic disorders associated with increased muscle tone. Its use is often associated with pain relief. METHODS: A possible direct analgesic effect of BTXA on C and Adelta fibers was studied on 16 healthy volunteers receiving 30 U BTXA into one forearm and pure saline into the other. To exclude the secondary effect due to muscular tone reduction, BTXA was injected intradermally. Thermal sensory testing of heat pain (threshold and tolerance) and neuroselective current sensory testing of current pain threshold/tolerance were performed at baseline and 3, 14, and 28 days after treatment. Thereafter, on day 28, capsaicin was administered simultaneously into both forearms to evaluate a possible peripheral effect and central effect on pain processing and on the axon reflex flare. RESULTS: The authors observed no significant difference in any of the perception outcome measures between BTXA and placebo pretreated areas. Flare areas as a result of the release of neuropeptides after capsaicin application showed no differences. CONCLUSIONS: The results suggest that pain reduction after BTXA treatment is mediated through its effect on muscle tone rather than a direct analgesic effect.

Muscle fatigue and fatigue-related biomechanical changes during a cyclic lifting task.

STUDY DESIGN: Electromyographic and biomechanical methods were utilized to investigate correlations between indexes of localized muscle fatigue and changes in the kinematics and kinetics of motion during a cyclic lifting task. SUMMARY OF BACKGROUND DATA: Recent advances in time-frequency analysis procedures for electromyographicic signal processing provide a new way of studying localized muscle fatigue during dynamic contractions. These methods provide a means to investigate fatigue-related functional impairments in patients with low back pain. OBJECTIVES: To study the relationship between localized muscle fatigue and the biomechanics of lifting and lowering a weighted box. Fatigue-related changes in the electromyographicic signal of trunk and limb muscles were evaluated and compared to kinematic and kinetic measures in order to determine whether lifting strategy is modified with fatigue. METHODS: A total of 14 healthy male subjects (26 +/- 5 years) cyclically lifted and lowered a 13 kg box (12 lifts/min) for 4.5 minutes. A 5-second static maximum lifting task was included immediately before and after the cyclic lifting task to measure changes in lifting strength and static electromyographicic fatigue indexes. Electromyographic signals from 14 muscle sites (including paravertebral and limb muscles) were measured. Changes in the electromyographicic Instantaneous Median Frequency, a fatigue index, were computed using time-frequency analysis methods. This index was compared with more standardized measures of fatigue, such as those based on electromyographicic median frequency acquired during a static trunk extension test, subjective fatigue measures, and maximal static lifting strength. Biomechanical measures were gathered using a motion analysis system to study kinematic and kinetic changes during the lifting task. RESULTS: During the cyclic lifting task, the electromyographic Instantaneous Median Frequency significantly decreased over time in the paravertebral muscles, but not in the limb muscles. Paravertebral electromyographicic Instantaneous Median Frequency changes were consistent with self-reports of fatigue as well as decreases in trunk extension strength. The magnitude of muscle-specific changes in electromyographicic Instantaneous Median Frequency was not significantly correlated with electromyographicic median frequency changes from the static trunk extension task. The load of the box relative to the maximal static lifting strength significantly affected the electromyographicic Instantaneous Median Frequency changes of paravertebral back muscles. Significant changes with fatigue during the task were found in the angular displacements at the knee, hip, trunk, and elbow. These biomechanical changes were associated with increased peak torque and forces at the L4-L5 vertebral segment. CONCLUSIONS: Our results demonstrate correlation between localized muscle fatigue and biomechanical adaptations that occur during a cyclic lifting task. This new technique may provide researchers and clinicians with a means to investigate fatigue-related effects of repetitive work tasks or assessment procedures that might be useful in improving education, lifting ergonomy, and back school programs. Although both the dynamic and static tasks resulted in spectral shifts in the electromyographicic data, the fact that these methods led to different muscle-specific findings indicates that they should not be considered as equivalent assessment procedures.

Characterization of risk factor differences in perimesencephalic subarachnoid hemorrhage.

OBJECTIVE: The non-aneurysmal perimesencephalic subarachnoid hemorrhage (p-SAH) has a favorable outcome. Our objective was to provide a reason for that assessing the association of putative risk factors with this different type of hemorrhage in comparison to the aneurysmal subarachnoid hemorrhage (a-SAH). METHODS: We selected 185 consecutive cases of subarachnoid hemorrhage between September 1994 and August 1998 in a hospital-based case-control study and investigated the influence of hypertension, body weight, smoking and the three interacting blood factors hematocrit, fibrinogen and leukocrit. RESULTS: 1) Risk factors for SAH: For the entire study group an association of hypertension with the incidence of SAH was found both in the univariate analysis of this variable alone (P=0.001) and in the multivariate logistic regression (P=0.0001), aside from the risk factors smoking (P=0.0047) and hematocrit (P=0.0001). As to the risk of hypertensive subjects to experience SAH for different reasons, the logistic regression yielded an odds ratio (OR) of 3.54 [CI (95 %): 2.21-5.56]. 2) Risk factors for p-SAH: Patients with p-SAH differ in their risk profile from typical aneurysmal SAH cases with respect to their blood pressure status (P=0.019) and the investigated hemorheological parameters. CONCLUSIONS: A different association between possible risk factors and the two types of subarachnoid hemorrhages was ascertained. In general there seem to be fewer putative risk factors in the perimesencephalic SAH cases compared to the typical aneurysmal hemorrhages.

Hepatopulmonary syndrome: prevalence and predictive value of various cut offs for arterial oxygenation and their clinical consequences.

BACKGROUND: The hepatopulmonary syndrome (HPS) is defined as the triad of liver disease, arterial deoxygenation, and pulmonary vascular dilatation. The reported prevalence of HPS in cirrhotic patients varies between 4% and 19%, and various threshold values defining arterial deoxygenation have been used and recommended previously. However, it is not known how the prevalence of HPS differs using different cut off values for arterial deoxygenation. METHODS: We studied 127 patients for the presence of HPS using transthoracic contrast echocardiography for detection of pulmonary vasodilation, pulmonary function tests, and blood gas analysis. RESULTS: Ninety eight patients were included in the study, of whom 33 (34%) had a positive contrast echocardiography. Using an increased alveolar-arterial difference for the partial pressure of oxygen (AaDO(2)) as an indication of hypoxaemia, the prevalence of HPS was considerably higher (>15 mm Hg, 32%; >20 mm Hg, 31%; and >age related threshold, 28%) than using reduced partial pressure of arterial oxygen (PaO(2)) as a threshold (<80 mm Hg, 19%; <70 mm Hg, 15%; and

Vascularity and uptake of photosensitizer in small human tumor nodules: implications for intraperitoneal photodynamic therapy.

PURPOSE: i.p. spread of cancers is a common clinical problem, with limited treatment options leading to morbidity and death. i.p. photodynamic therapy (IP-PDT) combines maximal surgical debulking of gross tumor with intraoperative light delivery to the peritoneum after preoperative i.v. injection of photosensitizer to treat residual disease. An issue of concern in IP-PDT is the potential lack of photosensitizer uptake by residual small tumor nodules (STNs) < or =5 mm in maximum diameter and by microscopic residual disease caused by incomplete development of a vascular supply. This study examined the existence of vasculature and Photofrin (PF) uptake in STNs in 12 patients in a Phase II clinical trial for IP-PDT. EXPERIMENTAL DESIGN: Patients received PF 2.5 mg/kg i.v. 48 h before surgery. STNs obtained during surgery were cryosectioned, immunostained for platelet/endothelial cell adhesion molecule 1, and analyzed by light microscopy. Mean vascular densities in STNs were determined by counting microvessels within a x200 field (0.28 mm(2) area). Sections were also examined for PF uptake by fluorescence image analysis using an epifluorescence microscope and IPLab Spectrum software. RESULTS: Data obtained showed that tumors as small as 1 mm in diameter stained positive for platelet/endothelial cell adhesion molecule 1 and contained PF. A negative control from a patient not given PF showed no detectable fluorescence. The average of all mean vascular densities in STNs was determined to be 100 +/- 29. CONCLUSIONS: We conclude that STNs, as small as 1 mm in diameter, have a functional vasculature, because these tumors show PF uptake after i.v. delivery. Both properties are crucial for the treatment of residual STNs by IP-PDT after surgical debulking.

A minimally invasive approach for treating postoperative seromas after incisional hernia repair.

BACKGROUND: The most frequent wound complication following repair of large incisional hernias is seroma formation, especially when the use of a mesh onlay requires extensive subcutaneous undermining. Treatment options for postoperative seromas include observation for spontaneous resolution, percutaneous aspiration, closed suction drainage, abdominal binders, and sclerosant. METHODS: A novel technique for treating persistent postoperative seromas is presented herein. This technique involves a 3-puncture minimally invasive approach that can be performed in an outpatient setting. Evacuation of serous fluid and fibrinous debris is followed by argon beam scarification of the seroma cavity lining. Talc slurry is then introduced into the cavity. Three patients have been treated with this technique. RESULTS: All 3 patients had successful ablation of seromas that had persisted despite standard treatment modalities. CONCLUSION: A minimally invasive approach is a reasonable and safe alternative for treating persistent postoperative seromas.

Insulin-regulated transcription factors: molecular link between insulin resistance and cardiovascular risk factors.

Patients with insulin resistance and/or type 2 diabetes have a 5-fold increase in cardiovascular mortality rate. Therefore, it is a current issue of discussion that arterial hypertension, lipid disorders as well as visceral obesity are coronary risk factors, which might belong to a syndrome that is caused by decreased insulin sensitivity. Concerning a possible molecular link between insulin resistance, atherosclerosis and obesity, we focus in our research on questions looking for a molecular link between lipid metabolism, insulin action, and obesity at a gene regulatory level. Alterations in the structure, function and regulation of transcription factors appear to be such signalling steps which might play an essential role in the pathogenesis and therapy of cardiovascular risk factors associated with insulin resistance, eg the so called metabolic syndrome. Recent examples are members of the nuclear hormone receptor superfamily, eg peroxisome proliferator-activated receptor (PPAR) isoforms and sterol regulatory element-binding proteins (SREBPs). Beside their regulation by different metabolites, these transcription factors are also targets of hormones, like insulin and leptin, growth factors, and inflammatory signals. Therefore, they appear to be a point of signalling convergence at a gene regulatory level. Major signalling pathways coupling receptors at the cell surface for hormones, growth factors as well as cytokines to gene regulatory events in the nucleus are the MAP-kinase cascades. We have recently defined different postreceptor defects in these pathways in patients with clinical phenotypes corresponding to congenital lipoatrophy. Therefore, these studies may identify novel pathways which play a role in the control of body weight, insulin sensitivity and cardiovascular risk.

Identification of post-translationally modified proteins in proteome studies.

Proteome studies are powerful tools to solve many different problems in metabolism, signal transduction, drug discovery, and other areas of interest in life sciences. Up to now, high-sensitive methods for protein identification after two-dimensional gel electrophoresis using mass spectrometry are available. However, the identification of post-translational modifications after two-dimensional gel electrophoresis is still an unsolved problem. In this paper, we want to give several examples for the successful identification of post-translational modifications and point mutations.

Intravenous magnesium sulfate for bronchial hyperreactivity: a randomized, controlled, double-blind study.

BACKGROUND: Magnesium has been shown to be helpful in the treatment of acute exacerbations of asthma. Conflicting data exist concerning the effect of magnesium on bronchial hyperreactivity. METHODS: We performed a randomized, double-blind, placebo-controlled study to investigate the effect of intravenous magnesium sulfate on bronchial reactivity to metacholine in 30 subjects with bronchial hyperreactivity. Two days after baseline metacholine provocation, 20 subjects received 0.3 mmol/kg/h of intravenous magnesium sulfate and 10 subjects received normal saline solution. Metacholine provocation was repeated 30 minutes after the initiation of the magnesium or placebo infusion. RESULTS: The difference of the postinterventional minus the baseline provocative dose of metacholine required to decrease the forced expiratory volume in 1 second by 20% (PC20) was significantly higher in the magnesium group compared with the placebo group (0.48 +/- 0.46 mg/mL versus 0.05 +/- 0.73 mg/mL, P = .028). In the magnesium group, the PC(20) significantly increased (from 0.83 +/- 0.54 mg/mL to 1.31 +/- 0.66 mg/mL, P = .0001), whereas there was no change in the placebo group (0.86 +/- 0.52 mg/mL to 0.91 +/- 0.54 mg/mL, P = .83). CONCLUSIONS: In the magnesium group, 30% of the subjects reached a normal PC(20) compared with 10% in the placebo group. We conclude that intravenous magnesium sulfate significantly improved bronchial hyperreactivity and may serve as an adjunct to standard treatment.