In situ investigation of production processes in a large chamber scanning electron microscope.

A large-chamber scanning electron microscope (LC-SEM) provides an ideal platform for the installation of large-scale in situ experiments. Our LC-SEM has internal chamber dimensions of 1,2 × 1,3 × 1,4 m3 (W × H × D) (Fig.1) and makes it possible to incorporate novel in situ experimental devices, which are reported on here. The present manuscript describes in detail the development of in situ test equipment for the study of a broad range of processes in production engineering. Direct observation of the materials modification mechanisms provides fundamental insight into the underlying process characteristics. An in situ turning device was developed, tested and used to observe the chip formation on the microstructure scale of a 43CrMo4-sample. Laser beam micro welding was integrated into the LC-SEM to achieve in situ analysis of the welding process on stainless steel 1.4310. A heating module was employed for in situ wetting experiments to observe the formation and solidification of the melt of a tin-copper brazing filler on an aluminium cast alloy.

Variations in truncal body circumferences affect fat mass quantification with bioimpedance analysis.

OBJECTIVE: To test the hypothesis that variations in trunk circumferences influence the accuracy of bioimpedance analysis (BIA) for assessment of percent fat mass (%FM). SUBJECTS AND METHODS: %FM was predicted with BIA, and compared with air-displacement plethysmography (ADP) in a small sample of 35 overweight (OW), 21 normal weight and 8 underweight volunteers. Waist and hip circumferences were assessed, and 15 of the OW subjects were measured before and after weight reduction. RESULTS: BIA and ADP provided similar cross-sectional estimates of group mean %FM (28.9±10.0 and 31.3±13.0%, respectively). However, within individuals, there were large between-method differences (Diff(BIA-ADP)) ranging from -13 to +13 %FM. Furthermore, we found a systematic bias of BIA related to the degree of adiposity. Consequently, %FM and fat mass loss during weight reduction in OW were underestimated with BIA when compared with ADP. Waist and hip circumferences were inversely associated with resistance (R) and reactance (P<0.01), and with Diff(BIA-ADP) (P<0.001). In women, the variability in hip circumference explained 76%, and in men, the variability in waist circumference explained 59% of Diff(BIA-ADP). CONCLUSION: Resistance changes associated with variations in trunk circumferences decrease resistance, and therefore impair the accuracy of BIA to assess %FM.

Neurohumoral and metabolic response to exercise in water.

Atrial natriuretic peptide (ANP) stimulates lipid mobilization and lipid oxidation in humans. The mechanism appears to promote lipid mobilization during exercise. We tested the hypothesis that water immersion augments exercise-induced ANP release and that the change in ANP availability is associated with increased lipid mobilization and lipid oxidation. In an open randomized and cross-over fashion we studied 17 men (age 31+/-3.6 years; body mass index 24+/-1.7 kg/m(2); body fat 17+/-6.7%) on no medication. Subjects underwent two incremental exercise tests on a bicycle ergometer. One test was conducted on land and the other test during immersion in water up to the xiphoid process. In a subset (n=7), we obtained electromyography recordings in the left leg. We monitored gas exchange, blood pressure, and heart rate. In addition, we obtained blood samples towards the end of each exercise step to determine ANP, norepinephrine, epinephrine, lactate, free fatty acids, insulin, and glucose concentrations. Heart rate, systolic blood pressure, and oxygen consumption at the anaerobic threshold and during peak exercise were similar on land and with exercise in water. The respiratory quotient was mildly reduced when subjects exercised in water. Glucose and lactate measurements were decreased whereas free fatty acid concentrations were increased with exercise in water. Water immersion attenuated epinephrine and norepinephrine and augmented ANP release during exercise. Even though water immersion blunts exercise-induced sympathoadrenal activation, lipid mobilization and lipid oxidation rate are maintained or even improved. The response may be explained by augmented ANP release.

Determinants of exercise-induced fat oxidation in obese women and men.

Endurance training at an intensity eliciting maximal fat oxidation may have a beneficial effect on body weight and glucose metabolism in obese patients. However, the exercise intensity at which maximal fat oxidation occurs and the factors limiting fat oxidation are not well studied in this population. Obese, otherwise healthy men (n=38) and women (n=91) performed an incremental exercise test up to exhaustion on a cycle ergometer. Substrate oxidation was estimated using indirect calorimetry. Magnetic resonance tomography and spectroscopy were conducted to assess body fat distribution and intramyocellular fat content. We determined the exercise intensity at which maximal body fat oxidation occurs and assessed whether body composition, body fat distribution, intramyocellular fat content, or oxidative capacity predict exercise-induced fat oxidation. Maximal exercise-induced fat oxidation was 0.30+/-0.02 g/min in men and 0.23+/-0.01 g/min in women (p<0.05). Exercise intensity at the maximum fat oxidation was 42+/-2.2% VO (2 max) in men and 43+/-1.7% VO (2 max) in women. With multivariate analysis, exercise-induced fat oxidation was related to fat-free mass, percent fat mass, and oxidative capacity, but not to absolute fat mass, visceral fat, or intramyocellular fat content. We conclude that in obese subjects the capacity to oxidize fat during exercise appears to be limited by skeletal muscle mass and oxidative capacity rather than the availability of visceral or intramyocellular fat.

Integrin-actin interactions.

The integrin family of extracellular matrix receptors regulates many aspects of cell life, in particular cell adhesion and migration. These two processes depend on organization of the actin cytoskeleton into adhesive and protrusive organelles in response to extracellular signals. Integrins are important switch points for the spatiotemporal control of actin-based motility in higher eukaryotes. Ligands of integrin cytoplasmic tails are central elements of signalling pathways involving small GTPases as well as protein and lipid kinases in the regulation of Factin crosslinking, actin treadmilling and de novo nucleation of actin filaments. We present an overview of common pathways and discuss recent evidence for their differential use by individual integrin receptors.

Chemical development of latent fingerprints: 1,2-indanedione has come of age.

The performance of 1,2-indanedione as a latent fingerprint reagent on some types of paper was found to exceed that of DFO, the leading fluorogenic fingerprint reagent. It even exceeds the performance of the sequence, DFO, followed by ninhydrin. No new prints could be observed when ninhydrin was applied after indanedione. On a large number of actual exhibits (used checks) indanedione developed 46% more identifiable prints than the sequence DFO-ninhydrin. A standard procedure for fingerprint development by indanedione is proposed. Best results are obtained with a 0.2% indanedione solution in HFE7100 solvent containing 7% ethyl acetate, but no acetic acid. It can be recommended to start using 1,2-indanedione, which is already commercially available, in actual fingerprint casework.

Gastrointestinal colonization by Candida albicans mutant strains in antibiotic-treated mice.

Antibiotic-treated mice orally inoculated with one of three Candida albicans strains (including two mutant strains) or indigenous Candida pelliculosa showed levels of candidal gastrointestinal colonization that were strain specific. However, regardless of strain, the numbers of viable candida were intermediate to high in the stomach, were consistently lowest in the upper small intestine, and increased progressively down the intestinal tract.

Clostridium difficile toxins A and B can alter epithelial permeability and promote bacterial paracellular migration through HT-29 enterocytes.

Clostridium difficile toxins A and B are the widely recognized etiologic agents of antibiotic-associated diseases ranging from diarrhea to pseudomembranous colitis. We hypothesized that C. difficile toxins may alter intestinal epithelial permeability and facilitate bacterial penetration of the intestinal epithelial barrier. Experiments were designed to clarify the effects of C. difficile toxins A and B on the flux of inert particles across HT-29 enterocyte monolayers, and to correlate these results with bacteria-enterocyte interactions. In all experiments, mature, confluent HT-29 cultures were preincubated 16 h with toxin A or B (1-100 ng/mL). To study alterations in epithelial permeability, toxin-treated enterocytes were incubated with 5 pM solutions of 10- and 40-kD inert dextran particles. Toxin A, but not toxin B, was associated with increased dextran flux through enterocyte monolayers. To study bacteria-enterocyte interactions, toxin-treated enterocytes were incubated with 10(8) Salmonella typhimurium, Proteus mirabilis, or Escherichia coli. Although numbers of internalized bacteria were generally unaffected, both toxins were associated with increased bacterial adherence, as well as increased bacterial transmigration through enterocyte monolayers. Bacterial transmigration was significantly greater using toxin A- compared to toxin B-treated enterocytes, consistent with the observation that dextran flux was significantly greater using toxin A- compared to toxin B-treated enterocytes. Thus intestinal colonization with toxigenic C. difficile may facilitate bacterial penetration of the intestinal epithelium by a mechanism involving increased permeability of the intestinal epithelial barrier.

Indications for a novel muscular dystrophy pathway. gamma-filamin, the muscle-specific filamin isoform, interacts with myotilin.

gamma-Filamin, also called ABP-L, is a filamin isoform that is specifically expressed in striated muscles, where it is predominantly localized in myofibrillar Z-discs. A minor fraction of the protein shows subsarcolemmal localization. Although gamma-filamin has the same overall structure as the two other known isoforms, it is the only isoform that carries a unique insertion in its immunoglobulin (Ig)-like domain 20. Sequencing of the genomic region encoding this part of the molecule shows that this insert is encoded by an extra exon. Transient transfections of the insert-bearing domain in skeletal muscle cells and cardiomyocytes show that this single domain is sufficient for targeting to developing and mature Z-discs. The yeast two-hybrid method was used to identify possible binding partners for the insert-bearing Ig-like domain 20 of gamma-filamin. The two Ig-like domains of the recently described alpha-actinin-binding Z-disc protein myotilin were found to interact directly with this filamin domain, indicating that the amino-terminal end of gamma-filamin may be indirectly anchored to alpha-actinin in the Z-disc via myotilin. Since defects in the myotilin gene were recently reported to cause a form of autosomal dominant limb-girdle muscular dystrophy, our findings provide a further contribution to the molecular understanding of this disease.

Reagents for the chemical development of latent fingerprints: scope and limitations of benzo[f]ninhydrin in comparison to ninhydrin.

Benzo[f]ninhydrin was compared to ninhydrin for fingerprint development on paper. Overall, the performance of ninhydrin on exhibits was slightly better than that of benzo[f]ninhydrin. The significant advantages of the benzo[f]ninhydrin over ninhydrin were the much stronger fluorescence it gave after treatment with zinc salts and a slightly quicker reaction under ambient conditions. This fluorescence is, however, similar to that obtained with other reagents, such as DFO or ninhydrin analogs. These advantages apparently are not sufficient to justify regular usage of benzo[f]ninhydrin, especially when one considers its low solubility and high cost.

Water molecules in the binding cavity of intestinal fatty acid binding protein: dynamic characterization by water 17O and 2H magnetic relaxation dispersion.

The hydration of intestinal fatty acid binding protein (IFABP) in apo-form and complexed with palmitate, oleate, and 1-anilino-8-naphthalene sulfonate (ANS) has been studied by water 17O and 2H magnetic relaxation dispersion (MRD) measurements. These ligands bind in a large internal cavity, displacing most of the crystallographically identified cavity water molecules. Unlike most other proteins, IFABP gives rise to MRD profiles with two dispersion steps. The low-frequency dispersion yields a correlation time of 7 ns at 300 K, matching the known tumbling time of IFABP. The dispersion amplitude requires only three (apo) or four (holo) long-lived and ordered water molecules (residence time 0.01-4 microseconds at 300 K). Comparison of MRD profiles from the different complexes indicates that the displaced cavity water molecules are short-lived. The few long-lived (>10 ns) water molecules required by the MRD data are tentatively assigned to crystallographic hydration sites on the basis of accessibility, positional order, and H-bonding. The amplitude of the high-frequency dispersion corresponds to 10-20 moderately ordered water molecules, with a correlation time of ca. 1 ns that may reflect a transient opening of the cavity required for exchange with external water.

Hypnotically-induced vasodilation in the treatment of repetitive strain injuries.

The study examined the effectiveness of behaviorally-induced vasodilation (hypnosis with biofeedback and autogenics) in the treatment of upper extremity repetitive strain injuries (RSI). Thirty patients with recent onset of upper extremity RSI symptoms were randomly assigned to 1 of 2 treatment conditions, i.e., hypnotically-induced vasodilation or a waiting-list control. Treatments were given on an individual basis, once a week for 6 weeks. Patients in the treatment condition showed highly significant increases in hand temperature between pre- and post-treatment. Patients in the treatment condition also showed highly significant reductions in pain in comparison to the waiting list condition.

Existence of the Frank-Starling mechanism in the failing human heart. Investigations on the organ, tissue, and sarcomere levels.

BACKGROUND: The Frank-Starling mechanism is one of the most important physiological principles for regulation of contractile performance. We therefore studied the question of whether this mechanism may be absent or attenuated in end-stage failing human left ventricular myocardium. METHODS AND RESULTS: Different methodological approaches were used to analyze the effects of this mechanism on the organ, tissue, and sarcomere levels: (1) In excised human whole left ventricles (2 donor hearts, 5 failing hearts), diastolic and systolic pressure-volume relationships were obtained. (2) In isolated muscle strip preparations from the left ventricular wall of donor hearts (n = 14) and failing hearts from patients with idiopathic dilated cardiomyopathy (n = 21) and ischemic cardiomyopathy (n = 11), peak developed force was measured at different muscle lengths of the preparation. (3) Skinned fiber preparations were obtained from failing right and left ventricles (n = 12). In all three studies, we clearly observed the existence of the Frank-Starling mechanism: (1) In isolated failing human left ventricles, peak developed isometric pressure is increased when the preload is elevated. (2) Peak developed tension is increased by approximately 50% to 70% (P < .01) in left ventricular preparations of failing and nonfailing ventricles when the muscles are stretched from 90% to 100% optimum length. (3) An increase in sarcomere length leads to a sensitization of contractile proteins of ventricular skinned fiber preparations from failing human hearts. At 1.9-microns sarcomere length, the EC50 value was 5.56 +/- 0.06, and at 2.3 microns it was 5.70 +/- 0.05 (P < .01; n = 7). CONCLUSIONS: The Frank-Starling mechanism is maintained in end-stage failing human hearts, whereas significant alterations of diastolic myocardial distensibility are evident in chronic heart failure.

The national jockey injury study: an analysis of injuries to professional horse-racing jockeys.

While there have been numerous reports in the literature of accidents in equestrian sports, no comprehensive study has been conducted to ascertain the nature and incidence of injuries incurred by professional horse-racing jockeys. A survey was conducted to determine the types of injuries to jockeys and racing-related health concerns, including weight reduction methods. The questionnaire was completed by 706 professional jockeys actively competing at United States racetracks between July and October 1990 about injuries they had sustained in their careers. More than 1,700 injuries were reported. Fractures (n = 1,113) accounted for 64% of the total. The most common cause of injury was becoming unseated, followed by the horse falling. Relationships between characteristics of jockeys and injuries were evaluated. There were significant numbers of serious injuries with prolonged periods of not riding. Recommendations for improving jockey safety are made.

Prevention: conditioning and orthotics.

The hand and wrist of both athletes and musicians are very susceptible to injuries. Proper conditioning of the muscles, tendons, ligaments, and joints of the hand and wrist are crucial for injury prevention. Often, orthotic devices, from simple taping to polyethylene casts, can provide adequate protection and support to allow injuries to heal properly or prevent injuries from recurring in susceptible tissues.

Brainstem auditory evoked potentials in migraine--evidence of increased side differences during the pain-free interval.

Brainstem auditory evoked potentials (BAEPs) were recorded in 38 unselected patients (6 m./32 f.; age 18-54 yrs.) with various forms of migraine, during the pain-free interval. The values were compared to those of 50 control persons (18 m./32 f.; age 17-75 yrs.). Peak latencies (PL I-VI), interpeak latencies (IPL I-III, III-V, I-V) and side differences of all peaks (delta I-VI) were calculated. In contrast to PL's of migraine patients, all PL's of control persons were normally distributed. In 6 migraine patients (2 with basilar migraine) PL's were pathologically delayed. Statistical analysis did not show any significant difference in regard to PL's and IPL's between migraine patients and controls. However, side differences of all peaks (except peak IV,VI) were significantly increased in migraine patients as compared to controls. Our results indicate a slight but permanent impairment of brainstem function in migraine.