Separation of Forearm Hemodynamics Into Skin and Muscle Components by Means of I-Epinephrine Iontophoresis

By a combination of iontophoresis of I-epinephrine into the skin of one arm and simultaneous venous occlusion plethysmography in both treated (muscle only) and untreated forearms (muscle plus skin), we examined in 16 normal volunteers forearm blood flow, capillary filtration coefficient and venous capacity at cuff pressure of 40 mm of mercury (VC₄₀) at rest, during tonic finger exercise and after interrupted repetitive finger exercise. Blood pressure did not change during the testing procedure. Forearm muscle conductance was about 60% to 70% of total conductance and was positively correlated with total conductance during rest and exercise. With standard exercises muscle conductance rises to 1½ to 2½ times resting level, and skin conductance rises to 2½ to 4½ times resting level. The capillary filtration coefficient is almost entirely in the muscle. It doubles in value with tonic exercise but decreases to half its resting value after interrupted repetitive exercise despite greatly increased conductance. Therefore, repetitive exercise-induced dissociation between conductance and filtration surface occurs in striated muscle. The mechanism is yet unknown. VC₄₀ in muscle is about 84% of total forearm VC₄₀. During tonic exercise muscle VC₄₀ was reduced, and during interrupted repetitive exercise the values for muscle and skin returned to resting values. A high correlation between muscle only and muscle-plus-skin for forearm blood flow and the identify between arms for measuring capillary filtration coefficient makes iontophoresis unnecessary for determining these values in forearm striated muscle under these experimental conditions.     

NDRG4 is downregulated in glioblastoma and inhibits cell proliferation

NDRG4 is a member of the N-myc downregulated gene family (NDRG) belonging to the alpha/beta hydrolase superfamily. We have previously documented discrepancy between our analysis of the expression and function of NDRG4 in glioblastoma multiforme (GBM) and a recent publication by Schilling et al., who reported that NDRG4 is upregulated in GBM compared to human cortex tissues and knock down of NDRG4 reduced the viability of GBM cells. In the present study, we found that NDRG4 is indeed downregulated, at both RNA and protein levels, by quantitative RT-PCR and Western blot analysis, in GBM compared to normal tissues, and that over expression of NDRG4 inhibited proliferation of GBM cells. These new observations can inform the selection of lead molecular compounds for drug discovery as well as novel diagnostics for GBM. They also lend evidence to NDRG4 a role of tumor suppressor.     

Chemical-ionization mass spectrometry of carbohydrates: behavior of the antibiotic celesticetin under various conditions of ionization

The chemical-ionization mass spectrum of the polyfunctional carbohydrate antibiotic, celesticetin, is remarkably simple, in contrast to its electron-impact mass spectrum. With ammonia as the ionizing gas the protonated molecular ion is the principal species, and simple fragmentation modes reflect eleavage between principal moietics of the molecule. Substantial modifications of the fragmentation pathway are observed when isobutane is used as the ionizing gas. The different pathways of fragmentation according to the mode of ionization are interpreted in terms of differential reactivity of various heteroatomic centers in the molecule toward each of the ionizing agents. Use of a combination of ionizing modes may provide a useful general method for identification on a micro scale of complex carbohydrates isolated from fermentation or other mixtures.     

Effects of EDTA treatment upon the protein subunit composition and mechanical properties of mammalian single skeletal muscle fibers

Considerable interest has been focused on the role of myosin light chain LC(2) in the contraction of vertebrate striated muscle. A study was undertaken to further our investigations (Moss, R.L., G.G. Giulian, and M.L. Greaser, 1981, J. Biol. Chem., 257:8588-8591) of the effects of LC(2) removal upon contraction in skinned fibers from rabbit psoas muscles. Isometric tension and maximum velocity of shortening, V(max), were measured in fiber segments prior to LC(2) removal. The segments were then bathed at 30 degrees C for up to 240 min in a buffer solution containing 20 mM EDTA in order to extract up to 60 percent of the LC(2). Troponin C (TnC) was also partially removed by this procedure. Mechanical measurements were done following the EDTA extraction and the readditions of first TnC and then LC(2) to the segments. The protein subunit compositions of the same fiber segments were determined following each of these procedures by SDS PAGE of small pieces of the fiber. V(max) was found to decrease as the LC(2) content of the fiber segments was reduced by increasing the duration of extraction. EDTA treatment also resulted in substantial reductions in tension due mainly to the loss of TnC, though smaller reductions due to the extraction of LC(2) were also observed. Reversal of the order of recombination of LC(2) and TnC indicated that the reduction in V(max) following EDTA treatment was a specific effect of LC(2) removal. These results strongly suggest that LC(2) may have roles in determining the kinetics and extent of interaction between myosin and actin.     

Chromatographic and mass spectrometric methods for determination of lysergic acid diethylamide (LSD) and metabolites in body fluids

Continued illicit use of the potent psychedelic drug lysergic acid diethylamide (LSD) has stimulated efforts to develop effective analytical methods for detection of the drug and its metabolites in body fluids from suspected LSD users. Recently reported methods based on gas and liquid chromatography, combined with single- and multiple-stage mass spectral analysis, now permit accurate detection and quantitation of LSD at sub-nanogram/milliliter concentrations.     

Body condition scoring: comparing newly trained scorers and micro-computed tomography imaging

A Body Condition Scoring (BCS) protocol is an easily learned tool that can be used as a means of body condition assessment for random populations of mice. Here, the authors use X-ray computed tomography technology to show that BCS is a quick and effective method for evaluating parameters such as muscle thickness and fat, and that the method is equally as accurate when employed by newly trained or expert scorers.     

The steroid glycoside H.g.-12 from Hoodia gordonii activates the human bitter receptor TAS2R14 and induces CCK release from HuTu-80 cells

Steroid glycosides extracted from the succulent plant Hoodia gordonii are suggested to have appetite-suppressant effects both in animals and humans. Yet, the mechanisms underlying the putative satiety action of Hoodia steroid glycosides are not fully understood. We found that H.g.-12, a steroid glycoside purified from H. gordonii extract, initiated cholecystokinin (CCK) secretion both ex vivo in rat intestine and in vitro in the human enteroendocrine (EC) cell line HuTu-80. CCK is known to exert central effects on appetite suppression via the vagus nerve which afferents terminate in the gut wall. Recent data show that G protein-coupled receptors signaling bitter taste (T2Rs) are expressed in both rodent and human gastrointestinal tract. It was further demonstrated that bitter sensing is functional in mouse STC-1 EC cells and leads to CCK secretion via increased intracellular Ca2(+) concentrations. Based on the bitter taste of H. gordonii purified extracts, we assessed whether H.g.-12 could activate human bitter receptors. The steroid glycoside activated selectively TAS2R7 and TAS2R14, both heterologously expressed in HEK 293 cells. Removing an essential structural feature from the steroid glycoside inhibited H.g.-12-induced Ca2(+) increase in TAS2R14-expressing HEK cells and abolished H.g.-12-induced CCK secretion from human EC cells. Similarly, a nonspecific bitter receptor antagonist abolished H.g.-12-induced CCK secretion in HuTu-80 cells. These results point to a potential route of action by which components of Hoodia might influence appetite control. Our data also provide additional evidence that bitter taste-sensing mechanisms are coupled to hormone release from EC cells in the intestine. Moreover, we identified a natural agonist of TAS2R7 and TAS2R14 for further studies on the role of bitter receptors in satiety control and food intake.