Effects of NAD or NADP on the stability of liver and pectoral muscle enzymes in 3-acetylpyridine treated quail by heat and trypsin

(1) The effects of long term treatment with 3-acetylpyridine on the stability of enzymes towards heat and trypsin treatment were studied. (2) In the liver NAD or NADP provided a similar degree of protection against heat inactivation at 55 degrees C for 6-phosphogluconate dehydrogenase (24%), glyceraldehyde-3-phosphate dehydrogenase (24%) and malic enzyme (20%), low level of protection of lactate dehydrogenase (13%) but didn't affect acetylcholinesterase at all. In the muscle, however, there was substantial protection against heat inactivation by coenzyme of glyceraldehyde-3-phosphate dehydrogenase (52%), an intermediate level of protection of lactate dehydrogenase (25%), low level of protection of 6-phosphogluconate dehydrogenase (17%) and malic enzyme (17%) and almost no protection of acetylcholinesterase. (3) In the susceptibility towards trypsin a low but similar degree of protection for dehydrogenases by coenzymes was observed in the liver whereas in the muscle there was substantial protection against trypsin inactivation by NAD of glyceraldehyde-3-phosphate dehydrogenase, an intermediate level of protection of 6-phosphogluconate dehydrogenase and malic enzyme and very little protection of lactate dehydrogenase but no protection of acetylcholinesterase. Among enzymes tested, glyceraldehyde-3-phosphate dehydrogenase showed the greatest protection against heat and trypsin inactivation by NAD. (4) The results suggest that the effect of 3-acetylpyridine treatment on the stability of muscle glyceraldehyde-3-phosphate dehydrogenase appears to be quite specific and selective.     

Plasma membrane cholesterol is a key molecule in shear stress-dependent activation of extracellular signal-regulated kinase

Shear stress, the dragging force generated by fluid flow, differentially activates extracellular signal-regulated kinase (ERK) and c-Jun NH2-terminal kinase (JNK) in bovine aortic endothelial cells (BAEC) (Jo, H., Sipos, K., Go, Y. M., Law, R., Rong, J., and McDonald, J. M. (1997) J. Biol. Chem. 272, 1395-1401). Here, we examine whether cholesterol-enriched compartments in the plasma membrane are responsible for such differential regulation. Pretreatment of BAEC with a cholesterol-binding antibiotic, filipin, did not inhibit shear-dependent activation of JNK. In contrast, filipin and other membrane-permeable cholesterol-binding agents (digitonin and nystatin), but not the lipid-binding agent xylazine, inhibited shear-dependent activation of ERK. The effect of cholesterol-binding drugs did not appear to be due to membrane permeabilization, since treatment of BAEC with a detergent, Triton X-100 which also permeabilizes membranes, did not inhibit shear-dependent activation of ERK. Furthermore, shear-dependent activation of ERK, but not JNK, was inhibited by cyclodextrin, a membrane-impermeable cholesterol-binding agent, which removes cell-surface cholesterol. Moreover, the effects of cyclodextrin were prevented by adding cholesterol during the incubation. These results indicate that cholesterol or cholesterol-sensitive compartments in the plasma membrane play a selective and essential role in activation of ERK, but not JNK, by shear stress. Although exposure to shear stress (1 h) increased the number of caveolae by 3-fold, treatment with filipin had no effect in either control or shear-exposed cells suggesting that caveolae density per se is not a crucial determinant in shear-dependent ERK activation. In summary, the current study suggests that cholesterol-sensitive microdomains in the plasma membrane, such as caveolae-like domains, play a critical role in differential activation of ERK and JNK by shear stress.     

Sleep in relation to age, sex, and chronotype in Japanese workers

The Morningness-Eveningness Questionnaire and Life Habits Inventory were administered to 622 Japanese workers matched for sex and age. We investigated the distributions of the scores on the Morningness-Eveningness Questionnaire and sleep-wake habits by age and sex. Subjects were classified into five age groups and three chronotypes. The distributions and mean scores on the questionnaire advanced slightly toward the Morning type from the young to the aged group. The habitual bedtimes and waking times were significantly earlier in all the chronotypes from the young to the aged group, and the preferred bedtimes and waking times were also clearly earlier from the young to the aged group. The length of sleep was shorter for the Evening than the Morning types, especially in the group below 24 yr. The differences in habitual and preferred sleep length were greater than 1 hour for all age groups, especially the two groups under 34 yr. The number of awakenings during night sleep increased from the young to the aged group for all chronotypes. The older Evening type tended more toward frequent napping and longer naptime. The variabilities of bedtime and sleep length were larger for the young and Evening type than for the old group and Morning types. Further, the mood upon waking and satisfaction with sleep length were better in the aged Evening type than the young Morning type. The women under 44 yr. woke up earlier than the men of the same age, and the women of the 35-54 yr. groups had a shorter length of sleep than others. These may be related to childcare and housework. These results indicated that the phase of circadian rhythms had moved forward from the young to the aged group, and the individual's rhythm, of those that were aged Morning types, showed better agreement with sleep-wake rhythms than did others.     

Tumor cell cytotoxicity of a novel metal chelator

We have synthesized a novel six-coordinate metal chelator from the triamine cis-1,3,5-triaminocyclohexane by the addition of a 2-pyridylmethyl pendant arm on each nitrogen, which we term tachpyr. The experiments described here were designed to explore whether this compound exhibits potential antitumor activity. When added to MBT2 or T24 cultured bladder cancer cells, tachpyr was profoundly cytotoxic, with an IC50 of approximately 4.6 micromol/L compared with 70 micromol/L for desferioxamine. To explore the mode of action of tachpyr, several metal complexes were prepared, including Fe(II), Ca(II), Mn(II), Mg(II), Cu(II), and Zn(II) tachpyr complexes. Of these, the Zn(II), Cu(II), and Fe(II) complexes were without toxic effect, whereas the Ca(II), Mn(II), and Mg(II) complexes remained cytotoxic. To further probe the role of Zn(II) and Cu(II) chelation in the cytotoxicity of tachpyr, sterically hindered tachpyr derivatives were prepared through N-alkylation of tachpyr. These derivatives were unable to strongly bind Fe(III) or Fe(II) but were able to bind Zn(II) and Cu(II). When added to cells, these sterically hindered tachpyr derivatives were nontoxic, consistent with a role of iron depletion in the cytotoxic mechanism of tachpyr. Further, the addition of tachpyr to proliferating cultures resulted in an early and selective inhibition of ferritin synthesis, an iron storage protein whose translation is critically dependent on intracellular iron pools. Taken together, these experiments suggest that tachpyr is a cytotoxic metal chelator that targets intracellular iron, and that the use of tachpyr in cancer therapy deserves further exploration.     

Synthesis and comparative molecular field analysis (CoMFA) of antitumor 3-arylisoquinoline derivatives

In this study a series of 3-arylisoquinoline derivatives were synthesized and cytotoxicity against human melanoma tumor cell evaluated, and a three dimensional quantitative structure-activity relationship was investigated using the comparative molecular field analysis (CoMFA). The results suggested that the electrostatic, steric and hydrophobic factors of 3-arylisoquinolines were strongly correlated with the antitumor activity. Considerable predictive ability (cross-validated r2 as high as 0.721) was obtained through CoMFA.     

PilT mutations lead to simultaneous defects in competence for natural transformation and twitching motility in piliated Neisseria gonorrhoeae

Neisseria gonorrhoeae, the Gram-negative aetiological agent of gonorrhoeae, is one of many mucosal pathogens of man that expresses competence for natural transformation. Expression of this phenotype by gonococci appears to rely on the expression of type IV pili (Tfp), but the mechanistic basis for this relationship remains unknown. During studies of gonococcal pilus biogenesis, a homologue of the PilT family of proteins, required for Tfp-dependent twitching motility in Pseudomonas aeruginosa and social gliding motility in Myxococcus xanthus, was discovered. Like the findings in these other species, we show here that gonococcal PilT mutants constructed in vitro no longer display twitching motility. In addition, we demonstrate that they have concurrently lost the ability to undergo natural transformation, despite the expression of structurally and morphologically normal Tpf. These results were confirmed by the findings that two classes of spontaneous mutants that failed to express twitching motility and transformability carried mutations in PilT. Piliated PilT mutants and a panel of pilus assembly mutants were found to be deficient in sequence-specific DNA uptake into the cell, the earliest demonstrable step in neisserial competence. The PilT-deficient strains represent the first genetically defined mutants that are defective in DNA uptake but retain Tfp expression.     

Service-oriented smart home applications: composition, code generation, deployment, and execution

A smart home usually has a variety of devices or home appliance, instead of designing software for a specific home, this paper proposes a service-oriented framework with a set of ontology systems to support service and device publishing, discovery of devices and their services, composition of control software using existing control services that wrap devices, deployment, and execution of the composed service in an computing environment, monitoring the execution, and recovery from device failure. The ontology systems specify semantic information about devices, services, and workflows used in various smart home, and users can compose and recompose services for their specific needs. New devices, workflows, and services can be added into ontology. Most of the steps in this process can be automated including code generation. For example, service composition will be carried out in three steps: abstract workflow design, function construction, and device discovery, and different codes can be generated for different computing platforms such as Java and Open Services Gateway initiative environments. In this way, a variety of smart home can be constructed rapidly using the framework by discovery and composition using existing services and workflows. This paper illustrates this framework using a media control example to illustrate the ontology, discovery, composition, deployment, execution, monitoring, and recovery.     

Interaction behavior of nitrogen in liquid niobium

The kinetics and mechanism of absorption/desorption of nitrogen in liquid Nb were investigated in the temperature range of 2470 °C to 2670 °C in samples levitated in a N₂/Ar stream with various nitrogen partial pressures. The nitrogen solution reaction in liquid Nb was found to be exothermic, with the standard enthalpy and entropy of solution of −236.4 ± 23.3 kJ/mol and −-5.3 ± 8.3 J/K · mol, respectively. Above the threshold flow rate of the N₂/Ar stream, the absorption process was determined to be second order with respect to nitrogen concentration, indicating that the rate-controlling step is either the adsorption of nitrogen molecules on the liquid surface or dissociation of adsorbed nitrogen molecules into surface-adsorbed atoms. The desorption process was found to be second order as well. At lower flow rates, however, the absorption rate was found to depend on the gas-phase mass transfer rate. The rate equation for nitrogen absorption in the range of 2470 °C to 2670 °C is given by with the value ofQ calculated to be −327.2 ± 20.6 kJ/mol, while nitrogen desorption at 2670 °C follows the relation      

Simple systolic arrays for discrete cosine transform

in this paper, simple 1-D and 2-D systolic array for realizing the discrete cosine transform (DCT) based on the discrete Fourier transform (DFT) fo an input sequence are presented. The proposed arrays are obtained by a simple modified DFT (MDFT) and an inverse DFT (IDFT) version of the Goertzel algorithm combined with Kung's approach. The 1-D array requiresN cells, one multiplier and takesN clock cycles to produce a completeN-point DCT. The 2-D array takes N clock cycles, faster than the 1-D array, but the area complexity is larger. A continuous flow of input data is allowed and no idle time is required between the input sequences.