Factors that affect the type of cell death induced by chemicals

Surveying about 1000 compounds, we found that several low molecular weight alpha, beta-unsaturated ketones induced non-apoptotic cell death characterized by the formation of autophagosomes, occasionally accompanied by mitochondrial shrinkage. The cytotoxic activity of these compounds was significantly reduced by the addition of N-acetyl-L-cysteine, suggesting their interaction with SH groups of intracellular targeted molecules (the so-called "non-sterically hindered Michael acceptor"). This suggests that the nature of the chemical structure as well as the type of target cells is another factor that determines the type of cell death induced by chemicals.     

The effect of a 12-week combined exercise intervention program on physical performance and gait kinematics in community-dwelling elderly women

This study aimed to determine if combined exercise intervention improves physical performance and gait joint-kinematics including the joint angle and dynamic range of motion (ROM) related to the risk of falling in community-dwelling elderly women. A 12-week combined exercise intervention program with extra emphasis on balance, muscle strength, and walking ability was designed to improve physical performance and gait. Twenty participants attended approximately two-hour exercise sessions twice weekly for 12 weeks. Participants underwent a physical performance battery, including static balance, sit and reach, whole body reaction time, 10 m obstacle walk, 10 m maximal walk, 30-second chair stand, to determine a physical performance score, and received quantitative gait kinematics measurements at baseline and in 12 weeks. Significant lower extremity strength improvement 13.5% (p<.001) was observed, which was accompanied by significant decreases in time of the 10 m obstacle walk (p<.05) and whole body reaction time (p<.001) in this study. However, no significant differences were seen for static balance and flexibility from baseline. For gait kinematics, in the mid-swing phase, knee and hip joint angle changed toward flexion (p<.01, p<.05, respectively). Ankle dynamic ROM significantly increased (p<.05) following exercise intervention. The plantar flexion angle of the ankle in the toe-off phase was increased significantly (p<.01). However, other gait parameters were not significantly different from baseline. These findings from the present investigation provide evidence of significant improvements in physical performance related to the risk factors of falling and safe gait strategy with a combined exercise intervention program in community-dwelling elderly women. The results suggest this exercise intervention could be an effective approach to ameliorate the risk factors for falls and to promote safer locomotion in elderly community-dwelling women.     

Histidine-44 of the A subunit of Escherichia coli enterotoxin is involved in its enzymatic and biological activities

We examined the role in toxicity of histidine-44 of the A subunit of Escherichia coli enterotoxin, which is located in the active site cavity close to glutamic acid-112. Although amino acid substitution of histidine-44 usually renders a mutant toxin unstable to trypsin, one mutant, alanine-44 (His44Ala) was found to be stable. His44Ala did not show any agmatine:ADP-ribosyltransferase activity in the presence or absence of recombinant ADP-ribosylation factor. It showed no diarrheal or rabbit skin permeability activity and was a competitor in enterotoxin-ADP-ribosyltransferase assays containing recombinant ADP-ribosylation factor. These results suggest that like glutamic acid-112, histidine-44 plays an essential role in toxicity. A tentative model, which explains NAD⁺ catalysis and the transfer of the ADP-ribosyl moiety to a target amino acid, is proposed for histidine-44 and glutamic acid-112.     

A unique DNA sequence of human enterotoxigenic Escherichia coli enterotoxin encoded by chromosomal DNA

We detected Ent plasmids in 300 strains of human enterotoxigenic Escherichia coli, but one strain, E. coli 240-3, had neither a small nor a large plasmid and encoded the heat-labile enterotoxin (LTh(240-3)) gene on its chromosome. DNA sequences showed that LTh(240-3) differed by 12 and 14 base pairs from LT (LTh) and LT (LTp) from human H10407 and porcine EWD299 strains, respectively. In deduced precursor toxins, LTh(240-3), LTh and LTp differed from LTh, LTp and LTh(240-3) at nine, eight and eleven positions, respectively. These data suggest that although LTh(240-3) encoded in the chromosome is antigenically similar to LTh, it cannot be grouped with LTh due to differences in its DNA and amino acids sequences.     

QOL models constructed for the community-dwelling elderly with ikigai (purpose in life) as a composition factor, and the effect of habitual exercise

The purpose of this study was to construct QOL models for the elderly that included ikigai as a composition factor and to clarify differences in two kinds of models, one constructed for the elderly with habitual exercise and the other for those without it.The subjects were 1,566 healthy community-dwelling independent people aged 60 years or more (752 males, 814 females). First, the ratio of subjects with ikigai was calculated. The ratios of subjects with different kinds of objects of ikigai were also calculated. Next, structural equation models (SEM) were constructed on the basis of social, physical, and mental QOL and ikigai. Fits of the models were evaluated. To examine whether the presence or absence of habitual exercise caused any difference in the QOL model, subjects were divided into 4 groups according to whether they were male or female and whether they had or did not have an exercise habit. Multi-population group simultaneous analysis was then performed among the four groups.More than 85% of the subjects had objects of ikigai. Ikigai is an important factor for comprehending the QOL of the elderly. It was possible to construct QOL models for the elderly with ikigai as a composition factor. The effect of physical QOL on mental QOL was negligible in females irrespective of whether they had an exercise habit. The effect of social QOL on mental QOL was profound in aged females with an exercise habit. The effect of the living situation on mental QOL was profound in aged females without an exercise habit. The effect of mental QOL on ikigai was more marked in subjects without an exercise habit than in those with an exercise habit.     

Crystal structure of M-Ras reveals a GTP-bound "off" state conformation of Ras family small GTPases

Although some members of Ras family small GTPases, including M-Ras, share the primary structure of their effector regions with Ras, they exhibit vastly different binding properties to Ras effectors such as c-Raf-1. We have solved the crystal structure of M-Ras in the GDP-bound and guanosine 5'-(beta,gamma-imido)triphosphate (Gpp(NH)p)-bound forms. The overall structure of M-Ras resembles those of H-Ras and Rap2A, except that M-Ras-Gpp(NH)p exhibits a distinctive switch I conformation, which is caused by impaired intramolecular interactions between Thr-45 (corresponding to Thr-35 of H-Ras) of the effector region and the gamma-phosphate of Gpp(NH)p. Previous 31P NMR studies showed that H-Ras-Gpp(NH)p exists in two interconverting conformations, states 1 and 2. Whereas state 2 is a predominant form of H-Ras and corresponds to the "on" conformation found in the complex with effectors, state 1 is thought to represent the "off" conformation, whose tertiary structure remains unknown. 31P NMR analysis shows that free M-Ras-Gpp(NH)p predominantly assumes the state 1 conformation, which undergoes conformational transition to state 2 upon association with c-Raf-1. These results indicate that the solved structure of M-Ras-Gp-p(NH)p corresponds to the state 1 conformation. The predominance of state 1 in M-Ras is likely to account for its weak binding ability to the Ras effectors, suggesting the importance of the tertiary structure factor in small GTPase-effector interaction. Further, the first determination of the state 1 structure provides a molecular basis for developing novel anti-cancer drugs as compounds that hold Ras in the state 1 "off" conformation.     

IFN-gamma-induced SOCS-1 regulates STAT6-dependent eotaxin production triggered by IL-4 and TNF-alpha

The production of eotaxin, which is a critical mediator for airway inflammation, is inhibited by IFN-gamma. Here, we investigated the precise mechanisms underlying IFN-gamma-dependent inhibition of eotaxin production using mouse embryonic fibroblasts (MEF). MEF produced high levels of eotaxin in STAT6-dependent manner when they were cultured with both IL-4 and TNF-alpha. However, the eotaxin production by MEF was strongly inhibited by addition of IFN-gamma. Western-blotting analysis demonstrated that IFN-gamma downmodulated STAT6 phosphorylation induced by IL-4 and TNF-alpha. Moreover, IFN-gamma did not exhibit its inhibitory effect on both STAT6-phosphorylation and eotaxin production in MEF obtained from deficient mice in STAT1, a key molecule of IFN-gamma signaling. We also demonstrated that SOCS-1, a potent inhibitory molecule of IL-4 signaling, was induced by IFN-gamma in STAT1-dependent manner. This indicated that SOCS-1 might be involved in IFN-gamma-mediated STAT1-dependent inhibition of eotaxin production. In SOCS-1(-/-) MEF, IFN-gamma inhibited neither STAT6 phosphorylation nor eotaxin production induced by IL-4 and TNF-alpha. Conversely, retroviral transduction of SOCS-1 into MEF inhibited STAT6 phosphorylation and eotaxin production induced by IL-4 and TNF-alpha, in the absence of IFN-gamma. Thus, we demonstrated that IFN-gamma-induced inhibition of STAT6 phosphorylation and eotaxin production were mediated by SOCS-1 induced in STAT1-dependent manner.     

Selective accumulation and growth inhibition of hybrid liposomes to human hepatocellular carcinoma cells in relation to fluidity of plasma membranes

Hybrid liposomes (HLs), composed of l-α-dimyristoylphosphatidylcholine (DMPC) and polyoxyethylene(23) dodecyl ether, have selectively inhibited the growth of human hepatocellular carcinoma (HCC) cells without affecting normal hepatocytes to trigger apoptosis via caspase-3 activation. Furthermore, HLs distinguished between the HCC and normal cells which had higher and lower membrane fluidities respectively, then fused and accumulated preferentially into the membranes of HCC cells. It is noteworthy that the anti-cancer activity of HLs correlated well with the fluidity of cell membranes for HCC and other cancer cells. These results suggest that HLs could target cancer cell-membranes in relation to their lipid fluidity that provide the possibility of novel nanotherapy for intractable cancer.     

Replacing the cyclohexene-linker of FR181157 leading to novel IP receptor agonists: orally active prostacyclin mimetics. Part 6

The synthesis and biological activity of novel derivatives of our previously reported IP receptor agonist FR181157 is described. SAR studies to replace the cyclohexene-linker of FR181157 led to the discovery of compound 1i (FR207845) as a potent non-prostanoid PGI2 mimetic with good oral bioavailability.