Synthesis and anti-influenza virus evaluation of triterpene-sialic acid conjugates

We are interested in new non-natural glycosides with sialic acid conjugates and their biological activities. We report the synthesis of eleven non-natural occurring glycosides, which are triterpene (glycyrrhetinic acid and its derivatives)-sialic acid conjugates, and their inhibitory activities against influenza virus sialidases and influenza virus multiplication in MDCK host cells. Deoxoglycyrrhetol-sialic acid conjugates (6d and 6e) and oleanolic acid-sialic acid conjugates (7d and 7e) showed strong inhibitory activities against three subtypes of influenza virus sialidases. These four compounds (6d, 6e, 7d and 7e) showed clear inhibition to influenza virus multiplication but not to MDCK host cell survival.     

Discovery of anti-influenza A virus activity of a corynanthe-type indole alkaloid, hirsutine, in vitro and the structure-activity relationship of natural and synthetic analogs

An indole alkaloid, hirsutine (1), was found to exhibit potent inhibitory effect against influenza A virus in vitro, and the essential structural feature for revealing the activity was elucidated by study of the structure-activity relationship using natural and synthetic derivatives of 1.     

A histochemical study on the correlation between secretory activity and the TCA cycle in the gland cell

Summary The activity of succinic dehydrogenase and malic dehydrogenase was observed histochemically in the gland stomach of rats, and also the relationship between the secretory activity of the gastric gland cells and the process of the TCA cycle in the cells was studied.Histochemically, enzyme activity is plainly visible in the gastric parietal cells but in the gastric chief cells and mucous neck cells.The secretory activity of the cells was promoted by the administration of food, the sub-cutaneous injection of histamine, histidine, acetylcholine or eserin.The activity of succinic dehydrogenase appears to be constant regardless of secretory activity except in a few cases. The activity of malic dehydrogenase increases as secretory activity is promoted. It seems very unlikely that one step in the cycle (the transformation of malic acid into oxalacetic acid) would be accelerated while the other step (the transformation of succinic acid into fumaric acid) is not. This inconsistency of activity may be attributed to the histochemical reaction. Thus the increase of malic dehydrogenase activity is seen as an acceleration of the whole TCA cycle. It is our conclusion, therefore, that the source of energy within the cell, i.e. the TCA cycle, is a process which parallels secretory activity.     

Production of acetic acid by Clostridium thermoaceticum in batch and continuous fermentations

Batch and continuous fermentations with Clostridium thermoaceticum (ATCC 39073) using automatic pH control were conducted. The value of mu(max) obtained from batch fermentation was about 0.14 h(-1); acetate yield, which was both growth and non-growth associated, was about 2 mole of acetic acid/mole of glucose, compared with a theoretical maximum value of 3. This low yield, compared with literature data, may be explained by glucose loss through a combination of degradation routes. Continuous fermentation could be sustained for 1600 h or more without contamination problems. Continuous fermentation at high dilution rates indicates that mu(max) may be well above 0.17 h(-1) when fresh feed medium is used. Acetate yields in continuous fermentation were about 77% of theoretical or 2.3 mole of acetic acid/mole of glucose.     

Photoperiodic induction of diapause in normal and allatectomized precocious pupae of Papilio machaon

Diapause in the swallowtail, Papilio machaon occurs at the pupal stage in response to short days during larval development. The precocious pupae obtained by allatectomy also entered diapause in response to short days. The photoperiodic induction of diapause in precocious pupae was very similar to that in normal pupae in spite of the abbreviation of the last-larval instar. The critical photoperiod for diapause in the precocious pupae was 14.5 h, being about the same as that in normal pupae. Occurrence of diapause in the swallowtail is closely associated with the pupal stage and the induction process may be triggered by the cessation of juvenile hormone secretion.     

Apodemus sylvaticus, a new host for Acanthocheilonema viteae (Nematoda: filarioidea)

Susceptibility of Apodemus sylvaticus and A. agrarius to infection with Acanthocheilonema viteae was compared with that of hamsters and jirds. Microfilaremia in A. sylvaticus was first noted on day 52 post-infection (p.i.) and lasted during the course of the study (up to day 150 p.i.). Maximum microfilaremic levels (female worm basis) of A. sylvaticus [mean +/- S.D. (n) = 690 +/- 1288(6)] were considerably higher than those of hamsters [16 +/- 18(6)] and jirds [51 +/- 25(5)]. Adult worm recovery in A. sylvaticus ranged from 2 to 40% of the number of infective larvae inoculated. Worm development in A. sylvaticus resembled that in hamsters and jirds. In contrast, microfilaremia was not detected in, nor adult worms recovered from A. agrarius throughout the study.     

Interactions between cytoskeletal components during myoplasm rearrangement in ascidian eggs

Ooplasmic segregation in ascidian eggs consists of two phases of cytoplasmic movement, the first phase is mediated by the microfilament system and the second is mediated by the microtubule system. Recently, two novel proteins, p58 and myoplasmin-C1, which are localized to the myoplasm, were suggested to have important roles in muscle differentiation. In order to analyze the molecular mechanisms underlying ooplasmic segregation, the interactions between actin, tubulin, p58 and myoplasmin-C1 were examined. During the first segregation, microtubule meshwork in the unfertilized egg disappeared. At the second segregation, a novel structure of the microtubules that extended from the sperm aster and localized in the cortical region of the myoplasm was found. Moreover, uniform distribution of the cortical actin filament was observed at the second segregation. During the course of myoplasm rearrangement, p58 and myoplasmin-C1 are colocalized and can form a molecular complex in vitro. This complex of p58 and myoplasmin-C1 is a good candidate for a cytoskeletal component of the myoplasm, and is likely to be involved in the correct distribution of cytoplasmic determinants.     

Differential Expression of NeuroD in Primary Cultures of Cerebral Cortical Neurons

We have investigated the expression patterns of a basic helix–loop–helix regulatory gene,neuroD,in primary cultures of murine cerebral cortical neurons. The differentiation states of neurons in primary cultures were determined by the sensitivity of neurons to glutamate toxicity and the expression of specific proteins such as the phosphorylated form of a 200-kDa neurofilament, HPC-1/syntaxin 1A, and cell adhesion molecule L1. The expression of neuroD was determined by RT-PCR analysis andin situhybridization. The experimental results thus obtained revealed that neuronal maturation is initiated between Day 7 and Day 11 in the culture as already known, and that the expression of neuroD decreases with increasing days in culture. Based on these findings, it was concluded that neuroD is expressed in immature neurons but not in mature ones.     

Fenofibrate reduces cisplatin-induced apoptosis by inhibiting the p53/Puma/Caspase-9 pathway and the MAPK/Caspase-8 pathway rather than by promoting autophagy in murine renal proximal tubular cells

The main lesion of cisplatin nephrotoxicity is damage to proximal tubular cells due to increased apoptosis via the mitochondrial and death receptor pathways, which may be alleviated by appropriate promotion of autophagy. Fenofibrate, a peroxisome proliferator-activated receptor-alpha (PPAR-α) activator, is recently reported to promote autophagy as well as protect against cisplatin nephrotoxicity, although the mechanisms were only partially analyzed. Here, the detailed mechanisms of these putative protective effects were investigated in a murine renal proximal tubular (mProx) cell line. Fenofibrate attenuated cisplatin-induced apoptosis of mProx cells based on flow cytometry. As for the mitochondrial apoptotic pathway, the reagent reduced cisplatin-stimulated caspase-3 activation by decreasing the phosphorylation of p53, JNK, and 14-3-3, cytosolic and mitochondrial Puma accumulation, cytochrome C release to the cytosol, and resulting cytosolic caspase-9 activation. Fenofibrate also decreased cisplatin-stimulated activation of caspases-8 by suppressing MAPK and NFkB pathways and reducing the gene expression of TNF-α, TL1A, and Fas, main mediators of the death receptor apoptotic pathway. Autophagy defined by p62 reduction and an increase in LC3 II/I was promoted by fenofibrate in mProx cells under starvation. Autophagy inhibition using 3-MA further increased basal and cisplatin-induced caspase-3 and -8 activation, but had no influence on the inhibitory effects of fenofibrate on caspase activation. In conclusion, our study suggests fenofibrate to be a candidate agent to mitigate cisplatin nephrotoxicity by inhibiting the mitochondrial and death apoptotic pathways rather than by promoting autophagy.