How a DNA polymerase clamp loader opens a sliding clamp

Processive chromosomal replication relies on sliding DNA clamps, which are loaded onto DNA by pentameric clamp loader complexes belonging to the AAA+ family of adenosine triphosphatases (ATPases). We present structures for the ATP-bound state of the clamp loader complex from bacteriophage T4, bound to an open clamp and primer-template DNA. The clamp loader traps a spiral conformation of the open clamp so that both the loader and the clamp match the helical symmetry of DNA. One structure reveals that ATP has been hydrolyzed in one subunit and suggests that clamp closure and ejection of the loader involves disruption of the ATP-dependent match in symmetry. The structures explain how synergy among the loader, the clamp, and DNA can trigger ATP hydrolysis and release of the closed clamp on DNA.     

1 -Antitrypsin deficiency: a variant with no detectable 1 -antitrypsin

No alpha(1)-antitrypsin could be detected in the serum of a 24-year-old man with advanced pulmonary emphysema by agarose electrophoresis, immnuno-electrophoresis, double diffusion in agarose gel, or alpha(1)-antitrypsin genetic typing by a combination of starch-gel electrophoresis and crossed antigen-antibody electrophoresis. A circulating alpha(1)-antitrypsin inactivator could not be demonstrated. Evidence was obtained in family members of genetic transmission of this new alpha(1)-antitrypsin variant.     

Starch degradation by alpha-amylase in tobacco leaves during the curing process

Tobacco (Nicotiana tabacum L.) leaf starch is degraded to sugars through curing (42°C/82.3% relative humidity/72 h). Total carbohydrate content remained almost constant, starch content decreased markedly, and soluble sugar content (mostly glucose) increased. α-Amylase and starch phosphorylase activities increased sixfold and threefold, respectively, whereas β-amylase activity was unaltered and isoamylase activity decreased. Increased α-amylase activity was accompanied by increased α-amylase protein levels. Although tobacco has four α-amylase gene members, only NtAMY1 mRNA levels increased. For other starch degradation genes, such as NtBAM1 and NtBMY2 (β-amylase), NtISO1 and NtISO2 (isoamylase) and NtGWD1 and NtGWD3 (glucan water dikinase), the mRNA levels remained unaltered during the first 48 h of curing. NtAMY1 expression was induced by osmotic stress but was unaffected by high temperature and/or injury stresses. Similarly, soluble sugar contents were largely increased by osmotic stress. This suggests that starch is degraded by α-amylase during curing and that α-amylase is coded by NtAMY1, induced by osmotic stress.     

Thioacetamide-induced Hepatocellular Necrosis Is Attenuated in Diet-induced Obese Mice

To assess modification of thioacetamide-induced hepatotoxicity in mice fed a high-fat diet, male C57BL/6J mice were fed a normal rodent diet or a high-fat diet for 8 weeks and then treated once intraperitoneally with thioacetamide at 50 mg/kg body weight. At 24 and 48 hours after administration, massive centrilobular hepatocellular necrosis was observed in mice fed the normal rodent diet, while the necrosis was less severe in mice fed the high-fat diet. In contrast, severe swelling of hepatocytes was observed in mice fed the high-fat diet. In addition, mice fed the high-fat diet displayed more than a 4-fold higher number of BrdU-positive hepatocytes compared with mice fed the normal rodent diet at 48 hours after thioacetamide treatment. To clarify the mechanisms by which the hepatic necrosis was attenuated, we investigated exposure to thioacetamide and one of its metabolites, the expression of CYP2E1, which converts thioacetamide to reactive metabolites, and the content of glutathione S-transferases in the liver. However, the reduced hepatocellular necrosis noted in mice fed the high-fat diet could not be explained by the differences in exposure to thioacetamide or thioacetamide sulfoxide or by differences in the expression of drug-metabolizing enzymes. On the other hand, at 8 hours after thioacetamide administration, hepatic total glutathione in mice fed the high-fat diet was significantly lower than that in mice fed the normal diet. Hence, decreased hepatic glutathione amount is a candidate for the mechanism of the attenuated necrosis. In conclusion, this study revealed that thioacetamide-induced hepatic necrosis was attenuated in mice fed the high-fat diet.     

Steroid 5α-reductase inhibitory activity of condensed tannins from woody plants

In humans, steroid 5α-reductase is involved in the development of benign prostatic hyperplasia (BPH). We tested the steroid 5α-reductase inhibitory activity of the 70% acetone extract of woody plants. The tannin polymer prepared from the 70% acetone extract showed the highest steroid 5α-reductase inhibitory activity. Furthermore, the steroid 5α-reductase inhibitory activity of tannins was not affected by the addition of superoxide dismutase or catalase. It was concluded that the steroid 5α-reductase inhibitory activity of condensed tannin was caused by binding to the steroid 5α-reductase, rather than from the peroxide/superoxide produced by tannins. The tannins, prepared from tree barks, with potential for steroid 5α-reductase inhibitory activity might be advantageous in therapy for steroid 5α-reductase diseases such as BPH or prostate cancer. Part of this report was presented at the 54th Annual Meeting of the Japan Wood Research Society, Sapporo, August 2004     

Supplementing transglucosidase with a high-fiber diet for prevention of postprandial hyperglycemia in streptozotocin-induced diabetic dogs

Indigestible oligosaccharides have been shown to normalize blood glucose and insulin concentration thereby promoting good health and preventing diseases, such as diabetes. Transglucosidase (TG, α-glucosidase, enzyme code (EC) 3.2.1.20) is an enzyme capable of converting starch to oligosaccharides, such as iso-malto-oligosaccharides from maltose, via the action of amylase. The aim of this study was to evaluate whether oral administration of TG with maltose or dextrin is capable of reducing post-prandial serum glucose concentration in experimentally streptozotocin (STZ)-induced diabetic dogs fed on a high-fiber diet. Five healthy and five STZ-induced diabetic dogs were employed in this study. TG supplementation with dextrin or maltose had no detrimental effect in healthy dogs. In fact, TG and dextrin exhibited a flatlined serum glucose pattern, while reducing mean post-prandial serum insulin and glucose concentration as compared to control diet alone. When TG supplementation was tested in STZ-induced diabetic dogs under the context of a high fiber diet, a 13.8% and 23.9% reduction in mean glucose concentration for TG with maltose and dextrin, respectively was observed. Moreover, TG with dextrin resulted in a 13% lower mean post-prandial glucose concentration than TG with maltose, suggesting that dextrin may be a more efficient substrate than maltose when used at the same concentration (1 g/kg). Our results indicate that TG supplementation with diet can lead to lower postprandial glucose levels versus diet alone. However, the efficacy of TG supplementation may depend on the type of diet it is supplemented with. As such, TG administration may be useful for preventing the progression of diabetes mellitus and in its management in dogs.     

Bioeconomic assessment of size separators in Pacific saury fishery

In the middle of the 1990s, Pacific saury fishery vessels began to install “size separators” to selectively land large-size class fish with a higher price. Contrary to expectations, this resulted in removal of the separators in 2006 because fishers believed the separators had contributed to price collapses in the 2000s. The intent of this paper is to investigate the effects of separators on both the fishery economy and stock of Pacific saury by simulating population and economic models under a single framework. For this purpose, we developed (1) an age-structured population dynamics model with stochasticity, and (2) an economic model spanning both price and inventory dynamics with stochasticity, in which each set of model parameters were estimated on the basis of time series data. In a 10-year simulation, the harvest quota was set constant from 20,000 to 400,000 t at intervals of 20,000 t, and the effects of separators were incorporated by controlling the catchability of 0-year-old fish. We found that separators increase the expected yield and reduce the deficit risk for harvests of 140,000 t and smaller. However, separators have the opposite effects for harvests of 160,000 t and larger.     

Attenuation of inorganic arsenic and cadmium in rice grains using by-product iron materials from the casting industry combined with different water management practices

We examined the effect of two types of iron (Fe) material produced by the casting industry (spent steel shot [SSS] and residual iron material from steel shot production) on the mobility of arsenic (As) and cadmium (Cd) in soils. We also examined the uptake of these elements by rice plants (Oryza Sativa L.) under continuously flooded (CF) and water-saving (WS) cultivation. The application of both Fe materials (at 10 and 30 t ha^?1) strictly limited As mobilization in soils under CF cultivation. As a result, As uptake by rice plants declined, along with the total and inorganic As (iAs) concentration in rice grains. In comparison, As immobilization caused by the application of Fe material was less clear under WS cultivation. The rate of Fe material application was negatively correlated with As uptake by rice plants. It was also negatively correlated with total and iAs concentration in rice grains under both water management practices. The combination of applying Fe materials and WS cultivation decreased iAs concentration in rice grains to approximately one-fifth of that in rice grains produced from plants grown on soils without Fe material application under CF cultivation. CF cultivation strictly decreased dissolved Cd in soils, as well as Cd in rice grains with and without Fe material application. The application of Fe materials decreased Cd mobility and, hence, Cd uptake in rice plants, ultimately reducing the accumulation of Cd in rice grains under WS cultivation. Residual Fe material had a statistically greater effect at attenuating Cd accumulation in rice grains than SSS. The present study demonstrated the potential of combining by-product Fe material application and water management practices to attenuate iAs and/or Cd concentrations in rice grains. Practical countermeasures should be carefully adopted that consider the existing risks of iAs and Cd on each paddy field, and the combined effect of Fe material application and water management practices.