Cloning and characterization of a NADP+-malic enzyme gene from Bradyrhizobium japonicum USDA110

Malic enzymes have been considered to play a key role in energy metabolism for nitrogenase reaction in bacteroids. To elucidate the physiological role of the malic enzymes in Bradyrhizobium japonicum bacteroids, a putative malic enzyme gene Bjtme1 was cloned by polymerase chain reaction (PCR) using degenerated primers from conserved regions of the protein sequences of bacterial malic enzymes and draft sequence data of the Bradyrhizobium japonicum USDA110 genome sequence project. To confirm the characteristics of the Bjtme1 gene, the protein encoded by this gene was over-expressed using a pET32a(+) system and it exhibited a NADP⁺-malic enzyme (EC 1.1.1.40) activity, indicating that Bjtme1 was the gene of the NADP⁺-malic enzyme. This is the first report on the cloning and characterization of the NADP⁺-malic enzyme gene from B. japonicum, and the gene structure was compared with that of NADP⁺-malic enzyme genes of other rhizobia.     

Malignant Peritoneal Mesothelioma with a Sarcomatoid Growth Pattern and Signet-Ring-Like Structure in a Female F344 Rat

We report a biphasic malignant mesothelioma in an aged female F344/DuCrlCrlj rat. Macroscopically, multiple pale brown nodules were observed in the abdominal cavity with retention of bloody ascites. Histopathologically, the tumor cells spread over the peritoneum and formed masses on the surface and underlying adipose tissues. The tumor cells dominantly proliferated in a solid, nodular or nest-like pattern with modest amount of fibrillar connective tissues, which contained hyaluronan. The tumor consisted of ovoid, polygonal or spindle-shaped cells that possessed eosinophilic cytoplasms including glycogen; some tumor cells showed a signet-ring-like structure. Multinucleated cells and mitosis were found frequently, and direct invasion to intra-abdominal organs and intravascular metastasis to the liver were observed. Immunohistochemically, keratin and mesothelin were strongly positive in most of tumor cells, while vimentin was mainly positive in spindle-shaped cells. Podoplanin was also positive, particularly in the cell membrane of tumor cells. Electron microscopically, tumor cells showed an intercellular desmosome-like structure, basement membrane and microvillus. We diagnosed the case as a malignant peritoneal mesothelioma with a sarcomatoid growth pattern and signet-ring-like structure.     

The importance of riparian subtropical lucidophyllous forest to odonate conservation

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Exogenous leptin advances puberty in domestic hen

The present study was undertaken to examine the effect of recombinant chicken leptin administered to fed ad libitum and feed-restricted immature chickens of a layer strain on ovarian development and the timing of sexual maturity. In the first experiment 11-week-old pullets (77 days of age) fed ad libitum were injected daily with leptin at four dose levels (4, 16, 64 and 256 microg/kg body weight) until sexual maturity (lay of the first egg). Leptin treatment at the highest dose significantly (P<0.05) advanced the onset of puberty (day 116.3+/-1.0) in comparison to controls (day 121.3+/-1.2). The rises of luteinizing hormone, estradiol and progesterone in blood plasma were also advanced by leptin treatment. In the second experiment, both full-fed and feed-restricted pullets (79 days of age) were injected daily with leptin (256 microg/kg body weight). In birds fed ad libitum, exogenous leptin again significantly (P<0.05) advanced first ovipostion (day 118.4+/-1.4 versus day 124.4+/-1.7), while abolishing the significant (P<0.05) delay caused by feed restriction (day 131.5+/-1.6) and restoring the normal onset of sexual maturity (day 125.7+/-1.6). Analysis of the ovaries in 106-day-old pullets revealed that leptin injections advanced follicular development, particularly in birds fed ad libitum, and significantly (P<0.01) reduced follicular apoptosis both in full-fed and feed-restricted birds. In conclusion, we have shown that in female chickens exogenous leptin advances the onset of puberty by attenuation of ovarian apoptosis and enhancement of folliculogenesis.     

Effect of nitrogen application at the flowering stage on the quality of soybean seeds

Application of nitrogen (N) fertilizer at the flowering stage changed the contents of storage compounds in seeds of Soybean (Glycine max L. cvs Enrei and Tamahomare). The effects of the N application on the maturation of soybean seeds were examined by comparing changes in the contents of amino acids, sugars, water, protein, and oil in seeds from N‐dressed plants (NDS) with those from undressed plants (UDS) during maturation. The application resulted in a decrease of contents of total and some amino acids (glutamine and asparagine) in developing seeds except for at the early maturation stage and in a decreased protein content of mature seeds. On the other hand, the N application led to faster accumulation of oil in developing seeds and to an increased oil content of mature seeds. Based on these results, it was concluded that the N application at the flowering stage changed the composition of solutes imported by developing seeds and resulted in variations in the contents of storage compounds. The results indicate that it is possible to improve seed quality by fine control of N application.