Expression of zebrafish btg-b, an anti-proliferative cofactor, during early embryogenesis

BTG/tob family proteins are thought to be a potential tumor suppressor due to their anti-proliferative activity. We cloned zebrafish btg-b, a member of the BTG1/2 subfamily, using in situ hybridization screening. The tissue-specific expression of btg-b is first observed in the organizer region at the early gastrula stage. Later in development, the forebrain, the hindbrain, the polster and the paraxial mesoderm transiently express btg-b. Recently, mouse Btg1 and Btg2 have been shown to be a cofactor for Hoxb9. Double in situ hybridization with zebrafish btg-b and hoxb9a indicates that the expression domains of these two genes overlap in the posterior paraxial mesoderm.     

Microcalorimetric study of iodized and noniodized cells and C-phycocyanin of Spirulina platensis

It was shown that eight stages of transition are observed in the heating process of Spirulina platensis cells in temperature range 5-140 degrees C. The first stage covers the temperature range 5-53 degrees C with maximum approximately 45 degrees C. The heat evolved in this temperature range is equal to 380 +/- 20 J/g of dry biomass, it does not change at scanning rate lower than 0.083 degrees C/min and belongs, mainly, to cell respiration in a stationary regime, in the dark. It was shown that endotherm approximately 66 degrees C belongs to denaturation of C-phycocyanin which denaturates in solutions with Td = 64.2 degrees C, deltaHd = 34.7 +/- 2.1 J/g and for it deltaHd(cal)/deltaH(V.H) is equal to 10.8 +/- 1.2. The endotherms with Td equal to 58 and 88 degrees C are connected with denaturation of phycobilisome proteins and endotherm with Td = 48 degrees C and deltaHd = 4.2J/g of dry biomass-with denaturation of protein which, apparently, is connected with cell respiration.     

OsSEND-1: a new RAD2 nuclease family member in higher plants

A novel endonuclease, a new member of the RAD2 nuclease family, has been identified from the higher plant, rice (Oryza sativa L. cv. Nipponbare), and designated as OsSEND-1. The open reading frame of the OsSEND-1 cDNA encoded a predicted product of 641 amino acid residues with a molecular weight of 69.9 kDa. The encoded protein showed a relatively high degree of sequence homology with the RAD2 nuclease family proteins, especially RAD2 nuclease, but it differed markedly from FEN-1, XPG or HEX1/EXO1. The N- and I-domains in the family were highly conserved in the OsSEND-1 sequence. The protein was much smaller than XPG, but larger than HEX1/EXO1 and FEN-1. The genome sequence was composed of 14 exons, and was localized at the almost terminal region of the short arm of chromosome 8. Northern blotting and in situ hybridization analyses demonstrated preferential expression of OsSEND-1 mRNA in proliferating tissues such as meristem. The mRNA level of OsSEND-1 was induced by UV and DNA-damaging agent such as MMS or H₂O₂, indicating that OsSEND-1 has some roles in the repair of many types of damaged DNA. The recombinant peptide showed endonuclease activity.     

Requirement of negative residues,Asp 95 and Asp 105, in S2 on membrane integration of a voltage-dependent K+ channel,KAT1

Voltage-dependent K+ channels consist of a voltage-sensing region and a pore-forming region. Here we have identified the negative residues of the second transmembrane segment in the plant voltage-dependent K+ channel, KAT1, which involves the function of voltage sensing. Point mutations at D95 and D105 but not D89 and D116 failed to complement the K+ uptake deficient properties of the mutant yeast. In vitro translation and translocation experiments showed that the membrane integration of the third and fourth segments involving voltage sensor were impaired by the replacement of D95 or D105 by serine. These data show that both the residues play a crucial role in the membrane topogenesis of the voltage sensor in KAT1.     

A rapid BOD sensing system using luminescent recombinants of Escherichia coli

A biochemical oxygen demand (BOD) sensing system based on bacterial luminescence from recombinant Escherichia coli containing lux A-E genes from Vibrio fischeri has been developed. It was possible to use frozen cells of luminescent recombinants of E. coli as the bacterial reagents for measurement. Steady bioluminescence was observed during the incubation time between 90 and 150 min in the presence of a sole carbon source such as glucose, acetate, L-glutamate and BOD standard solution (GGA solution). This disposable bacterial reagent was applied to measure and detect organic pollution due to biodegradable substances in various wastewaters. The obtained values of this study showed a similar correlation with that of the conventional method for BOD determination (BOD5). Bacterial luminescence that was visualized with an imaging system using a charge coupled device (CCD) camera and a photomulti-counter demonstrated that this method could also be used for multi-sample detection of organic pollution due to biodegradable substances by using a microtiter plate. These results suggested for successful achievement of high-though-put detection of BOD in practical.     

Role of the furrow of the proximal colon in the production of soft and hard feces in nutrias, Myocastor coypus

The bacterial level of soft feces is higher than that of hard feces in nutrias. This suggests the heterogeneity of bacterial density in the large intestine. To show the heterogeneity of bacteria in the contents of the large intestine in nutrias, we divided the contents of the large intestine into 12 regions, then measured the nitrogen (N), total amino acids (TAA) and diaminopimelic acid (DAP), a bacterial marker, of these regions. Levels of N, TAA and DAP varied along the cross section of the proximal colon. The greater curvature of the main lumen and furrow had higher N, TAA and DAP concentrations than the lesser curvature. We also examined the involvement of the furrow in producing two types of feces differing in bacterial nitrogen content by surgically preventing the flow of the furrow contents. We compared the concentrations of N, TAA and DAP between soft and hard feces among operated, sham-operated and intact animals. Surgical closure of the furrow abolished the difference in levels of N, TAA and DAP between soft and hard feces, suggesting that the furrow of the proximal colon is responsible for making the bacterial density higher in soft feces than in hard feces. Accepted: 13 July 2000