Occurrence,horizontal transfer and degeneration of VDE intein family in Saccharomycete yeasts

VDE is a homing endonuclease gene originally discovered as an intervening element in VMA1s of Saccharomyces cerevisiae. There have been two independent subfamilies of VDE, one from S. cerevisiae strain X2180-1A and the other from Saccharomyces sp. DH1-1A in the host VMA1 gene, and they share the identity of 96.3%. In order to search the occurrence, intra/interspecies transfer and molecular degeneration of VDE, complete sequences of VMA1 in 10 strains of S. cerevisiae, eight species of saccharomycete yeasts, Candida glabrata and Kluyveromyces lactis were determined. We found that six of 10 S. cerevisiae strains contain VDEs 99.7-100% identical to that of the strain X2180-1A, one has no VDE, whereas the other three harbour VDEs 100% identical to that of the strain DH1-1A. S. carlsbergensis has two VMA1s, one being 99.8% identical to that of the strain X2180-1A with VDE 100% identical to that of the strain DH1-1A and the other containing the same VMA1 in S. pastorianus with no VDE. This and other evidence indicates that intra/interspecies transmissions of VDEs have occurred among saccharomycete yeasts. Phylogenetic analyses of VMA1 and VDE suggest that the S. cerevisiae VDEs had branched earlier than other VDEs from an ancestral VDE and had invaded into the host loci as relatively late events. The two VDEs seemed to degenerate in individual host loci, retaining their splicing capacity intact. The degeneration of the endonuclease domains was distinct and, if compared, its apparent rate was much faster than that of the protein-splicing domains.     

MgATP-induced conformational changes in a single myosin molecule observed by atomic force microscopy: periodicity of substructures in myosin rods

This paper discusses the conformational changes in a single myosin molecule directly observed using atomic force microscopy (AFM). The myosin molecules were pretreated in rigor solutions without MgATP or in relaxed solutions with various concentrations of MgATP. The images of these molecules were obtained using a tapping mode AFM. The results indicate that the orientation of the myosin's heads and tail strongly depend on the MgATP concentration. Without using MgATP, almost all of the myosin molecules are in the extended form; however, when MgATP is used, the molecules bend according to the level of MgATP concentration. The mean-square end-to-end distance of the myosin molecules is significantly shorter with p[MgATP] = 4 than with p[MgATP] = 6. The rod region did not show the same level of intensity along their length in the extended form. The rods exhibited clusters of discontinuity, which were identified as substructures. The size of these substructures change at intervals that are multiples of 14.3-14.5 nm, which reflects the periodicity of the alpha-helical coiled coils. The substructure clusters also correspond to the myosin crossbridge spacing in muscles (14.3 or 43 nm). These results suggest that the myosin's head bends in conjunction with the bending or tilting in the helical substructures. Conformational changes of the myosin molecule induced by MgATP seem to mimic the molecular motions in a muscle's force generation process.     

Coagulation mechanism of salt solution-extracted active component in Moringa oleifera seeds

This study focuses on the coagulation mechanism by the purified coagulant solution (MOC-SC-PC) with the coagulation active component extracted from M. oleifera seeds using salt solution. The addition of MOC-SC-PC tap water formed insoluble matters. This formation was responsible for kaolin coagulation. On the other hand, insoluble matters were not formed when the MOC-SC-PC was added into distilled water. The formation was affected by Ca2+ or other bivalent cations which may connect each molecule of the active coagulation component in MOC-SC-PC and form a net-like structure. The coagulation mechanism of MOC-SC-PC seemed to be an enmeshment of Kaolin by the insoluble matters with the net-like structure. In case of Ca2+ ion (bivalent cations), at least 0.2 mM was necessary for coagulation at 0.3 mgC l-1 dose of MOC-SC-PC. Other coagulation mechanisms like compression of double layer, interparticle bridging or charge neutralization were not responsible for the coagulation by MOC-SC-PC.     

Superfluid-Normal Interface of 4He Near a Wall

We observed a profile of nonequilibrium superfluid-normal (SN) interface of ⁴He near a vertical wall. A glass, brass or copper wall was used. The SN interface was produced by cooling liquid ⁴He in a bath from the bottom, where liquid ⁴He was pumped through a flow impedance in order to cool down the liquid. Superfluid (Normal fluid) occupied the lower (upper) part of the bath. The SN interface was visualized by three methods: simple visualization, shadowgraphy and schlieren method. The interface touched a vertical glass wall at almost 90°. A large hollow was observed near a brass wall which had intermediate thermal conductivity. Downward flow was observed on a copper wall due to the very good thermal conductivity of the wall. Various types of interface profile were observed depending on the thermal conductivity of the wall used.      

Entirely VPE-grown 1-5?m DFB lasers with low threshold currents

Entirely hydride VPE-grown 1.5?m DFB lasers have been obtained by means of high controllability in film thickness and alloy composition for the GalnAsP/InP system. A low threshold current of 13 mA was achieved by improving the growth method for the layer burying the grating. High uniformity in threshold current and lasing wavelength (Ith = 27.3 ± 9.7 mA, ? = 15571 ± 12?) was obtained.     

A 250-Mb/s/pin, 1-Gb double-data-rate SDRAM with a bidirectionaldelay and an interbank shared redundancy scheme

This paper describes three circuit technologies indispensable for high-bandwidth multibank DRAM's. (1) A clock generator based on a bidirectional delay (BDD) eliminates the output skew. The BDD measures the cycle time as the quantity charged or discharged of an analog quantity, and replicates it in the next cycle. This achieves a 0.18-mm ², two-cycle-lock clock generator operating from 25 to 167 MHz with a 30-ps resolution. (2) A quad-coupled receiver eliminates the internal skew caused by the difference between a rise input and a fall input by 40%. (3) An interbank shared redundancy scheme (ISR) with a variable unit redundancy (VUR) efficiently increases yield in multibank DRAM's. The ISR allows redundancy match circuits to be shared with two or more banks. The VUR allows the number of units replaced to be variable. These circuit technologies achieved a 250-Mb/s/pin, 8-bank, 1-Gb double-data-rate synchronous DRAM     

Osteogenic activity of human periosteal sheets cultured on salmon collagen-coated ePTFE meshes

Our animal implantation studies have demonstrated that, after osteogenic processing, cultured human periosteal sheets form osteoid tissue ectopically without the aid of conventional scaffolding materials. To improve the osteogenic activity of these periosteal sheets, we have tested the effects of including a scaffold made of salmon collagen-coated ePTFE mesh. Periosteal sheets were produced with minimal manipulation without enzymatic digestion. Outgrown cells penetrated into the coated mesh fiber networks to form complex multicellular layers and increased expression of alkaline phosphatase activity in response to the osteoinduction. In vitro mineralization was notably enhanced in the original tissue segment regions, but numerous micro-mineral deposits were also formed on the coated-fiber networks. When implanted subcutaneously into nude mice, periosteal sheets efficiently form osteoid around the mineral deposits. These findings suggest that the intricate three-dimensional mesh composed of collagen-coated fibers substantially augmented the osteogenic activity of human periosteal sheets both in vitro and in vivo.