Physical Mapping of Rice Chromosomes 4 and 7 Using YAC Clones

Physical maps of rice chromosomes 4 and 7 were constructed bylanding yeast artificial chromosomes (YACs) along our high-densitymolecular linkage map. Using 114 DNA markers, 258 individualYACs were located on chromosome 4. Sixty-two out of 258 YACscarried two or more DNA marker positions and formed 16 contigswhich covered a total length of 17.1 cM. The other YACs werearranged to 23 positions. On chromosome 7, 203 individual YACswere landed on 109 DNA markers. Sixty-four out of 203 YACs formed15 contigs which covered a total length of 21.8 cM and 139 YACswere localized to 26 positions. Chromosomes 4 and 7 were coveredwith minimum tiling paths of 45 and 48 YACs, respectively. Takingthe average size of YAC insert DNA to be 350 kb and the entiregenome size to be 430 Mb, about 16–18 Mb of each chromosomeor an estimated 50% of their total lengths have been coveredwith YACs. Physical maps of these 2 chromosomes should be ofgreat help in identifying useful trait genes and unravelinggenetic and biological characteristics in rice.     

Induction of interdigitating reticulum cell-like differentiation in human monocytic leukemia cells by conditioned medium from IL-2-stimulated helper T-cells.

Monocytic leukemia (MoL) cells were obtained from the peripheral blood of a patient in whom the leukemic cells infiltrating various lymphoreticular organs exhibited features intermediate between interdigitating reticulum cells (IDC) and ordinary phagocytic macrophages, whereas the leukemic cells in the peripheral blood were essentially monocytic and lacked such features. Peripheral blood CD4+ T-cells were established as an interleukin-2-dependent T-cell line. When the MoL cells were exposed for a few days to conditioned medium from the T-cell line, they extended several dendritic cytoplasmic projections and became intensely positive for HLA-DR antigen, cytoplasmic S-100 beta protein, and CD1 antigen. Functionally, the conditioned medium significantly down-regulated Fc-mediated and Fc-independent phagocytic activities, and the levels of lysosomal enzymes such as lysozyme and nonspecific esterase in the MoL cells. Moreover, the conditioned medium significantly up-regulated the accessory cell function of the MoL cells as measured by the primary allogenic mixed leukocyte reaction (MLR). Furthermore, the conditioned medium significantly down-regulated the expression of CD14 antigen. Biochemical analysis indicated that the factor responsible for these changes is a protein which is distinct from known human cytokines and whose molecular weight is approximately 31 kDa. These findings suggest that IDC are closely related the monocytic lineage and that helper T-cells play an important role in constructing the microenvironment of T-lymphoid tissues which is necessary for the differentiation and maturation of IDC.     

Studies on the neutralization of herpes simplex virus. X. Demonstration of complement-requiring neutralizing (CRN) and slow-reacting CRN (s-CRN) antibodies in late IgG.

A sample of late IgG from a rabbit hyperimmunized with herpes simplex virus was analyzed for neutralizing (N) and complement-requiring neutralizing (CRN) antibodies. In a usual endpoint test, N and CRN titers were 1: 40 and 1: 160, respectively, but when virus-IgG mixtures were incubated at 0 C overnight before addition of complement (C), an endpoint of 1:1280 was obtained. Virus sensitized at 0 C overnight required more C for inactivation than did sensitized virus formed earlier. Sensitization kinetic curve experiments employing a proper initial virus concentration, which permitted differentiation of sensitized viruses requiring different amounts of C, indicated that formation of sensitized virus detectable only with a relatively large amount of C proceeded slowly at IgG dilutions where the ordinary CRN antibody requiring a smaller amount of C was negligible. The results strongly suggested that the IgG sample contained slow-reacting CRN (s-CRN) antibody in excess of the hitherto known CRN antibody. As to the mechanism of formation of s-CRN complexes, experiments failed to prove the occurrence of complexes initially insensitive to C, and it appears more likely that s-CRN antibody has a comparatively low avidity for virus.     

Regulatory role of polyamine in the acid phosphatase from potato tubers.

Effects of polyamine and metal ions on the new type of acid phosphatase purified from potato (Solanum tuberosum L. Irish Cobbler) tubers were analyzed. The enzyme belongs to nonspecific acid phosphatase family (EC 3.1.3.2), which hydrolyzes various phosphorylated substrates. The enzyme hydrolyzed inorganic pyrophosphate as a preferred substrate, and exhibited the hyperbolic kinetics with respect to the substrate, inorganic pyrophosphate in the absence of metal cations. Polyamine activated the enzyme effectively by lowering the K(m) value without appreciable changes in the maximal velocity. The most effective polyamines as activators were spermine and spermidine. Mg(2+) ion increased the K(m) value without affecting the maximal velocity of the enzyme, but Ca(2+) ion decreased both the K(m) and V(max) values. Increasing concentrations of spermine also decreased the K(m) value irrespective of Mg(2+) ion included, but gave a constant K(m) and V(max) values in the absence and presence of Ca(2+) ion. Action of spermine and metal ions can be explained by the complex formation with the substrate pyrophosphate. The acid phosphatase from potato can utilize the pyrophosphate-spermine or pyrophosphate-Ca(2+) complex as the preferred substrates. However, the enzyme can use the pyrophosphate-Mg complex with a weak affinity for the active site. Polyamine activates acid phosphatase in the absence and presence of metal cations, and activation by polyamine of the enzyme may contribute to the stimulation of starch biosynthesis and the control of glycolysis/gluconeogenesis by regulating PPi levels in growing potato tubers.     

Coloration and ontogenetic features of fluviatile species of <Emphasis Type="Italic">Rhinogobius</Emphasis> (Gobioidei: Gobiidae) in Amami-oshima Island,Ryukyu Islands,Japan

 The fluviatile Rhinogobius sp. 1 (hereinafter R. sp. 1) (Japanese name kibara-yoshinobori or English name yellow belly medium-egg type) is broadly distributed in the Ryukyu Islands ranging from Amami-oshima Island to Iriomote Island. However, based on the molecular evidence, this species has probably had multiple origins from amphidromous species. We compared the coloration of specimens at Amami-oshima I. with that of the other specimens (islands) in the Ryukyu Is. Amami-oshima I. specimens of R. sp. 1 were different from the specimens of other populations in having five to six cross lines on the caudal fin. We examined the early life history of fluviatile Rhinogobius at Amami-oshima I., based on rearing experiments, and estimated the age of wild specimens by counting otolith increments. Spawned egg capsules of “Rhinogobius sp. 1” at Amami-oshima I. were larger than those of amphidromous species. Their developmental stage in newly hatched larvae was similar to that of Okinawan R. sp. 1 in having a straight notochord; however, it had a flexed notochord earlier than that of Rhinogobius sp. 2 (hereinafter R. sp. 2) (aobara-yoshinobori or blue belly medium-egg type). The ontogenetic trajectory of “Rhinogobius sp. 1” in Amami-oshima I. was similar to Okinawan R. sp. 1 (30 days in larval duration) but different from R. sp. 2 (15 days). In amphidromous relatives of R. sp. 2, developmental timing during larval period was earlier than in the amphidromous relatives of R. sp. 1. It has been suggested that the phenomenon of fluvial landlocking has been accompanied somewhat by egg size increase (on the basis of a few examples). However, the ontogenetic trajectories of fluviatile Rhinogobius species may be a reflection of those of ancestral amphidromous species. Received: March 18, 2002 / Revised: November 7, 2002 / Accepted: December 10, 2002     

The Dictyostelium developmental cDNA project: generation and analysis of expressed sequence tags from the first-finger stage of development.

In an effort to identify and characterize genes expressed during multicellular development ill Dictyostelium, we have undertaken a cDNA sequencing project. Using size-fractionated subsets of cDNA from the first finger stage, two sets of gridded libraries were constructed for cDNA sequencing. One, library S, consisting of 9984 clones, carries relatively short inserts, and the other, library L, which consists of 8448 clones, has longer inserts. We sequenced all the selected clones in library S from their 3'-ends, and this generated 3093 non-redundant, expressed sequence tags (ESTs). Among them, 246 ESTs hit known Dictyostelium genes and 910 showed significant similarity to genes of Dictyostelium and other organisms. For library L, 1132 clones were randomly sequenced and 471 non-redundant ESTs were obtained. In combination, the ESTs from the two libraries represent approximately 40% of genes expressed in late development, assuming that the non-redundant ESTs correspond to independent genes. They will provide a useful resource for investigating the genetic networks that regulate multicellular development of this organism.