Identification and characterization of a dextranase gene of Streptococcus criceti

The dextranase gene, dex, was identified in Streptococcus criceti strain E49 by degenerate PCR and sequenced completely by the gene-walking method. A sequence of 3,960 nucleotides was determined. The dex gene encodes a 1,200-amino acid protein, which has a calculated molecular mass of 128,129.91 and pI of 4.15 and is predicted to be a cell-surface protein. The deduced amino acid sequence of dex showed homology to S. downei dextranase (63.9% identity). Phylogenetic analysis revealed the similarity of the deduced amino acid sequence of dextranases in S. criceti, S. sobrinus, and S. downei. A recombinant form of the protein with six histidine residues tagged in the C-terminus was partially purified and showed dextranase activity on blue-dextran sodium dodecyl sulfate-polyacrylamide gel electrophoresis (BD-SDSPAGE) followed by renaturation. We also detected dextranase activity in S. criceti cell extracts and culture supernatant by renatured BD-SDS-PAGE, whereas no dextranase activity of the cells was observed on blue-dextran brain heart infusion (BD-BHI) agar plates. Furthermore, PCR-based mutations of dextranase indicated that a deletion mutant of the C-terminal region could hydrolyze blue dextrans and that the D453E mutation, W793L mutation, and double mutations (W793L and deletion of the C-terminal region) resulted in a loss of dextranase activity. These findings suggest that Asp-453 and Trp-793 residues of S. criceti dextranase are critical to the enzyme's activity.     

Mutations in the helix termination motif of mouse type I IRS keratin genes impair the assembly of keratin intermediate filament

Two classical mouse hair coat mutations, Rex (Re) and Rex wavy coat (Re(wc)), are linked to the type I inner root sheath (IRS) keratin genes of chromosome 11. An N-ethyl-N-nitrosourea-induced mutation, M100573, also maps close to the type I IRS keratin genes. In this study, we demonstrate that Re and M100573 mice bear mutations in the type I IRS gene Krt25; Re(wc) mice bear an additional mutation in the type I IRS gene Krt27. These three mutations are located in the helix termination motif of the 2B alpha-helical rod domain of a type I IRS keratin protein. Immunohistological analysis revealed abnormal foam-like immunoreactivity with an antibody raised to type II IRS keratin K71 in the IRS of Re/+ mice. These results suggest that the helix termination motif is essential for the proper assembly of types I and II IRS keratin protein complexes and the formation of keratin intermediate filaments.     

Quantitative analysis of a stand of Pinus densiflora undergoing succession to Quercus mongolica ssp. crispula : II. Growth and population dynamics of Q. mongolica ssp. crispula under the P. densiflora canopy

We report here the results of an 8-year study of the growth and population dynamics of Quercus mongolica ssp. crispula in a Pinus densiflora stand in a state of succession. In 1998, there were 169 Q. mongolica ssp. crispula individuals in a 400-m² plot under the P. densiflora canopy. This number remained nearly constant between 1998 and 2005. Mean recruitment of new individuals was 11 year⁻¹, while mean mortality was 12 year⁻¹. Of the 35 individuals ≥60 cm in height existing in 1998, 30 were still surviving in 2005. We were able to represent the height growth of Q. mongolica ssp. crispula individuals as H=30 [1+21.96 exp(−0.0839t)]⁻¹, with t = years since 1998 and H = height in meters. Using this equation we predict that by 2015 the mean height of Q. mongolica ssp. crispula trees in the stand will exceed those of understory trees, such as Rhus trichocarpa and Prunus maximowiczii. Once above the understory stratum, the Q. mongolica ssp. crispula trees can be expected to grow more rapidly due to the better light conditions, thereby rapidly reaching the canopy stratum of the P. densiflora stand.     

Molecular cloning and characterization of an alpha-amylase from Pichia burtonii 15-1

An alpha-amylase secreted by Pichia burtonii 15-1 isolated from a traditional starter murcha of Nepal, named Pichia burtonii alpha-amylase (PBA), was studied. The gene was cloned and its nucleotide sequence was determined. PBA was deduced to consist of 494 amino acid residues. It shared certain degrees of amino acid sequence identity with other homologous proteins: 60% with Schwanniomyces occidentalis alpha-amylase, 58% with Saccharomycopsis sp. alpha-amylase, and 47% with Taka-amylase A from Aspergillus oryzae. A three-dimensional structural model of PBA generated using the known three-dimensional structure of Taka-amylase A as a template suggested high structural similarity between them. Kinetic analysis revealed that the K(m) values of PBA were lower than those of Taka-amylase A for the oligosaccharides. Although the k(cat) values of PBA were lower than those of Taka-amylase A for the oligosaccharide substrates, the k(cat)/K(m) values of PBA were higher.     

Plural light chains in a single plasma cell of a monoclonal gammopathy undetermined significance case: An ultrastructural study

A 44-year-old man was found to have M-proteins of IgG consisting of kappa- and lambda-chains in serum without lymphadenopathy or splenomegaly. The serum concentrations of IgG, IgA and IgM were within normal limits. Bone marrow examination showed normal cellular marrow containing 6.3% of plasma cells with no abnormal features. No chromosomal abnormality was observed at all. The patient was diagnosed as having monoclonal gammopathy of undetermined significance. The bone marrow plasma cells possessed free kappa- and lambda-chains in Golgi apparatus, rough endoplasmic reticula and cytoplasmic matrices. Plural light chains were simultaneously produced with the same heavy chain in a plasma cell by immunoelectron microscopy. This is the first report in the world of a monoclonal gammopathy of undetermined significance producing plural light chains with the same heavy chain.     

Triiodothyronine (T3) and fructose coordinately enhance expression of the GLUT5 gene in the small intestine of rats during weaning period

Jejunal GLUT5 is elevated with triiodothyronine (T(3)) during weaning of rats. A perfusion of fructose into the small intestine of T(3)-injected rats at 21 d induced expression of the GLUT5 gene, but one into that of vehicle-injected rats did not. These results suggest that T(3) and fructose coordinately enhance jejunal expression of the GLUT5 gene in rats during weaning period.     

Effects of leaf quality and microhabitat on the survival of a leaf-rolling weevil (Attelabidae)

Female Cycnotrachelus roelofsi (Coleoptera: Attelabidae) construct two types of leaf roll, i.e., cut-off cradles (CCs) and suspended cradles (SCs), in which to lay eggs; these cradles are generally constructed using older and younger leaves, respectively. We conducted two experiments to determine whether the quality of cradle leaves affects egg and larval survival. In the first experiment, we severed all SCs from a tree and placed them on the ground with unmanipulated CCs in the early breeding season. In the second experiment, we resuspended all CCs in a tree with unmanipulated SCs in the late breeding season. We also compared leaf mass per area (LMA), polyphenol content, and nitrogen content between the two cradle types to determine whether there were any differences in leaf quality. Larval mortality, probably caused by cradle herbivory, was significantly greater in severed SCs than in intact CCs in the early season, suggesting that leaf quality had a profound effect on larval mortality in the terrestrial microhabitat. In contrast, larval mortality did not differ between resuspended CCs and intact SCs in the late season, suggesting that leaf quality had little effect on larval mortality in the arboreal microhabitat. LMA was higher in CCs than in SCs, but there were no differences in the nitrogen and polyphenol contents. These results suggest that cradles constructed using mature, tough leaves were more effective against terrestrial cradle herbivores than those constructed using new, soft leaves.