Water-soluble glucans from the seed of Coix lacryma-jobi var. ma-yuen

The water-soluble major polysaccharides from the seed of Coix lacryma-jobi var. ma-yuen eluted as a broad peak by gel filtration on Sepharose CL-2B. The mixture (CS-Glucan) was resolved into 7 glucans by HPLC on the column of Asahi-Pak GS-510 + GS-320. Similarities were observed between M, shown in the gel filtration profile and the elution volume in HPLC. Methylation analysis indicated that the ethanol-fractionated CS-glucan contained 4-O- and 4,6-di-O-substituted glucosyl residues. ¹H and ¹³C NMR data accorded with the results of methylation analysis, and the glycosidic linkages were shown to have an α-configuration. Thus, CS-glucan contained (1 → 4) linked α-d-glucans to which are attached glucosyl side chains at O-6 of the main chain in a similar way to amylopectin. Each purified glucan was shown to have different absorption maxima ( > 550 nm or 530 nm) in the iodine reaction. The results of the methylation analysis and of the pullulanase digestion suggest that the 550 nm-glucan has a lower branching frequency and shorter side chains than the 530 nm-glucan. Although CS-glucan was found to have weak anti-complementary activity, HPLC-purified > 550 nm-glucan was found to be more potent than the 530 nm-glucan. Thus CS-glucan is highly heterogeneous, and the glucans which form a tight complex when tested with iodine, generally tend to have considerable anti-complementary activity.     

Differences in hair density of Japanese monkeys (Macaca fuscata fuscata) with locality and age

The hair density of free-ranging Japanese monkeys (Macaca fuscata fuscata) living in three different areas was investigated. The Japanese monkeys had thicker hair than other macaques. The hair density in the Japanese monkeys varied with locality: the northern monkeys had thicker hair than the southern ones. The density did not vary markedly with age up to 3 years of age, but then decreased gradually up to adult age (≧7 years old). The remarkable growth of the trunk suggested that the total number of hairs increased with age, especially during the period as a juvenile.     

Purification and characterization of an antibacterial factor from snail mucus

The antibacterial factor from the body surface of the African giant snail, Achatina fulica Férussac, was isolated by DEAE-Toyopearl 650M ion exchange chromatography. The isolated preparation exhibited highly positive antibacterial activity both for the Gram-positive bacteria, Bacillus subtilis and Staphylococcus aureus and for the Gram-negative bacteria, Escherichia coli and Pseudomonas aeruginosa, but it lost such activity when heated at 75 degrees C for 5 min. The antibacterial factor of the snail mucus was a glycoprotein whose molecular weight (MW) was about 160,000. It was composed of two subunits of MW 70,000-80,000.     

The mating system ofTokunagayusurika akamusi (Diptera; Chironomidae): II. Experimental analysis of male mating behaviour at the resting place

Mating behaviour ofTokunagayusurika akamusi searching at the resting place (lakeside vegetation, although mating also occurs in the air by swarming) was investigated at the shore of Lake Biwa. Coinciding with the arrival of newly emerged adults, previously emerged males walked about to search for mates at the resting place. When a searching male came into contact with a fresh (newly emerged) individual, he attempted to copulate with it regardless of its sex. Consequently, males copulated with fresh females, which were more likely to be virgin at the resting place, suggesting that the first male might contribute best to the fertilization.     

Mating system ofTokunagayusurika akamusi (Diptera: Chironomidae): I. Copulation in the air by swarming and on the ground by searching

Observations on the mating system of the midge,Tokunagayusurika akamusi, revealed mating to occur both in the air by swarming and on the ground by searching. At the shores of Lake Biwa, midges appeared from November to early December. Newly emerged adults arrived at the resting place, lakeside vegetation, in the morning, during which time a number of males also walked about in search of mates. Many copulating pairs were observed at the resting place. Huge swarms occurred chiefly before sunset but the frequency of copulation observed in the swarm was extremely low. It is likely that, in the Lake Biwa population, the proportion of females inseminated by searching males at the resting place was much larger than that by swarming males in the air. Furthermore, by searching, males copulated with younger females than by swarming. The differences between the searching and swarming tactics are discussed.     

Nucleotide sequence of the herpes simplex virus type 2 (HSV-2) thymidine kinase gene and predicted amino acid sequence of thymidine kinase polypeptide and its comparison with the HSV-1 thymidine kinase gene

To analyze the boundaries of the functional coding region of the HSV-2(333) thymidine kinase gene (TK gene), deletion mutants of hybrid plasmid pMAR401 H2G, which contains the 17.5 kbp BglII-G fragment of HSV-2 DNA, were prepared and tested for capacity to transform LM(TK-) cells to the thymidine kinase-positive phenotype. These studies showed that hybrid plasmids containing 2.2-2.4 kbp subfragments of HSV-2 BglII-G DNA transformed LM(TK-) cells to the thymidine kinase-positive phenotype and suggested that the region critical for transformation might be less than 2 kbp. That the activity expressed in the transformants was HSV-2 thymidine kinase was shown by experiments with type-specific enzyme-inhibiting rabbit antisera and by disc-polyacrylamide gel electrophoresis analyses. DNA fragments of the HSV-2 TK gene were subcloned in phage M13mp9 and M13mp8. A sequence of 1656 bp containing the entire coding region of the TK gene and the flanking sequences was determined by the dideoxynucleotide chain termination method. Comparisons with the HSV-1(Cl 101) TK gene revealed that PstI, PvuII, and EcoRI cleavage sites had homologous locations as did promoter, translational start and stop, and polyadenylation signals. Extensive homology was observed in the nucleotide sequence preceding the ATG translational start signal and in portions of the coding region of the genes. Comparisons of the predicted amino acid sequences of the HSV-1 and HSV-2 thymidine kinase polypeptides revealed that both were enriched in alanine, arginine, glycine, leucine, and proline residues and that clear, but interrupted homology existed within several regions of the polypeptide chains. Stretches of 15-30 amino acid residues were identical in conserved regions. The possibility is suggested that domains containing some of the conserved amino acid sequences might have a role in substrate binding and as major antigenic determinants.     

Clinical Use of a New Mitral Disc Valve

A disc valve of new design was used successfully for the replacement of the mitral valve in patients with rheumatic mitral valve disease. This valve would appear to have the following advantages over the mitral ball valve prosthesis:• Lower left atrial pressure after replacement.• Elimination of the hazard of left ventricular outflow tract obstruction with mitral valve replacement.• Decreased incidence of thromboembolization.• Abolition of possibility of ventricular septal irritation.Despite the better outlook for this valve compared with the ball valve for mitral valve substitution, the mitral valve should always be repaired whenever feasible. Repair is possible in the majority of patients.     

Cell proliferation and abnormal nuclei induced by radiation in renal tubule epithelium as an early manifestation of late damage

The time of appearance and the dose response of radiation effects in the mouse kidney were assessed from the determination of increases in labeling index, the appearance of proximal tubule cells with abnormally large nuclei, and kidney weight loss. Increased labeling indices and abnormally large nuclei were observed in the irradiated proximal tubule cells before any other histological changes were seen. The labeling index increased with dose (from 3 to 15 Gy) but not with time (from 1 to 12 months after irradiation). Increased labeling was evident as soon as 1 month after irradiation. Cell depletion as measured by a decrease in kidney weights compared to those of age-matched controls was not significant until 6 or more months after 11-, 13-, or 15-Gy irradiation. The frequency of cells with large nuclei increased steadily during the first 9 months after 15 Gy and tended to decline between 9 and 12 months, coincident with accelerating renal weight loss. These findings are consistent with the hypothesis that the production of these cells is a result of an abortive mitotic division and their loss is an eventual result of such an aberration. The increased proliferation induced by irradiation increases the chance for an abortive mitosis and death, presumably at a subsequent mitosis, of radiation-damaged proximal tubule cells, which is a major factor in the appearance of late radiation damage in the kidney.     

Increased plasma level of endothelin with vasoconstriction after dextran infusion.

Our previous study demonstrated that volume expansion with dextran produced blood pressure elevation due to vasoconstriction 3 hours after the cessation of infusion. To examine whether endogenous endothelin contributes to this vasoconstriction, we measured plasma level of endothelin before, immediately after, and 3 hours after the administration of dextran. Plasma level of endothelin decreased immediately after the administration (from 1.5 +/- 0.3 pg/ml to 1.1 +/- 0.2 pg/ml, P less than 0.05), and increased 3 hours after the administration (2.1 +/- 0.3 pg/ml, P less than 0.05). However, the changes in the plasma level of endothelin did not significantly correlated with those in blood pressure or total peripheral resistance. Thus, vasoconstriction after dextran infusion was accompanied by an increase in the plasma level of endothelin, but further evaluation is needed for the direct role of this peptide in the vasoconstrictive blood pressure elevation.     

Alkaline serine protease produced from citric acid by Bacillus alcalophilus subsp. halodurans KP 1239

Summary Maximum production of alkaline serine protease by Bacillus alcalophilus subsp. halodurans KP 1239 was achieved after 24 h cultivation, at an initial pH of 7.6, on a medium containing 1.0% sodium citrate, 0.3% yeast extract, and 0.3% KH₂PO₄. The enzyme was purified to crystalline form from culture broth. The enzyme was most active at 60° C and at pH 11.5. The molecular weight, isoelectric point and sedimentation coefficient in water at 20° C were estimated as 29 000, 8.8 and 3.3S, respectively. The N-terminal amino acid sequence was Ala-Gln-Ser-Val-Pro-Trp-Gly-Ile-Ser-Arg-Val-Gln-Ala-Pro-Ala-Ala-His-Asn-Arg-Gly-. The enzyme shared its antigenic determinants with B. alcalophilus ATCC 21522 serine protease, but not with the subtilisins Carlsberg and BPN. Offprint requests to: Yuzuru Suzuki