Treatment with YIGSR peptide ameliorates mouse tail lymphedema by 67 kDa laminin receptor (67LR)-dependent cell-cell adhesion.

Impaired microcirculation can cause lymphatic leakage which leads to a chronic swelling in the tissues of the body. However, no successful treatment gives any protection against lymphedema due to the lack of well-revealed pathophysiology of secondary lymphedema. Binary image of laminin immunohistochemical expression revealed that distribution of laminin expression localized during surgically induced lymphedema. 67 kDa laminin receptor (67LR) mRNA expression showed a peak at during lymphedema exacerbation. Since the response of 67LR molecules may affect the prevention of inflammation and edema, here we have hypothesized that 67LR ligand of YIGSR peptide could permit reconstructive environment for amelioration of lymphedema and evaluated the effect of YIGSR in a mouse tail model of lymphedema. Indeed, intra-abdominal injections of YIGSR for the first 3 days after inducing lymphedema in the mouse tail model reduced the tail lymphedema on day 14 by 27% (P = 0.035). Histology showed that YIGSR treatment protected lymphedema impairment in epidermis and dermis, and it also inhibited the expansion of intercellular spaces and enhanced especially cell adhesion in the basement membrane as revealed by transmission electron microscopy. Interestingly, the treatment also reduced the local expression of transforming growth factor (TGF)ß. Further elucidation of the mechanisms of 67LR-facilitated lymphangiogenesis contributes to find potential targets for the treatment of lymphedema.

New conformational search method using genetic algorithm and knot theory for proteins.

We have proposed a parallel simulated annealing using genetic crossover as one of powerful conformational search methods, in order to find the global minimum energy structures for protein systems. The simulated annealing using genetic crossover method, which incorporates the attractive features of the simulated annealing and the genetic algorithm, is useful for finding a minimum potential energy conformation of protein systems. However, when we perform simulations by using this method, we often find obviously unnatural stable conformations, which have "knots" of a string of an amino-acid sequence. Therefore, we combined knot theory with our simulated annealing using genetic crossover method in order to avoid the knot conformations from the conformational search space. We applied this improved method to protein G, which has 56 amino acids. As the result, we could perform the simulations, which avoid knot conformations.

Relationship between alcoholism and CYP2E1 C/D polymorphism.

The genotypes of the CYP2E1 loci of 35 alcoholic and 130 nonalcoholic (healthy controls) Japanese were investigated to determine the relationship between CYP2E1 (C/D) polymorphism and susceptibility to alcohol dependence. There was a significant (d.f. = 1, chi2 = 11.0, p < 0.001) difference in the CYP2E1 C gene frequency between alcohol dependents (0.41) and controls (0.28), suggesting that the C gene of CYP2E1 may be related to the risk of developing alcoholism in Japanese, whereas the frequency of the D gene was higher among controls than alcoholics. Healthy Japanese controls showed a significantly (d.f. = 1, chi2 = 30.1, p < 0.001) higher frequency of the C allele than Swedish healthy controls (0. 10) did.

[Relationship between alcoholism and CYP2E1 genotypes].

The genotype of the CYP2E1 loci in 36 alcoholic and 42 non-alcoholic (healthy) Japanese were investigated to examine the relationship between the polymorphisms of CYP2E1 (C/D) and the susceptibility to alcohol dependence. There was a significant (df = 1, chi 2 = 4.39, p < 0.05) difference in CYP2E1 CD (heterozygote) genotype frequency between alcohol dependents (56%) and controls (33%), suggesting that the CD (heterozygote) genotype of CYP2E1 may have something to the risk of developing alcoholism in Japanese, whereas DD (homozygote) genotype was high among controls.

Immunological detection of the GerA spore germination proteins in the spore integuments of Bacillus subtilis using scanning electron microscopy.

To clarify the molecular mechanisms that trigger spore germination of Bacillus subtilis, the location of GerA proteins (GerAA, GerAB and GerAC), which were reported to be putative gene products of a receptor for one of the germinants, L-alanine, was investigated by immunological techniques using anti-GerA peptide antibodies. Four antibodies were raised against the corresponding epitopes, two in GerAA, one in GerAB and the other in GerAC molecules. The binding of all four antibodies to the inner surface of the cortex-less spore coat fragments could be seen by scanning immunoelectron microscopy with colloidal gold particles. The result agreed with the fact, previously reported, that the colloidal gold particles were visualized just inside the spore coat layer by transmission immunoelectron microscopy using another anti-GerAB peptide antibody.

Immunoelectron microscopic localization of one of the spore germination proteins, GerAB, in Bacillus subtilis spores.

Ultrastructural localization of GerAB, one of the proteins of Bacillus subtilis spores related to L-alanine-initiated germination, was investigated by immunoelectron microscopy with antipeptide (residues 61 to 80 of GerAB) antiserum and a colloidal gold-immunoglobulin G complex. Immunogold particles were visualized in the boundary region between the cortex and coat of dormant spores, and they were broadly dispersed into the cortex region after germination.

[Experimental and clinical studies of bestatin as an immunomodulator].

The experimental and clinical effects of bestatin were examined and the results obtained were as follows. Bestatin which is an immunomodulator discovered by Umezawa did not increase the bone marrow stem cells examined by the method of the spleen colony assay in the 60Co irradiated mouse. However, it prolonged the survival time slightly when it was administered 10-25mg/kg intraperitoneally. Clinically, bestatin was administered to the patients with gastrointestinal cancer. It did not influence on the PHA-induced lymphocyte blastformation rate, but it increased the peripheral lymphocyte count and PPD skin reaction in the cases of curative resection and increased the peripheral lymphocyte counts in the cases of nonresection after 1 month of the operation.