Induction of antibody responses in mice immunized intranasally with Type I interferon as adjuvant and synergistic effect of chitosan

Type I IFNs are a range of host-derived molecules with adjuvant potential; they have been used for many years in the treatment of cancer and viral hepatitis. Therefore, the safety of IFNs for human use has been established. In this study, we evaluated the mucosal adjuvanticity of IFN-β administered intranasally to mice with diphtheria toxoid, and suggested a method to improve its adjuvanticity. When IFN-β alone was used as a mucosal adjuvant, no clear results were obtained. However, simultaneous administration of IFN-β and chitosan resulted in an enhancement of the specific serum immunoglobulin G (IgG) and IgA antibody responses, the mucosal IgA antibody response, and antitoxin titers. Furthermore, the intranasal administration of IFN-α alone resulted in a greater increase in antibody titer than IFN-β, and a synergistic effect with chitosan was also observed. These findings suggest that intranasal administration of chitosan and Type I IFNs may display an effective synergistic mucosal adjuvant activity.     

Response of Merkel cells in the palatal rugae to the continuous mechanical stimulation by palatal plate

The aim of the present study was to investigate the responses of Merkel cells that are numerous in the palatine rugae, due to the continuous mechanical stimulation exerted by the palatal plate. Forty golden hamsters were used in this experiment. The palatal plate was made of adhesive resin and it was set on the palate of the animal. To exert a continuous pressure, a 0.8?mm elevation on the internal surface of the palatal plate was created at the middle portion of the fourth palatine ruga. Thereafter, the number of Merkel cells in the mucosa was calculated by immunohistochemical observation. Morphological changes of Merkel cells were examined by electron microscopy. There was significant difference among the control and any of the treated groups on the number of CK20 positive Merkel cells (p?<?0.05) and that numbers were decreased at the sites where continuous mechanical stimulation was exerted. Degeneration of the cytoplasm mitochondria and nerve endings, and a decrease in both the number of neurosecretory granules and cytoplasmic processes were observed. Furthermore, the presence of nuclear chromatin aggregation and fragmentation was recognized. The continuous mechanical stimulation by the palatal plate affected the responses of Merkel cells and nerve endings, thus inducing a decrease in the number of Merkel cells. A portion of these changes was also associated with the expression of apoptosis.     

The role of the helper lipid dioleoylphosphatidylethanolamine (DOPE) for DNA transfection cooperating with a cationic lipid bearing ethylenediamine

Gene therapy is expected to treat various incurable diseases including viral infections, autoimmune disorders, and cancers. Cationic lipids (CL) have been used as carriers of therapeutic DNAs for gene therapy because they can form a complex with DNA and such a complex can be incorporated into cells and transport the bound DNA to cytosol. The CL/DNA complexes are called lipoplexes and categorized as a non-viral vector. Lipoplexes are often prepared by adding a neutral phospholipid dioleoylphosphatidylethanolamine (DOPE) to CL in order to enhance transfection. However, the role of DOPE is not fully understood. We synthesized a new CL having an ethylenediamine cationic head group, denoted by DA, and found that addition of DOPE to DA achieved a good efficiency, almost in the similar level of commonly used transfection reagent Lipofectamine 2000 (Invitrogen). The composition of DA:DOPE = 1:1 showed the highest efficiency. This lipoplex showed structural transition when pH was changed from 7 to 4, corresponding pH lowering in late endosome, while DOPE itself showed structural transition at more basic pH around 8. The present data showed that the DOPE/DA composition determines the structural transition pH and choosing a suitable pH, i.e., a suitable composition, is essential to increase the transfection efficiency.     

The Structures of Diethylaminoethylated Glucose and Oligosaccharides Derived from Cationic Starch

Phosphatidylethanolamine N-methyltransferase, which catalyzes all the three-step methylation from phosphatidylethanolamine to phosphatidylcholine, was purified to homogeneity from the membrane fraction of Zymomonas mobilis. The purified enzyme exhibited a single band on SDS- polyacrylamide gel electrophoresis and its molecular weight was estimated to be 42,000 on comparison with those of marker proteins. The three activities dependent on phosphatidylethanolamine, phosphatidyl-N monomethylethanolamine and phosphatidyl-N Af-dimethylethanolamine of the purified enzyme showed similar pH profiles with an optimum of pH 8.5, and were enhanced in the same manner by Triton X-100 and l-cysteine. The maximal velocities of the three reactions for S-adenosyl-l-methionine were 0.04, 1.36 and 0.69 nmol/mg protein/min with apparent Michaelis constant values of 3.6, 1.9 and 3.9 fiM, respectively, indicating that the first-step methylation is rate-limiting for the pathway in the organism.     

Isolation and Chemical Characterization of a Mating Pheromone Produced by Saccharomyces kluyveri

The pullulanase gene (pul) of Klebsiella aerogenes was transferred in vivo to Escherichia coli by using RP4:: Mu cts. The pul gene was expressed in E. coli, although the level of pullulanase activity in E. coli was lower than that in K. aerogenes, and the Pul⁺ transconjugants were relatively unstable in an unselective medium. Production of pullulanase, which is used to make maltose from starch, was induced in E. coli by pullulan, waxy maize amylopectin, soluble starch and maltose. When the transconjugant cells of E. coli were grown with pullulan or maltose, most pullulanase was produced intracellularly, whereas K. aerogenes produced pullulanase extracellularly. Retransfer of the pul_k gene from E. coli to K. aerogenes by conjugation resulted in an increase of the production of extracellular pullulanase.     

Structures of Ezomycins A1 and A2

The structures of ezomycins A₁. and A₂, antifungal antibiotics produced by a strain of Streptomyces, were determined as 1 and 2, respectively, by degradative and spectrometric studies.     

Isolation and Identification of a Sexual Hormone in Yeast

5-Fluorotryptophan (5FT), indolmycin (IM), 4-fluorotryptophan and 7-azatryptophan were found on screening to be tryptophan antagonists among various chemically synthesized and naturally occurring tryptophan analogues for the isolation of l-tryptophan (l-Trp) producing mutants of Bacillus subtilis K.

From among 5FT resistant mutants, potent l-Trp producers were obtained using an improved isolation medium. Growth of the isolated 5FT-resistant l-Trp producer, AJ 11709, was inhibited by IM. From among 5FT and IM resistant mutants, the best strain, AJ 11979, which produced 9.0 g/liter of l-Trp from 13% glucose on 120hr cultivation, was selected.     

Occurrence of Castasterone,Brassinolide and Methyl 4-Chloroindole- 3-acetate in Immature Vicia faba Seeds

The rice lamina inclination test indicated the presence of brassinosteroid-like active substances in immature Vicia faba seeds. Two of these were identified as castasterone and brassinolide by GC/MS and GC/SIM, respectively. Another active principle was identified as methyl 4- chloroindole-3-acetate by GC/MS and HPLC.     

Synthesis of Dihydrohelminthosporic and Allodihydrohelminthosporic Acids and their Plant Growth-regulating Activity

Li-NH₃ reduction of helminthosporic acid gave dihydrohelminthosporic acid (III) and allodihydrohelminthosporic acid (IV). III and IV are the epimers with respect to the carboxyl and III is labil form. The stereochemistry of III and IV is elucidated from the chemical and spectroscopic evidences. III showed a marked plant growth promoting activity but IV did not.     

Studies on Flavor Components in Soybean

Aliphatic carbonyl compounds in soybean were studied. Volatile carbonyl compounds in defatted soybean flour were identified as methanal, ethanal, n-hexanal, 2-propanone, 2- pentanone, 2-heptanone, 2-heptenal, and 2,4-decadienal, while those in raw soybean as ethanal, n-hexanal, and 2-propanone. Four kinds of non-volatile carbonyl compounds were found in defatted soybean, two of which seemed to be carbonyl ester and carbonylic acid. It is probable that the compounds in defatted soybean are mostly the secondary products derived from autoxidation of the residual fatty acids and esters in the defatting process and/or during the storage thereafter. n-Hexanal in raw soybean amounts to approximately 10 p.p.m., which is, owing to its extremely low flavoring threshold, likely to be one of the main components of the green bean flavor.