Behavioral responses of malePeriplaneta americana L. to female sex pheromone components,periplanone-A and periplanone-B

Bioassays were performed on malePeriplaneta americana L. using synthetic (–)-periplanone-B (P-B) and Hauptmann's (–)-periplanone-A (P-A), and their mixtures at various ratios to estimate the roles of both periplanones for the sexual communication of the species. Both P-A and P-B elicited qualitatively the same responses, such as antennal movement, rapid locomotion, wing raising, and homosexual behavior of male cockroaches, but the threshold of the pheromone activities for P-B was two orders of magnitude lower than that of P-A. Neither synergistic nor inhibitory but only a simple integrated effect on the responses was observed when mixtures of P-A and P-B were applied.     

Synthesis of two biologically active insulin analogues with modifications at the N-terminal and N- and C-terminal amino acid residues

The synthesis and isolation in purified form of two analogues of insulin is described. [21-Isoasparagine-A] ([Iasn21-A]) insulin differs from the parent molecule in that the amino acid residue, asparagine, found at the C terminus of the A chain (A21) has been replaced by isoasparagine. [Sar1, Iasn21-A] insulin differs from insulin in that both the A1 and A21 amino acid residues, glycine and asparagine, have been substituted by sarcosine and isoasparagine, respectively. The synthesis of these analogues followed the pattern employed in this laboratory for the synthesis of insulin and its analogues. The S-sulfonated derivatives of the A chain analogues were chemically synthesized, converted to their sulfhydryl forms, and then combined with the S-sulfonated B chain to produce the respective insulin analogues. Isolation of the insulin analogues from the combination mixtures was effected by chromatography on a carboxymethylcellulose column with an exponential sodium chloride gradient. By the mouse convulsion assay method [Iasn21-A]insulin possessed a potency of 21 IU/mg and [Sar1, Iasn21-A] insulin 15 IU/mg. The radioimmunoassay method gave values of 16 IU/mg for the former and 7IU/mg for the altter analogue. The natural hormone has a potency of 23-25 IU/mg by both assay methods. These data indicate that the alpha- and beta-carboxyl groups of the A21 amino acid residue are nearly equivalent in terms of their contribution to the expression of the biological activity of insulin. Furthermore, these data strengthen the speculation (Cosmatos, A., Johnson, S., Breier, B., and Katsoyannis, P. G. (1975), J. Chem. Soc. Perkin Trans. 1, 2157) that the change in the relative positive charge at the N-terminal amino acid residue of the A chain is responsible for the considerable decrease in the immunoreactivity observed in such modified insulins.     

枯草芽孢杆菌糖化型α-淀粉酶的研究(Ⅲ)——酶的催化作用

糖化型α-淀粉酶(BSA)作用于淀粉时与液化型α-淀粉酶(BLA)具有相同的基本特性——随机切割α-1.4葡萄糖苷键,产生小分子糊精、低聚糖。BSA能水解低聚糖(G7-G4),且对G3有一定的分解力,主要产物为G2和G。其水解淀粉程度比BLA彻底,同一蓝值时产生的还原端比BLA多。BSA不能切割1.6-葡萄糖苷键,作用于支链淀粉时残留异麦芽糖苷麦芽糖BSA还有一定的分子间糖基转移催化活性。     

Growth and poliovirus production of Vero cells on a novel microcarrier with artificial cell adhesive protein under serum-free conditions

A microcarrier is used for the three-dimensional (3D) culture of adhesion-dependent mammalian cells. We developed a novel microcarrier by binding ProNectin F, an artificial cell adhesive protein synthesized by genetically engineered Escherichia coli to a polyacrylic superabsorbent polymer. The microcarrier is characterized by containing no animal-derived components. The serum-free culture of Vero cells for vaccine production using the microcarrier increased the number of Vero cells by approximately 30% compared with the existing dextran beads coated with porcine Type I collagen, which resulted in approximately a 30% to 40% increase in the infectivity titer of the Sabin 2 strain of poliovirus. These results suggested that the developed microcarrier should be unprecedented in permitting high-yield vaccine production by means of a serum-free culture.     

An outbreak of food-borne listeriosis due to cheese in Japan, during 2001

Food-borne outbreaks caused by Listeria monocytogenes have been recognized in US and European countries. Only sporadic cases, of neonatal listeriosis, have been reported in Japan. Since L. monocytogenes has been often isolated from foods in Japan, food-borne outbreaks potentially could have occurred. In February 2001, L. monocytogenes serotype 1/2b was isolated from a washed-type cheese during routine Listeria monitoring of 123 domestic cheeses. Further samples from products and the environments at the plant that produced the contaminated cheese were examined for L. monocytogenes. L. monocytogenes serotype 1/2b was detected in 15 cheese samples, at most probable number that ranged from <30 to 4.6 x 10(9)/100 g, and in environmental samples. Studies with people who had consumed cheese from the plant revealed 86 persons who had been infected with L. monocytogenes. Thirty-eight of those people had developed clinical symptoms of gastroenteritis or the common cold type after the consumption of cheese. Isolates from those patients exhibited the same serotype, pathogenicity for mice and HeLa cells, DNA fingerprinting patterns and PCR amplification patterns. From the epidemiological and genetic evidence, it appeared that the outbreak was caused by cheese. This is the first documented incidence of food-borne listeriosis in Japan.     

Utilization of adzuki bean extract as a natural antioxidant in cured and uncured cooked pork sausages

A commercial adzuki bean extract (AE) was evaluated for antioxidant effectiveness in cured and uncured cooked pork sausages. TBARS values, instrumental color evaluation and sensory panel scores were assessed. For uncured sausages, AE at 0.2% was equally effective as 0.1% butylated hydroxytoluene (BHT) in reducing TBARS values. Similarly, AE at 0.2% significantly (P<0.01) reduced the TBARS in cured sausages. Incorporation of 0.2% AE into sausages produced higher (P<0.05) CIE lab color a* value and lower (P<0.05) L* and b* values. Sensory panels did not detect any difference in color, odor, taste, flavor, and overall acceptance in uncured pork sausages with addition of 0.2% AE. However, there were adverse changes in the color and odor of cured sausages, even though the taste, flavor, and overall acceptance were similar. Therefore, the results suggest that AE is a potential antioxidant.     

In vivo visual reporter system for detection of estrogen-like substances by transgenic medaka

Detection of endocrine disrupting chemicals, in particular, environmental estrogens with living organisms, has many advantages if compared to chemical analysis. The screening of novel pollutants with meaningful endpoints, the integration of uptake, bioconcentration, and excretion as well as the evaluation of endocrine disrupting effects with respect to toxicity require in vivo biotests for estrogen-like substances (ELSs). Critical disadvantages of whole organism biotests are their low sensitivity and the need for laborious and time-consuming work. To overcome these problems, we have developed a transgenic medaka strain harboring the green fluorescence protein (GFP) gene driven by choriogenin H gene regulatory elements. Choriogenin H is an egg envelope protein induced by estrogens in the liver. With yolk sac larvae of this strain, GFP induction in liver was observed 24 h after onset of aqueous exposure to 0.63 nM 17beta-estradiol (E2), 0.34 nM ethynylestradiol, or 14.8 nM estrone. Furthermore, concentrated sewage treatment effluent induced GFP expression. Comparison of E2 equivalents estimated by GFP-induction in transgenic medaka, a YES assay, and GC/MS showed detection limits in the same order of magnitude. These results indicated that the sensitivity of the transgenic medaka strain was sufficient for application as an alternative model in monitoring environmental water samples for ELSs.     

Cloning and characterization of a β-N-acetylglucosaminidase (BmFDL) from silkworm Bombyx mori

In insects, β-N-acetylglucosaminidase (GlcNAcase) participates in critical physiological processes such as fertilization, metamorphosis, and glycoconjugate degradation. Insects produce glycoproteins carrying paucimannosidic-type N-glycans, the terminal GlcNAc residue of which is cleaved by a GlcNAc-linkage specific GlcNAcase, also known as the fused lobes (FDL) protein. To obtain information on the structure of GlcNAcases and insight into their contribution to physiological processes, we cloned Bombyx mori FDL (BmFDL) from silkworm larvae. The full-length cDNA (1.9 kb) encoded a protein of 633 amino acids with 42% amino acid sequence identity to Drosophila melanogaster FDL (DmFDL). Recombinant BmFDL cleaved only β-1,2-linked GlcNAc residues from the α-1,3 branch of biantennary N-glycan. This substrate specificity was similar to that of DmFDL. Microsomal FDL activity was inhibited by anti-BmFDL antibodies. Taken together, our results suggest that BmFDL is a N-glycan-processing GlcNAcase in B. mori.     

Mumefural and related HMF derivatives from Japanese apricot fruit juice concentrate show multiple inhibitory effects on pandemic influenza A (H1N1) virus

Fruit-juice concentrate of Japanese apricot (Prunus mume Sieb. et Zucc.) has been shown to be effective against influenza A infection in MDCK cells. In this study, we isolated five components from the fruit-juice concentrate of Japanese apricot, 5-(hydroxymethyl)-2-formylfuran (HMF), 1-[5-(2-formylfuryl)methyl]dihydrogen 2-hydroxypropane-1,2,3-tricarboxylate (mumefural, MF), 2-[5-(2-formylfuryl)methyl]dihydrogen 2-hydroxypropane-1,2,3-tricarboxylate (MF‘), 1-[5-(2-formylfuryl)methyl]hydrogen 1-hydroxyethane-1,2-dicarboxylate (MA1) and 2-[5-(2-formylfuryl)methyl]hydrogen 1-hydroxyethane-1,2-dicarboxylate (MA2), and investigated their inhibitory activities against the novel influenza A/Narita/1/2009 (H1N1) pandemic virus hemagglutinin and neuraminidase functions, which are essential for viral attachment and budding, respectively. An hemagglutination inhibition assay indicated that MF and MF‘ were effective at minimum hemagglutination concentrations of 3.1 and 6.3 mM, respectively. An inhibition study for sialidase activity of the neuraminidase spike showed that MF was the most active anti-sialidase compound with an IC₅₀ value of 0.21 ± 0.01 mM, followed by MA2 (IC₅₀, 0.71 ± 0.09 mM), MA1 (IC₅₀, 1.64 ± 0.31 mM) and MF‘(IC₅₀, 1.62 ± 0.22 mM). Furthermore, MF was shown to inhibit the growth of the pandemic virus in a dose-dependent manner (62 ± 3% inhibition at 5 mM). The results suggest that MF, a citric acid ester linked to HMF at the 1-position of the propane backbone, might be a lead compound for the development of anti-influenza A inhibitors.