Reactivity of serum immunoglobulin E to bullfrog Rana catesbeiana parvalbumins in fish-allergic patients

ABSTRACT:   Parvalbumin, a calcium-binding sarcoplasmic protein of approximately 12 kDa, represents the cross-reactive, major allergen in fish. In consideration of the fact that parvalbumin is contained at high levels not only in fish muscle but also in frog muscle, the present study was undertaken to clarify whether fish-allergic patients react to two parvalbumins (α- and β-parvalbumins) purified from the bullfrog Rana catesbeiana , which is sometimes consumed as a delicacy in Japan. In enzyme-linked immunosorbent assays (ELISA), sera from 12 of the 14 patients tested reacted equally to both parvalbumins purified from the Pacific mackerel Scomber japonicus and the bigeye tuna Thunnus obesus . Of the 12 sera positive to fish parvalbumins, eight sera also reacted to α- and β-parvalbumins of the bullfrog with different spectra: one serum reacted strongly to α-parvalbumin, six sera reacted strongly to β-parvalbumin and one serum reacted equally to both α- and β-parvalbumins. In addition, inhibition ELISA experiments revealed cross-reactivity between fish and bullfrog parvalbumins. Based on these results, it is proposed that fish-allergic patients should avoid the consumption of frog meat unless they are accurately diagnosed as lacking immunoglobulin E against frog.     

Isolation and characterization of the interspecific fusants from Streptomycetes obtained using a liquid regeneration method

ABSTRACT: Protoplast fusion between different species of Streptomyces was performed using a liquid regeneration method developed for a rapid and simple preparation of the fusants. Consequently, new clones, which could not be obtained using the conventional agar regeneration method, were obtained. In the crosses between S. griseus and S. durhamensis , and between S. californicus and S. catenulae , eight and two recombinants, respectively, were obtained using the liquid regeneration method. Conversely, in the case of crosses between S. ornatus and S. catenulae , and between S. ornatus and S. vendargensis , seven recombinants each were obtained using only the agar method. The physiological characteristics, such as the assimilation of carbohydrate and antibiotic resistance, of these fusants differed considerably from those of their parental strains. Using the proposed liquid regeneration method, a simpler and quicker procedure for protoplast fusion is described.     

Starch Properties of the Sago Palm (<Emphasis Type="Italic">Metroxylon sagu</Emphasis> Rottb.) in Different Soils

We investigated the relationships between starch concentrations and activities of starch synthetic enzymes in sago palms (Metroxylon sagu Rottb.) under acid sulfate and mineral soil conditions. Plants grown naturally that had reached their maturated stage were sampled. We found that the growth in acid sulfate soil is lower than that in mineral soil and that starch granules were larger and there was more amylase activity in acid sulfate soil than in mineral soil. Lower amylase activity in mineral soil could eliminate the degradation of starch, making the smaller granules suitable for storing large amounts of starch in a limited space inside cells.     

Antioxidative effects of the acetone fraction and vanillic acid from Chenopodium murale on tomato plants

From the active acetone fraction of Chenopodium murale , vanillic acid was isolated and identified, based on ¹H and ¹³C nuclear magnetic resonance spectral analyses. Free phenolic compounds inside the active acetone were qualified and quantified by using high performance liquid chromatography analysis, which revealed the presence of seven compounds with an abundance of vanillic and p -hydroxybenzoic acids. The allelopathic potential of the acetone fraction and vanillic acid was evaluated through laboratory bioassays against tomato plants. Our results showed that the allelopathic potential induced by low concentrations of the acetone fraction and vanillic acid stimulated the germination and growth of tomato and had stimulating effects on the activity of some antioxidant enzymes. We observed an enhancement in the activity of catalase, peroxidase, superoxide dismutase, and polyphenol oxidase, as well as the content of soluble protein and phenolic glycoside. Meanwhile, the levels of free phenolic compounds, H₂O₂, and lipid peroxidation decreased. The highest stimulations were recorded at 50 p.p.m. of acetone fraction and 0.5 p.p.m. of vanillic acid. In contrast, the highest concentrations exerted negative effects on all the measured parameters to record the maximum value of inhibition at 400 p.p.m. of acetone fraction and 4 p.p.m. of vanillic acid. These results proved the antioxidative effects of active acetone and vanillic acid at low concentrations and their potent use as a stimulator for tomato germination and growth.     

Association between antimicrobial treatment and resistance of pathogenic Escherichia coli isolated from diseased swine in Kagoshima Prefecture,Japan

Pathogenic Escherichia coli is an important cause of diarrhea, edema disease, and septicemia in swine. In Japan, the volume of antimicrobial drugs used for animals is highest in swine, but information about the prevalence of antimicrobial-resistant bacteria is confined to apparently healthy animals. In the present study, we determined the O serogroups, virulence factors, and antimicrobial resistance of 360 E. coli isolates from swine that died of disease in Kagoshima Prefecture, Japan, between 1999 and 2017. The isolates of the predominant serogroups O139, OSB9, O149, O8, and O116 possessed virulence factor genes typically found in diarrheagenic E. coli. We further found five strains of third-generation cephalosporin-resistant E. coli that each produced an extended-spectrum β-lactamase encoded by bla_CTX-M-14, bla_CTX-M-15, bla_CTX-M-24, bla_CTX-M-61, or bla_SHV-12. In 218 swine with a clear history of antimicrobial drug use, we further analyzed associations between the use of antimicrobials for the treatment of diseased swine and the isolation of resistant E. coli. We found significant associations between antimicrobial use and selection of resistance to the same class of antimicrobials, such as the use of ceftiofur and resistance to cefotaxime, cefazolin, or ampicillin, the use of aminoglycosides and resistance to streptomycin, and the use of phenicols and resistance to chloramphenicol. A significant association between antimicrobial use and the resistance of E. coli isolates to structurally unrelated antimicrobials, such as the use of ceftiofur and resistance to chloramphenicol, was also observed.     

Porphyromonas gulae 41-kDa fimbriae induced osteoclast differentiation and cytokine production

Porphyromonas gulae is considered to be associated with canine periodontitis. We have previously reported that the P. gulae American Type Culture Collection (ATCC) 51700 comprised 41-kDa fimbriae. The purpose of the present study was to demonstrate the roles of 41-kDa fimbrial protein in periodontal disease. In this study, we examined the involvement of the 41-kDa fimbrial protein in osteoclast differentiation and cytokine production in murine macrophages. Furthermore, alveolar bone resorption induced by P. gulae infection in rats was evaluated. To estimate osteoclast differentiation, bone marrow cells and MC3T3-G2/PA6 cells were cultured with or without the 41-kDa fimbrial protein for 7 days. BALB/c mouse peritoneal macrophages were stimulated with the 41-kDa fimbrial protein, and the levels of interleukin (IL)-1β and tumor necrosis factor (TNF)-α production were determined by enzyme-linked immunosorbent assay. Osteoclast differentiation was significantly enhanced by treatment with the 41-kDa fimbrial protein in a dose-dependent manner. The total area of pits formed on the dentine slices with osteoclasts incubated with the 41-kDa fimbrial protein was significantly greater than that of the control. The purified 41-kDa fimbrial protein induced IL-1β and TNF-α production in BALB/c mouse peritoneal macrophages after 6 hr of incubation in a dose-dependent manner. The bone loss level in rats infected with P. gulae was significantly higher than that of the sham-infected rats. These results suggest that P. gulae 41-kDa fimbriae play important roles in the pathogenesis of periodontal disease.     

Detection and molecular characterization of bovine leukemia virus in beef cattle presented for slaughter in Egypt

Bovine leukemia virus (BLV) is the etiological agent of enzootic bovine leukosis, the most common neoplastic disease of cattle worldwide and a serious problem for the cattle industry. Previous studies have shown the molecular prevalence of BLV and the coexistence of BLV genotype-1 and -4 in Egyptian dairy cattle; however, the molecular characteristics of BLV in Egyptian beef cattle are unknown. Therefore, we collected blood samples of 168 beef cattle from slaughterhouses in three governorates in Egypt. Based on BLV-CoCoMo-qPCR-2 targeting long terminal repeats and nested PCR targeting the env-gp51 gene, the BLV provirus infection rates were found to be 47/168 (28.0%) and 42/168 (25.0%), respectively. Phylogenetic analysis based on 501 bp of the BLV env-gp51 gene from 42 BLV isolates revealed that at least six distinctive strains (b, e, f, g, x, and z) were prevalent in cattle across the examined regions. Furthermore, phylogenetic analysis of the 420 bp sequence of the BLV env-gp51 region of the six strains against 11 known genotypes showed that the strains b, e, f, and g were clustered into genotype-1, and strains x and z were clustered into genotype-4. Our results also indicated that strains b and x exist in both dairy and beef cattle in Egypt. The present study is the first to detect and genotype BLV among beef cattle in Egypt.