Antibacterial activity of antagonistic bacterium Bacillus subtilis DJM-51 against phytopathogenic Clavibacter michiganense subsp. michiganense ATCC 7429 in vitro.

To investigate antibacterial activity against the tomato pathogen Clavibacter michiganense subsp. michiganense ATCC 7429 (Cmm ATCC 7429), Bacillus subtilis DJM-51 was isolated from rhizosphere soil. For isolation of bacteria, samples were taken from rhizosphere soil. The isolate, DJA-51, had strong antagonistic ability against Tomato pathogen Cmm ATCC 7429 on nutrient-broth yeast extract agar (NBYA) as indicated by inhibition zones around colonies. On the basis of the nucleotide sequence of a conserved segment of the 16S rRNA gene, the bacterium has been identified as B. subtilis DJM-51. The growth of Cmm ATCC 7429 on NBYA plates was inhibited by culture broth of B. subtilis DJM-51 including cells, by the supernatant of culture broth of B. subtilis DJM-51, and by the liquid material resulting from butanol extract of bacterial cultures. The OD value in co-culture mixture was lower than the control throughout the entire incubation period. Antibiotics obtained from B. subtilis DJM-51 inhibited the growth of Tomato pathogen Cmm ATCC 7429. These results provide potentially information about the protection of tomato from pathogen Cmm ATCC 7429 under greenhouse conditions in Quebec.

Association between interleukin-27 polymorphisms and pulmonary tuberculosis.

OBJECTIVE: To investigate the effect of interleukin (IL) 27 -964A/G, 2095T/G, 4603G/A and 4730T/C gene polymorphisms on the development of pulmonary tuberculosis (PTB), radiographic characteristics and severity. DESIGN: Differences in the allele and genotype distributions of the -964A/G, 2095T/G, 4603G/A and 4730T/C polymorphisms between 224 PTB patients and 233 healthy controls, between patients with single- and multi-lobe involvement, and between patients with and without cavitation, were investigated. Serum IL-27 concentration was measured using an enzyme-linked immunosorbent assay. RESULTS: There were no significant differences in the allele or genotype distributions between PTB patients and healthy controls. However, the -964A/A genotype was more prevalent in patients with single-lobe involvement than the -964A/G or -964G/G genotype in patients with multi-lobe involvement (50.0% vs. 31.3%, P = 0.01). There was no difference between patients with and without cavitation (P > 0.05). Serum median IL-27 concentration was significantly higher in patients with single-lobe involvement than in those with multi-lobe involvement (P = 0.03) and in those with -964A/A genotypes than in those with -964A/G or -964G/G genotypes (P = 0.02). CONCLUSIONS: In terms of serum IL-27 levels, the -964 A/A genotype may be associated with a protective role that prevents the intrapulmonary spread of PTB rather than its development.

Selection of a high affinity angiogenin-binding peptide from a peptide library displayed on phage coat protein.

Although the phage-displayed peptide libraries have been widely used for the biopanning of peptide specific for various types of target molecules, the selected peptides often have affinities too low for practical purposes. In this report, selection of a high affinity peptide ligand specific for human angiogenin is described. We constructed a random dodecapeptide library displayed on the gene III protein of filamentous bacteriophage by use of a phagemid. The peptide insert was flanked by two cysteines to constrain the peptide structure by a disulfide bond. Phages were collected from 1.5 x 10(3) independent transformants. After the library phages were allowed to bind to the human angiogenin coated on a plate, the phages bound to the actin-binding site of angiogenin were selectively eluted with actin. The activity of each phage clone was estimated and three high affinity phage clones were identified. The peptides displayed by the three phage clones were synthesized as fusion proteins with Escherichia coli maltose-binding protein, and used for the quantitative estimation of their affinities. By this procedure, we were able to select a peptide having a dissociation constant of about 60 nM.

Localization of mesial temporal lobe seizures with sphenoidal electrodes.

Few studies have compared sphenoidal electrodes with scalp electrodes to determine their localizing value in temporal lobe seizures. We reviewed 76 ictal recordings with the standard International 10-20 System and T1/2 and sphenoidal electrodes from 31 patients whose subsequent intracranial EEG studies precisely localized seizures in one temporal lobe. Ictal EEGs were reviewed in a blinded fashion in both longitudinal bipolar and referential montages. Of the 23 seizures in 11 patients with mesial temporal lobe epilepsy (MTLE), all seven seizures in 3 patients were localized exclusively to one sphenoidal electrode, before involvement of T1/2 and temporal scalp electrodes. The remaining 16 seizures from 8 MTLE patients and all 53 seizures in the 20 patients with neocortical temporal lobe epilepsy (NTLE) had simultaneous involvement of sphenoidal, T1/2, and temporal scalp electrodes at seizure onset when analyzed with referential montages. Sphenoidal electrodes may provide valuable localization data for presurgical evaluation of patients with possible TLE. In particular, seizure rhythms confined to the sphenoidal electrode at ictal onset, without involvement of scalp electrodes, occurred only in patients with MTLE (p < 0.04). Further studies comparing these electrodes with other types of surface electrodes are needed.

Application of surface modified polypropylene membranes to an anaerobic membrane bioreactor.

In order to increase hydrophilicity and thereby to reduce membrane fouling caused by hydrophobic adsorption, the surface of a hydrophobic 0.2 microm polypropylene (PP) membrane was modified by ozone treatment followed by graft polymerization with 2-hydroxy-ethyl methacrylate (HEMA). The modified PP (MPP) membranes were characterized in terms of contact angle, morphology and degree of grafting (DG). The contact angle was reduced from 112 degrees for a PP membrane to nearly 0 degrees for MPP membranes by introducing functional groups such as hydroxyl (-OH) and carbonyl groups (C=O) on the membrane surface. As the DG increased, the O/C ratio and membrane resistance of the MPP membrane increased. Using the MPP membrane in the crossflow operation of an anaerobic membrane bioreactor (MBR), the membrane permeability was enhanced although it was largely dependent on the DG of MPP.

Extraction of chitin from red crab shell waste by cofermentation with Lactobacillus paracasei subsp. tolerans KCTC-3074 and Serratia marcescens FS-3.

For one-step extraction of chitin from red crab shell waste, cofermentation with Lactobacillus paracasei subsp. tolerans KCTC-3074, a lactic-acid-producing bacterium, and Serratia marcescens FS-3, a protease-producing bacterium, was conducted. Fermentation with single strain (L. 3074 or FS-3) was also conducted. At day 7, the pH in L. 3074, FS-3, and L. 3074+FS-3 (1:1) treatment decreased from 6.90 to 3.30, 5.88, and 3.48, respectively. Ash content in the residue after fermentation treatment of crab shells in L. 3074 and L. 3074+FS-3 (1:1) treatment drastically decreased from 41.2% to 3.19 and 1.15%, respectively. In L. 3074+FS-3 (1:1) cofermentation, the level of demineralization was the highest value of 97.2%, but the level of deproteinization in the cofermentation was 52.6% at day 7. Protein content in the treatment of FS-3 alone reduced from 22.4 to 3.62%. These results indicate that cofermentation of the shells using the two strains is efficient and applicable for the one-step extraction of crude chitin from red crab shell waste.

Demineralization of red crab shell waste by lactic acid fermentation.

Lactic acid fermentation was applied to demineralize red crab shell waste using Lactobacillus paracasei subsp. tolerans KCTC-3074. Various concentrations (0, 2.5, 5.0, 10.0%) of glucose were supplemented as an initial carbon source and various amounts (2.5, 5.0, 10.0%) of the bacterial culture were inoculated as a starter. Microbial growth was very dependent on glucose concentration but little dependent on inoculum level. The pH decreased rapidly from pH 8 to pH 6 during the first day, at all three inoculum levels. At day 5 of fermentation, the 2.5, 5.0, and 10.0% inoculum levels with 10% glucose supply gave pH 5.5, 5.1, and 4.6, respectively, i.e. a decrease in pH concomitant with an increase in the inoculum level. The total titratable acidities (TTA) at day 5 for 2.5, 5.0, and 10.0% inoculum levels with 10% glucose supplement were 3.1, 4.5, and 8.3%, and the relative residual ash contents were 26.6, 25.9, and 19.0%, respectively. A negative relationship was found between pH and demineralization level (r2 = 0.8571), but there was a positive relationship between TTA and demineralization level (r2 = 0.5532).

Production of N-acetyl-beta-D-glucosamine from chitin by Aeromonas sp. GJ-18 crude enzyme.

A bacterium, GJ-18, having strong chitinolytic activity was isolated from coastal soil. The isolated strain was identified as Aeromonas sp. by morphological and biochemical properties along with 16S rRNA gene sequence. The crude chitinolytic activity of culture supernatants was maximal on the 5th day of culture. Below 45 degrees C, chitin was effectively hydrolyzed to N-acetyl-beta-D-glucosamine (GlcNAc) by Aeromonas sp. GJ-18 crude enzymes, but hydrolysis decreased markedly above 50 degrees C. The optimum pH for enzyme activity was 5.0. TLC and HPLC analysis revealed that, below 45 degrees C, the major reaction product was GlcNAc with a small amount of (GlcNAc)(2) and (GlcNAc)(3), whereas above 50 degrees C the major product was (GlcNAc)(2). When swollen chitin (100 mg) was incubated with crude enzyme preparations (10 U) at 40 degrees C, chitin was hydrolyzed to 83.0 and 94.9% yield of GlcNAc within 5 and 9 days, respectively.

Hepatic involvement in hypereosinophilia: sonographic findings.

Hypereosinophilic syndrome may cause eosinophil-related tissue damage to various organs. The purpose of this paper is to describe sonographic findings in 13 patients with hypereosinophilia in whom the liver was involved. The diagnosis in these 13 patients was based on liver biopsy in seven patients with bone marrow biopsy in six patients. Eight patients had hypereosinophilic syndrome and five patients had clonorchiasis. All 13 patients had mild to marked hepatomegaly. Seven of 13 patients showed multiple round or oval hypoechoic (n = 6) or variably echogenic (n = 1) lesions measuring 1 to 2 cm with poorly defined margins in both lobes of the liver. Four patients had one or two hypoechoic lesions 3 to 4 cm in size, with geographic pattern and poorly defined margins. Two patients showed diffuse hepatomegaly with increased parenchymal echogenicity. The number of lesions and the extent of diffuse lesions seem to be proportional to the degree of eosinophilia. Hypereosinophilia may produce multiple small focal hepatic lesions or diffuse segmental or lobar echogenic lesions simulating primary or metastatic tumor of the liver.