Identification of a distinct lineage of Cacopsylla chinensis (Hemiptera: Psyllidae) in Japan on the basis of two mitochondrial DNA sequences.

Psyllids are a menacing pests of pears (Pyrus spp.) grown in temperate and subtropical regions of the world, including Taiwan and China. Pear psyllids belong to the large genus Cacopsylla (Psyllidae: Psyllinae). Among the 28 psyllid species that infest pear trees, Cacopsylla chinensis (Yang and Li, 1981) is considered the most harmful. Two psyllid outbreaks involving exotic species affected Japanese pear (Pyrus pyrifolia variety culta) orchards in the Saga Prefecture in July and November 2011. The psyllids were morphologically identical to the summer and winter forms of C. chinensis. In this study, we performed DNA sequence analysis of the mitochondrial cytochrome oxidase I-leucine tRNA-cytochrome oxidase II (COI-tRNA(Leu)-COII) and 16S rDNA regions to elucidate the phylogenetic relationships among 82 summer-form psyllids, five winter-form psyllids from several orchards in Japan, and those reported from Taiwan and China. The sequences of the COI-tRNA(LEU)-COII and 16S rDNA regions were identical among all 87 psyllids from Japan, regardless of summer/winter forms or orchards in Saga, Japan. Comparison of nucleotide sequences and phylogenetic analysis differentiated Japanese psyllids from the Taiwanese and Chinese C. chinensis isolates, with approximately 8 and 7% nucleotide difference in the COI-tRNA(LEU)-COII and 16S rDNA regions, respectively. The results suggest that C. chinensis possess a high level of genetic variability and that the psyllids responsible for the outbreak in Saga, Japan belong to a distinct lineage of C. chinensis.

[Postoperative recurrence of spontaneous pneumothorax].

Few reports on recurrence after thoracoscopic bullectomy for spontaneous pneumothorax specify the follow-up period and follow-up ratio. Because of the variation in follow-up periods, many reported recurrence rates were not comparable. Some reports compared simple recurrence rate (number of recurrent cases/number of operated cases) of different groups with different follow-up periods. In this study, we employ the Kaplan-Meier method along with a set of optimal follow-up periods and ratios in order to determine a more reliable recurrence rate. Consecutive 68 patients (74 surgical procedures) underwent thoracoscopic bullectomy for spontaneous pneumothorax at our institution between November 2000 and December 2005. A follow-up survey was conducted by phone to determine the rate of recurrent pneumothorax. The follow-up ratio and the mean follow-up period were 92.6% and 1,316 +/- 481 days, respectively. Postoperative recurrence was confirmed for 4 patients. The interval up to recurrence was 144, 345, 476 and 616 days after the bullectomy, respectively. All cases of recurrent pneumothorax occurred within 2 years following the bullectomy. The 1-year, 2-year and 3-year cumulative recurrence rate was 3.0%, 6.3% and 6.3%, respectively. In light of these findings, we feel that comparison analysis of pneumothorax recurrence rates should be evaluated using the Kaplan-Meier method, furthermore, our data suggests that a follow-up period of 2 or more years is advisable.

Nucleotide sequence of the coat protein genes of citrus mosaic virus.

The sequence of the 3'-terminal 2,201 nucleotides of RNA2 of citrus mosaic virus (CiMV) was determined. The sequence contains a long open reading frame (ORF) of 1989 nucleotides in the virus-sense strand and at 3' untranslated region of 212 nucleotides upstream of a poly(A) tail. The N-terminal amino acid sequence of the two coat proteins was determined by Edman degradation and the corresponding coding regions were identified in the polyprotein. The larger coat protein with Mr 48,122 is encoded upstream of the smaller one with Mr 24,172. The coat proteins are apparently cleaved from the polyprotein at an Arg-Gly and a Thr-Asn bond. Although CiMV has properties in common with the comoviruses and nepoviruses, there is no significant sequence homology between the coat proteins of CiMV and those of either group. Furthermore, the coat proteins of CiMV lack homology with those of strawberry latent ringspot virus (SLRSV), which has been reported to be more distantly related to the como- and nepoviruses. This lack of homology distinguishes CiMV from the como- and nepoviruses, SLRSV and other viruses.

Post-thymectomy myasthenia gravis with an episode of Osserman stage III.

Here, we present the first case of post-thymectomy Myasthenia Gravis with onset at Osserman stage III. An 81-year-old woman was admitted for an abnormal shadow seen in a chest radiograph. She had no symptoms of Myasthenia Gravis. Acetylcholine receptor antibody was within the normal range. Chest computed tomography (CT) showed a bulky anterior mediastinal tumor. She was diagnosed as having thymoma by tissue biopsy under CT guidance. The tumor was completely resected by performing thymothymectomy, left upper lobectomy, pericardial resection, and phrenicectomy. Pathological examination of the tumor identified it as a thymoma (type B2, Masaoka stage II). Two months after the surgery, she experienced the onset of post-thymectomy myasthenia gravis with Osserman stage III. The acetylcholine receptor antibody level was remarkably elevated (220 nmol/L); however, there was no evidence of tumor recurrence.

Microbial changes in patients with acute periodontal abscess after treatment detected by PadoTest.

AIMS: To investigate changes in bacterial counts in subgingival plaque from patients with acute periodontal abscess by IAI-PadoTest. MATERIALS AND METHODS: Ninety-one patients were randomly allocated to either test or control groups. In all the patients, pockets with acute periodontal abscess were irrigated with sterilized physiological saline, and in the test group, 2% minocycline hydrochloride ointment was applied once into the pocket in addition. Subgingival plaque samples were collected by paper point before treatment and 7 days after treatment. The total bacterial count was determined and Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia, were detected using IAI-PadoTest, a DNA/RNA probe method. RESULTS: The total bacterial count decreased in both groups, with a significant decrease in the test group. The counts and number of sites positive for P. gingivalis, T. forsythia and T. denticola significantly decreased in the test group after treatment, compared with those in the control group. Pocket depth decreased in the both groups, with a statistically significant decrease in the test group. CONCLUSION: Topical treatment with minocycline in pockets with acute periodontal abscess was effective in reducing the bacterial counts as shown by the microbiological investigation using PadoTest 4.5.

Cheravirus and Sadwavirus: two unassigned genera of plant positive-sense single-stranded RNA viruses formerly considered atypical members of the genus Nepovirus (family Comoviridae).

The genus Nepovirus (family Comoviridae) was known both for a good level of homogeneity and for the presence of atypical members. In particular, the atypical members of the genus differed by the number of capsid protein (CP) subunits. While typical nepoviruses have a single CP subunit with three structural domains, atypical nepoviruses have either three small CP subunits, probably corresponding to the three individual domains, or a large and a small subunit, probably containing two and one structural domains, respectively. These differences are corroborated by hierarchical clustering based on sequences derived from both genomic RNAs. Therefore, these atypical viruses are now classified in two distinct genera, Cheravirus (three CP subunits; type species Cherry rasp leaf virus) and Sadwavirus (two CP subunits; type species Satsuma dwarf virus).

Nucleotide sequence of melon yellow spot virus M RNA segment and characterization of non-viral sequences in subgenomic RNA.

The nucleotide sequence of melon yellow spot virus (MYSV) M RNA segment was determined. The M RNA segment contains one open reading frame (ORF) encoding 308 amino acids (aa) in the sense orientation and another ORF encoding 1,127 aa in the complementary orientation, which were homologous to the NSm protein and G1/G2 glycoprotein precursor (Gp) protein, respectively. Amino acid sequences identities with the other tospovirus suggested that MYSV is closely related to groundnut bud necrosis virus and watermelon silver mottle virus. To analyze subgenomic RNA of the M RNA segment, RNA transcripts corresponding to the NSm and Gp genes were specifically amplified, and the nucleotide sequence of the 5' terminal region was determined. Sequence analysis of the NSm and Gp transcripts showed that they had a non-viral sequence 12-18 and 10-18 nucleotides long, respectively. Although these sequences varied considerably, in more than half of the cases, a cytosine residue was observed at the 3' end of the non-viral leader sequence, which suggests that the viral transcriptase prefers certain cap-donor sequences harboring a 3'CA dinucleotide.

Nucleotide sequences and taxonomy of satsuma dwarf virus.

The nucleotide sequences of genomic RNA1 (6795 nt) and RNA2 (5345 nt) of satsuma dwarf virus (SDV), a tentative member of the genus Nepovirus, were determined. The deduced genome organization of SDV showed similarities to the organization in como-, faba- and nepoviruses. There is extensive amino acid sequence similarity in the N-terminal regions of the proteins encoded by RNA1 and RNA2, as reported previously only for tomato ringspot nepovirus. However, unlike definitive nepoviruses, which have a single coat protein, SDV has two coat proteins. SDV RNA2 does not contain the long (> 1300 nt) 3' non-coding region characteristic of some nepoviruses. Phylogenetic analysis of SDV RNA polymerase placed SDV apart from como-, faba- and nepoviruses. These unique features suggest that SDV is distinct from the Comovirus, Fabavirus and Nepovirus genera, and needs to be separated into a new genus, probably within the family Comoviridae.

Satsuma dwarf and related viruses belong to a new lineage of plant picorna-like viruses.

Satsuma dwarf virus (SDV) and two closely related viruses, Citrus mosaic (CiMV), and Naval orange infectious mottling (NIMV), seriously affect citrus varieties grown in Japan and East Asia. All three viruses have icosahedral particles built of two proteins encapsidating two single-stranded genomic RNAs. The natural mode of transmission of these SDV-like viruses is unknown, and they were previously placed among tentative members of the family Comoviridae. Recently, a complete genome of SDV was sequenced, and its replication-related proteins were found only distantly related to those of viruses from the family Comoviridae (Iwanami T., Kondo Y., and Karasev A.V. J Gen Virol 80, 793-797, 1999). Here we present a partial genome sequence for another SDV-like virus, NIMV, and a thorough phylogenetic analysis of the gene products encoded by SDV, CiMV, and NIMV to assess their relationships with picorna-like viruses infecting plants, insects, and vertebrates. The RdRp's of SDV-like viruses form a new lineage, separate from members of Como- and Sequiviridae families. Phylogenetic analysis suggests that SDV-like viruses may represent a new family of plant picorna-like viruses. Sequence analysis of the capsid proteins (CPs) encoded by the SDV-like viruses revealed a region of similarity to CPs of animal calici- and picornaviruses that encompasses the structural core of the eight-strand beta-barrel characteristic of picornaviral CPs. These data suggest that SDV and related bipartite viruses evolved separately from the viruses in the family Comoviridae and that the split of an ancestor, monopartite picorna-like virus genome might have occurred more than once.

Increased serum concentration of urinary trypsin inhibitor with asthma exacerbation.

The aim of the study was to determine whether the amount of urinary trypsin inhibitor (UTI) in serum, a degenerate induced by neutrophil elastase (NE), reflects the degree of bronchial inflammation in children with acute asthma exacerbation. The involvement of neutrophil-mediated inflammation plays as important a role as eosinophil-mediated inflammation in the pathogenesis of acute asthma exacerbation. However, no measurable marker is sensitive enough to assess neutrophil-mediated inflammation in the airways. The pre-alpha-/inter-alpha-trypsin inhibitors are assumed to be precursors of UTI. NE degrades pre-alpha-/inter-alpha-trypsin inhibitors to liberate UTI. UTI concentrations in 25 childhood patients admitted with asthma exacerbation and 15 control subjects were measured by means of one-step sandwich-type enzyme immunoassay. Serum UTI concentrations in the patients at admission were significantly higher than control values (10.597+/-0.649 and 6.136+/-0.303 U x mL(-1), respectively (mean+/-SEM)). These levels returned to baseline values with improvement in the asthmatic symptoms. However, serum NE and alpha1 antitrypsin concentrations were not significantly different between patients and controls, even during acute exacerbation in the former. The findings suggest that neutrophil-mediated inflammatory events are involved in exacerbation of childhood asthma. The monitoring of urinary trypsin inhibitor concentrations might be useful for evaluating the neutrophil-mediated inflammation in childhood asthma attack.

Nucleotide sequence of a Japanese isolate of Squash mosaic virus. Brief report.

The complete nucleotide sequence of the RNA-1 of Squash mosaic virus (SqMV) was determined using a Japanese isolate (Y-SqMV). The sequence consisted of 5865 nucleotides excluding the poly (A) at the 3' terminus and contained a single long open reading frame with a coding capacity for a protein of Mr209971. Analysis of the deduced amino acid sequence suggested a genomic organization typical of comoviruses. The nucleotide sequence of the RNA-2 of Y-SqMV was also determined and compared with the SqMV isolates from the United States. The larger and smaller capsid protein (CP) coding region was compared to those of K-SqMV and Z-SqMV, which represent two subgroups of SqMV. The larger CP gene of Y-SqMV showed 93.0% and 88.0% identities with those of K-SqMV and Z-SqMV, respectively at the nucleotide level. The smaller CP gene of Y-SqMV was 94.1% and 88.4% identical with those of K-SqMV and Z-SqMV. The results suggested that the Japanese SqMV isolate (Y-SqMV) is distinct from those in the United States, and might represent a third subgroup.

Complete nucleotide sequences of genome segments 1 and 3 of Rosellinia anti-rot virus in the family Reoviridae.

The nucleotide sequences of genome segments 1 and 3 of Rosellinia anti-rot virus (RArV) from a hypovirulent isolate, W370, of the plant pathogen Rosellinia necatrix were determined. The complete nucleotide sequence of the genome segment 1 encoded a putative RNA-dependent RNA polymerase (RDRP). The deduced amino acid sequence of RDRP of RArV showed 29% identity with RDRPs of Colorado tick fever virus (CTFV) and European Eyach virus (EYAV) in the genus Coltivirus, and identities of 23-21% with members of the genera Fijivirus and Cypovirus. Both RArV and the Coltivirus member might have originated from a common virus ancestor.

A new virus related to Satsuma dwarf virus: the nucleotide sequence of the 3'-terminal regions of Hyuganatsu virus RNAs 1 and 2. Brief Report.

The sequences of the 3'-terminal 1306 and 2160 nucleotides of RNAs 1 and 2 of a virus serologically related to Satsuma dwarf virus (SDV) from Hyuganatsu ( Citrus tamurana Hort. ex Tan.) were determined, respectively. We found that the partial RNA-dependent RNA polymerase region in RNA1 and the coat proteins (CPs) region in RNA2 of the virus tentatively named Hyuganatsu virus (HV) have 78.3-84.0% and 76.9-80.7% amino acid sequence identities to those of known SDV-related viruses (SDV-RVs), i.e., SDV, Citrus mosaic virus, and Navel orange infectious mottling virus. Sequence analyses show that HV is classifiable as a new SDV-RV species.

Purification and certain properties of a bacteriocin from Streptococcus mutans.

An inhibition factor from Streptococcus mutans strain C3603 (serotype c) was purified and isolated, and its properties indicated that it was a bacteriocin. Bacteriocin C3603 is a basic protein with a pI value of 10 and a molecular weight of 4,800. The activity of this bacteriocin was not affected by pH over a range of 1.0 to 12.0 or by storage at 100 degrees C for 10 min at pH 2.0 to 7.0 or storage at 121 degrees C for 15 min at pH 4.0. Pronase; papain, phospholipase C, trypsin, and alpha-amylase had no effect on the activity of the bacteriocin, whereas alpha-chymotrypsin and pancreatin were partially active against it. Bacteriocin activity was greater against certain S. mutans strains of serotypes b, c, e, and f than against certain S. mutans strains of serotypes a, d, and g. Bacteriocin C3603 was also effective against selected strains of S. sanguis, S. salivarius, S. bovis, S. faecium, S. lactis, Lactobacillus casei, L. plantarum, L. fermentum, Bifidobacterium bifidum, Bifidobacterium longum, Propionibacterium acnes, and Bacteroides melaninogenicus, but it was not effective against certain strains of Escherichia coli, Klebsiella pneumoniae, Corynebacterium parvum, and Candida albicans. The inhibition of S. mutans strains BHT and PS-14 by bacteriocin C3603 was found to be due to the bacteriocidal activity of the bacteriocin. When water or a diet containing bacteriocin C3603 was consumed by gnotobiotic and specific pathogen-free rats infected with S. mutans PS-14, the caries score was found to be significantly reduced.

The nucleotide sequence of the coat protein genes of satsuma dwarf virus and naval orange infectious mottling virus.

The sequence of the 3'-terminal 4320 and 2409 nucleotides were determined for RNA2 of satsuma dwarf virus (SDV) and navel infectious mottling virus (NIMV). Both sequences contained a part of a long open reading frame which encodes larger and smaller coat proteins (CPs) at the 3'-terminus followed by a 3'non-coding region upstream of a poly (A) tail. Amino acid sequence identity for larger and smaller CPs ranged 81-84% and 68-78%, respectively, among SDV, NIMV and the previously sequenced citrus mosaic virus (CiMV). No significant sequence similarity was found between the CPs of SDV or NIMV and those of the como-, nepo- or other viruses. The nucleotide sequence identity of the 3' non-coding region of RNA2 were 68%-78% among SDV, CiMV and NIMV. These results suggest that SDV, CiMV and NIMV are distinct, though related, viruses. They may be assigned as members of the new genus, which is close to the genera of Comovirus and Nepovirus.

Nucleotide sequence and taxonomy of Cycas necrotic stunt virus. Brief report.

Cycas necrotic stunt virus (CNSV) is the only well-characterized virus from gymnosperm. cDNA segments corresponding to the bipartite genome RNAs (RNA1, RNA2) were synthesized and sequenced. Each RNA encoded a single polyprotein, flanked by the 5' and 3' non-coding regions (NCR) and followed by a poly (A) tail. The putative polyproteins encoded by RNA1 and RNA2 had sets of motifs, which were characteristic of viruses in the genus Nepovirus. The polyproteins showed higher sequence identities to Artichoke Italian latent virus, Grapevine chrome mosaic virus and Tomato black ring virus, all of which belong to subgroup b of the genus Nepovirus, than to other nepoviruses. Phylogenetic analysis of RNA dependent RNA polymerase and coat protein also showed closer relationships with these viruses than other viruses. The data obtained supported the taxonomical status of CNSV as a definitive member of the genus Nepovirus, subgroup b.

Sequence diversity and interrelationships among isolates of satsuma dwarf-related viruses.

The 3'-region of RNA2 of three viruses (Natsudaidai dwarf virus (isolate ND-1), and two unidentified isolates (LB-1, Az-1)), which were related to Satsuma dwarf virus (SDV), were sequenced. Phylogenetic analysis including the previously reported SDV-related viruses (Citrus mosaic virus (CiMV, Ci-968), Navel orange infectious mottling virus (NIMV, NI-1)) showed that they were classified into three groups, SDV (S-58), CiMV (Ci-968, LB-1, Az-1, ND-1), and NIMV (NI-1). The results suggested these groups might correspond to the three distinct virus species. ND-1, LB-1, and Az-1 were considered strains of CiMV, although they do not induce citrus mosaic on the fruit rind.

Effect of mutacin administration on Streptococcus mutans-induced dental caries in rats.

A bacteriocin from serotype c Streptococcus mutans strain C3603 was examined for its inhibitory effect on experimental dental caries in rats infected with S. mutans MT8148R (serotype c). Significant reduction in the incidence of dental caries was found only when bacteriocin was incorporated both in the drinking water and in the diet at a high concentration. However, caries reduction was not as great as expected and the addition of bacteriocin to drinking water alone had no effect on the recovery of S. mutans, plaque deposition or caries incidence. The bacteriocin activity must have been reduced in the oral cavity of rats, and the reasons were examined. Bacteriocin-resistant mutants were not detected and the bacteriocin was not inactivated by saliva. Whereas the bacteriocin did not kill the S. mutans cells grown in a sucrose-containing medium, it completely killed the cells grown in a sucrose-free medium.

Structural similarity of ghrelin derivatives to peptidyl growth hormone secretagogues.

Ghrelin is a 28-amino acid residue endogenous growth hormone secretagogue. Intensive investigations revealed that the N-terminus tetrapeptide, having octanoyl group at Ser(3), is the minimum active core. In this study, we further explored the structure-function relationships of the active N-terminus portion of ghrelin using a Ca(2+) mobilization assay. The smallest and most potent ghrelin derivative we have found so far is 5-aminopentanoyl-Ser(Octyl)-Phe-Leu-aminoethylamide, showing comparable activity to the natural molecule. In the process of modifying the active core, the ghrelin-derived short analogues emerged structurally close to peptidyl growth hormone secretagogues. The N-terminus modification suggested that Gly(1)-Ser(2) unit works as a spacer, forming adequate distance between N(alpha)-amino group and n-octanoyl group. Replacement of 3rd and 4th amino acid residues to D-isomer suggested that the N-terminal dipeptide contributes to shape the biologically active geometry by effecting conformation of residues in positions 3 and 4.