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1.
J Periodontal Res ; 53(4): 536-544, 2018 Aug.
Article in English | MEDLINE | ID: mdl-29603738

ABSTRACT

BACKGROUND AND OBJECTIVE: Full-mouth scaling and root planing (FM-SRP) acts as a potent inflammatory stimulus immediately after treatment; however, systemic inflammation typically improves in the long term. The contribution of FM-SRP to systemic biological and acute-phase responses is largely unknown. The purpose of this prospective intervention study was to assess the systemic and local biological responses after FM-SRP. MATERIAL AND METHODS: Thirty-one patients with generalized moderate-to-severe chronic periodontitis received 1-stage FM-SRP. Measurement of clinical parameters and body temperature as well as collection of subgingival plaque, peripheral blood and gingival crevicular fluid was performed before and after treatment 2 or 3 times. Quantification of periodontopathic bacteria in the sulcus and measurement of corresponding serum IgG titers were performed. Systemic and local inflammatory markers such as endotoxin, high-sensitive C-reactive protein (hs-CRP) and 6 inflammatory cytokines were assessed using high-sensitivity assays. RESULTS: Compared to baseline values, FM-SRP resulted in a substantial improvement in clinical parameters (P < .05), lower bacterial counts (P < .01) and a significant decrease of IgG titers against Porphyromonas gingivalis (P < .001) 6 weeks after treatment. Comparing baseline parameters to those at 1 day post-treatment, there was a statistically significant elevation in body temperature (P = .007). In addition, a 5-fold increase in hs-CRP (P < .001), a remarkable increase in interferon-γ (P < .001) and a slight increase in interleukin (IL)-12p70 (P = .001) were detected in serum samples. In the gingival crevicular fluid, marked increases in hs-CRP (P < .001), IL-5 (P = .001), IL-6, IL-12p70 and tumor necrosis factor-α (P < .001 for the latter 3 markers) were noted 1 day after treatment. Endotoxin levels were below measurable limits for most time points. CONCLUSION: FM-SRP resulted in clinical and microbiological improvement 6 weeks post-treatment, but produced a moderate systemic acute-phase response including elevated inflammatory mediators 1 day post-treatment.


Subject(s)
Chronic Periodontitis/therapy , Dental Scaling , Inflammation Mediators/metabolism , Root Planing , Chronic Periodontitis/microbiology , Endotoxins/blood , Female , Follow-Up Studies , Gingival Crevicular Fluid/chemistry , Humans , Immunoglobulin G/metabolism , Japan , Male , Middle Aged , Prospective Studies , Treatment Outcome
2.
Arch Oral Biol ; 79: 35-41, 2017 Jul.
Article in English | MEDLINE | ID: mdl-28285126

ABSTRACT

OBJECTIVE: Although the pathogeneses of Alzheimer's disease (AD) and periodontal diseases have overlapping features, including ageing and chronic inflammation, the association between AD and periodontitis remains unclear. To explore the pathogenesis of periodontitis, a comprehensive gene expression/transcriptome analysis in periodontitis-affected gingival tissues found that the AD pathway was significantly up-regulated in periodontitis-affected gingival tissues. AD-related genes, amyloid beta precursor protein (APP), interleukin-1 beta and compliment 1QA, were significantly elevated in periodontitis. In the present study, balance between mRNA expression of APP and a potent amyloid degradation enzyme, neprilysin (NEP), as well as protein localisation of APP and NEP were analysed. DESIGN: Eighteen periodontitis-affected and 18 clinically healthy control gingival tissues were taken from patients with severe chronic periodontitis or undergoing tooth extraction. Total RNA was purified and used for quantitative reverse transcription real-time polymerase chain reaction (qRT-PCR). The localisation of APP and NEP was analysed by immunohistochemistry (IHC). RESULTS: Both APP and NEP genes were up-regulated in periodontitis-affected gingival tissues. APP-expressing macrophages and NEP-expressing neutrophils and fibroblasts, reflecting inflammatory stages, were detected in inflamed gingival tissues by IHC. CONCLUSION: The up-regulation of APP and NEP mRNA levels in periodontitis-affected gingival tissues compared with healthy controls was confirmed by qRT-PCR analyses. Since NEP is one of the primary enzymes that degrades amyloid beta, increased NEP mRNA levels in periodontitis may act as an inhibitor of amyloid beta accumulation in gingival tissues, balancing increased APP mRNA expression. However, NEP has several effects including degradation of vasoactive substances; therefore, further sresearch is needed.


Subject(s)
Chronic Periodontitis/metabolism , Gingiva/drug effects , Neprilysin/biosynthesis , Neprilysin/pharmacology , Adult , Aged , Alzheimer Disease , Amyloid beta-Protein Precursor/genetics , Amyloid beta-Protein Precursor/metabolism , Chronic Periodontitis/diagnostic imaging , Chronic Periodontitis/genetics , Chronic Periodontitis/pathology , Female , Fibroblasts/metabolism , Gene Expression Regulation , Gingiva/diagnostic imaging , Gingiva/pathology , Humans , Immunohistochemistry , Inflammation , Interleukin-1beta/genetics , Interleukin-1beta/metabolism , Japan , Macrophages/metabolism , Male , Middle Aged , Neprilysin/genetics , Neutrophils/metabolism , RNA, Messenger/biosynthesis , Real-Time Polymerase Chain Reaction , Up-Regulation
3.
BMC Oral Health ; 17(1): 46, 2017 Jan 16.
Article in English | MEDLINE | ID: mdl-28093069

ABSTRACT

BACKGROUND: The diagnosis of the progression of periodontitis presently depends on the use of clinical symptoms (such as attachment loss) and radiographic imaging. The aim of the multicenter study described here was to evaluate the diagnostic use of the bacterial content of subgingival plaque recovered from the deepest pockets in assessing disease progression in chronic periodontitis patients. METHODS: This study consisted of a 24-month investigation of a total of 163 patients with chronic periodontitis who received trimonthly follow-up care. Subgingival plaque from the deepest pockets was recovered and assessed for bacterial content of Porphyromonas gingivalis, Prevotella intermedia, and Aggregatibacter actinomycetemcomitans using the modified Invader PLUS assay. The corresponding serum IgG titers were measured using ELISA. Changes in clinical parameters were evaluated over the course of 24 months. The sensitivity, specificity, and prediction values were calculated and used to determine cutoff points for prediction of the progression of chronic periodontitis. RESULTS: Of the 124 individuals who completed the 24-month monitoring phase, 62 exhibited progression of periodontitis, whereas 62 demonstrated stable disease. The P. gingivalis counts of subgingival plaque from the deepest pockets was significantly associated with the progression of periodontitis (p < 0.001, positive predictive value = 0.708). CONCLUSIONS: The P. gingivalis counts of subgingival plaque from the deepest pockets may be associated with the progression of periodontitis.


Subject(s)
Chronic Periodontitis/diagnosis , Chronic Periodontitis/microbiology , Dental Plaque/microbiology , Saliva/microbiology , Aged , Antigens, Bacterial/blood , Chronic Periodontitis/therapy , Colony Count, Microbial , Disease Progression , Enzyme-Linked Immunosorbent Assay , Female , Humans , Immunoglobulin G/blood , Japan , Male , Middle Aged , Prospective Studies
4.
J Periodontal Res ; 51(6): 768-778, 2016 Dec.
Article in English | MEDLINE | ID: mdl-26791469

ABSTRACT

BACKGROUND AND OBJECTIVE: A diagnosis of periodontitis progression is presently limited to clinical parameters such as attachment loss and radiographic imaging. The aim of this multicenter study was to monitor disease progression in patients with chronic periodontitis during a 24-mo follow-up program and to evaluate the amount of bacteria in saliva and corresponding IgG titers in serum for determining the diagnostic usefulness of each in indicating disease progression and stability. MATERIAL AND METHODS: A total of 163 patients with chronic periodontitis who received trimonthly follow-up care were observed for 24 mo. The clinical parameters and salivary content of Porphyromonas gingivalis, Prevotella intermedia and Aggregatibacter actinomycetemcomitans were assessed using the modified Invader PLUS assay, and the corresponding serum IgG titers were measured using ELISA. The changes through 24 mo were analyzed using cut-off values calculated for each factor. One-way ANOVA or Fisher's exact test was used to perform between-group comparison for the data collected. Diagnostic values were calculated using Fisher's exact test. RESULTS: Of the 124 individuals who completed the 24-mo monitoring phase, 62 exhibited periodontitis progression, whereas 62 demonstrated stable disease. Seven patients withdrew because of acute periodontal abscess. The ratio of P. gingivalis to total bacteria and the combination of P. gingivalis counts and IgG titers against P. gingivalis were significantly related to the progression of periodontitis. The combination of P. gingivalis ratio and P. gingivalis IgG titers was significantly associated with the progression of periodontitis (p = 0.001, sensitivity = 0.339, specificity = 0.790). CONCLUSIONS: It is suggested that the combination of P. gingivalis ratio in saliva and serum IgG titers against P. gingivalis may be associated with the progression of periodontitis.


Subject(s)
Antibodies, Bacterial/blood , Chronic Periodontitis/pathology , Immunoglobulin G/blood , Saliva/microbiology , Aggregatibacter actinomycetemcomitans , Bacterial Load , Bacteroidaceae Infections/microbiology , Bacteroidaceae Infections/pathology , Chronic Periodontitis/blood , Chronic Periodontitis/metabolism , Disease Progression , Enzyme-Linked Immunosorbent Assay , Female , Humans , Male , Middle Aged , Pasteurellaceae Infections/microbiology , Pasteurellaceae Infections/pathology , Porphyromonas gingivalis , Prevotella intermedia , Prospective Studies
5.
J Microbiol Methods ; 117: 4-10, 2015 Oct.
Article in English | MEDLINE | ID: mdl-26159910

ABSTRACT

UNLABELLED: An important goal for the improved diagnosis and management of infectious and inflammatory diseases, such as periodontitis, is the development of rapid and accurate technologies for the decentralized detection of bacterial pathogens. The aim of this prospective multicenter study was to evaluate the clinical use of a novel immunochromatographic device with monoclonal antibodies for the rapid point-of-care detection and semi-quantification of Porphyromonas gingivalis in subgingival plaque. Sixty-three patients with chronic periodontitis and 28 periodontally healthy volunteers were subjected to clinical and microbiological examinations. Subgingival plaque samples were analyzed for the presence of P. gingivalis using a novel immunochromatography based device DK13-PG-001, designed to detect the 40k-outer membrane protein of P. gingivalis, and compared with a PCR-Invader method. In the periodontitis group, a significant strong positive correlation in detection results was found between the test device score and the PCR-Invader method (Spearman rank correlation, r=0.737, p<0.0001). The sensitivity, specificity, and positive and negative predictive values of the test device were 96.2%, 91.8%, 90.4% and 96.7%, respectively. The detection threshold of the test device was determined to be approximately 10(4) (per two paper points). There were significant differences in the bacterial counts by the PCR-Invader method among groups with different ranges of device scores. With a cut-off value of ≥0.25 in device score, none of periodontally healthy volunteers were tested positive for the subgingival presence of P. gingivalis, whereas 76% (n=48) of periodontitis subjects were tested positive. There was a significant positive correlation between device scores for P. gingivalis and periodontal parameters including probing pocket depth and clinical attachment level (r=0.317 and 0.281, respectively, p<0.01). The results suggested that the DK13-PG-001 device kit can be effectively used for rapid, chair-side detection and semi-quantification of P. gingivalis in subgingival plaque. TRIAL REGISTRATION: UMIN Clinical Trials Registry (UMIN-CTR) UMIN000011943.


Subject(s)
Bacteriological Techniques/instrumentation , Chromatography, Affinity/instrumentation , Dental Plaque/microbiology , Porphyromonas gingivalis/isolation & purification , Adult , Aged , Aged, 80 and over , Antibodies, Bacterial , Antibodies, Monoclonal , Bacteriological Techniques/methods , Chromatography, Affinity/methods , Equipment Design , Female , Humans , Male , Middle Aged , Periodontitis/microbiology , Point-of-Care Systems , Porphyromonas gingivalis/immunology , Predictive Value of Tests , Prospective Studies
6.
Arch Oral Biol ; 59(6): 586-94, 2014 Jun.
Article in English | MEDLINE | ID: mdl-24690593

ABSTRACT

OBJECTIVE: Periodontitis involves periodontal tissue destruction and is associated with chronic inflammation and ageing. Periodontitis has recently been recognised as a risk factor for Alzheimer's disease (AD). We showed upregulation of molecules in the AD pathway including amyloid beta (A4) precursor protein (APP), a key gene in AD, interleukin-1 beta (IL-1ß), and complement component 1 (q subcomponent, A chain) (C1QA) in periodontitis compared to healthy tissues. Here, we quantitatively analysed the expression levels of APP, IL-1ß, and C1QA and determined the localisation of APP in gingival tissues. DESIGN: Fourteen chronic periodontitis patients and 14 healthy participants were enrolled. Six samples of total RNA from two distinct sites of healthy and periodontitis-affected gingival tissues from three randomly selected patients were used for microarray analyses, and significant biological pathways in periodontitis were identified. Differential gene expression of APP, IL-1ß, and C1QA, which belong to the AD pathway, were analysed with quantitative reverse transcription real-time polymerase chain reaction (qRT-PCR) using samples from these 14 chronic periodontitis patients and 14 healthy controls. APP localisation was analysed with immunohistochemistry. RESULTS: APP, IL-1ß, and C1QA mRNA levels were significantly upregulated in periodontitis-affected gingival tissues. APP was mainly localised in macrophages in gingival connective tissues underneath the epithelial layers. CONCLUSIONS: An association between AD and periodontitis was detected with microarray and computer-aided data mining analyses. qRT-PCR identified differential gene expression in periodontitis-affected gingival tissue that may be related to AD pathogenesis. Elevated APP, IL-1ß, and C1QA transcripts and APP-expressing macrophages in periodontitis-affected gingival tissues were observed, suggesting a relationship between periodontitis and AD pathogenesis.


Subject(s)
Amyloid beta-Peptides/metabolism , Gingiva/metabolism , Periodontitis/metabolism , Adult , Aged , Amyloid beta-Protein Precursor/metabolism , Case-Control Studies , Complement C1/metabolism , Female , Gene Expression , Humans , Immunoenzyme Techniques , Interleukin-1beta/metabolism , Male , Microarray Analysis , Middle Aged , Periodontitis/genetics , Real-Time Polymerase Chain Reaction
7.
J Periodontal Res ; 46(3): 345-53, 2011 Jun.
Article in English | MEDLINE | ID: mdl-21382035

ABSTRACT

BACKGROUND AND OBJECTIVE: Gene expression is related to the pathogenesis of periodontitis and plays a crucial role in local tissue destruction and disease susceptibility. The aims of the present study were to identify the expression of specific genes and biological pathways in periodontitis-affected gingival tissue using microarray and quantitative real-time RT-PCR analyses. MATERIAL AND METHODS: Healthy and periodontitis-affected gingival tissues were taken from three patients with severe chronic periodontitis. Total RNAs from six gingival tissue samples were used for microarray analyses. Data-mining analyses, such as comparisons, gene ontology and pathway analyses, were performed and biological pathways with a significant role in periodontitis were identified. In addition, quantitative real-time RT-PCR analysis was performed on samples obtained from 14 patients with chronic periodontitis and from 14 healthy individuals in order to confirm the results of the pathway analysis. RESULTS: Comparison analyses found 15 up-regulated and 13 down-regulated genes (all of which showed a change of more than twofold in expression levels) in periodontitis-affected gingival tissues. Pathway analysis identified 15 up-regulated biological pathways, including leukocyte transendothelial migration, and five down-regulated pathways, including cell communication. Quantitative real-time RT-PCR verified that five genes in the leukocyte transendothelial migration pathway were significantly up-regulated, and four genes in the cell communication pathway were significantly down-regulated, which was consistent with pathway analysis. CONCLUSION: We identified up-regulated genes (ITGB-2, MMP-2, CXCL-12, CXCR-4 and Rac-2) and down-regulated genes (connexin, DSG-1, DSC-1 and nestin) in periodontitis-affected gingival tissues; these genes may be related to the stimulation of leukocyte transendothelial migration and to the the impairment of cell-to-cell communication in periodontitis.


Subject(s)
Chronic Periodontitis/immunology , Gene Expression/genetics , Gingiva/immunology , Leukocytes/immunology , Transendothelial and Transepithelial Migration/immunology , Adult , CD18 Antigens/genetics , Cell Communication/immunology , Chemokine CXCL12/genetics , Chemotaxis, Leukocyte/immunology , Chronic Periodontitis/pathology , Connexins/genetics , Desmocollins/genetics , Desmoglein 1/genetics , Down-Regulation/genetics , Endothelial Cells/immunology , Endothelial Cells/pathology , Female , Gene Expression Regulation/genetics , Gingiva/pathology , Humans , Intermediate Filament Proteins/genetics , Male , Matrix Metalloproteinase 2/genetics , Microarray Analysis , Middle Aged , Nerve Tissue Proteins/genetics , Nestin , RNA/genetics , Receptors, CXCR4/genetics , Reverse Transcriptase Polymerase Chain Reaction , Up-Regulation/genetics , rac GTP-Binding Proteins/genetics , RAC2 GTP-Binding Protein
8.
Anim Genet ; 39(1): 22-7, 2008 Feb.
Article in English | MEDLINE | ID: mdl-18254733

ABSTRACT

The porcine MX1 and MX2 promoters were characterized in this study. Sequencing of the 332-bp MX1 promoter region identified 15 substitutions and insertions at three positions in 21 pigs from 15 breeds, in which nine genotypes were classified. Among the nine genotypes, no statistically significant differences in the promoter activities were observed after interferon (IFN-alpha 2b) treatment of transiently transfected cells containing constructs with luciferase reporter plasmids. The 341-bp MX2 promoter region contained regulatory sequences for ISRE, GC box, Sp1 and AP-1, as well as a TATA box. Nucleotide sequences of the MX2 promoter region revealed four substitutions and one deletion, in which six genotypes were classified. Among the six genotypes, a statistically significant difference (P < 0.05) in MX2 promoter activities after IFN-alpha 2b treatment was detected in transiently transfected cells.


Subject(s)
GTP-Binding Proteins/genetics , Polymorphism, Genetic , Promoter Regions, Genetic , Swine/genetics , Animals , Base Sequence , DNA/genetics , DNA Primers/genetics , Genetic Variation , Genotype , Interferon alpha-2 , Interferon-alpha/pharmacology , LLC-PK1 Cells , Molecular Sequence Data , Mutagenesis, Insertional , Myxovirus Resistance Proteins , Polymorphism, Single Nucleotide , Promoter Regions, Genetic/drug effects , Recombinant Proteins , Sequence Deletion , Species Specificity , Sus scrofa/classification , Sus scrofa/genetics , Swine, Miniature/classification , Swine, Miniature/genetics , Transfection
9.
Anim Genet ; 37(1): 10-6, 2006 Feb.
Article in English | MEDLINE | ID: mdl-16441290

ABSTRACT

To clarify the structure of the porcine genomic region that contains quantitative trait loci (QTL) related to fat, we constructed a bacterial artificial chromosome (BAC) contig of the region from DST to SRPK1 on porcine chromosome 7 and performed low-redundancy 'skim' shotgun sequencing of the clones that composed a minimum tiling path of the contig. This analysis revealed that the gene order from VPS52 to SRPK1 is conserved between human and swine and that comparison with the human sequence identified a rearrangement in the swine genome at the proximal end of VPS52. Analysis of the nucleotide sequences of three BAC clones that included the rearrangement point demonstrated that COL21A1 and DST, which were not present in the corresponding human region, were located adjacent to the rearrangement point. These results provide useful information about the genomic region containing QTL for fat in pigs and help to clarify the structure of the so-called 'extended-class II' region distal to the porcine major histocompatibility complex class II region.


Subject(s)
Chromosomes, Mammalian/genetics , Gene Order/genetics , Genome/genetics , Swine/genetics , Adipose Tissue , Animals , Base Sequence , Chromosomes, Artificial, Bacterial , Conserved Sequence/genetics , Genes, MHC Class II/genetics , Humans , In Situ Hybridization, Fluorescence , Molecular Sequence Data , Quantitative Trait Loci , Sequence Analysis, DNA
10.
J Anim Sci ; 83(10): 2247-54, 2005 Oct.
Article in English | MEDLINE | ID: mdl-16160033

ABSTRACT

The objective of the research was to identify QTL affecting the number of vertebrae in swine, one of the major determining factors of growth and body composition. Previously, we reported a QTL for the number of vertebrae located on SSC1qter (terminal band of the q arm of SSC 1) in an F2 family produced by crossing a Göttingen miniature male with two Meishan females. Eight other swine families were subsequently produced by crosses between different breeds of European, Asian, and miniature pigs. In these families, the QTL on SSC1qter for the number of vertebrae was detected. Unlike the Asian alleles, all European alleles in this study had the effect of increasing the number of vertebrae by 0.44 to 0.69 and acted additively without dominance. The Göttingen miniature sire, for which we previously reported a smaller additive effect, seemed to be heterozygous at the QTL. In the present study, another QTL was found for the number of vertebrae on SSC7. This QTL was not fixed in the European pigs used as parents in our experimental families, and some of the European alleles increased the number of vertebrae. A half-sib analysis confirmed that this QTL was segregating in a commercial Large White population. Analysis in an F2 family in which the parental pigs were fixed for alternative alleles revealed that the effects of the QTL on SSC1 and on SSC7 were additive and similar in size. The two QTL acted independently without epistatic effects and explained an increase of more than two vertebrae.


Subject(s)
Chromosome Mapping/veterinary , Quantitative Trait Loci/physiology , Spine/physiology , Sus scrofa/genetics , Animals , Chromosomes/genetics , Female , Heterozygote , Inbreeding , Male , Quantitative Trait Loci/genetics , Spine/anatomy & histology , Sus scrofa/anatomy & histology , Sus scrofa/physiology
11.
Anim Genet ; 35(6): 445-50, 2004 Dec.
Article in English | MEDLINE | ID: mdl-15566466

ABSTRACT

Several quantitative trait loci (QTL) have been detected on SSC1qter (Sus scrofa chromosome 1qter), including QTL for the number of vertebrae, as reported in our previous study. To provide the tools for analysis of QTLs on SSC1qter, we constructed a comparative map of swine and human. In addition, we identified 26 swine STSs and mapped 16 of them on SSC1qter using the INRA - University of Minnesota porcine radiation hybrid (IMpRH) panel. We screened a BAC library using these swine STSs and developed 35 new polymorphic microsatellite markers from the BAC clones, of which 26 were informative in our reference family. We also mapped nine microsatellite markers we had isolated previously. Consequently a total of 44 new polymorphic microsatellite markers were located within a 60-cM region of SSC1qter, spanning from SW1092 to the telomere.


Subject(s)
Chromosome Mapping , Chromosomes, Mammalian/genetics , Microsatellite Repeats/genetics , Quantitative Trait Loci , Sus scrofa/genetics , Animals , Chromosomes, Artificial, Bacterial
12.
Anim Genet ; 34(3): 216-20, 2003 Jun.
Article in English | MEDLINE | ID: mdl-12755823

ABSTRACT

We have constructed a radiation hybrid (RH) map of the porcine genome using an RH panel generated by an irradiation dose of 5000-rad (Sus scrofa radiation hybrid map, SSRH map). Normal porcine aortic endothelial cells were irradiated and fused with a thymidine kinase-deficient mouse cell line, L-M (TK-). A total of 110 cell lines were selected and used for further analysis. Among 1091 microsatellite (MS) markers selected for mapping, 842 markers (77%) could be typed on the panel. The framework map comprised 342 MS markers and an additional 247 MS markers were then added to generate the whole-genome map. The average retention frequency for the data set was 30.6%. The total map length was 5596.2 centiRay (cR). Using an estimated physical length of 2718 Mbp, the average ratio between cR and physical distance over the porcine genome was estimated to be 0.49 Mb/cR.


Subject(s)
Genome , Radiation Hybrid Mapping , Sus scrofa/genetics , Animals , Microsatellite Repeats
13.
Phys Rev Lett ; 89(23): 231601, 2002 Dec 02.
Article in English | MEDLINE | ID: mdl-12484996

ABSTRACT

We study the correlation between CP violation in neutrino oscillations and leptogenesis in the framework with two heavy Majorana neutrinos and three light neutrinos. Among three unremovable CP phases, a heavy Majorana phase contributes to leptogenesis. We show how the heavy Majorana phase contributes to Jarlskog determinant J as well as neutrinoless double beta decay by identifying a low energy CP-violating phase which signals the CP-violating phase for leptogenesis. For some specific cases of the Dirac mass term of neutrinos, a direct relation between lepton number asymmetry and J is obtained. We also study the effect coming from the phases which are not related to leptogenesis.

14.
Anal Sci ; 17(6): 763-8, 2001 Jun.
Article in English | MEDLINE | ID: mdl-11707948

ABSTRACT

An application of capillary electrophoresis (CE) using sulfated beta-cyclodextrin (SCD) has been investigated for separating alkylphenols with different chain lengths, as well as bisphenol A and bisphenol S. In the absence of SCD in running buffer, all the phenols migrated at the same velocity as the electroosmotic flow (EOF), whereas the addition of SCD effectively led to the baseline separation of alkylphenols on the basis of the difference in the abilities to bind into the hydrophobic cavity of CD. The host-guest binding constants between analyte phenols and SCD were evaluated from Benesi-Hildebrand plots of the data obtained by two independent methods, CE and UV-visible measurements, demonstrating that the greater the hydrophobicity of the phenols, the larger the binding constants. The effects of organic solvents on the resolution for alkylphenols and bisphenols were also examined. This system using SCD was effective for the separation of 4-octylphenol and 4-nonylphenol isomers having longer alkyl chains.

15.
Biochem Genet ; 39(7-8): 251-60, 2001 Aug.
Article in English | MEDLINE | ID: mdl-11590831

ABSTRACT

Much is known about the antiviral activity of Mx proteins in species such as mouse and human. In the mouse, loss of resistability to influenza virus has been shown to be due to specific polymorphisms in the Mx gene. This gene is therefore an interesting candidate gene for disease resistance in farm animals. The porcine Mx1 gene has already been identified and characterized based on its homology with mouse Mx1; however, until now no evidence of polymorphisms in the porcine gene has been reported. In this study, we have found two new polymorphisms in exon 14 of porcine Mx1 by DNA sequencing and confirmed their presence in different breeds, using polymerase chain reaction (PCR)-restriction fragment length polymorphisms (RFLP) with NarI and NaeI restriction enzymes. On the basis of the deduced amino acid sequence, one allele contains a deletion that may result in a frameshift to yield several amino acid substitutions and extension of the carboxyl terminal region of Mx1 protein. The deletion allele, Mx1c, was found to be segregating in Landrace, Berkshire, Duroc, Hampshire, and Yucatan miniature pig. A second point mutation, Mx1b, was detected in Meishan and two Vietnamese native pig breeds. All other breeds tested were fixed for the Mx1a allele that is identical to the sequence reported previously. It will be interesting to determine if the Mx1c deletion is associated with variation in resistance to the myxovirus family in the pig.


Subject(s)
GTP-Binding Proteins , Polymorphism, Restriction Fragment Length , Proteins/genetics , Swine/genetics , Amino Acid Sequence , Animals , Chromosomes, Artificial, Bacterial , Exons , Immunity, Innate/genetics , Mice , Molecular Sequence Data , Myxovirus Resistance Proteins , Polymerase Chain Reaction , Proteins/physiology , Species Specificity
16.
Jpn Circ J ; 65(9): 783-7, 2001 Sep.
Article in English | MEDLINE | ID: mdl-11548876

ABSTRACT

The left ventricle's morphological adaptation to high blood pressure is classified into 4 patterns based on mass and wall thickness. The geometric changes caused by maladaptation to pressure overload possibly relate to progression of contractile dysfunction with abnormal energy metabolism. The present study assessed whether the geometric adaptation of the left ventricle (LV) to high blood pressure relates to changes in myocardial energy metabolism, especially free fatty acid (FFA) utilization. Thirty-five patients with essential hypertension underwent echocardiography and dual isotopes myocardial scintigraphy using iodine-123 labeled 15-p-iodophenyl-3-(R,S)-methylpentadecanoic acid (BMIPP, an analogue of a FFA) and thallium-201 (Tl-201). Systolic (endocardial fractional shortening; %FS) and diastolic indices (the ratio of early to atrial filling waves; E/A) of LV function were also assessed. Quantitative myocardial BMIPP uptake was evaluated by the BMIPP/TI-201 myocardial uptake ratio (B/T). The subjects were divided into 4 groups based on LV mass and wall thickness: (1) concentric hypertrophy (CH), (2) eccentric hypertrophy (EH), (3) concentric remodeling (CR), and (4) normal geometry (N). The %FS was lower in the EH group than in the other groups. The mitral E/A ratio in the CH group was lowest. B/T was significantly decreased in the EH group compared with the N group (p < 0.05). B/T correlated with the mitral E/A ratio significantly (p < 0.05, r = 0.42), whereas there was no relationship between %FS and B/T. These results indicate that the geometric changes occurring in hypertensive hearts strongly correlate with alternations in cardiac function and with abnormal myocardial FFA metabolism, and that the latter is associated with diastolic abnormality, but not with systolic function.


Subject(s)
Fatty Acids, Nonesterified/metabolism , Hypertension/metabolism , Hypertension/pathology , Myocardium/metabolism , Myocardium/pathology , Aged , Echocardiography , Energy Metabolism/physiology , Fatty Acids/pharmacokinetics , Female , Heart Ventricles/diagnostic imaging , Heart Ventricles/metabolism , Heart Ventricles/pathology , Hemodynamics , Humans , Hypertension/diagnostic imaging , Hypertrophy, Left Ventricular/diagnostic imaging , Hypertrophy, Left Ventricular/metabolism , Hypertrophy, Left Ventricular/pathology , Iodine Radioisotopes , Iodobenzenes/pharmacokinetics , Male , Middle Aged , Radionuclide Imaging , Thallium Radioisotopes/pharmacokinetics , Ventricular Remodeling
17.
J Periodontal Res ; 36(3): 160-8, 2001 Jun.
Article in English | MEDLINE | ID: mdl-11453114

ABSTRACT

Differential gene expression was examined in human neutrophils stimulated with lipopolysaccharide from Porphyromonas gingivalis (P. gingivalis-LPS) using RNA fingerprinting by arbitrarily primed polymerase chain reaction (RAP-PCR). LPS from Escherichia coli (E. coli-LPS) was used as control. More than 200 differently expressed transcripts were found in 8 subjects showing differential regulation at the transcriptional level. Densitometric analyses revealed that 42-100 genes were upregulated, while 53-116 genes were downregulated by P. gingivalis-LPS compared with E. coli-LPS. The results of sequencing identification showed the presence of supervillin (SVIL) and vascular endothelial growth factor (VEGF) genes in the clones which were upregulated by P. gingivalis-LPS. Consequently, semiquantitative analyses proved that the level of mRNA for SVIL and VEGF were significantly higher in P. gingivalis-LPS-stimulated neutrophils than in other bacterial (E. coli, Actinobacillus actinomycetemcomitans, Prevotella intermedia) LPS- or synthetic lipid A-stimulated neutrophils. Our findings suggest that an elevated mRNA expression for SVIL could be associated with impaired function of neutrophils when stimulated by P. gingivalis-LPS. Further, the VEGF mRNA over-expression might be related to the pathogenesis of P. gingivalis-associated periodontitis. The RAP-PCR technique used in this study enabled us to identify a number of P. gingivalis-LPS regulated genes which hitherto have not been reported.


Subject(s)
Endothelial Growth Factors/genetics , Lipopolysaccharides/pharmacology , Lymphokines/genetics , Membrane Proteins/genetics , Microfilament Proteins/genetics , Neutrophils/metabolism , Porphyromonas gingivalis/physiology , Protein Isoforms/genetics , RNA, Messenger/genetics , Adult , Aggregatibacter actinomycetemcomitans/physiology , DNA Fingerprinting , Down-Regulation/genetics , Escherichia coli/physiology , Gene Expression Regulation , Humans , Lipid A/pharmacology , Male , Periodontitis/genetics , Periodontitis/microbiology , Prevotella intermedia/physiology , Reverse Transcriptase Polymerase Chain Reaction , Sequence Analysis, DNA , Statistics, Nonparametric , Transcription, Genetic , Up-Regulation/genetics , Vascular Endothelial Growth Factor A , Vascular Endothelial Growth Factors
18.
Appl Environ Microbiol ; 67(1): 484-9, 2001 Jan.
Article in English | MEDLINE | ID: mdl-11133487

ABSTRACT

The prevalence of Shiga toxin-producing Escherichia coli (STEC) in Japan was examined by using stool samples from 87 calves, 88 heifers, and 183 cows on 78 farms. As determined by screening with stx-PCR, the prevalence was 46% in calves, 66% in heifers, and 69% in cows; as determined by nested stx-PCR, the prevalence was 100% in all animal groups. Of the 962 isolates picked by colony stx hybridization, 92 isolates from 54 farms were characterized to determine their O serogroups, virulence factor genes, and antimicrobial resistance. Of these 92 isolates, 74 (80%) could be classified into O serogroups; 50% of these 74 isolates belonged to O serogroups O8, O26, O84, O113, and O116 and 1 isolate belonged to O serogroup O157. Locus of enterocyte effacement genes were detected in 24% of the isolates, and enterohemorrhagic E. coli (EHEC) hlyA genes were detected in 72% of the isolates. Neither the bundle-forming pilus gene nor the enteropathogenic E. coli adherence factor plasmid was found. STEC strains with characteristics typical of isolates from human EHEC infections, which were regarded as potential EHEC strains, were present on 11.5% of the farms.


Subject(s)
Cattle/microbiology , Escherichia coli/classification , Escherichia coli/isolation & purification , Shiga Toxins/biosynthesis , Animals , Escherichia coli/metabolism , Escherichia coli/pathogenicity , Escherichia coli Infections/epidemiology , Escherichia coli Infections/microbiology , Feces/microbiology , Female , Humans , Japan/epidemiology , Male , Polymerase Chain Reaction , Prevalence , Shiga Toxins/genetics , Virulence
19.
Ann Nucl Med ; 15(5): 411-6, 2001 Oct.
Article in English | MEDLINE | ID: mdl-11758945

ABSTRACT

This study investigated the clinical value of I-123 MIBG pulmonary accumulation and washout in patients with chronic heart failure (CHF). Nineteen patients with CHF and 15 normal volunteers (NL) were included. The uptake ratio of heart to mediastinum (H/M), that of lung fields to mediastinum (L/M), and washout rate (WR) of the heart and lung fields were calculated in anterior planar images and compared with results of echocardiography and cardiac catheterization. In the CHF group, the lung uptake in delayed images increased and lung WR was decreased, suggesting pulmonary endothelial lesions. Furthermore, there was a negative correlation between right and left lung WR and pulmonary arterial diastolic pressure (PA(D)) and pulmonary arterial systolic pressure (PA(s)) in the CHF group. Since the WR of MIBG reflected PA, it may be used as an index of severity of cardiac dysfunction.


Subject(s)
3-Iodobenzylguanidine/pharmacokinetics , Cardiomyopathy, Dilated/diagnostic imaging , Lung/metabolism , Adult , Cardiomyopathy, Dilated/complications , Chronic Disease , Female , Humans , Hypertrophy, Left Ventricular/complications , Hypertrophy, Left Ventricular/diagnostic imaging , Lung/diagnostic imaging , Male , Middle Aged , Myocardial Ischemia/complications , Myocardial Ischemia/diagnostic imaging , Myocardium/metabolism , Radionuclide Imaging , Radiopharmaceuticals/pharmacokinetics , Stroke Volume , Ultrasonography
20.
J Periodontal Res ; 36(6): 390-7, 2001 Dec.
Article in English | MEDLINE | ID: mdl-11762875

ABSTRACT

Differential gene expression was investigated in neutrophils stimulated with N-formyl-methionyl-leucyl-phenylalanine using RNA fingerprinting by arbitrarily primed polymerase chain reaction (RAP-PCR). The cells were isolated from 3 groups of subjects: patients with generalized aggressive periodontitis (Aggressive-P. n = 6), generalized chronic periodontitis (Chronic-P, n = 6) and healthy controls (H, n = 8). Our results show that 37 genes were upregulated. while 27 genes were down-regulated in all Aggressive-P neutrophils by using RAP-PCR with 45 primer pairs. Reverse transcription-PCR analyses revealed that mRNA levels were significantly different (p<0.05) for heat shock transcription factor 4b (HSF4b) gene. Kruppel-like zinc finger transcription factor 9 (Zf9) and muskelin genes. HSF4b was greater in neutrophils from Aggressive-P compared to groups H and Chronic-P. Zf9 and muskelin genes were lower in Aggressive-P compared to the H groups, but no significant difference was noted compared to the Chronic-P group. The control genes, IL-1beta and VEGF genes, were expressed at a significantly higher level in Aggressive-P and Chronic-P than H (p<.01, p<0.05). In conclusion, the RAP-PCR technique used in this study enabled us to identify 3 Aggressive-P related genes, which had not been reported previously. Neutrophil functions in Aggressive-P patients are suggested to be altered by regulatory factors of the immune system including HSF4b (transcription factor), Zf9 (activator of TGF-beta) and muskelin (cellular adhesion).


Subject(s)
Intercellular Signaling Peptides and Proteins , Periodontitis/genetics , Periodontitis/immunology , Proto-Oncogene Proteins , Adult , Aggressive Periodontitis/genetics , Aggressive Periodontitis/immunology , Aggressive Periodontitis/metabolism , Case-Control Studies , Cell Adhesion Molecules , DNA-Binding Proteins/biosynthesis , DNA-Binding Proteins/genetics , Dual-Specificity Phosphatases , Female , Gene Expression , Heat Shock Transcription Factors , Humans , Intracellular Signaling Peptides and Proteins , Kruppel-Like Factor 6 , Kruppel-Like Transcription Factors , Male , Mitogen-Activated Protein Kinase Phosphatases , Neutrophils/metabolism , Periodontitis/metabolism , Phosphofructokinases/biosynthesis , Phosphofructokinases/genetics , Protein Biosynthesis , Protein Tyrosine Phosphatases/biosynthesis , Protein Tyrosine Phosphatases/genetics , Proteins/genetics , RNA/analysis , Random Amplified Polymorphic DNA Technique , Reverse Transcriptase Polymerase Chain Reaction , Sequence Analysis, DNA , Trans-Activators/biosynthesis , Trans-Activators/genetics , Transcription Factors/biosynthesis , Transcription Factors/genetics
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