Crystallization and Preliminary X-ray Crystallographic Analysis of Malonamidase E2, an Amidase Signature Family Member

Acta Crystallographica. Section D, Biological Crystallography. Mar, 2002  |  Pubmed ID: 11856856

Malonamidase E2 from Bradyrhizobium japonicum catalyzes the hydrolysis of malonamate. The enzyme belongs to an amidase signature family which has a highly conserved serine- and glycine-rich sequence over a stretch of approximately 45 amino acids. More than 100 known or predicted members belonging to this family, whose biological functions vary widely, can be identified in sequence databases. Although urgently needed, no three-dimensional structure of any protein of this family is yet available. The crystallization of malonamidase E2 was undertaken as a first step toward the goal of providing information on the canonical structure of the amidase signature family. The enzyme was crystallized using the hanging-drop vapour-diffusion method at 277 K under two different conditions. One crystal form, which is easier to work with than the other form, belongs to the orthorhombic space group P2(1)2(1)2, with unit-cell parameters a = 104.29, b = 95.58, c = 74.90 A. The unit cell is likely to contain two molecules of MAE2, with a crystal volume per protein mass (V(M)) of 2.045 A(3)Da(-1) and solvent content of about 39.9% by volume. A native data set to 1.8 A resolution was obtained from a flash-cooled crystal using synchrotron radiation.

Refolding of the Catalytic and Hinge Domains of Human MT1-mMP Expressed in Escherichia Coli and Its Characterization

Molecules and Cells. Feb, 2002  |  Pubmed ID: 11911461

The catalytic and hinge domain (Tyr112-Ile318) of the human membrane type-1 matrix metalloproteinase (MT1-MMP; MMP-14), containing hexa-histidines at the C-terminus (chMT1-MMP), was overexpressed in Escherichia coli. The expressed polypeptide was almost exclusively found in the inclusion body, and then purified by a single Ni2+-NTA agarose column chromatography after solubilization with 6 M urea. During refolding, the 26.9 kDa chMT1-MMP was processed to a 24.3 kDa intermediate form and then to a 22.2 kDa mature form. By Western blot analysis and mass spectrometry combined with N-terminal sequencing, the intermediate form was identified as a mixture of the Tyr112-Thr299 with a translation-initiating methionine and Ile114-Thr299, and that the mature form corresponds to Ile114-Pro290. These results demonstrate that the mature form was generated by successive autoproteolysis of the N- and C-terminal sites between Thr299-Thr300, Ala113-Ile114, and Pro290-Thr291 during refolding. Catalytic activity of the mature chMT1-MMP was demonstrated by a peptide cleavage assay. In addition, it has gelatinolytic activity and is able to activate proMMP-2 to the mature MMP-2. These results indicate that the refolded chMT1-MMP retains characteristics of MT1-MMP.

Effects of Compounds Found in Propolis on Streptococcus Mutans Growth and on Glucosyltransferase Activity

Antimicrobial Agents and Chemotherapy. May, 2002  |  Pubmed ID: 11959560

Propolis, a resinous bee product, has been shown to inhibit the growth of oral microorganisms and the activity of bacterium-derived glucosyltransferases (GTFs). Several compounds, mainly polyphenolics, have been identified in this natural product. The present study evaluated the effects of distinct chemical groups found in propolis on the activity of GTF enzymes in solution and on the surface of saliva-coated hydroxyapatite (sHA) beads. Thirty compounds, including flavonoids, cinnamic acid derivatives, and terpenoids, were tested for the ability to inhibit GTFs B, C, and D from Streptococcus mutans and GTF from S. sanguinis (GTF Ss). Flavones and flavonols were potent inhibitors of GTF activity in solution; lesser effects were noted on insolubilized enzymes. Apigenin, a 4',5,7-trihydroxyflavone, was the most effective inhibitor of GTFs, both in solution (90.5 to 95% inhibition at a concentration of 135 microg/ml) and on the surface of sHA beads (30 to 60% at 135 microg/ml). Antibacterial activity was determined by using MICs, minimum bactericidal concentrations (MBCs), and time-kill studies. Flavanones and some dihydroflavonols, as well as the sesquiterpene tt-farnesol, inhibited the growth of S. mutans and S. sobrinus; tt-farnesol was the most effective antibacterial compound (MICs of 14 to 28 microg/ml and MBCs of 56 to 112 microg/ml). tt-Farnesol (56 to 112 microg/ml) produced a 3-log-fold reduction in the bacterial population after 4 h of incubation. Cinnamic acid derivatives had negligible biological activities. Several of the compounds identified in propolis inhibit GTF activities and bacterial growth. Apigenin is a novel and potent inhibitor of GTF activity, and tt-farnesol was found to be an effective antibacterial agent.

High Molecular Weight Water-soluble Chitosan Protects Against Apoptosis Induced by Serum Starvation in Human Astrocytes

The Journal of Nutritional Biochemistry. Apr, 2002  |  Pubmed ID: 11988407

The effect of high molecular weight water-soluble chitosan (WSC) on serum starvation-induced apoptosis in human astrocytes (CCF-STTG1 Cells) was investigated. WSC, having an average molecular weight of 300 kDa and a degree of deacetylation over 90%, can be produced using a simple multi-step membrane separation process. Serum starvation led to growth arrest, rounding up of cells and appearance of p53 bands. Prolonged (48 h) incubation in serum starved medium led to cell detachment and death. WSC significantly protected the serum starvation-induced cellular rounding up and protected the serum starvation-induced cell death as tested by flow cytometry. WSC also protected serum starvation-induced p53 activation as determined by Western blot. These results suggest that WSC may prevent serum starvation-induced apoptosis of CCF-STTG1 cells via p53 inactivation.

Structure of Malonamidase E2 Reveals a Novel Ser-cisSer-Lys Catalytic Triad in a New Serine Hydrolase Fold That is Prevalent in Nature

The EMBO Journal. Jun, 2002  |  Pubmed ID: 12032064

A large group of hydrolytic enzymes, which contain a conserved stretch of approximately 130 amino acids designated the amidase signature (AS) sequence, constitutes a super family that is distinct from any other known hydrolase family. AS family enzymes are widespread in nature, ranging from bacteria to humans, and exhibit a variety of biological functions. Here we report the first structure of an AS family enzyme provided by the crystal structure of malonamidase E2 from Bradyrhizobium japonicum. The structure, representing a new protein fold, reveals a previously unidentified Ser-cisSer-Lys catalytic machinery that is absolutely conserved throughout the family. This family of enzymes appears to be evolutionarily distinct but has diverged to acquire a wide spectrum of individual substrate specificities, while maintaining a core structure that supports the catalytic function of the unique triad. Based of the structures of the enzyme in two different inhibited states, an unusual action mechanism of the triad is proposed that accounts for the role of the cis conformation in the triad.

[Biliary Papillomatosis]

Taehan Kan Hakhoe Chi = The Korean Journal of Hepatology. Sep, 2002  |  Pubmed ID: 12499794

Effect of a Novel Type of Propolis and Its Chemical Fractions on Glucosyltransferases and on Growth and Adherence of Mutans Streptococci

Biological & Pharmaceutical Bulletin. Apr, 2003  |  Pubmed ID: 12673037

Flavonoids have been considered the main biologically active components in propolis. However, a new variety of flavonoid-free propolis was recently found and chemically classified as type 6. Because it showed activity against oral microorganisms, this study evaluated the effects of the crude ethanolic extract of this propolis and its chemical fractions on the activity of purified glucosyltransferases (GTFs) and on the growth and adherence of mutans streptococci. The inhibitory effect of propolis extracts on GTF activities was determined either in solution or adsorbed onto saliva-coated hydroxyapatite. Streptococcus mutans Ingbritt 1600, Streptococcus sobrinus 6715, and two clinical isolates of each species were used for antibacterial assays. Susceptibilities to the test extracts were analyzed using the agar diffusion method and by determining the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC); the effect on bacterial adherence to a glass surface was also assessed. The activity of GTFs in solution was effectively inhibited by the ethanolic extract of propolis type 6 (EEP) (>80% inhibition at 0.5 mg/ml), hexane, and chloroform fractions (60-90% inhibition at 100 microg/ml); their inhibitory effects on surface enzymes were less pronounced. The EEP, hexane, and chloroform fractions also showed significant antibacterial activity. The data showed that propolis type 6 remarkably reduced GTF activity and inhibited mutans streptococci growth and adherence; these biological activities are associated with its nonpolar components.

Characterization of a Novel Ser-cisSer-Lys Catalytic Triad in Comparison with the Classical Ser-His-Asp Triad

The Journal of Biological Chemistry. Jul, 2003  |  Pubmed ID: 12711609

Amidase signature family enzymes, which are widespread in nature, contain a newly identified Ser-cisSer-Lys catalytic triad in which the peptide bond between Ser131 and the preceding residue Gly130 is in a cis configuration. In order to characterize the property of the novel triad, we have determined the structures of five mutant malonamidase E2 enzymes that contain a Cys-cisSer-Lys, Ser-cisAla-Lys, or Ser-cisSer-Ala triad or a substitution of Gly130 with alanine. Cysteine cannot replace the role of Ser155 due to a hyper-reactivity of the residue, which results in the modification of the cysteine to cysteinyl sulfinic acid, most likely inside the expression host cells. The lysine residue plays a structural as well as a catalytic role, since the substitution of the residue with alanine disrupts the active site structure completely. The two observations are in sharp contrast with the consequences of the corresponding substitutions in the classical Ser-His-Asp triad. Structural data on the mutant containing the Ser-cisAla-Lys triad convincingly suggest that Ser131 plays an analogous catalytic role as the histidine of the Ser-His-Asp triad. The unusual cis configuration of Ser131 appears essential for the precise contacts of this residue with the other triad residues, as indicated by the near invariance of the preceding glycine residue (Gly130), structural data on the G130A mutant, and by a modeling experiment. The data provide a deep understanding of the role of each residue of the new triad at the atomic level and demonstrate that the new triad is a catalytic device distinctively different from the classical triad or its variants.

A Case of ACTH-producing Pheochromocytoma Associated with Pregnancy

Endocrine Journal. Dec, 2003  |  Pubmed ID: 14709846

Ectopic ACTH syndrome is rarely caused by pheochromocytoma. We report a case of a 28-year-old woman with Cushing's syndrome due to ACTH-producing adrenal pheochromocytoma. She had delivered preterm baby at 32nd week of gestation with 'severe preeclampsia'. After delivery, persistent hypertension accompanied by severe headache led her to being misdiagnosed as Cushing's syndrome due to right adrenal adenoma (normal plasma ACTH level) and cerebral vasculitis of unknown etiology. She was referred to our hospital for surgical treatment. Repeated biochemical studies suggested coexistence of ectopic ACTH syndrome and pheochromocytoma. To reverse her clinical deterioration, right total and left subtotal adrenalectomy was performed with presumptive diagnosis of 1) right adrenal pheochromocytoma causing ectopic ACTH syndrome or 2) coexistence of ACTH-dependent Cushing's syndrome and right adrenal pheochromocytoma. Pathologic examination of right adrenal mass revealed pheochromocytoma which showed strong immunostaining for ACTH. Plasma ACTH and urinary cortisol excretion normalized after surgery, but she succumbed to multiple cerebral infarcts and disseminated intravascular coagulation. Pregnancy and inappropriately low plasma ACTH at initial evaluation might have hampered early diagnosis. To our knowledge, this is the first description of a case with ectopic ACTH syndrome due to pheochromocytoma associated with pregnancy.

Effects of Behavior Modification on Body Image, Depression and Body Fat in Obese Korean Elementary School Children

Yonsei Medical Journal. Feb, 2004  |  Pubmed ID: 15004870

This study was performed to investigate the effects of behavior modification on body image, depression and body fat in obese elementary school children. Sixty-two elementary students of the 4th to 6th grade were selected from two different Seoul schools. Thirty-four children in one school were designated as the experimental group, and 28 children from the other school as the control group. The experimental group received 60 - 70 minutes of behavior modification, once a week, for 8 weeks. The control group received neither management nor treatment. The results indicated a significant improvement of body image and a reduction in the increase rate of body fat for the experimental group. This finding strongly supports the theory that behavior modification can be used as an effective strategy in the treatment of obese children.

The Influence of Mutanase and Dextranase on the Production and Structure of Glucans Synthesized by Streptococcal Glucosyltransferases

Carbohydrate Research. Aug, 2004  |  Pubmed ID: 15280057

Glucanohydrolases, especially mutanase [alpha-(1-->3) glucanase; EC 3.2.1.59] and dextranase [alpha-(1-->6) glucanase; EC 3.2.1.11], which are present in the biofilm known as dental plaque, may affect the synthesis and structure of glucans formed by glucosyltransferases (GTFs) from sucrose within dental plaque. We examined the production and the structure of glucans synthesized by GTFs B (synthesis of alpha-(1-->3)-linked glucans) or C [synthesis of alpha-(1-->6)- and alpha-(1-->3)-linked glucans] in the presence of mutanase and dextranase, alone or in combination, in solution phase and on saliva-coated hydroxyapatite beads (surface phase). The ability of Streptococcus sobrinus 6715 to adhere to the glucan, which was formed in the presence of the glucanohydrolases was also explored. The presence of mutanase and/or dextranase during the synthesis of glucans by GTF B and C altered the proportions of soluble to insoluble glucan. The presence of either dextranase or mutanase alone had a modest effect on total amount of glucan formed, especially in the surface phase; the glucanohydrolases in combination reduced the total amount of glucan. The amount of (1-->6)-linked glucan was reduced in presence of dextranase. In contrast, mutanase enhanced the formation of soluble glucan, and reduced the percentage of 3-linked glucose of GTF B and C glucans whereas dextranase was mostly without effect. Glucan formed in the presence of dextranase provided fewer binding sites for S. sobrinus; mutanase was devoid of any effect. We also noted that the GTFs bind to dextranase and mutanase. Glucanohydrolases, even in the presence of GTFs, influence glucan synthesis, linkage remodeling, and branching, which may have an impact on the formation, maturation, physical properties, and bacterial binding sites of the polysaccharide matrix in dental plaque. Our data have relevance for the formation of polysaccharide matrix of other biofilms.

Estimated Exposure to Phthalates in Cosmetics and Risk Assessment

Journal of Toxicology and Environmental Health. Part A. Dec, 2004  |  Pubmed ID: 15513891

Some phthalates such as di(2-ethylhexyl) phthalate (DEHP) and dibutyl phthalate (DBP) and their metabolites are suspected of producing teratogenic or endocrine-disrupting effects. To predict possible human exposure to phthalates in cosmetics, the levels of DEHP, diethyl phthalate (DEP), DBP, and butylbenzyl phthalate (BBP) were determined by high-performance liquid chromatography (HPLC) in 102 branded hair sprays, perfumes, deodorants, and nail polishes. DBP was detected in 19 of the 21 nail polishes and in 11 of the 42 perfumes, and DEP was detected in 24 of the 42 perfumes and 2 of the 8 deodorants. Median exposure levels to phthalates in cosmetics by dermal absorption were estimated to be 0.0006 g/kg body weight (bw)/d for DEHP, 0.6 g/kg bw/d for DEP, and 0.103 g/kg bw/d for DBP. Furthermore, if phthalates in cosmetics were assumed to be absorbed exclusively via 100% inhalation, the median daily exposure levels to phthalates in cosmetics were estimated to be 0.026 g/kg bw/d for DEHP, 81.471 g/kg bw/d for DEP, and 22.917 g/kg bw/d for DBP, which are far lower than the regulation levels set buy the Scientific Committee on Toxicity, Ecotoxicity, and the Environment (CSTEE) (37 g/kg bw/d, DEHP), Agency for Toxic Substances and Disease Registry (ATSDR) (7000 g/kg bw/d, DEP), and International Programme on Chemical Safety (IPCS) (66 g/kg bw/d, DBP), respectively. Based on these data, hazard indices (HI, daily exposure level/regulation level) were calculated to be 0.0007 for DEHP, 0.012 for DEP, and 0.347 for DBP. These data suggest that estimated exposure to-phthalates in the cosmetics mentioned are relatively small. However, total exposure levels from several sources may be greater and require further investigation.

Genotoxicity Evaluation of Isaria Sinclairii (ISE) Extract

Journal of Toxicology and Environmental Health. Part A. Dec, 2004  |  Pubmed ID: 15513901

The mutagenic potential Isaria sinclairii, a traditional Chinese medicine composed of the fruiting bodies of I. sinclairii and its parasitic host larva, was evaluated using short-term genotoxicity tests, namely, the Ames test, chromosome aberration (CA), and micronuclei (MN) tests. In a Salmonella typhimurium assay, I. sinclairii extract (ISE) did not produce any mutagenic response in the absence or presence of 59 mix with TA98, TA100, TA1535, and TA1537. In the chromosome aberration (CA) test, ISE induced no significant effect on Chinese hamster ovary (CHO) cells compared with control. In the MN test, no significant change in the occurrence of micronucleated polychromatic erythrocytes was observed in male ICR mice intraperitoneally administered ISE at doses of 15, 150, or 1500 mg/kg. These results indicate that ISE has no mutagenic potential in these in vitro and in vivo systems.

The Putative Autolysin Regulator LytR in Streptococcus Mutans Plays a Role in Cell Division and is Growth-phase Regulated

Microbiology (Reading, England). Feb, 2005  |  Pubmed ID: 15699211

Streptococcus mutans is the primary odontopathogen present in supragingival plaque and causes the oral disease known as dental caries. Colonization of the oral cavity by S. mutans requires the bacteria to adhere to the tooth surface and occurs by both sucrose-dependent and -independent mechanisms. Sucrose-independent adhesion of S. mutans in vitro has been shown to involve an ORF (ORF0317) encoding a homologue (39 %) to LytR, a regulator of autolysin activity in Bacillus subtilis. The protein encoded by ORF0317, LytR, belongs to the LytR/CpsA/Psr protein family. This family has a putative role in cell-wall structural maintenance, possibly through autolysin regulation. Autolysins have also been shown to be important in surface adhesion in Lactococcus lactis and in the pathogenic properties of Streptococcus pneumoniae. To investigate the role of autolysins in the adhesion and pathogenesis of S. mutans, a LytR mutant was constructed. The mutant grows in long chains, which may indicate a defect in cell division. Further experiments with the mutant strain show increased autolytic activity, indicating that LytR attenuates S. mutans autolytic activity, possibly through regulation of the expression of autolytic enzymes. No defect in cell-to-surface adherence or biofilm growth was seen in the LytR mutant. However, a connection between cell growth phase and transcription of lytR was found.

In Vitro and in Vivo Effects of Isolated Fractions of Brazilian Propolis on Caries Development

Journal of Ethnopharmacology. Oct, 2005  |  Pubmed ID: 15913934

Recently, two chemically different types of Brazilian propolis (type-3 and -12) were shown to have cariostatic properties. This study aimed to evaluate the influence of their isolated fractions on mutans streptococci viability, glucosyltransferases (GTFs) activity and caries development in rats. The ethanolic extracts of propolis (EEPs) were serially fractionated into hexane (H-fr), chloroform, ethyl acetate, and ethanol. The ability of the four fractions and EEP to inhibit Streptococcus mutans and Streptococcus sobrinus growth and adherence to a glass surface was examined. The effect on GTFs B and C activity was also determined. For the caries study, 60 Wistar rats infected with Streptococcus sobrinus were treated topically twice daily as follows: (1) EEP type-3, (2) H-fr type-3, (3) EEP type-12, (4) H-fr type-12, and (5) control. In general, the H-fr from both types of propolis showed the highest antibacterial activity and GTFs inhibition. Furthermore, the EEP and H-fr type-3 and -12 were equally effective in reducing dental caries in rats. The data suggest that the putative cariostatic compounds of propolis type-3 and -12 are mostly non-polar; and H-fr should be the fraction of choice for identifying further potentially novel anti-caries agents.

Human Monitoring of Phthalates and Risk Assessment

Journal of Toxicology and Environmental Health. Part A. Aug, 2005  |  Pubmed ID: 16009652

Some phthalates, such as di(2-ethylhexyl) phthalate (DEHP) and dibutyl phthalate (DBP), and their metabolites are suspected of producing teratogenic and endocrino-disrupting effects. In this study, urinary levels of phthalates (DEHP, DBP, diethyl phthalate (DEP), butylbenzyl phthalate BBP), and monoethylhexyl phthalate (MEHP, a major metabolite of DEHP) were measured by high performance liquid chromatography (HPLC) in human populations (women [hospital visitors], n = 150, and children, n = 150). Daily exposure level of DEHP in children was estimated to be 12.4 microg/kg body weight/d (male 9.9 microg/kg body weight/d, female 17.8 microg/kg body weight/d), but, in women was estimated to be 41.7 microg/kg body weight/d, which exceeded the tolerable daily intake (TDI, 37 microg/kg body weight/day) level established by the European Union (EU) Scientific Committee for Toxicity, Ecotoxicity, and the Environment (SCTEE) based on reproductive toxicity. Based on these data, hazard indices (HIs) were calculated to be 1.12 (41.7/37 TDI) for women and 0.33 (12.4/37 TDI) for children, respectively. These data suggest that Koreans (women and children) were exposed to significant levels of phthalates, which should be reduced to as low a level as technologically feasible to protect Koreans from the exposure to toxic phthalates.

Predictors of Suicidal Ideation for Adolescents by Gender

Taehan Kanho Hakhoe Chi. Dec, 2005  |  Pubmed ID: 16415624

The purposes of this study were 1) to examine the differences in suicidal ideation and psychological variables by gender, 2) compare the contribution of demographic-behavioral variables and psychosocial variables in explaining the variance in suicidal ideation, and 3) identify the most important predictors of suicidal ideation for male adolescents and female adolescents.

Influence of Apigenin on Gtf Gene Expression in Streptococcus Mutans UA159

Antimicrobial Agents and Chemotherapy. Feb, 2006  |  Pubmed ID: 16436708

Apigenin, a potent inhibitor of glucosyltransferase activity, affects the accumulation of Streptococcus mutans biofilms in vitro by reducing the formation of insoluble glucans and enhancing the soluble glucan content of the polysaccharide matrix. In the present study, we investigated the influence of apigenin on gtfB, gtfC, and gtfD expression in S. mutans UA159. Apigenin (0.1 mM) significantly decreased the expression of gtfB and gtfC mRNA (P < 0.05); in contrast, it increased the expression of gtfD in S. mutans growing in the planktonic state. The protein levels of GTF B, GTF C, and GTF D in culture supernatants were also affected; less GTF B and C were detected, whereas the level of GTF D was significantly elevated (P < 0.05). A similar profile of gtf expression was obtained with biofilms, although an elevated concentration (1 mM) of apigenin was required to elicit the effects. The influence of apigenin on gtf gene expression was independent of any effect on GTF activity, did not involve inhibition of growth or effects on pH, and was not affected by addition of sucrose. The data show that apigenin modulates the genetic expression of virulence factors in S. mutans.

Co-operative Inhibition by Fluoride and Zinc of Glucosyl Transferase Production and Polysaccharide Synthesis by Mutans Streptococci in Suspension Cultures and Biofilms

FEMS Microbiology Letters. Jan, 2006  |  Pubmed ID: 16451191

Fluoride and zinc, alone or in combination at concentrations of 0.2 mM, inhibited production-secretion of glucosyltranferases by Streptococcus mutans UA159 growing in suspension cultures. Inhibition did not involve growth inhibition or starvation. Fluoride and zinc also inhibited glucan production, especially insoluble glucan, in fed-batch biofilms. Inhibition of biofilms appeared to be associated with starvation as indicated by markedly decreased ATP pools and iodophilic polysaccharide levels in biofilm cells. As insoluble glucans are important for virulence of mutans streptococci, the inhibitory actions of fluoride and zinc could significantly affect cariogenicity.

[Satisfaction with Life and It's Predictors of Korean Adolescents]

Taehan Kanho Hakhoe Chi. Feb, 2006  |  Pubmed ID: 16520574

This study was done to investigate the satisfaction with life in adolescents, and to identify factors affecting satisfaction with life for them.

Inhibitory Effects of Cranberry Polyphenols on Formation and Acidogenicity of Streptococcus Mutans Biofilms

FEMS Microbiology Letters. Apr, 2006  |  Pubmed ID: 16553831

Cranberry fruit is a rich source of polyphenols, and has shown biological activities against Streptococcus mutans. In the present study, we examined the influence of extracts of flavonols (FLAV), anthocyanins (A) and proanthocyanidins (PAC) from cranberry on virulence factors involved in Streptococcus mutans biofilm development and acidogenicity. PAC and FLAV, alone or in combination, inhibited the surface-adsorbed glucosyltransferases and F-ATPases activities, and the acid production by S. mutans cells. Furthermore, biofilm development and acidogenicity were significantly affected by topical applications of PAC and FLAV (P<0.05). Anthocyanins were devoid of any significant biological effects. The flavonols are comprised of mostly quercetin glycosides, and the PAC are largely A-type oligomers of epicatechin. Our data show that proanthocyanidins and flavonols are the active constituents of cranberry against S. mutans.

Predictors of Suicidal Ideation Among High School Students by Gender in South Korea

The Journal of School Health. May, 2006  |  Pubmed ID: 16635202

The purpose of this study was to examine the evidence to determine if there are gender differences in suicidal ideation of adolescents. This study examined the main effect of risk factors from 5 domains and protective factors from 1 domain in relation to suicidal ideation by gender and identified the most important predictors of suicidal ideation for males (N = 654) and females (N = 658). This study was a cross-sectional survey, and data were collected through self-report questionnaires. In the univariate analysis, especially, risk factors from behavioral variables and psychosocial-environmental variables appeared to be gender skewed. For males, all behavioral variables were predictive of suicidal ideation. For the females, unlike the males, Wang-tta or victim of bullying behavior and sexual orientation as behavioral variables were predictive of suicidal ideation. For males, parental divorce and parental alcohol abuse as psychosocial-environmental variables were predictive of suicidal ideation. For the females, again unlike for the males, all the psychosocial-environmental variables were not predictive of suicidal ideation. The most important predictors of suicidal ideation for males as a result of the multivariate analysis were history of suicidal attempt, depression, hostility, smoking, parental alcohol abuse, communication with friends, and self-esteem. The most important predictors of suicidal ideation for females as a result of the multivariate analysis were depression, hostility, sexual orientation, and self-esteem. These results would indicate that an effective suicide screening and prevention program for adolescents should consider gender differences.

A Giant Colonic Hamartoma and Multiple Colonic Hamartomatous Polyps in a Middle-aged Man

Yonsei Medical Journal. Oct, 2006  |  Pubmed ID: 17066524

Colonic hamartomas are rare polypoid lesions. We report an unusual case of multiple colonic hamartomatous polyps, including a giant hamartoma, unrelated to hereditary or familial polyposis syndromes, in a 48-year-old man. The diameter of the largest polyp was 9.5 cm, and endoscopy revealed that the lesion caused colonic obstruction. The clinical, endoscopic and histological aspects of this case are discussed.

The Influence of a Novel Propolis on Mutans Streptococci Biofilms and Caries Development in Rats

Archives of Oral Biology. Jan, 2006  |  Pubmed ID: 16054589

A flavonoids-free Brazilian propolis (type 6) showed biological effects against mutans streptococci and inhibited the activity of glucosyltransferases. This study evaluated the influence of the ethanolic extract of a novel type of propolis (EEP) and its purified hexane fraction (EEH) on mutans streptococci biofilms and the development of dental caries in rats. The chemical composition of the propolis extracts were examined by gas chromatography/mass spectrometry. The effects of EEP and EEH on Streptococcus mutans UA159 and Streptococcus sobrinus 6715 biofilms were analysed by time-kill and glycolytic pH drop assays. Their influence on proton-translocating F-ATPase activity was also tested. In the animal study, the rats were infected with S. sobrinus 6715 and fed with cariogenic diet 2000. The rats were treated topically twice a day with each of the extracts (or control) for 5 weeks. After the experimental period, the microbial composition of their dental plaque and their caries scores were determined. The results showed that fatty acids (oleic, palmitic, linoleic and stearic) were the main compounds identified in EEP and EEH. These extracts did not show major effects on the viability of mutans streptococci biofilms. However, EEP and EEH significantly reduced acid production by the biofilms and also inhibited the activity of F-ATPase (60-65%). Furthermore, both extracts significantly reduced the incidence of smooth surface caries in vivo without displaying a reduction of the percentage of S. sobriuns in the animals' plaque (P < 0.05). However, only EEH was able to reduce the incidence and severity of sulcal surface caries (P < 0.05). The data suggest that the cariostatic properties of propolis type 6 are related to its effect on acid production and acid tolerance of cariogenic streptococci; the biological activities may be attributed to its high content of fatty acids.

Metabolic Profiling and Phylogenetic Analysis of Medicinal Zingiber Species: Tools for Authentication of Ginger (Zingiber Officinale Rosc)

Phytochemistry. Aug, 2006  |  Pubmed ID: 16169024

Phylogenetic analysis and metabolic profiling were used to investigate the diversity of plant material within the ginger species and between ginger and closely related species in the genus Zingiber (Zingiberaceae). In addition, anti-inflammatory data were obtained for the investigated species. Phylogenetic analysis demonstrated that all Zingiber officinale samples from different geographical origins were genetically indistinguishable. In contrast, other Zingiber species were significantly divergent, allowing all species to be clearly distinguished using this analysis. In the metabolic profiling analysis, the Z. officinale samples derived from different origins showed no qualitative differences in major volatile compounds, although they did show some significant quantitative differences in non-volatile composition, particularly regarding the content of [6]-, [8]-, and [10]-gingerols, the most active anti-inflammatory components in this species. The differences in gingerol content were verified by HPLC. The metabolic profiles of other Zingiber species were very different, both qualitatively and quantitatively, when compared to Z. officinale and to each other. Comparative DNA sequence/chemotaxonomic phylogenetic trees showed that the chemical characters of the investigated species were able to generate essentially the same phylogenetic relationships as the DNA sequences. This supports the contention that chemical characters can be used effectively to identify relationships between plant species. Anti-inflammatory in vitro assays to evaluate the ability of all extracts from the Zingiber species examined to inhibit LPS-induced PGE(2) and TNF-alpha production suggested that bioactivity may not be easily predicted by either phylogenetic analysis or gross metabolic profiling. Therefore, identification and quantification of the actual bioactive compounds are required to guarantee the bioactivity of a particular Zingiber sample even after performing authentication by molecular and/or chemical markers.

Toxicokinetic Relationship Between Di(2-ethylhexyl) Phthalate (DEHP) and Mono(2-ethylhexyl) Phthalate in Rats

Journal of Toxicology and Environmental Health. Part A. Mar, 2007  |  Pubmed ID: 17454563

The toxicokinetic relationship between di(2-ethylhexyl) phthalate (DEHP) and mono(2-ethylhexyl) phthalate (MEHP), a major metabolite of DEHP, was investigated in Sprague-Dawley rats orally treated with a single dose of 14C-DEHP. Urinary excretion of total 14C-DEHP and of its metabolites was followed by liquid scintillation counting (LSC). Concentrations of DEHP and MEHP were determined 6, 24, and 48 h after treatment in rat serum and 6, 12, 24, and 48 h after treatment in urine by high-performance liquid chromatography (HPLC). After 24 h, peak concentrations of MEHP in both urine and serum were observed in animals treated with 40, 200, or 1000 mg DEHP/kg. HPLC showed that general toxicokinetic parameters, such as Tmax (h), Cmax (microg/ml), Ke (1/h), and AUC (microg-h/ml/) were greater for MEHP than DEHP in both urine and serum. In contrast, the half-lives (t1/2 [h]) of DEHP were greater than those of MEHP. The AUC ratios between DEHP and MEHP were relatively smaller in serum than in urine, suggesting the important role of urinary DEHP data for exposure assessment of DEHP. The toxicokinetic relationship between DEHP and MEHP in rats suggests that DEHP exposure assessment should be based on DEHP and MEHP in urine and serum for risk assessment applications.

[The Study on Predictors of Depression for Korean Female Adolescents]

Taehan Kanho Hakhoe Chi. Aug, 2007  |  Pubmed ID: 17804938

The purposes of this study were 1) to compare the contribution of demographic-behavioral variables and psychological variables in explaining the variance of depression, 2) identify the most important predictors of depression for Korean female adolescents.

[Relationships Among Cybersex Addiction, Gender Egalitarianism, Sexual Attitude and the Allowance of Sexual Violence in Adolescents]

Taehan Kanho Hakhoe Chi. Dec, 2007  |  Pubmed ID: 18182882

This study was done to investigate cybersex addiction, gender egalitarianism, sexual attitude and the allowance of sexual violence in adolescents, and to identify the relationships among these variables.

Role of Unsaturated Fatty Acid Biosynthesis in Virulence of Streptococcus Mutans

Infection and Immunity. Mar, 2007  |  Pubmed ID: 17220314

An insertionally inactivated fabM strain of Streptococcus mutans does not produce unsaturated membrane fatty acids and is acid sensitive (E. M. Fozo and R. G. Quivey, Jr., J. Bacteriol. 186:4152-4158, 2004). In this study, the strain was shown to be poorly transmissible from host to host. Animals directly infected with the fabM strain exhibited fewer and less severe carious lesions than those observed in the wild-type strain.

Extraction and Purification of Total RNA from Streptococcus Mutans Biofilms

Analytical Biochemistry. Jun, 2007  |  Pubmed ID: 17475197

RNA isolation from Streptococcus mutans within biofilms is challenging because of the presence of extracellular polysaccharide matrix that interferes with RNA extraction procedures. In an effort to solve this difficult problem, we examined several protocols to extract and purify RNA from S. mutans biofilms. A combination of sonication (three times using a 30-s pulse at 7 W) with washing in phosphate-buffered saline removed most of the extracellular polysaccharides from the biofilms and provided the highest RNA yield. Further homogenization-mechanical cells disruption in NAES buffer (50 mM sodium acetate buffer, 10 mM EDTA, and 1% SDS, pH 5.0) and acid phenol/chloroform yielded 547.2+/-23.4 microg RNA/100 mg of biofilm dry weight. An additional acid phenol/chloroform extraction further improved the purification of RNA without significantly affecting the RNA yield. The combination of DNase I in silica gel-based column and recombinant DNase I in solution effectively removed the genomic DNA as determined by real-time quantitative reverse transcriptase PCR (RT-PCR), resulting in 92.0+/-0.6 microg of purified RNA per 100 mg of biofilm dry weight. The complementary DNAs generated from the purified RNA sample were efficiently amplified using gtfB S. mutans-specific primers. The results demonstrated a method that yields high-quality RNA from biofilms of S. mutans in sufficient quantity for real-time RT-PCR analyses, and our data have relevance for isolation of RNA from other biofilm-forming microorganisms.

Hershberger Assay for Antiandrogenic Effects of Phthalates

Journal of Toxicology and Environmental Health. Part A. Aug, 2007  |  Pubmed ID: 17654256

The antiandrogenic effects of seven phthalates, di(2-ethylhexyl) phthalate (DEHP), dibutyl phthalate (DBP), butyl benzyl phthalate (BBP), di-isononyl phthalate (DINP), di-isodecyl phthalate (DIDP), di-n-heptyl phthalate (DnHP), and mono-2-ethyhexyl phthalate (MEHP), were investigated by Hershberger assay in castrated male SD rats. An androgen agonist, testosterone (0.4 mg/kg/d), was administered for 10 consecutive days by subcutaneous (s.c.) injection as a positive control. Additionally, 20, 100, or 500 mg/kg body weight (bw)/d of 6 phthalates (DEHP, DBP, BBP, DINP, DIDP, or DnHP) or 10, 50, or 250 mg/kg bw/d of MEHP, the primary metabolite of DEHP, were also administered orally in combination with testosterone (0.4 mg/kg/d, s.c.) for 10 consecutive days, respectively. In the testosterone-treated groups, glans penis, seminal vesicles, ventral prostate, and levator ani/bulbocavernosus muscles (LABC) weights were found to be significantly increased. Ventral prostate weights were significantly decreased in animals treated with DEHP or DBP at doses of 20 mg/kg bw/d or above, 500 mg/kg bw/d DIDP, and 250 mg/kg bw/d MEHP. Seminal vesicles weights were also significantly decreased by DEHP at > 100 mg/kg bw/d, DINP at > 20 mg/kg bw/d, DIDP at 500 mg/kg bw/d, or MEHP at 50 or 250 mg/kg bw/d, respectively. In addition, LABC weights were decreased by DEHP at 500 mg/kg bw/d, DINP at 500 mg/kg bw/d, and MEHP at 50 or 100 mg/kg bw/d. These data suggest that some phthalates possess antiandrogenic activity, and that multiple cross-talk between androgen, estrogen, and steroid hormone receptors occurs.

Chemical Characterization of Red Wine Grape (Vitis Vinifera and Vitis Interspecific Hybrids) and Pomace Phenolic Extracts and Their Biological Activity Against Streptococcus Mutans

Journal of Agricultural and Food Chemistry. Dec, 2007  |  Pubmed ID: 17999462

Grapes are rich sources of potentially bioactive polyphenols. However, the phenolic content is variable depending on grape variety, and may be modified during vinification. In this study, we examined the chemical composition and biological activity of phenolic extracts prepared from several red wine grape varieties and their fermented byproduct of winemaking (pomace) on some of the virulence properties of Streptococcus mutans a well-known dental pathogen. Grape phenolic extracts were obtained from Vitis vinifera varieties Cabernet Franc and Pinot Noir and Vitis interspecific hybrid varieties Baco Noir and Noiret. The anthocyanins and flavan-3-ols content were highly variable depending on grape variety and type of extract (whole fruit vs fermented pomace). Nevertheless, all grape phenolic extracts remarkably inhibited glucosyltransferases B and C (70-85% inhibition) at concentrations as low as 62.5 microg/mL (P < 0.01). Furthermore, the glycolytic pH-drop by S. mutans cells was inhibited by the grape extracts without affecting the bacterial viability; an effect that can be attributed to partial inhibition of F-ATPase activity (30-65% inhibition at 125 microg/mL; P < 0.01). The biological activity of fermented pomace was either as effective as or significantly better than whole fruit grape extracts. The results showed that grape phenolic extracts, especially from pomace, are highly effective against specific virulence traits of S. mutans despite major differences in their phenolic content.

Isolation and Purification of Total RNA from Streptococcus Mutans in Suspension Cultures and Biofilms

Brazilian Oral Research. Jul-Sep, 2008  |  Pubmed ID: 18949306

The presence of extracellular polysaccharides matrix makes extraction and purification of RNA from Streptococcus mutans within biofilms challenging. In this study, several approaches to purify RNA extracted from S. mutans in suspension cultures and biofilms were examined. The combination of sonication (3 pulses of 30 s at 7 W), suspension in NAES buffer (50 mM sodium acetate buffer, 10 mM EDTA and 1% SDS; pH 5.0) and homogenization-mechanical cells disruption in NAES- acid phenol:chloroform, yielded 9.04 mg (or 0.52 mg) of crude preparation of RNA per 100 mg of total cell (or biofilm) dry-weight. The crude RNA preparations were subjected to various DNAse I treatments. The combination of DNAse I in silica-gel based column followed by recombinant DNase I in solution provided the best genomic DNA removal, resulting in 4.35 mg (or 0.06 mg) of purified RNA per 100 mg of total cell (or biofilm) dry-weight. The cDNAs generated from the purified RNA sample were efficiently amplified using gtfB S. mutans-specific primers. The results showed a method that yields high-quality RNA from both planktonic cells and biofilms of S. mutans in sufficient quantity and quality for real-time RT-PCR analyses.

Inhibitory Effects of 7-epiclusianone on Glucan Synthesis, Acidogenicity and Biofilm Formation by Streptococcus Mutans

FEMS Microbiology Letters. May, 2008  |  Pubmed ID: 18371068

The aim of this study was to examine the effects of 7-epiclusianone, a new prenylated benzophenone isolated from the plant Rheedia gardneriana, on some of the virulence properties of Streptococcus mutans associated with biofilm development and acidogenicity. The synthesis of glucans by glucosyltransferases B (GTF B) and C (GTF C) was markedly reduced by 7-epiclusianone showing more than 80% inhibition of enzymatic activity at a concentration of 100 microg mL(-1). Double-reciprocal analysis (Lineweaver-Burk plots) revealed that the inhibition of GTF B activity was noncompetitive (mixed) while GTF C was inhibited uncompetitively. The glycolytic pH drop by S. mutans cells was also disrupted by 7-epiclusianone without affecting the bacterial viability, an effect that can be attributed, in part, to inhibition of F-ATPase activity (61.1+/-3.0% inhibition at 100 microg mL(-1)). Furthermore, topical applications (1-min exposure, twice daily) of 7-epiclusianone (at 250 microg mL(-1)) disrupted biofilm formation and physiology. The biomass (dry-weight), extracellular insoluble polysaccharide concentration and acidogenicity of the biofilms were significantly reduced by the test agent (P<0.05). The data show that 7-epiclusianone disrupts the extracellular and intracellular sugar metabolism of S. mutans, and holds promise as a novel, naturally occurring compound to prevent biofilm-related oral diseases.

Molecular Mass Sorting of Proteome Using Hollow Fiber Flow Field-flow Fractionation for Proteomics

Journal of Proteomics. Apr, 2008  |  Pubmed ID: 18541480

Hollow fiber flow field-flow fractionation (HF FlFFF) has been demonstrated as a tool for pre-fractionating proteomes by differences in molecular mass (Mr), where the resulting protein fractions are subsequently digested and analyzed by shotgun proteomics using two-dimensional liquid chromatography-electrospray ionization-tandem mass spectrometry (2D-LC-ESI-MS/MS). HF FlFFF is a separation device capable of fractionating proteins or cells by hydrodynamic radius, and protein fraction can be readily collected as intact conditions in aqueous buffer solutions. In this study, HF FlFFF was applied to fractionate the proteome of Corynebacterium glutamicum, a well known soil bacterium that has been widely used in bioindustry due to its remarkable ability to secrete high amounts of glutamic acid. The collected HF FlFFF fractions of different MW intervals were enzymatically digested for protein identification by 2D-LC-ESI-MS/MS. Experiments showed improvements in protein identification when HF FlFFF pre-fractionation was applied, due to decreases in the ionization suppression effect and the MS exclusion effect by spectral congestion. Pre-fractionation of C. glutamicum proteome allowed us to find 90 additional proteins by 2D-LC-ESI-MS/MS that were not found by a direct shotgun analysis without pre-fractionation. A total of 415 proteins were found overall with 203 proteins commonly found from experiments with and without pre-fractionation.

Adsorption of Salivary and Serum Proteins, and Bacterial Adherence on Titanium and Zirconia Ceramic Surfaces

Clinical Oral Implants Research. Aug, 2008  |  Pubmed ID: 18705809

OBJECTIVES: The aim of this study was to determine the pattern of salivary and serum proteins present in pellicles formed on titanium (Ti) and zirconia ceramic (ZrO(2)) surfaces, and the ability of bacterial cells to adhere to the experimental pellicles. In addition, the protein profiles and bacterial binding properties of pellicles on Ti and ZrO(2) were compared to those formed on hydroxyapatite (HA) surface. METHODS: The pellicles were formed in vitro by incubating the materials with whole saliva, serum or saliva+serum. Protein composition in each of the pellicles was investigated by SDS-PAGE and immunodetection. The adherence of radiolabeled Streptococcus mutans and Actinomyces naeslundii to uncoated surfaces and experimental pellicles was determined by means of scintillation counting. Statistical analyses were done using ANOVA and Tukey's test at significance level at P<0.05. In general, the electrophoretic analysis of the pellicles formed on HA, Ti and ZrO(2) revealed few qualitative differences of the composition of proteins of the pellicles formed on HA, Ti and ZrO(2) surfaces. Pellicle components identified included amylase, IgA, IgG, albumin, fibronectin and fibrinogen. The number of S. mutans cells adhered to uncoated Ti and ZrO(2) was significantly higher than those adhered to HA (P<0.05). In contrast, lower number of A. naeslundii cells adhered to uncoated Ti and ZrO(2) than to HA (P<0.05). However, the presence of saliva and saliva+serum pellicles greatly reduced the number of S. mutans cells bound to each of the surfaces. The data showed that Ti and ZrO(2) display similar pellicle protein composition and bacterial binding properties; however, significant differences were observed when both materials were compared to HA.

Molecular Characterization of ORFs 2 to 7 of Korean Porcine Reproductive and Respiratory Syndrome Virus (CA) and Its Protein Expression by Recombinant Baculoviruses

Journal of Microbiology (Seoul, Korea). Dec, 2008  |  Pubmed ID: 19107402

To determine the characteristics of the Korean porcine reproductive and respiratory syndrome virus (PRRSV), CA, which was isolated from the serum of an infected pig in 2006, we investigated the nucleotide sequence and expression of the structural ORFs (ORFs 2 to 7) using the bApGOZA system. We found that the structural ORFs 2 to 7 of CA consisted of 3188 nucleotides that were the same as those formed from VR-2332. Comparison of the CA with the other strains revealed nucleotide sequence identity ranging from 89.8 to 99.5%. To better understand the genetic relationships between other strains, phylogenetic analyses were performed. The CA strain was closely related to the other North American genotype strains but formed a distinct branch with high bootstrap support. Additionally, expression levels of the PRRSV proteins in insect cells were strong or partially weak. The results of this study have implications for both the taxonomy of PRRSV and vaccine development.

Role of Clp Proteins in Expression of Virulence Properties of Streptococcus Mutans

Journal of Bacteriology. Apr, 2009  |  Pubmed ID: 19181818

Mutational analysis revealed that members of the Clp system, specifically the ClpL chaperone and the ClpXP proteolytic complex, modulate the expression of important virulence attributes of Streptococcus mutans. Compared to its parent, the DeltaclpL strain displayed an enhanced capacity to form biofilms in the presence of sucrose, had reduced viability, and was more sensitive to acid killing. The DeltaclpP and DeltaclpX strains displayed several phenotypes in common: slow growth, tendency to aggregate in culture, reduced autolysis, and reduced ability to grow under stress, including acidic pH. Unexpectedly, the DeltaclpP and DeltaclpX mutants were more resistant to acid killing and demonstrated enhanced viability in long-term survival assays. Biofilm formation by the DeltaclpP and DeltaclpX strains was impaired when grown in glucose but enhanced in sucrose. In an animal study, the average number of S. mutans colonies recovered from the teeth of rats infected with the DeltaclpP or DeltaclpX strain was slightly lower than that of the parent strain. In Bacillus subtilis, the accumulation of the Spx global regulator, a substrate of ClpXP, has accounted for the DeltaclpXP phenotypes. Searching the S. mutans genome, we identified two putative spx genes, designated spxA and spxB. The inactivation of either of these genes bypassed phenotypes of the clpP and clpX mutants. Western blotting demonstrated that Spx accumulates in the DeltaclpP and DeltaclpX strains. Our results reveal that the proteolysis of ClpL and ClpXP plays a role in the expression of key virulence traits of S. mutans and indicates that the underlying mechanisms by which ClpXP affect virulence traits are associated with the accumulation of two Spx orthologues.

The Utility of a Multibending Endoscope for Selective Cannulation During ERCP in Patients with a Billroth II Gastrectomy (with Video)

Gastrointestinal Endoscopy. Apr, 2009  |  Pubmed ID: 19327479

Selective cannulation in patients with a Billroth II gastrectomy is still challenging.

Overtube-balloon-assisted Direct Peroral Cholangioscopy by Using an Ultra-slim Upper Endoscope (with Videos)

Gastrointestinal Endoscopy. Apr, 2009  |  Pubmed ID: 19327480

The "mother-baby" endoscope system currently used for peroral cholangioscopy (POC) has several limitations. Endoscopic direct cholangioscopy when using an ultra-slim upper endoscope with a guidewire to maintain access has been reported, but appropriate accessories are required to increase the success rate. Herein, we describe a novel method, overtube-balloon-assisted endoscopy, for direct POC.

Adjunctive Chemotherapeutic Suppression of Mutans Streptococci in the Setting of Severe Early Childhood Caries: an Exploratory Study

Journal of Public Health Dentistry. 2009  |  Pubmed ID: 19486465

This investigational study assessed the suppressive effect of 10 percent povidone iodine (PI) coupled with elimination of active carious lesions on salivary mutans streptococci (MS) populations in children with severe early childhood caries (S-ECC).

Effect of Biliary Stenting Combined with Ursodeoxycholic Acid and Terpene Treatment on Retained Common Bile Duct Stones in Elderly Patients: a Multicenter Study

The American Journal of Gastroenterology. Oct, 2009  |  Pubmed ID: 19568225

For frail, elderly patients with large impacted common bile duct (CBD) stones, long-term treatment with biliary stenting provides palliation. Biliary stenting with choleretic agents such as ursodeoxycholic acid (UDCA) and terpene preparations may promote CBD stone size reduction. We studied the effectiveness of biliary stenting combined with UDCA and a terpene preparation as a medical treatment for difficult-to-remove CBD stones in patients older than 65 years in this multicenter, prospective study.

Combined Endoscopic Stent-in-stent Placement for Malignant Biliary and Duodenal Obstruction by Using a New Duodenal Metal Stent (with Videos)

Gastrointestinal Endoscopy. Oct, 2009  |  Pubmed ID: 19595319

Self-expandable metal stents (SEMSs) are an effective palliative treatment for malignant biliary or duodenal strictures, but endoscopic stenting for combined malignant biliary and duodenal obstruction remains technically difficult.

Direct Peroral Cholangioscopy Using an Ultra-slim Upper Endoscope for the Treatment of Retained Bile Duct Stones

The American Journal of Gastroenterology. Nov, 2009  |  Pubmed ID: 19623165

Intracorporeal lithotripsy must be used under direct visual control with an additional endoscope in order to ensure safety and precise targeting during the removal of difficult-to-extract bile duct stones using conventional methods. The currently available "mother-baby" scope system has several disadvantages. We evaluated the feasibility and efficacy of direct peroral cholangioscopy (POC) using an ultra-slim upper endoscope for electrohydraulic lithotripsy (EHL) or laser lithotripsy (LL) for the treatment of difficult-to-extract bile duct stones.

[Development of a Cell Phone Addiction Scale for Korean Adolescents]

Journal of Korean Academy of Nursing. Dec, 2009  |  Pubmed ID: 20071895

This study was done to develop a cell phone addiction scale for Korean adolescents.

Extracellular Polysaccharides Matrix--an Often Forgotten Virulence Factor in Oral Biofilm Research

International Journal of Oral Science. Dec, 2009  |  Pubmed ID: 20690427

Control of Spin Precession in a Spin-injected Field Effect Transistor

Science (New York, N.Y.). Sep, 2009  |  Pubmed ID: 19762637

Spintronics increases the functionality of information processing while seeking to overcome some of the limitations of conventional electronics. The spin-injected field effect transistor, a lateral semiconducting channel with two ferromagnetic electrodes, lies at the foundation of spintronics research. We demonstrated a spin-injected field effect transistor in a high-mobility InAs heterostructure with empirically calibrated electrical injection and detection of ballistic spin-polarized electrons. We observed and fit to theory an oscillatory channel conductance as a function of monotonically increasing gate voltage.

Influences of Naturally Occurring Agents in Combination with Fluoride on Gene Expression and Structural Organization of Streptococcus Mutans in Biofilms

BMC Microbiology. 2009  |  Pubmed ID: 19863808

The association of specific bioactive flavonoids and terpenoids with fluoride can modulate the development of cariogenic biofilms by simultaneously affecting the synthesis of exopolysaccharides (EPS) and acid production by Streptococcus mutans, which enhanced the cariostatic effectiveness of fluoride in vivo. In the present study, we further investigated whether the biological actions of combinations of myricetin (flavonoid), tt-farnesol (terpenoid) and fluoride can influence the expression of specific genes of S. mutans within biofilms and their structural organization using real-time PCR and confocal fluorescence microscopy.

Renal Hilar Control During Laparoscopic Partial Nephrectomy: to Clamp or Not to Clamp

Journal of Endourology / Endourological Society. Aug, 2010  |  Pubmed ID: 20629571

To decrease intraoperative and perioperative morbidity that is associated with laparoscopic partial nephrectomy (LPN), clamping of the renal hilum has been advocated. It has been suggested, however, that renal hilar control is not necessary in all patients. We compared the perioperative and pathologic results of 21 consecutive patients who underwent LPN with or without renal hilar clamping at our institution.

Endoscopic Removal of an Enterolith Causing Afferent Loop Syndrome Using Electrohydraulic Lithotripsy

Digestive Endoscopy : Official Journal of the Japan Gastroenterological Endoscopy Society. Jul, 2010  |  Pubmed ID: 20642613

Electrohydraulic lithotripsy is a very useful method for fragmenting biliary stones and it can be used for endoscopic removal of difficult biliary stones. Acute afferent loop syndrome induced by enterolith is very rare, and surgical treatment is the usual choice for this condition. We describe a patient with acute afferent loop syndrome, which was induced by an enterolith after a Billroth II gastrectomy. We used electrohydraulic lithotripsy to endoscopically remove the enterolith.

Protocols to Study the Physiology of Oral Biofilms

Methods in Molecular Biology (Clifton, N.J.). 2010  |  Pubmed ID: 20717780

The oral cavity harbors several hundred different bacterial species that colonize both hard (teeth) and soft tissues, forming complex populations known as microbial biofilms. It is widely accepted that the phenotypic characteristics of bacteria grown in biofilms are substantially different from those grown in suspensions. Because biofilms are the natural habitat for the great majority of oral bacteria, including those contributing to oral diseases, a better understanding of the physiology of adherent populations is clearly needed to control oral microbes in health and disease. In this chapter, we use oral streptococci as examples for studying the physiology of oral biofilms.

Human Common Salivary Protein 1 (CSP-1) Promotes Binding of Streptococcus Mutans to Experimental Salivary Pellicle and Glucans Formed on Hydroxyapatite Surface

Journal of Proteome Research. Dec, 2010  |  Pubmed ID: 20858015

The saliva proteome includes host defense factors and specific bacterial-binding proteins that modulate microbial growth and colonization of the tooth surface in the oral cavity. A multidimensional mass spectrometry approach identified the major host-derived salivary proteins that interacted with Streptococcus mutans (strain UA159), the primary microorganism associated with the pathogenesis of dental caries. Two abundant host proteins were found to tightly bind to S. mutans cells, common salivary protein-1 (CSP-1) and deleted in malignant brain tumor 1 (DMBT1, also known as salivary agglutinin or gp340). In contrast to gp340, limited functional information is available on CSP-1. The sequence of CSP-1 shares 38.1% similarity with rat CSP-1. Recombinant CSP-1 (rCSP-1) protein did not cause aggregation of S. mutans cells and was devoid of any significant biocidal activity (2.5 to 10 μg/mL). However, S. mutans cells exposed to rCSP-1 (10 μg/mL) in saliva displayed enhanced adherence to experimental salivary pellicle and to glucans in the pellicle formed on hydroxyapatite surfaces. Thus, our data demonstrate that the host salivary protein CSP-1 binds to S. mutans cells and may influence the initial colonization of this pathogenic bacterium onto the tooth surface.

Inhibition of Streptococcus Mutans Biofilm Accumulation and Development of Dental Caries in Vivo by 7-epiclusianone and Fluoride

Biofouling. Oct, 2010  |  Pubmed ID: 20938851

7-Epiclusianone (7-epi), a novel naturally occurring compound isolated from Rheedia brasiliensis, effectively inhibits the synthesis of exopolymers and biofilm formation by Streptococcus mutans. In the present study, the ability of 7-epi, alone or in combination with fluoride (F), to disrupt biofilm development and pathogenicity of S. mutans in vivo was examined using a rodent model of dental caries. Treatment (twice-daily, 60s exposure) with 7-epi, alone or in combination with 125 ppm F, resulted in biofilms with less biomass and fewer insoluble glucans than did those treated with vehicle-control, and they also displayed significant cariostatic effects in vivo (p < 0.05). The combination 7-epi + 125 ppm F was as effective as 250 ppm F (positive-control) in reducing the development of both smooth- and sulcal-caries. No histopathological alterations were observed in the animals after the experimental period. The data show that 7-epiclusianone is a novel and effective antibiofilm/anticaries agent, which may enhance the cariostatic properties of fluoride.

Dynamics of Streptococcus Mutans Transcriptome in Response to Starch and Sucrose During Biofilm Development

PloS One. 2010  |  Pubmed ID: 20976057

The combination of sucrose and starch in the presence of surface-adsorbed salivary α-amylase and bacterial glucosyltransferases increase the formation of a structurally and metabolically distinctive biofilm by Streptococcus mutans. This host-pathogen-diet interaction may modulate the formation of pathogenic biofilms related to dental caries disease. We conducted a comprehensive study to further investigate the influence of the dietary carbohydrates on S. mutans-transcriptome at distinct stages of biofilm development using whole genomic profiling with a new computational tool (MDV) for data mining. S. mutans UA159 biofilms were formed on amylase-active saliva coated hydroxyapatite discs in the presence of various concentrations of sucrose alone (ranging from 0.25 to 5% w/v) or in combination with starch (0.5 to 1% w/v). Overall, the presence of sucrose and starch (suc+st) influenced the dynamics of S. mutans transcriptome (vs. sucrose alone), which may be associated with gradual digestion of starch by surface-adsorbed amylase. At 21 h of biofilm formation, most of the differentially expressed genes were related to sugar metabolism, such as upregulation of genes involved in maltose/maltotriose uptake and glycogen synthesis. In addition, the groEL/groES chaperones were induced in the suc+st-biofilm, indicating that presence of starch hydrolysates may cause environmental stress. In contrast, at 30 h of biofilm development, multiple genes associated with sugar uptake/transport (e.g. maltose), two-component systems, fermentation/glycolysis and iron transport were differentially expressed in suc+st-biofilms (vs. sucrose-biofilms). Interestingly, lytT (bacteria autolysis) was upregulated, which was correlated with presence of extracellular DNA in the matrix of suc+st-biofilms. Specific genes related to carbohydrate uptake and glycogen metabolism were detected in suc+st-biofilms in more than one time point, indicating an association between presence of starch hydrolysates and intracellular polysaccharide storage. Our data show complex remodeling of S. mutans-transcriptome in response to changing environmental conditions in situ, which could modulate the dynamics of biofilm development and pathogenicity.

Effect of Transdermal Fentanyl Patches on the Motility of the Sphincter of Oddi

Gut and Liver. Sep, 2010  |  Pubmed ID: 20981215

Pain is one of the most troublesome symptoms of pancreatitis. Transdermal fentanyl patches (TFPs) are long-acting analgesics with a reduced risk of dependency. This prospective study evaluated the effect of TFPs on sphincter of Oddi (SO) motility for the management of pain in pancreatitis.

Identification of Gene Targets Eliciting Improved Alcohol Tolerance in Saccharomyces Cerevisiae Through Inverse Metabolic Engineering

Journal of Biotechnology. Aug, 2010  |  Pubmed ID: 20600383

The economic production of biofuels from renewable biomass using Saccharomyces cerevisiae requires tolerance to high concentrations of sugar and alcohol. Here we applied an inverse metabolic engineering approach to identify endogenous gene targets conferring improved alcohol tolerance in S. cerevisiae. After transformation with a S. cerevisiae genomic library, enrichment of the transformants exhibiting improved tolerance was performed by serial subculture in the presence of iso-butanol (1%). Through sequence analysis of the isolated plasmids from the selected transformants, four endogenous S. cerevisiae genes were identified as overexpression targets eliciting improved tolerance to both iso-butanol and ethanol. Overexpression of INO1, DOG1, HAL1 or a truncated form of MSN2 resulted in remarkably increased tolerance to high concentrations of iso-butanol and ethanol. Overexpression of INO1 elicited the highest ethanol tolerance, resulting in higher titers and volumetric productivities in the fermentation experiments performed with high glucose concentrations. In addition, the INO1-overexpressing strain showed a threefold increase in the specific growth rate as compared to that of the control strain under conditions of high levels of glucose (10%) and ethanol (5%). Although alcohol tolerance in yeast is a complex trait affected by simultaneous interactions of many genes, our results using a genomic library reveal potential target genes for better understanding and possible engineering of metabolic pathways underlying alcohol tolerance phenotypes.

Effects of 7-epiclusianone on Streptococcus Mutans and Caries Development in Rats

Planta Medica. Jan, 2011  |  Pubmed ID: 20665370

The aim of this study was to evaluate the effects of 7-epiclusianone (7-epi) on specific virulence attributes of Streptococcus mutans in vitro and on development of dental caries in vivo. 7-Epi was obtained and purified from fruits of Rheedia brasiliensis. We investigated its influence on surface-adsorbed glucosyltransferase (Gtf) B activity, acid production, and viability of S. MUTANS in biofilms, as well as on caries development using a rodent model. 7-Epi (100 µg/mL) significantly reduced the activity of surface-adsorbed GtfB (up to 48.0 ± 1.8 of inhibition at 100 µg/mL) and glycolytic pH-drop by S. mutans in biofilms (125 and 250 µg/mL) (vs. vehicle control, p < 0.05). In contrast, the test compound did not significantly affect the bacterial viability when compared to vehicle control (15 % ethanol, p > 0.05). Wistar rats treated topically with 7-epi (twice daily, 60-s exposure) showed significantly smaller number of and less severe smooth- and sulcal-surface carious lesions (p < 0.05), without reducing the S. mutans viable population from the animals' dental biofilms. In conclusion, the natural compound 7-epiclusianone may be a potentially novel pharmacological agent to prevent and control dental caries disease.

Improved Galactose Fermentation of Saccharomyces Cerevisiae Through Inverse Metabolic Engineering

Biotechnology and Bioengineering. Mar, 2011  |  Pubmed ID: 21246509

Although Saccharomyces cerevisiae is capable of fermenting galactose into ethanol, ethanol yield and productivity from galactose are significantly lower than those from glucose. An inverse metabolic engineering approach was undertaken to improve ethanol yield and productivity from galactose in S. cerevisiae. A genome-wide perturbation library was introduced into S. cerevisiae, and then fast galactose-fermenting transformants were screened using three different enrichment methods. The characterization of genetic perturbations in the isolated transformants revealed three target genes whose overexpression elicited enhanced galactose utilization. One confirmatory (SEC53 coding for phosphomannomutase) and two novel targets (SNR84 coding for a small nuclear RNA and a truncated form of TUP1 coding for a general repressor of transcription) were identified as overexpression targets that potentially improve galactose fermentation. Beneficial effects of overexpression of SEC53 may be similar to the mechanisms exerted by overexpression of PGM2 coding for phosphoglucomutase. While the mechanism is largely unknown, overexpression of SNR84, improved both growth and ethanol production from galactose. The most remarkable improvement of galactose fermentation was achieved by overexpression of the truncated TUP1 (tTUP1) gene, resulting in unrivalled galactose fermentation capability, that is 250% higher in both galactose consumption rate and ethanol productivity compared to the control strain. Moreover, the overexpression of tTUP1 significantly shortened lag periods that occurs when substrate is changed from glucose to galactose. Based on these results we proposed a hypothesis that the mutant Tup1 without C-terminal repression domain might bring in earlier and higher expression of GAL genes through partial alleviation of glucose repression. mRNA levels of GAL genes (GAL1, GAL4, and GAL80) indeed increased upon overexpression of tTUP. The results presented in this study illustrate that alteration of global regulatory networks through overexpression of the identified targets (SNR84 and tTUP1) is as effective as overexpression of a rate limiting metabolic gene (PGM2) in the galactose assimilation pathway for efficient galactose fermentation in S. cerevisiae. In addition, these results will be industrially useful in the biofuels area as galactose is one of the abundant sugars in marine plant biomass such as red seaweed as well as cheese whey and molasses.

Elevated Incidence of Dental Caries in a Mouse Model of Cystic Fibrosis

PloS One. 2011  |  Pubmed ID: 21304986

Dental caries is the single most prevalent and costly infectious disease worldwide, affecting more than 90% of the population in the U.S. The development of dental cavities requires the colonization of the tooth surface by acid-producing bacteria, such as Streptococcus mutans. Saliva bicarbonate constitutes the main buffering system which neutralizes the pH fall generated by the plaque bacteria during sugar metabolism. We found that the saliva pH is severely decreased in a mouse model of cystic fibrosis disease (CF). Given the close relationship between pH and caries development, we hypothesized that caries incidence might be elevated in the mouse CF model.

Expression of Aldehyde Dehydrogenase 6 Reduces Inhibitory Effect of Furan Derivatives on Cell Growth and Ethanol Production in Saccharomyces Cerevisiae

Bioresource Technology. May, 2011  |  Pubmed ID: 21421300

Yeast dehydrogenases and reductases were overexpressed in Saccharomyces cerevisiae D452-2 to detoxify 2-furaldehyde (furfural) and 5-hydroxymethyl furaldehyde (HMF), two potent toxic chemicals present in acid-hydrolyzed cellulosic biomass, and hence improve cell growth and ethanol production. Among those enzymes, aldehyde dehydrogenase 6 (ALD6) played the dual roles of direct oxidation of furan derivatives and supply of NADPH cofactor to their reduction reactions. Batch fermentation of S. cerevisiae D452-2/pH-ALD6 in the presence of 2g/L furfural and 0.5 g/L HMF resulted in 20-30% increases in specific growth rate, ethanol concentration and ethanol productivity, compared with those of the wild type strain. It was proposed that overexpression of ALD6 could recover the yeast cell metabolism and hence increase ethanol production from lignocellulosic biomass containing furan-derived inhibitors.

Influences of Trans-trans Farnesol, a Membrane-targeting Sesquiterpenoid, on Streptococcus Mutans Physiology and Survival Within Mixed-species Oral Biofilms

International Journal of Oral Science. Apr, 2011  |  Pubmed ID: 21485314

Trans-trans farnesol (tt-farnesol) is a bioactive sesquiterpene alcohol commonly found in propolis (a beehive product) and citrus fruits, which disrupts the ability of Streptococcus mutans (S. mutans) to form virulent biofilms. In this study, we investigated whether tt-farnesol affects cell-membrane function, acid production and/or acid tolerance by planktonic cells and biofilms of S. mutans UA159. Furthermore, the influence of the agent on S. mutans gene expression and ability to form biofilms in the presence of other oral bacteria (Streptococcus oralis (S. oralis) 35037 and Actinomyces naeslundii (A. naeslundii) 12104) was also examined. In general, tt-farnesol (1 mmol x L(-1)) significantly increased the membrane proton permeability and reduced glycolytic activity of S. mutans in the planktonic state and in biofilms (P < 0.05). Moreover, topical applications of 1 mmol x L(-1) tt-farnesol twice daily (1 min exposure/treatment) reduced biomass accumulation and prevented ecological shifts towards S. mutans dominance within mixed-species biofilms after introduction of 1% sucrose. S. oralis (a non-cariogenic organism) became the major species after treatments with tt-farnesol, whereas vehicle-treated biofilms contained mostly S. mutans (>90% of total bacterial population). However, the agent did not affect significantly the expression of S. mutans genes involved in acidogenicity, acid tolerance or polysaccharide synthesis in the treated biofilms. Our data indicate that tt-farnesol may affect the competitiveness of S. mutans in a mixed-species environment by primarily disrupting the membrane function and physiology of this bacterium. This naturally occurring terpenoid could be a potentially useful adjunctive agent to the current anti-biofilm/anti-caries chemotherapeutic strategies.

A Case of Crystalline Keratopathy in Monoclonal Gammopathy of Undetermined Significance (MGUS)

Korean Journal of Ophthalmology : KJO. Jun, 2011  |  Pubmed ID: 21655047

A 62-year-old female visited our clinic with progressively decreased vision in both eyes beginning 12 years prior. Idiopathic corneal opacity in all layers of the cornea was found in both eyes. One year later, we performed penetrating keratoplasty on the undiagnosed right eye. During post-surgical follow-up, corneal edema and stromal opacity recurred, and penetrating keratoplasty was performed two more times. The patient's total serum protein level, which had previously been normal, was elevated prior to the final surgery. She was diagnosed with monoclonal gammopathy of undetermined significance. We made a final diagnosis of monoclonal gammopathy-associated crystalline keratopathy after corneal biopsy. Monoclonal gammopathy-associated crystalline keratopathy is difficult to diagnose and may lead to severe visual loss. A systemic work-up, including serologic tests like serum protein or cholesterol levels, is needed in patients with unexplainable corneal opacity.

Characterization of Beauveria Bassiana MsW1 Isolated from Pine Sawyers, Monochamus Saltuarius

Journal of Basic Microbiology. Oct, 2011  |  Pubmed ID: 21656803

Monochamus saltuarius is a vector for pine wilt disease that causes enormous damage to native pine trees in Korea. To develop a biological control method for this pine wilt disease vector, an entomopathogenic fungus was isolated from the cadaver of an adult M. saltuarius supporting fungal conidiation. This fungus was named MsW1 and identified as Beauveria bassiana by microscopic examination, PCR amplification using B. bassiana -specific primers and genetic sequencing of the ITS and EF1-α regions. Virulence tests against M. saltuarius were conducted with conidial suspensions (1 × 10(8) conidia/ml) of B. bassiana MsW1 in laboratory conditions. The median lethal times (LT(50)) of adults and larvae were 7.2 and 7 days, and 100% mortality was observed at 11 and 13 days after inoculation, respectively. This is the first characterization of B. bassiana from M. saltuarius.

Tobacco Mitochondrial Small Heat Shock Protein NtHSP24.6 Adopts a Dimeric Configuration and Has a Broad Range of Substrates

BMB Reports. Dec, 2011  |  Pubmed ID: 22189686

There is a broad range of different small heat shock proteins (sHSPs) that have diverse structural and functional characteristics. To better understand the functional role of mitochondrial sHSP, NtHSP24.6 was expressed in Escherichia coli with a hexahistidine tag and purified. The protein was analyzed by non-denaturing PAGE, chemical cross-linking and size exclusion chromatography and the H6NtHSP24.6 protein was found to form a dimer in solution. The in vitro functional analysis of H6NtHSP24.6 using firefly luciferase and citrate synthase demonstrated that this protein displays typical molecular chaperone activity. When cell lysates of E. coli were heated after the addition of H6NtHSP24.6, a broad range of proteins from 10 to 160 kD in size remained in the soluble state. These results suggest that NtHSP24.6 forms a dimer and can function as a molecular chaperone to protect a diverse range of proteins from thermal aggregation.

A Case of Intrapancreatic Accessory Spleen Mistaken As a Pancreatic Mass Due to Different Enhancing Pattern from Normal Spleen

The Korean Journal of Gastroenterology = Taehan Sohwagi Hakhoe Chi. Dec, 2011  |  Pubmed ID: 22198236

Most cases of accessory spleen show similar features as normal spleen in imaging studies. However, some accessory spleen has unusual scan feature which can be misdiagnosed. We present a case of intrapancreatic accessory spleen that was discovered incidentally during a workup for abdominal pain in a 47-year-old woman. CT and MRI revealed a different enhancing pattern from that of the spleen. Further evaluation with endoscopic ultrasonography failed to identify the pancreatic mass. Therefore, it was surgically removed and diagnosed pathologically as an accessory spleen.

Production of Classical Swine Fever Virus Envelope Glycoprotein E2 As Recombinant Polyhedra in Baculovirus-infected Silkworm Larvae

Molecular Biotechnology. Mar, 2012  |  Pubmed ID: 21706129

Although, classical swine fever virus (CSFV) envelope glycoprotein E2 subunit vaccine has been developed using the baculovirus expression system, the expression of viral antigens in baculovirus-infected insect cells is often ineffective. Therefore, an alternative strategy to the traditional baculovirus expression system is needed that is more productive and effective. Here, we report a novel strategy for the large-scale production of a CSFV E2 in the larvae of a baculovirus-infected silkworm, Bombyx mori. We constructed a recombinant B. mori nucleopolyhedrovirus (BmNPV) that expressed recombinant polyhedra together with the N-terminal 179 amino acids of CSFV E2 (E2ΔC). BmNPV-E2ΔC-infected silkworm larvae expressed native polyhedrin and approximately 44-kDa fusion protein that was detected using both anti-polyhedrin and anti-CSFV E2 antibodies. Electron and confocal microscopy both demonstrated that the recombinant polyhedra contained both the fusion protein and native polyhedrin were morphologically normal and contained CSFV E2ΔC. The CSFV E2ΔC antigen produced in BmNPV-E2ΔC-infected silkworm larvae reached 0.68 mg/ml of hemolymph and 0.53 mg/larva at 6-days post-infection. Six-week-old female BALB/c mice that were immunized with the E2ΔC protein purified from solubilized recombinant polyhedra elicited CSFV E2 antibodies, which indicated that the CSFV E2ΔC protein from recombinant polyhedra was immunogenic. The virus neutralization test showed that the serum from mice that were treated with E2ΔC protein from recombinant polyhedra contained significant levels of virus neutralization activity. These results demonstrate that this strategy can be used for the large-scale production of CSFV E2 antigen.