Periodontal disease could be a potential risk factor for non-alcoholic fatty liver disease: An 11-year retrospective follow-up study.

OBJECTIVE: This study aimed to evaluate the association of periodontal disease with non-alcoholic fatty liver disease (NAFLD). MATERIALS AND METHODS: A retrospective follow-up study using the National Health Insurance Service-National Sample Cohort was performed from 2002 to 2015 in the Korean population. A total of 165,032 subjects were followed up for incident NAFLD during 11 years. Periodontal disease and NAFLD were defined by a diagnosis using the International Statistical Classification of Diseases and Related Health Problems, 10th revision (ICD-10) codes. Periodontal status was used as the severity of periodontal status and the number of dental visit due to PD. RESULTS: Periodontitis was associated with a 4% increase in risk for NAFLD after adjusting for socio-demographic factor, health behaviors, and systemic diseases (adjusted hazard ratio [aHR] = 1.04, 95% CI = 1.01 to 1.07). Between the number of dental visit due to PD and the risk for NAFLD was observed a dose-effect association (aHR = 1.02, 95% CI = 0.99 to 1.05 for once; aHR = 1.10, 95% CI = 1.06 to 1.15 for two times; aHR = 1.14, 95% CI = 1.06 to 1.24 for three times). CONCLUSIONS: Our data confirmed that periodontitis showed an association with a higher incidence of NAFLD. CLINICAL RELEVANCE: Prevention and management of periodontal disease could be beneficial for reducing the risk of NAFLD.

Association between diet quality and untreated dental caries: results from the Korea National Health and Nutrition Examination Survey.

BACKGROUND/OBJECTIVES: Few studies have provided evidence of the association between diet quality and dental caries. This study aimed to examine the association between diet quality and untreated dental caries in a Korean representative population. SUBJECTS/METHODS: The study population included a sample of 13,815 participants, aged ≥ 19 from the Korea National Health and Nutrition Examination Survey during 2013-2015. The explanatory variable was diet quality and the outcome variable was untreated dental caries. Untreated dental caries were defined by the number of decayed teeth recorded according to the criteria established by the World Health Organization. Diet quality was defined by using the Korean Healthy Eating Index (KHEI) through the 24-h recall methods. We assessed the association between diet quality and untreated dental caries while adjusting for age, sex, education, income, smoking status, dental visits, toothbrushing frequencies, obesity, and diabetes mellitus. RESULTS: The mean overall KHEI scores in the untreated dental caries group were significantly lower than those in the group without untreated dental caries. Significant differences were observed in the untreated dental caries group based on the KHEI quartiles (P < 0.001). After adjusting for potential confounders, the quartiles of KHEI scores showed an association with untreated dental caries, demonstrating a dose-effect trend (odds ratio [OR], 1.57; 95% confidence interval [CI], 1.35-1.84 for 1st quartile; OR, 1.38; 95% CI, 1.19-1.59 for 2nd quartile; OR, 1.32; 95% CI, 1.14-1.53 for 3rd quartile; reference quartile highest]). CONCLUSIONS: The findings indicated an inverse association between diet quality and untreated dental caries in Korean adults. Healthcare providers should take into account the significant role of diet quality in preventing and managing oral health.

Establishment of muscle mass-based indications for the cystatin C test in renal function evaluation.

Background: We aimed to suggest muscle mass-based criteria for using of the cystatin C test for the accurate estimated glomerular filtration rate (eGFR). Materials and methods: We recruited 138 Korean subjects and evaluated eGFRcr (derived from Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) based on creatinine) was compared to eGFRcys based on cystatin C as the reference value. The skeletal muscle mass index (SMI) by bioelectrical impedance analysis (BIA) was used as representative of muscle mass. Calf circumference (CC) was also evaluated. We defined the patients by eGFRcr as those with values of eGFRcr ≥ 60 mL/min/1.73 m2 but eGFRcys < 60 mL/min/1.73 m2 as the detection of hidden renal impairment (DHRI). Cut-off values were determined based on muscle mass for the cases of DHRI suggesting the criteria of cystatin C test in renal function evaluation. Results: We confirmed significant negative correlation between %difference of eGFRcr from eGFRcys and SMI (r, -0.592 for male, -0.484 for female) or CC (r, -0.646 for male, -0.351 for female). SMI of 7.3 kg/m2 for males and 5.7 kg/m2 for females were suggested to be significant cutoffs for indication of cystatin C test. We also suggested CC would be valuable for cystatin C indication. Conclusion: We suggested the muscle mass-based objective criteria relating to SMI and CC that would indicate the use of cystatin C to evaluate renal function test in sarcopenic cases. Our results highlight the importance of muscle mass-based selection of renal function.

Radiation-Inactivated S. gallinarum Vaccine Provides a High Protective Immune Response by Activating Both Humoral and Cellular Immunity.

Salmonella enterica subsp. enterica serovar Gallinarum (SG) is a common pathogen in chickens, and causes an acute systemic disease that leads to high mortality. The live attenuated vaccine 9R is able to successfully protect chickens older than six weeks by activating a robust cell-mediated immune response, but its safety and efficacy in young chickens remains controversial. An inactivated SG vaccine is being used as an alternative, but because of its low cellular immune response, it cannot be used as a replacement for live attenuated 9R vaccine. In this study, we employed gamma irradiation instead of formalin as an inactivation method to increase the efficacy of the inactivated SG vaccine. Humoral, cellular, and protective immune responses were compared in both mouse and chicken models. The radiation-inactivated SG vaccine (r-SG) induced production of significantly higher levels of IgG2b and IgG3 antibodies than the formalin-inactivated vaccine (f-SG), and provided a homogeneous functional antibody response against group D, but not group B Salmonella. Moreover, we found that r-SG vaccination could provide a higher protective immune response than f-SG by inducing higher Th17 activation. These results indicate that r-SG can provide a protective immune response similar to the live attenuated 9R vaccine by activating a higher humoral immunity and a lower, but still protective, cellular immune response. Therefore, we expect that the radiation inactivation method might substitute for the 9R vaccine with little or no side effects in chickens younger than six weeks.

An efficient method for the expression and reconstitution of thermostable Mn/Fe superoxide dismutase from Aeropyrum pernix K1.

The gene APE0743 encoding the superoxide dismutase (ApSOD) of a hyperthermophilic archaeon Aeropyrum pernix K1 was cloned and over-expressed as a GST fusion protein at a high level in Escherichia coli. The expressed protein was simply purified by the process of glutathione affinity chromatography and thrombin treatment. The ApSOD was a homodimer of 25 kDa subunits and a cambialistic SOD which was active with either Fe(II) or Mn(II) as a cofactor. The ApSOD was highly stable against high temperature. This thermostable ApSOD is expected to be applicable as a useful biocatalyst for medicine and bio-industrial processes.

Development of a Reporter System Monitoring Regulated Intramembrane Proteolysis of the Transmembrane bZIP Transcription Factor ATF6α.

When endoplasmic reticulum (ER) functions are perturbed, the ER induces several signaling pathways called unfolded protein response to reestablish ER homeostasis through three ER transmembrane proteins: inositol-requiring enzyme 1 (IRE1), PKR-like ER kinase (PERK), and activating transcription factor 6 (ATF6). Although it is important to measure the activity of ATF6 that can indicate the status of the ER, no specific cell-based reporter assay is currently available. Here, we report a new cell-based method for monitoring ER stress based on the cleavage of ATF6α by sequential actions of proteases at the Golgi apparatus during ER stress. A new expressing vector was constructed by using fusion gene of GAL4 DNA binding domain (GAL4DBD) and activation domain derived from herpes simplex virus VP16 protein (VP16AD) followed by a human ATF6α N-terminal deletion variant. During ER stress, the GAL4DBD-VP16AD(GV)-hATF6α deletion variant was cleaved to liberate active transcription activator encompassing GV-hATF6α fragment which could translocate into the nucleus. The translocated GV-hATF6α fragment strongly induced the expression of firefly luciferase in HeLa Luciferase Reporter cell line containing a stably integrated 5X GAL4 site-luciferase gene. The established double stable reporter cell line HLR-GV-hATF6α(333) represents an innovative tool to investigate regulated intramembrane proteolysis of ATF6α. It can substitute active pATF6(N) binding motif-based reporter cell lines.

Protective efficacy of a Brucella vaccine using a Salmonella-based delivery system expressing Brucella Omp3b, BCSP31, and SOD proteins against brucellosis in Korean black goats.

This study aimed to assess the protective efficacy of a novel Brucella vaccine formulation in goats. Twenty black goats were separated into 2 groups. Group A was injected with 3.0 × 109 CFU (colony-forming units)/mL of a Salmonella-based delivery system harboring only vector (pMMP65). Group B was immunized with 3.0 × 109 CFU/mL of the vaccine, a mixture of 3 Brucella vaccine strains (using a Salmonella-based delivery system) expressing each recombinant B. abortus Omp3b, BCSP31, and SOD protein. No Salmonella delivery strain was isolated from all tested lymph nodes and parenchymal organs. Serum immunoglobulin (Ig) G titers and interferon gamma concentrations were significantly higher in group B than those in group A. After intraconjunctival challenge with virulent B. abortus strain 544, 40% of the vaccinated animals in group B were protected against B. abortus infection. The infection index and colonization of B. abortus in tested tissues was significantly lower in group B than group A. We conclude that this Brucella vaccine induces significant antigen-specific immune responses and provides effective protection against B. abortus infection in goats. Further studies are needed to enhance the protection rate of this Brucella vaccine and to discover its practical application in small ruminants.

La présente étude visait à évaluer l'efficacité protectrice d'une nouvelle formulation de vaccin contre Brucella chez les chèvres. Vingt chèvres noires furent séparées en deux groupes. Le Groupe A reçut par injection 3,0 × 109 unités formatrices de colonies (UFC)/mL de Salmonella servant de système de livraison ne contenant seulement que le vecteur (pMMP65). Le groupe B fut immunisé avec 3,0 × 109 UFC/mL du vaccin, un mélange de trois souches vaccinales de Brucella (utilisant le système de livraison à base de Salmonella) exprimant chaque protéine recombinante Omp3b, BCSP31, et SOD de B. abortus. Aucune bactérie Salmonella du système de livraison ne fut isolée des ganglions lymphatiques et organes testés. Les concentrations sériques d'immunoglobulines G (IgG) et d'interféron gamma étaient significativement plus élevées dans le groupe B que dans le groupe A. À la suite d'une infection défi par voie intra-conjonctivale avec une souche virulente de B. abortus (544), 40 % des animaux vaccinés dans le groupe B étaient protégés contre l'infection par B. abortus. L'index d'infection et de colonisation par B. abortus dans les tissus testés étaient significativement plus faible dans le groupe B comparativement au groupe A. Nous avons conclu que ce vaccin contre Brucella induisait des réponses immunes spécifiques d'antigène significatives et fournissait une protection efficace contre l'infection par B. abortus chez les chèvres. Des études additionnelles sont requises afin d'augmenter le taux de protection de ce vaccin (Brucella) et pour découvrir son application pratique chez les petits ruminants.(Traduit par Docteur Serge Messier).

Protective efficacy of an inactivated Brucella abortus vaccine candidate lysed by GI24 against brucellosis in Korean black goats.

The efficacy of GI24-lysed Brucella abortus cells as a vaccine candidate against brucellosis in goats was evaluated on 2 groups of Korean black goats. Group A goats were immunized subcutaneously (SC) with sterile phosphate-buffered saline, whereas group B goats were immunized SC with approximately 3 × 109 lysed B. abortus cells. Subcutaneous immunization with the lysed cells did not cause any negative impact on the overall clinical status, such as behavior and appetite, throughout the study period. The enzyme-linked immunosorbent assay (ELISA) optical densities values for B. abortus lipopolysaccharide in serum were considerably higher in group B than those in group A. Also, the levels of the cytokines interleukin 4 (IL-4), tumor necrosis factor-alpha (TNF-α), and interferon gamma (IFN-γ) were significantly elevated in group B compared with those in group A. Following intraconjunctival challenge with B. abortus strain 544, the severity of brucellosis in terms of infection index and colonization of B. abortus in tissues was significantly lower in group B than in group A. The present study concluded that 3 of 5 goats immunized with GI24-lysed bacteria were completely protected against challenge. Future investigations are required to improve the protective efficacy offered by lysed B. abortus cells for practical applications in small ruminants.

L'efficacité de cellules lysées de Brucella abortus GI24 comme vaccin candidat contre la brucellose chez les chèvres a été évaluée chez deux groupes de chèvres noires coréennes. Les chèvres du groupe A ont été immunisées par voie sous-cutanée (SC) avec de la saline tamponnée stérile, alors que les chèvres du groupe B ont été immunisées SC avec environ 3 × 109 cellules lysées de B. abortus. L'immunisation sous-cutanée avec les cellules lysées n'a pas eu d'impact négatif sur l'état clinique général, tel que le comportement et l'appétit, tout au long de la période d'étude. Les valeurs de densité optique obtenues lors d'épreuves immunoenzymatiques (ELISA) utilisant le lipopolysaccharide de B. abortus étaient considérablement plus élevées avec le sérum des animaux du groupe B que celui des animaux du groupe A. De plus, les niveaux des cytokines interleukine-4 (IL-4), du facteur-alpha nécrosant de tumeur (TNF-α), d'interféron-gamma (IFN-γ) étaient significativement plus élevés dans le groupe B comparativement au groupe A. Pour donner suite à l'infection-défi intra-conjonctivale avec la souche 544 de B. abortus, la sévérité de brucellose en termes d'index d'infection et de colonisation des tissus par B. abortus était significativement moindre dans le groupe B que dans le groupe A. La présente étude a permis de conclure que 3 des 5 chèvres immunisées avec les bactéries GI24 lysées étaient complètement protégées contre l'infection. Des études ultérieures sont requises pour améliorer l'efficacité protectrice offerte par les cellules lysées de B. abortus pour une application pratique chez les petits ruminants.(Traduit par Docteur Serge Messier).

Periodontitis and the risk for non-fatal stroke in Korean adults.

BACKGROUND: The association between periodontal inflammation and non-fatal stroke is still controversial and limited to evidence in Western countries. The aim of this study was to investigate whether periodontitis is independently associated with non-fatal stroke in Korean adults. METHODS: A case-control study was conducted on 265 non-fatal chronic stroke cases at the National Rehabilitation Center, Seoul, Korea, and 214 non-stroke population controls. Medical specialists diagnosed stroke by using brain imaging from magnetic resonance imaging and/or computerized tomography. A dentist recorded the clinical attachment level (CAL), the distance between the cemento-enamel junction and the probed base of the periodontal pocket, using a University of North Carolina-15 manual probe. An interview assessed 17 sociodemographic, behavioral, systemic/oral health-related possible confounders. Multiple logistic regression analysis was used to evaluate the association between periodontitis and stroke while controlling for age, gender, income, education, smoking, drinking, history of systemic disease, body mass index, familial cardiovascular risk factors, and oral health behaviors. Subgroup analyses were also performed. RESULTS: Stroke was strongly associated with periodontitis (presence of CAL > or =6 mm): the odds ratio was 4.0 (95% confidence interval: 2.3 to 7.0) after controlling for all possible confounders. The association with periodontitis (tertiary percentage of CAL > or =5 mm) had a dose-response effect. The association between periodontitis and stroke was higher among adults younger than age 60 (6.0 versus 2.6) and normotensives (4.8 versus 3.2). CONCLUSION: Our data suggested that periodontitis is independently associated with non-fatal stroke, and its impact seems to be greater among younger or normotensive Korean adults.

Protective efficacy of a canine brucellosis vaccine candidate based on live attenuated Salmonella expressing recombinant Brucella BCSP31, Omp3b and SOD proteins in Beagles.

The aim of this study was to establish a proof-of-concept of protective efficacy of Salmonella-based B. abortus vaccine candidate in Beagles. Group A Beagles (n=10) were subcutaneously (SC) inoculated with S. Typhimurium delivery strain containing pMMP65 (vector to deliver antigens) only as vector control. Group B Beagles (n=10) were SC vaccinated with the mixture of the three Salmonella delivery strains expressing the recombinant B. abortus BCSP31, Outer membrane protein 3b (Omp3b), and superoxide dismutase (SOD) proteins, respectively. No Salmonella delivery strains were isolated from all tissues tested. Serum IgG, interleukin-4, tumor necrosis factor-alpha, and interferon-gamma concentrations were significantly higher in group B than in group A. Following intraconjunctival challenge with B. abortus 544, among 5 group B Beagles, the challenge strain was isolated from mandibular, and retropharyngeal lymph nodes of three Beagles, and no isolates were observed from all tissues of two Beagle. However, the challenge strains were detected from spleen, uterus (except two Beagles), and mandibular, prescapular, retropharyngeal, and superficial inguinal lymph nodes of all group A Beagles. These results suggest that the mixture of three S. Typhimurium delivery strains be a good vaccine candidate against brucellosis by B. abortus in dogs. Further investigations are needed to improve the protective efficacy of the Salmonella-based B. abortus vaccine candidate and explore its practical application in dogs.

Brucella abortus lysed cells using GI24 induce robust immune response and provide effective protection in Beagles.

The aim of the present study is to estimate the protective efficacy of Brucella abortus lysed cells by GI24 against brucellosis in Beagles. Group A was subcutaneously (sc) immunized with sterile phosphate-buffered saline, and group B was sc immunized with approximately 3 × 109 of the lysed cells. Brucella-LPS-specific serum IgG titers and IL-4, TNF-α and IFN-γ concentrations were investigated by enzyme linked immunosorbent assay. All dogs were intraconjunctivally challenged with B. abortus strain 544 at 6 weeks post-prime immunization. The serum IgG titers were considerably higher in group B than in group A. The levels of IL-4, TNF-α and IFN-γ in group B than in group A were significantly higher. Following challenge, no challenge strain was observed from all tissues of three dogs of group B. However, challenge strain was detected from spleen, uterus (except one Beagle) and inguinal and retropharyngeal lymph nodes of all group A Beagles. The results of this study demonstrated that sc immunization with the lysed cells induced robust antibody and cell-mediated immune responses in Beagles. The lysed cells also conferred protection against infection with B. abortus. These results suggest that sc immunization with B. abortus lysed cells by GI24 is a good vaccine candidate against brucellosis in dogs.

Expression and purification of a recombinant LL-37 from Escherichia coli.

Human cathelicidin-derived LL-37 is a 37-residue cationic, amphipathic alpha-helical peptide. It is an active component of mammalian innate immunity. LL-37 has several biological functions including a broad spectrum of antimicrobial activities and LPS-neutralizing activity. In order to determine the high-resolution three-dimensional structure of LL-37 using NMR spectroscopy, it is important to obtain the peptide with isotopic labels such as (15)N, (13)C and/or (2)H. Since it is less expensive to obtain such a peptide biologically, in this study, we report for the first time a method to express in E. coli and purify LL-37 using Glutathione S-transferase (GST) fusion system. LL-37 gene was inserted into vector pGEX-4T3 and expressed as a GST-LL-37 fusion protein in BL21(DE3) strain. The recombinant GST-LL-37 protein was purified with a yield of 8 mg/l by affinity chromatography and analyzed its biochemical and spectroscopic properties. Factor Xa was used to cleave a 4.5-kDa LL-37 from the GST-LL-37 fusion protein and the peptide was purified using a reverse-phase HPLC on a Vydac C(18) column with a final yield of 0.3 mg/l. The protein purified using reverse-phase HPLC was confirmed to be LL-37 by the analyses of Western blot and MALDI-TOF-Mass spectrometry. E. coli cells harboring the expression vector pGEX-4T3-LL-37 were grown in the presence of the (15)N-labeled M9 minimal medium and culture conditions were optimized to obtain uniform (15)N enrichment in the constitutively expressed LL-37 peptide. These results suggest that our production method will be useful in obtaining a large quantity of recombinant LL-37 peptide for NMR studies.

Protective efficacy by various doses of a new brucellosis vaccine candidate based on Salmonella strains expressing Brucella abortus BSCP31, Omp3b and superoxide dismutase against brucellosis in murine model.

Brucella species are important etiological agents of zoonotic diseases. Attenuated Salmonella strains expressing Brucella abortus BCSP31, Omp3b and superoxide dismutase proteins were tested as vaccine candidates in this study. In order to determine the optimal dose for intraperitoneal (IP) inoculation required to obtain effective protection against brucellosis, mice were immunized with various doses of a mixture of the three vaccine strains. Fifty BALB/c mice were divided into five equal groups (groups A-E). Group A mice were intraperitoneally inoculated with 100 µL of sterile phosphate-buffered saline. Group B, C, D and E mice were intraperitoneally immunized with approximately 1.2 × 105 colony-forming units (CFU) mL-1 of Salmonella containing pMMP65 in 100 µL and with 1.2 × 104 CFU mL-1, 1.2 × 105 CFU mL-1 and 1.2 × 106 CFU mL-1 of the mixture of the three strains in 100 µL, respectively. Serum IgG, tumor necrosis factor alpha and interferon gamma concentrations were significantly higher in group E than in groups A-D. Following challenge with B. abortus 544, the challenge strain was not detected in the spleen of any mouse from group E. Thus, IP immunization with 1.2 × 106 CFU mL-1 of the mixture of the three vaccine strains induced immune responses and provided effective protection against brucellosis in mice.

Protective efficacy of a Salmonella Typhimurium ghost vaccine candidate constructed with a recombinant lysozyme-PMAP36 fusion protein in a murine model.

A Salmonella Typhimurium ghost vaccine was constructed with the use of a recombinant fusion protein consisting of lysozyme and porcine myeloid antimicrobial peptide 36 expressed by the Escherichia coli overexpression system. After confirmation of its effectiveness by transmission electron microscopy the vaccine was evaluated in a murine model. Of the 60 BALB/c mice equally divided into 4 groups, group A mice were intramuscularly inoculated with 100 µL of sterile phosphate-buffered saline, and the mice in groups B, C, and D were intramuscularly inoculated with approximately 1.0 × 104, 1.0 × 105, or 1.0 × 106 cells of the S. Typhimurium ghost vaccine, respectively, in 100-µL amounts. The serum IgG titers against S. Typhimurium outer membrane proteins were significantly higher in groups B to D than in group A, as were the concentrations of interleukin-10 and interferon gamma in supernatants of harvested splenocytes. After challenge with wild-type S. Typhimurium, all the vaccinated groups showed significant protection compared with group A, notably perfect protection in groups C and D. Overall, these results show that intramuscular vaccination with 1.0 × 105 cells of this ghost vaccine candidate provided efficient protection against systemic infection with virulent S. Typhimurium.

Un vaccin fantôme dirigé contre Salmonella Typhimurium a été construit en utilisant une protéine de fusion recombinante composée de lysozyme et du peptide myéloïde antimicrobien 36 d'origine porcine exprimée par le système de surexpression d'Escherichia coli. Après confirmation de son efficacité par microscopie électronique à transmission, le vaccin a été évalué dans un modèle murin. Soixante souris BALB/c ont été séparées en quatre groupes. Les souris du groupe A ont été inoculées par voie intramusculaire (IM) avec 100 µL de saline tamponnée stérile, alors que les souris des groupes B, C, et D ont été inoculées IM avec approximativement 1,0 × 104, 1,0 × 105, ou 1,0 × 106 cellules du vaccin fantôme S. Typhimurium, respectivement, dans des volumes de 100 µL. Les titres d'IgG sériques contre les protéines de la membrane externe de S. Typhimurium étaient significativement plus élevés dans les groupes B à D que dans le groupe A, de même que les concentrations d'interleukine-10 et d'interféron gamma dans les surnageants de splénocytes récoltés. Suite à une infection défi avec une souche sauvage de S. Typhimurium, les animaux de tous les groupes vaccinés étaient protégés de manière significative comparativement à ceux du groupe A, notamment une protection parfaite pour les groupes C et D. De manière générale, ces résultats montrent que la vaccination IM avec 1,0 × 105 de ce vaccin fantôme candidat fourni une protection efficace contre une infection systémique par une souche virulente de S. Typhimurium.(Traduit par Docteur Serge Messier).

Yb3+,Er3+,Eu3+-codoped YVO4 material for bioimaging with dual mode excitation.

We propose an efficient bioimaging strategy using Yb3+,Er3+,Eu3+-triplet doped YVO4 nanoparticles which were synthesized with polymer as a template. The obtained particles possess nanoscale, uniform, and flexible excitation. The effect of Eu3+ ions on the luminescence properties of YVO4:Yb3+,Er3+,Eu3+ was investigated. The upconversion mechanism of the prepared material was also discussed. The structure and optical properties of the prepared material were characterized by using X-ray diffraction (XRD), Fourier-transform IR spectroscopy (FTIR), scanning electron microscopy (SEM), Transmission electron microscopy (TEM) upconversion and photoluminescence spectra. The Commission International de I'Eclairage (CIE) chromaticity coordinates was investigated to confirm the performance of color luminescent emission. The prepared YVO4:Yb3+,Er3+,Eu3+ nanoparticles could be easily dispersed in water by surface modification with cysteine (Cys) and glutathione (GSH). The aqueous dispersion of the modified YVO4:Yb3+,Er3+,Eu3+ exhibits bright upconversion and downconversion luminescence and has been applied for bioimaging of HeLa cells. Our developed material with dual excitation offers a promising advance in bioimaging.

Protection efficacy of the Brucella abortus ghost vaccine candidate lysed by the N-terminal 24-amino acid fragment (GI24) of the 36-amino acid peptide PMAP-36 (porcine myeloid antimicrobial peptide 36) in murine models.

Brucella abortus cells were lysed by the N-terminal 24-amino acid fragment (GI24) of the 36-amino acid peptide PMAP-36 (porcine myeloid antimicrobial peptide 36). Next, the protection efficacy of the lysed fragment as a vaccine candidate was evaluated. Group A mice were immunized with sterile PBS, group B mice were intraperitoneally (ip) immunized with 3 × 108 colony-forming units (CFUs) of B. abortus strain RB51, group C mice were immunized ip with 3 × 108 cells of the B. abortus vaccine candidate, and group D mice were orally immunized with 3 × 109 cells of the B. abortus vaccine candidate. Brucella lipopolysaccharide (LPS)-specific serum IgG titers were considerably higher in groups C and D than in group A. The levels of interleukin (IL)-4, IL-10, tumor necrosis factor alpha (TNF-α) and interferon gamma (IFN-γ) were significantly higher in groups B-D than in group A. After an ip challenge with B. abortus 544, only group C mice showed a significant level of protection as compared to group A. Overall, these results show that ip immunization with a vaccine candidate lysed by GI24 can effectively protect mice from systemic infection with virulent B. abortus.

Changes in antioxidant defense systems by 2,2',5,5'-tetrachlorobiphenyl exposure in neuronal SK-N-MC cells.

Polychlorinated biphenyls (PCBs) are known to alter the mammalian antioxidant defense system. To determine whether similar detoxification processes are activated in human neuronal cells, we investigated activities of antioxidant enzymes and the glutathione status (i.e., the levels of reduced and oxidized glutathione, GSH and GSSG) in human neuronal SK-N-MC cells exposed to 2,2',5,5'-tetrachlorobiphenyl (PCB 52). Upon PCB 52 treatment, time- and concentration-dependent inhibitions of cell viability were observed. PCB 52 did not affect GSH contents upon increasing the concentration up to 15 microg/ml, but significant depletions in GSH were observed at the concentrations of 20 and 25 microg/ml. PCB 52 exposure increased GSSG levels in the SK-N-MC cells, while GSH levels were decreased, and these changes naturally modified the GSSG/GSH ratios. Cytosolic glutathione S-transferase (GST) activity with 1-chloro-2,4-dinitrobenzene as substrate was enhanced by two-fold in neuronal cells after exposure to PCB 52 versus controls. In contrast, neuronal cells showed a sustained decrease in glutathione peroxidase activity with increasing concentrations of PCB 52, and a sustained decrease in Cu/Zn-superoxide dismutase (SOD) activity with increasing concentrations of PCB 52. Catalase activity was increased until 12 h after exposure to PCB 52, but was decreased 24 h after exposure. Overall, these results imply a major effect of PCB 52 on GSH status and upon the activities of antioxidant enzymes in human neuronal SK-N-MC cells, and upon the overall process of detoxification in human neuronal cells.

Protective effect of ginseng extract against apoptotic cell death induced by 2,2',5,5'-tetrachlorobiphenyl in neuronal SK-N-MC cells.

Oxidative stress plays an important role in the pathological processes of neurodegenerative diseases. Polychlorinated biphenyls (PCBs) are ubiquitous environmental contaminants, some of which may be neurotoxic. 2,2',5,5'-Tetrachlorobiphenyl (PCB 52) induces apoptotic death in human neuronal SK-N-MC cells, as demonstrated by gel electrophoresis, which demonstrates the proteolytic cleavage of beta-catenin and poly(ADP-ribose) polymerase (PARP) and the characteristic ladder patterns of DNA fragmentation. In the present study, we investigated whether Panax ginseng extract protect human neuronal SK-N-MC cells from PCB 52-induced apoptosis. The addition of 500 microg/ml of ginseng extract to a culture medium significantly protected neuronal cell from the apoptosis mediated by PCB 52 and remarkably attenuated lipid peroxidation, the generation of reactive oxygen species, and DNA fragmentation, and markedly reduced the PCB 52 induced proteolytic cleavage of beta-catenin and PARP. These results show that Panax ginseng extract protects human neuronal SK-N-MC cells from the apoptosis induced by PCB 52. We suggest that Panax ginseng extracts may protect neuronal cells from oxidative injury.

In vivo effects of Panax ginseng extracts on the cytochrome P450-dependent monooxygenase system in the liver of 2,3,7,8-tetrachlorodibenzo-p-dioxin-exposed guinea pig.

The effects of the subchronic administration of Panax ginseng extracts were examined on the hepatic cytochrome P450-dependent monooxygenase system of guinea pigs pre-exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). Panax ginseng extracts were intraperitoneally administered to guinea pigs at 100 mg/kg/day for 14 days from 1 week after a single intraperitoneal injection of 1 microg of TCDD/kg of body weight. TCDD treatment increased the total cytochrome P450 content 2.86-fold, and this was remarkably inhibited by the administration of Panax ginseng extracts. Treatment with ginseng extract alone also decreased the contents of cytochrome P450 by 33%, but both TCDD and ginseng extracts had no effect on cytochrome b(5) content. The administration of TCDD resulted in a 1.73-fold increase in microsomal NADPH-cytochrome P450 reductase activity in the guinea pig liver, and this was significantly inhibited by ginseng extracts, but treatment with ginseng extracts alone had no effect on its activity, and no statistical changes in the activity of NADPH-cytochrome b(5) reductase were observed in guinea pig liver due to TCDD and/or ginseng extract administration. Compared to the control, ECOD activity remarkably (1.76-fold) increased after TCDD administration, but this increase was completely inhibited by treatment with ginseng extract. Treatment with ginseng extract alone resulted in a 50% reduction of ECOD activity. TCDD administration remarkably induced benzphetamine demethylation (BPDM) activity, while ginseng extract also slightly increased the enzyme's activity, but the induction attributed to ginseng extracts was not statistically significant. Even though administration of ginseng extracts slightly inhibited TCDD-induced BPDM activity, the inhibition was not statistically significant. These results indicate that ginseng extract exerts different effect on the induction of P450 isozymes. From these results, we suggest that Panax ginseng extracts may act as an inhibitor of CYP1A rather than that of CYP2B.

Effects of tetramethylpyrazine on microglia activation in spinal cord compression injury of mice.

Secondary mechanisms, including inflammation and microglia activation, serve as targets for the development and application of pharmacological strategies in the management of spinal cord injury (SCI). Tetramethylpyrazine (TMP), an active ingredient of Ligusticum wallichii (chuanxiong), has shown anti-inflammatory and neuroprotective effects against SCI. However, it remains uncertain whether the inflammation-suppressive effects of TMP play a modulatory role over microglia activation in SCI. The present study investigated the effects of TMP on microglia activation and pro-inflammatory cytokines in spinal cord compression injury in mice. For a real-time PCR measurement of pro-inflammatory cytokines, SCI was induced in mice by the clip compression method (30 g force, 1 min) and TMP (15 or 30 mg/kg, i.p.) was administered once, 30 minutes before the SCI induction. For immunohistochemistry, TMP (30 mg/kg, i.p.) treatment was given three times during the first 48 hours after the SCI. 30 mg/kg of TMP treatment reduced the up-regulation of TNF-α, IL-1ß and COX-2 mRNA in the spinal tissue at four hours after the SCI induction. TMP also significantly attenuated microglia activation and neutrophil infiltration at 48 hours after the SCI induction. In addition, iNOS expression in the spinal tissue was attenuated with TMP treatment. These results suggest that TMP plays a modulatory role in microglia activation and may protect the spinal cord from or potentially delay secondary spinal cord injury.