Surface Roughness of Dentin and Formation of Early Cariogenic Biofilm after Silver Diamine Fluoride and Potassium Iodide Application / 대한소아치과학회지

This study aimed to evaluate the effect of silver diamine fluoride (SDF) and potassium iodide (KI) on the formation of cariogenic biofilm and surface roughness in vitro. A total of 48 bovine dentin specimens with artificially induced caries were prepared and divided into 3 groups of 16: untreated control, SDF-treated, and SDF-treated followed by KI (SDFKI). Ten specimens from each group were used to observe microbial adhesion. Multispecies cariogenic biofilms including Streptococcus mutans , Lactobacillus casei , and Candida albicans were cultured on the specimens. Microbes were cultured for 24 hours, and the colony-forming unit was calculated. The remaining specimens were observed by atomic force microscope and scanning electron microscope (SEM). The number of bacteria was significantly lower in the SDF and SDFKI groups. KI did not inhibit the antibacterial activity of SDF significantly. SEM images showed particles generated after SDF and SDFKI application were deposited on the dentin, but there was no significant difference in surface roughness between the 3 groups. This study confirmed that SDF and SDFKI application did not have a significant effect on the surface roughness of dentin, but effectively inhibited the formation of the early cariogenic bacterial film after 24 hours compared to the control.

Recovery of Streptococcus Mutans Biofilm after Photodynamic Therapy with Erythrosine and LED Light Source / 대한소아치과학회지

The aim of this study was to evaluate the effects of erythrosine-mediated photodynamic therapy (PDT) on Streptococcus mutans biofilm recovery by counting its colony-forming units (CFUs) and via confocal laser scanning microscopy analysis at different time points following PDT. In PDT, photosensitizer was an erythrosine. S. mutans ATCC25175 biofilms were irradiated using an LED curing light. Chlorhexidine (CHX) was used as positive control. After each antimicrobial treatment, samples were cultured to allow biofilm recovery. Viability was measured by calculating the CFU counts after treatment and after every 3 hours for up to 24 hours. Immediately after treatment, the PDT and CHX groups showed equally significant decreases in S. mutans CFU counts compared to the negative control. After 12 hours of reculture, the PDT group showed no significant difference in the decrease in CFU count compared to the negative control, whereas the CHX group showed significantly lower CFU counts throughout the 24-hour period. Erythrosine-mediated PDT can effectively inhibit S. mutans biofilm formation. However, biofilm recovery occurred earlier in the CHX group after PDT. This study provides insights into the clinical effectiveness of PDT in preventing dental caries.

Effect of Photothermal Therapy with Indocyanine Green in Multispecies Biofilm / 대한소아치과학회지

The purpose of this study is to investigate the antibacterial effects of indocyanine green (ICG) and near-infrared diode lasers on multispecies biofilms.Multispecies biofilms of Streptococcus mutans, Lactobacillus casei and Candida albicans were treated with different irradiation time using photosensitizer ICG and 808 nm near-infrared diode laser. Colony forming unit (CFU) was measured, and qualitative evaluation of biofilm was performed with confocal laser scanning microscopy (CLSM). Temperature measurement was conducted to evaluate photothermal effect.In the groups using ICG and diode laser, reduction in CFU was statistically significant, but the difference in antibacterial effect on L. casei and C. albicans with irradiation time was not significant, and similar results were confirmed with CLSM. Groups with ICG and diode laser showed higher temperature elevation than groups without ICG, and results of measured temperature were similar to the range of hyperthermia.In conclusion, ICG and near-infrared diode laser showed antibacterial effects on multispecies biofilms, but studies on protocol are necessary for clinical application.

The Progression of SARS Coronavirus 2 (SARS-CoV2): Mutation in the Receptor Binding Domain of Spike Gene

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV2) is a positive-sense singlestranded RNA (+ssRNA) that causes coronavirus disease 2019 (COVID-19). The viral genome encodes twelve genes for viral replication and infection. The third open reading frame is the spike (S) gene that encodes for the spike glycoprotein interacting with specific cell surface receptor – angiotensin converting enzyme 2 (ACE2) – on the host cell membrane. Most recent studies identified a single point mutation in S gene. A single point mutation in S gene leading to an amino acid substitution at codon 614 from an aspartic acid 614 into glycine (D614G) resulted in greater infectivity compared to the wild type SARS-CoV2. We were interested in investigating the mutation region of S gene of SARS-CoV2 from Korean COVID-19 patients. New mutation sites were found in the critical receptor binding domain (RBD) of S gene, which is adjacent to the aforementioned D614G mutation residue. This specific sequence data demonstrated the active progression of SARS-CoV2 by mutations in the RBD of S gene.The sequence information of new mutations is critical to the development of recombinant SARS-CoV2 spike antigens, which may be required to improve and advance the strategy against a wide range of possible SARS-CoV2 mutations.

The Progression of SARS Coronavirus 2 (SARS-CoV2): Mutation in the Receptor Binding Domain of Spike Gene

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV2) is a positive-sense singlestranded RNA (+ssRNA) that causes coronavirus disease 2019 (COVID-19). The viral genome encodes twelve genes for viral replication and infection. The third open reading frame is the spike (S) gene that encodes for the spike glycoprotein interacting with specific cell surface receptor – angiotensin converting enzyme 2 (ACE2) – on the host cell membrane. Most recent studies identified a single point mutation in S gene. A single point mutation in S gene leading to an amino acid substitution at codon 614 from an aspartic acid 614 into glycine (D614G) resulted in greater infectivity compared to the wild type SARS-CoV2. We were interested in investigating the mutation region of S gene of SARS-CoV2 from Korean COVID-19 patients. New mutation sites were found in the critical receptor binding domain (RBD) of S gene, which is adjacent to the aforementioned D614G mutation residue. This specific sequence data demonstrated the active progression of SARS-CoV2 by mutations in the RBD of S gene.The sequence information of new mutations is critical to the development of recombinant SARS-CoV2 spike antigens, which may be required to improve and advance the strategy against a wide range of possible SARS-CoV2 mutations.

Streptococcus Mutans Biofilm Inhibition Effect of Indocyanine Green and Near Infrared Diode Laser / 대한소아치과학회지

The purpose of this study was to evaluate the effect of Indocyanine Green (ICG) and near-infrared (NIR) diode laser on Streptococcus mutans biofilms depending on ICG concentrations.S. mutans biofilms were formed on a Hydroxyapatite disk, and 0.5, 1.0, 2.0, 3.0, 4.0, 5.0 mg/mL ICG solutions dissolved in sterile distilled water and a NIR diode laser having a power of 300 mW and a wavelength of 808 nm were applied to the biofilms. The temperature changes of the biofilm surface according to the concentrations of the ICG solution were measured using a 1-channel thermocouple thermometer.Compared to the control group, in the groups with only the 3.0, 4.0, 5.0 mg/mL ICG solution application, and in the groups with the 1.0, 2.0, 3.0, 4.0, 5.0 mg/mL ICG solution application and light irradiation, a statistically significant decrease in the bacterial counts were observed.The temperature increase according to the concentration of the ICG solutions was 9.53℃, 10.43℃, 11.40℃, 12.10℃, 12.67℃, and 13.63℃ in ICG solutions of 0.5, 1.0, 2.0, 3.0, 4.0, and 5.0 mg/mL respectively.This study presents the potential for clinical application of ICG and NIR diode lasers as a new method for preventing dental caries.

Antimicrobial Effect of Photodynamic Therapy Using Plaque Disclosing Agent / 대한소아치과학회지

The aim of this study was to evaluate the antimicrobial effect of photodynamic therapy (PDT) using plaque disclosing agent, 10 - 20 mM erythrosine, as a photosensitizer.Multispecies cariogenic biofilms containing Streptococcus mutans , Lactobacillus casei and Candida albicans were formed on hydroxyapatite disc. 20 μM, 10 mM and 20 mM erythrosine were applied as a photosensitizer for 3 minutes, and then light-emitting diode (LED) irradiated for 24 seconds. Colony-forming unit (CFU) were measured and biofilms were observed using confocal laser scanning microscopy (CLSM). CFU were significantly decreased in the PDT groups using 10 - 20 mM erythrosine (10 mM, 20mM) and the results were also confirmed by CLSM.This study confirms the high antimicrobial effect of photodynamic therapy using plaque disclosing agent as a photosensitizer.

Surface Roughness and Microbial Adhesion After Finishing of Alkasite Restorative Material / 대한소아치과학회지

This study is aimed to evaluate and compare the surface roughness and microbial adhesion to alkasite restorative material (Cention N), resin-modified glass ionomer (RMGI), and composite resin. And to examine the correlation between bacterial adhesion and surface roughness by different finishing systems.Specimens were fabricated in disk shapes and divided into four groups by finishing methods (control, carbide bur, fine grit diamond bur, and white stone bur). Surface roughness was tested by atomic force microscope and surface observation was performed by scanning electron microscope. Colony forming units were measured after incubating Streptococcus mutans biofilm on specimens using CDC biofilm reactor.Cention N surface roughness was less than 0.2 μm after finishing procedure. Control specimens of resin and Cention N specimens were significantly (p = 0.01) rougher. Pearson correlation coefficient (PCC = 0.13) indicated a weak correlation between surface roughness and S. mutans adhesion to the specimens.Compared with resin specimens, RMGI and Cention N showed lower microbial adhesion. Surface roughness and bacterial adhesion were not significantly different, regardless of the finishing systems.

Application of Teeth Whitening LED for Prevention of Dental Caries : Antimicrobial Photodynamic Therapy Approach / 대한소아치과학회지

The present study is aimed to assess the effect of antimicrobial photodynamic therapy (aPDT) on Streptococcus mutans biofilm through teeth whitening light emitting diode (LED).Planktonic and dynamic biofilm state cultures of S. mutans were used. Erythrosine 20 μM/L was used as the photosensitizer. Irradiation was performed by exposing cultures to clinic and homecare whitening LEDs for 15 minutes. The viability was measured through Colony Forming Unit counts and confocal laser scanning microscopy.aPDT using whitening LEDs and erythrosine significantly decreased the CFU count of S. mutans compared to that in the control group. Dynamic biofilm group showed more resistant features to aPDT compared with planktonic state. Clinic and homecare whitening LED device showed similar antimicrobial effect.The whitening LED, which could irradiate the entire oral arch, showed a significant photodynamic effect on cariogenic S. mutans biofilm. aPDT mediated by erythrosine and LEDs used for teeth whitening exhibited promising antimicrobial activity.

Susceptibility of Mutans streptococci in the Planktonic and Biofilm State to Erythrosine / 대한소아치과학회지

The aim of this study was to investigate the susceptibility of Mutans streptococci in both planktonic and biofilm states to erythrosine.S. mutans was cultured in brain-heart infusion (BHI) broth. Erythrosine was diluted in BHI broth and prepared at a concentration range of 0.02 – 10000 µg/L. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were measured using the microdilution method. After forming biofilms on 96-well plates, the minimum biofilm inhibitory concentration (MBIC) and minimum biofilm eradication concentration (MBEC) were measured.S. mutans was susceptible to erythrosine in both planktonic and biofilm states. MIC and MBC values were both 19.5 µg/L for the planktonic state, while MBIC and MBEC values were 313 µg/L and 2500 µg/L, respectively, for the biofilm state.Erythrosine (19.5 µg/L) exhibited a bactericidal effect on S. mutans (killing 99.9%) in the planktonic state. For biofilms, erythrosine inhibited biofilm growth and eradicated 99.9% of biofilm bacteria at higher concentrations than MIC and MBC. These MBIC and MBEC concentrations are much lower than known noxious doses, and the MIC, MBC, and MBIC values were even lower than clinical concentrations.

Structural Characteristics of Seven IL-32 Variants

IL-32 exists as seven mRNA transcripts that can translate into distinct individual IL-32 variants with specific protein domains. These translated protein domains of IL-32 variants code for specific functions that allow for interaction with different molecules intracellularly or extracellularly. The longest variant is IL-32γ possessing 234 amino acid residues with all 11 protein domains, while the shortest variant is IL-32α possessing 131 amino acid residues with three of the protein domains. The first domain exists in 6 variants except IL-32δ variant, which has a distinct translation initiation codon due to mRNA splicing. The last eleventh domain is common domain for all seven IL-32 variants. Numerous studies in different fields, such as inflammation, autoimmunity, pathogen infection, and cancer biology, have claimed the specific biological activity of individual IL-32 variant despite the absence of sufficient data. There are 4 additional IL-32 variants without proper transcripts. In this review, the structural characteristics of seven IL-32 transcripts are described based on the specific protein domains.

L1 Recombinant Proteins of HPV Tested for Antibody Forming Using Sera of HPV Quadrivalent Vaccine

Virus-like particles (VLPs) derived from human papillomavirus (HPV) L1 capsid proteins were used for HPV quadrivalent recombinant vaccine. The HPV quadrivalent vaccine is administrated in a 3-dose regimen of initial injection followed by subsequent doses at 2 and 6 months to prevent cervical cancer, vulvar, and vaginal cancers. The type 6, 11, 16, or 18 of HPV infection is associated with precancerous lesions and genital warts in adolescents and young women. The HPV vaccine is composed of viral L1 capsid proteins are produced in eukaryotic expression systems and purified in the form of VLPs. Four different the L1 protein of 3 different subtypes of HPV: HPV11, HPV16, and HPV18 were expressed in Escherichia coli divided into 2 fragments as N- and C-terminal of each protein in order to examine the efficacy of HPV vaccine. Vaccinated sera failed to recognize N-terminal L1 HPV type 16 and type 18 by western blot while they detected N-terminal L1 protein of HPV type 11. Moreover, the recombinant C-terminal L1 proteins of type 16 was non-specifically recognized by the secondary antibody conjugated with horseradish peroxidase. This expression and purification system may provide simple method to obtain robust recombinant L1 protein of HPV subtypes to improve biochemical analysis of antigens with immunized sera.

Study of Bacteria Associated with Dental Caries Using a 3 Tone Disclosing Agent / 대한소아치과학회지

The aim of this study was to determine the efficacy of a 3 tone plaque disclosing gel in assessing the risk of caries related to the population of Streptococcus mutans, Streptococcus sobrinus, and Lactobacillus spp. quantified using a quantitative real-time polymerase chain reaction (qRT-PCR).15 healthy children of ages 9 – 12 years were randomly examined. The 3 tone plaque disclosing gel was applied on teeth surfaces, which changed the color to pink or red, blue or purple and light blue. Plaque was divided into 3 groups based on staining. Genomic DNA from each sample was subjected to a qRT-PCR assay for quantitative detection of target bacteria. The Kruskal-Wallis test was conducted for correlation between the color of plaque and the number of bacterial species.The levels of S. mutans, S. sobrinus, and Lactobacillus spp. were significantly different in the plaque samples of the 3 groups (p < 0.05). The proportion of S. sobrinus to S. mutans showed correlation to the color of plaque.The different color-dyed plaque was related to the number of acidogenic bacteria. The 3 tone plaque disclosing gel could be used as one of the indicators to assess the clinical risk of caries associated with the population of S. mutans, S. sobrinus, and Lactobacillus spp.

Antimicrobial Effect on Streptococcus mutans in Photodynamic Therapy using Different Light Source / 대한소아치과학회지

In a photodynamic therapy, the difference of antibacterial capacity was compared according to the type of source of light when the same quantity of energy is irradiated.After S. mutans is formed in planktonic state and biofilm state, erythrosine diluted to 40 µM was treated for 3 minutes, and as the type of light source, Halogen, LED, and Plasma arc were used, which were irradiated for 30 seconds, 15 seconds and 9.5 seconds, respectively.After the completion of the experiment, CFU of each experiment arm was measured to compare the photodynamic therapeutic effects according to each condition.The CFU of each experiment arm had no statistically significant difference.Under the same quantity of energy, the photodynamic therapeutic effect can be said to be the same regardless of types of light source, which is a useful result in the clinical field with various light irradiators.

Erratum: Identification of Epithelial-Mesenchymal Transition-related Target Genes Induced by the Mutation of Smad3 Linker Phosphorylation

The original version of this article contained error in the URL of the SUPPLEMENTARY MATERIALS.

Identification of Epithelial-Mesenchymal Transition-related Target Genes Induced by the Mutation of Smad3 Linker Phosphorylation

BACKGROUND: Smad3 linker phosphorylation plays essential roles in tumor progression and metastasis. We have previously reported that the mutation of Smad3 linker phosphorylation sites (Smad3-Erk/Pro-directed kinase site mutant constructs [EPSM]) markedly reduced the tumor progression while increasing the lung metastasis in breast cancer. METHODS: We performed high-throughput RNA-Sequencing of the human prostate cancer cell lines infected with adenoviral Smad3-EPSM to identify the genes regulated by Smad3-EPSM. RESULTS: In this study, we identified genes which are differentially regulated in the presence of Smad3-EPSM. We first confirmed that Smad3-EPSM strongly enhanced a capability of cell motility and invasiveness as well as the expression of epithelial-mesenchymal transition marker genes, CDH2, SNAI1, and ZEB1 in response to TGF-β1 in human pancreatic and prostate cancer cell lines. We identified GADD45B, CTGF, and JUNB genes in the expression profiles associated with cell motility and invasiveness induced by the Smad3-EPSM. CONCLUSIONS: These results suggested that inhibition of Smad3 linker phosphorylation may enhance cell motility and invasiveness by inducing expression of GADD45B, CTGF, and JUNB genes in various cancers.

IL-32-induced Inflammatory Cytokines Are Selectively Suppressed by α1-antitrypsin in Mouse Bone Marrow Cells

The induction of interleukin (IL)-32 in bone marrow (BM) inflammation is crucial in graft versus host disease (GvHD) that is a common side effect of allogeneic BM transplantation. Clinical trials on α-1 antitrypsin (AAT) in patients with GvHD are based on the preliminary human and mouse studies on AAT reducing the severity of GvHD. Proteinase 3 (PR3) is an IL-32-binding protein that was isolated from human urine. IL-32 primarily induces inflammatory cytokines in myeloid cells, probably due to PR3 expression on the membrane of the myeloid lineage cells. The inhibitory activity of AAT on serine proteinases may explain the anti-inflammatory effect of AAT on GvHD. However, the anti-inflammatory activity of AAT on BM cells remains unclear. Mouse BM cells were treated with IL-32γ and different inflammatory stimuli to investigate the anti-inflammatory activity of AAT. Recombinant AAT-Fc fusion protein inhibited IL-32γ-induced IL-6 expression in BM cells, but failed to suppress that induced by other stimuli. In addition, the binding of IL-32γ to PR3 was abrogated by AAT-Fc. The data suggest that the specific anti-inflammatory effect of AAT in mouse BM cells is due to the blocking of IL-32 binding to membrane PR3.

Species Specific Antiviral Activity of Porcine Interferon-α8 (IFNα8)

Interferons (IFNs) have been known as antiviral genes and they are classified by type 1, type 2, and type 3 IFN. The type 1 IFN consists of IFNα, IFNβ, IFNτ, and IFNω whereas the type 2 IFN consists of only IFNγ, which is a key cytokine driving T helper cell type 1 immunity. IFNλ belongs to the type 3 IFN, which is also known as IL-28 and IL-29 possessing antiviral activities. Type 1 IFN is produced by viral infection whereas type 2 IFN is induced by mitogenic or antigenic T-cell stimuli. The IFNτ of bovine was first discovered in an ungulate ruminant recognition hormone. IFNτ belongs to the type 1 IFN with the common feature of type 1 IFN such as antiviral activity. IFNs have been mostly studied for basic research and clinical usages therefore there was no effort to investigate IFNs in industrial animals. Here we cloned porcine IFNα8 from peripheral blood mononuclear cells of Korean domestic pig (Sus scrofa domestica). The newly cloned IFNα8 amino acid sequence from Korean domestic pig shares 98.4% identity with the known porcine IFNα8 in databank. The recombinant porcine IFNα8 showed potent antiviral activity and protected bovine Madin-Darby bovine kidney epithelial (MDBK) cells from the cytopathic effect of vesicular stomatitis virus, but it failed to protect human Wistar Institute Susan Hayflick (WISH) cells and canine Madin-Darby canine kidney epithelial-like (MDCK) cells. The present study demonstrates species specific antiviral activity of porcine IFNα8.

Dendritic Cell Dysfunction in Patients with End-stage Renal Disease

End-stage renal disease (ESRD) with immune disorder involves complex interactions between the innate and adaptive immune responses. ESRD is associated with various alterations in immune function such as a reduction in polymorphonuclear leukocyte bactericidal activity, a suppression of lymphocyte proliferative response to stimuli, and a malfunction of cell-mediated immunity at the molecular level. ESRD also increases patients' propensity for infections and malignancies as well as causing a diminished response to vaccination. Several factors influence the immunodeficiency in patients with ESRD, including uremic toxins, malnutrition, chronic inflammation, and the therapeutic dialysis modality. The alteration of T-cell function in ESRD has been considered to be a major factor underlying the impaired adaptive cellular immunity in these patients. However, cumulative evidence has suggested that the immune defect in ESRD can be caused by an Ag-presenting dendritic cell (DC) dysfunction in addition to a T-cell defect. It has been reported that ESRD has a deleterious effect on DCs both in terms of their number and function, although the precise mechanism by which DC function becomes altered in these patients is unclear. In this review, we discuss the effects of ESRD on the number and function of DCs and propose a possible molecular mechanism for DC dysfunction. We also address therapeutic approaches to improve immune function by optimally activating DCs in patients with ESRD.

Cytokine-like Activity of Liver Type Fatty Acid Binding Protein (L-FABP) Inducing Inflammatory Cytokine Interleukin-6

It has been reported that fatty acid binding proteins (FABPs) do not act only as intracellular mediators of lipid responses but also have extracellular functions. This study aimed to investigate whether extracellular liver type (L)-FABP has a biological activity and to determined serum L-FABP levels in patients with end-stage renal disease (ESRD). We isolated L-FABP complementary deoxyribonucleic acid (cDNA) from the Huh7 human hepatocarcinoma cell line and expressed the recombinant L-FABP protein in Escherichia coli. A549 lung carcinoma and THP-1 monocytic cells were stimulated with the human recombinant L-FABP. Human whole blood cells were also treated with the human recombinant L-FABP or interleukin (IL)-1α. IL-6 levels were measured in cell culture supernatants using IL-6 enzyme-linked immunosorbent assay (ELISA). Human recombinant L-FABP induced IL-6 in a dose-dependent manner in A549, THP-1 cells, and whole blood cells. The blood samples of healthy volunteers and patients with ESRD were taken after an overnight fast. The serum levels of L-FABP in healthy volunteers and ESRD patients were quantified with L-FABP ELISA. The values of L-FABP in patients with ESRD were significantly lower than those in the control group. Our results demonstrated the biological activity of L-FABP in human cells suggesting L-FABP can be a mediator of inflammation.