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Immune checkpoint therapies, such as programmed cell death ligand 1 (PD-L1) blockade, have shown remarkable clinical benefit in many cancers by restoring the function of exhausted T cells. Hence, the identification of novel PD-L1 regulators and the development of their inhibition strategies have significant therapeutic advantages. Here, we conducted pooled shRNA screening to identify regulators of membrane PD-L1 levels in lung cancer cells targeting druggable genes and cancer drivers. We identified WNK lysine deficient protein kinase 3 (WNK3) as a novel positive regulator of PD-L1 expression. The kinase-dead WNK3 mutant failed to elevate PD-L1 levels, indicating the involvement of its kinase domain in this function. WNK3 perturbation increased cancer cell death in cancer cell–immune cell coculture conditions and boosted the secretion of cytokines and cytolytic enzymes, promoting antitumor activities in CD4+ and CD8+ T cells. WNK463, a pan-WNK inhibitor, enhanced CD8+ T-cell-mediated antitumor activity and suppressed tumor growth as a monotherapy as well as in combination with a low-dose anti-PD-1 antibody in the MC38 syngeneic mouse model. Furthermore, we demonstrated that the c-JUN N-terminal kinase (JNK)/c-JUN pathway underlies WNK3-mediated transcriptional regulation of PD-L1. Our findings highlight that WNK3 inhibition might serve as a potential therapeutic strategy for cancer immunotherapy through its concurrent impact on cancer cells and immune cells.

Laboratory mice have long been an invaluable tool in biomedical science and have made significant contributions in research into life-threatening diseases. However, the translation of research results from mice to humans often proves difficult due to the incomplete nature of laboratory animal-based research. Hence, there is increasing demand for complementary methods or alternatives to laboratory mice that can better mimic human physiological traits and potentially bridge the translational research gap. Under these circumstances, the natural/naturalized mice including “wild”, “dirty”, “wildling”, and “wilded” systems have been found to better reflect some aspects of human pathophysiology. Here, we discuss the pros and cons of the laboratory mouse system and contemplate how wild mice and wild microbiota are able to help in refining such systems to better mimic the real-world situation and contribute to more productive translational research.

Background: Evidence suggests that the human respiratory tract, as with the gastrointestinal tract, has evolved to its current state in association with commensal microbes. However, little is known as how the airway microbiome affected airway immune development. Here, we uncover a previously unidentified mode of interaction between host airway immunity and a unique strain of Staphylococcus epidermidis, a predominant species of the nasal microbiome. Results: Intranasal inoculation of S. epidermidis AIT01 accompanied the active recruitment of neutrophils and monocytes into mouse lungs on the murine model. The recruitment of the immune cells resulted in the complete protection of the murine host against Pseudomonas aeruginosa respiratory infection. Interestingly, an AIT01-secreted protein identified as glyceraldehyde-3-phosphate dehydrogenase (GAPDH), a well-known bacterial moonlighting protein, mediated this protective effect. Intranasal delivery of the purified protein conferred significant resistance against other additional airway infections of Gram-negative pathogens or influenza A virus. Moreover, single cell RNA-seq analysis using mouse lung cells revealed that transcription of genes involved in cytokine production, phagocytosis, chemotaxis, and immune cell migration was considerably upregulated in neutrophils and monocytes in response to the treatment with the protein. Conclusions: Our results illustrate how an indigenous nasal microbe and its secretory protein can extensively instigate an innate immune defense. We also suggest a promising prophylactic option for airway infections in the era of global pandemics. Keywords: Staphylococcus epidermidis, human nasal microbiome, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), innate immune defense

The human gastrointestinal tract has an enormous and diverse microbial community, termed microbiota, that is necessary for the development of the immune system and tissue homeostasis. In contrast, microbial dysbiosis is associated with various inflammatory and autoimmune diseases as well as neurological disorders in humans by affecting not only the immune system in the gastrointestinal tract but also other distal organs. FOXP3

Exposure to heightened inflammation in pregnancy caused by infections or other inflammatory insults has been associated with the onset of neurodevelopmental and psychiatric disorders in children. Rodent models have provided unique insights into how this maternal immune activation (MIA) disrupts brain development. Here, we discuss the key immune factors involved, highlight recent advances in determining the molecular and cellular pathways of MIA, and review how the maternal immune system affects fetal development. We also examine the roles of microbiomes in shaping maternal immune function and the development of autism-like phenotypes. A comprehensive understanding of the gut bacteria-immune-neuro interaction in MIA is essential for developing diagnostic and therapeutic measures for high-risk pregnant women and identifying targets for treating inflammation-induced neurodevelopmental disorders.

Children with autism spectrum disorders often display dysregulated immune responses and related gastrointestinal symptoms. However, the underlying mechanisms leading to the development of both phenotypes have not been elucidated. Here, we showed that mouse offspring exhibiting autism-like phenotypes due to prenatal exposure to maternal inflammation were more susceptible to developing intestinal inflammation following challenges later in life. In contrast to its prenatal role in neurodevelopmental phenotypes, interleukin-17A (IL-17A) generated immune-primed phenotypes in offspring through changes in the maternal gut microbiota that led to postnatal alterations in the chromatin landscape of naïve CD4(+) T cells. The transfer of stool samples from pregnant mice with enhanced IL-17A responses into germ-free dams produced immune-primed phenotypes in offspring. Our study provides mechanistic insights into why children exposed to heightened inflammation in the womb might have an increased risk of developing inflammatory diseases in addition to neurodevelopmental disorders.

Antibodies have been widely used to provide targeting ability and to enhance bioactivity owing to their high specificity, availability, and diversity. Recent advances in biotechnology and nanotechnology permit site-specific engineering of antibodies and their conjugation to the surfaces of nanoparticles (NPs) in various orientations through chemical conjugations and physical adhesions. This study proposes the conjugation of poly(lactic-co-glycolic acid) (PLGA) NPs with antibodies by using two distinct methods, followed by a comparison between the cell-targeting efficiencies of both techniques. Full-length antibodies were conjugated to the PLGA-poly(ethylene glycol)-carboxylic acid (PLGA-PEG-COOH) NPs through the conventional carbodiimide coupling reaction, and f(ab′)2 antibody fragments were conjugated to the PLGA-poly(ethylene glycol)-maleimide(PLGA-PEG-Mal) NPs through interactions between the f(ab′)2 fragment thiol groups and the maleimide located on the nanoparticle surface. The results demonstrate that the PLGA nanoparticles conjugated with the f(ab′)2 antibody fragments had a higher targeting efficiency in vitro and in vivo than that of the PLGA nanoparticles conjugated with the full-length antibodies. The results of this study can be built upon to design a delivery technique for drugs through biocompatible nanoparticles.

Cancer immunotherapy is a next-generation treatment strategy; however, its side effects limit its clinical translation. Here, a novel combination of a multi-functional nano-adjuvant (M-NA) prepared with an iron oxide/gold core and a cationic polymer shell via multilayer synthesis with CpG oligodeoxynucleotide (CpG-ODN) electrostatically complexed on its surface, and irreversible electroporation (IRE) technique was developed for effective image-guided in situ cancer vaccination. The M-NA can be retained long-term in the dense tumoral extracellular matrix after intratumoral injection and internalized by antigen-presenting cells (APCs). The IRE can induce immunogenic cell death. Indeed, in a mouse tumor model, the M-NA showed longer tumor retention time than free CpG-ODN. Compared with other treatments, the combined treatment significantly inhibited tumor growth with 100% survival rate for ∼60 days. The therapy induced the activation of cytotoxic lymphocytes and the maturation of APCs in vivo. This treatment could be effective in image-guided local cancer immunotherapy.

In article number 2001940, June Hong Ahn, Wonhwa Lee, Chun Gwon Park, and co‐workers demonstrate that NETosis factor can be a potential therapeutic target for the treatment of SARS‐CoV‐2‐induced sepsis. DNase‐I coated melanin‐like nanospheres (DNase‐I pMNSs) show excellent stability, modulation of sepsis‐associated NETosis dysregulation and prevention of further disease progression. [Image: see text]

The current outbreak of the beta‐coronavirus (beta‐Cov) severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) began in December 2019. No specific antiviral treatments or vaccines are currently available. A recent study has reported that coronavirus disease 2019 (COVID‐19), the disease caused by SARS‐CoV‐2 infection, is associated with neutrophil‐specific plasma membrane rupture, and release excessive neutrophil extracellular traps (NETs) and extracellular DNAs (eDNAs). This mechanism involves the activation of NETosis, a neutrophil‐specific programmed cell death, which is believed to play a crucial role in COVID‐19 pathogenesis. Further progression of the disease can cause uncontrolled inflammation, leading to the initiation of cytokine storms, acute respiratory distress syndrome (ARDS), and sepsis. Herein, it is reported that DNase‐I‐coated melanin‐like nanospheres (DNase‐I pMNSs) mitigate sepsis‐associated NETosis dysregulation, thereby preventing further progression of the disease. Recombinant DNase‐I and poly(ethylene glycol) (PEG) are used as coatings to promote the lengthy circulation and dissolution of NET structure. The data indicate that the application of bioinspired DNase‐I pMNSs reduce neutrophil counts and NETosis‐related factors in the plasma of SARS‐CoV‐2 sepsis patients, alleviates systemic inflammation, and attenuates mortality in a septic mouse model. Altogether, the findings suggest that these nanoparticles have potential applications in the treatment of SARS‐CoV‐2‐related illnesses and other beta‐CoV‐related diseases., It is reported here that DNase‐I‐coated melanin‐like nanospheres (DNase‐I pMNSs) are able to modulate COVID‐19‐associated NETosis dysregulation and prevent further progression of the disease. The DNase‐I pMNSs have potential applications in the treatment of SARS‐CoV‐2‐related illnesses and other beta‐CoV‐related diseases.

3′-Sialyllactose (3′-SL), a natural prebiotic, maintains immune homeostasis and exerts anti-inflammatory and anti-arthritic effects. Although regulatory T cells (Tregs) prevent excessive inflammation and maintain immune tolerance, the effect of 3′-SL on Treg regulation is unclear. This study aimed to investigate the effect of 3′-SL on Treg responses in atopic dermatitis (AD) pathogenesis. Oral administration of 3′-SL reduced AD-like symptoms such as ear, epidermal, and dermal thickness in repeated topical application of house dust mites (HDM) and 2,4-dinitrochlorobenzene (DNCB). 3′-SL inhibited IgE, IL-1β, IL-6, and TNF-α secretion and markedly downregulated AD-related cytokines including IL-4, IL-5, IL-6, IL-13, IL-17, IFN-γ, TNF-α, and Tslp through regulation of NF-κB in ear tissue. Additionally, in vitro assessment of Treg differentiation revealed that 3′-SL directly induced TGF-β-mediated Treg differentiation. Furthermore, 3′-SL administration also ameliorated sensitization and elicitation of AD pathogenesis by suppressing mast cell infiltration and production of IgE and pro-inflammatory cytokines in mouse serum by mediating the Treg response. Furthermore, Bifidobacterium population was also increased by 3′-SL administration as prebiotics. Our data collectively show that 3′-SL has therapeutic effects against AD progression by inducing Treg differentiation, downregulating AD-related cytokines, and increasing the Bifidobacterium population.

Background Cinnamomum cassia bark is the outer skin of an evergreen tall tree belonging to the family Lauraceae containing several active components such as essential oils (cinnamic aldehyde and cinnamyl aldehyde), tannin, mucus and carbohydrate. They have various biological functions including anti-oxidant, anti-microbial, anti-inflammation, anti-diabetic and anti-tumor activity. Previously, we have reported that anti-cancer effect of cinnamon extracts is associated with modulation of angiogenesis and effector function of CD8+ T cells. In this study, we further identified that anti-tumor effect of cinnamon extracts is also link with enhanced pro-apoptotic activity by inhibiting the activities NFκB and AP1 in mouse melanoma model. Methods Water soluble cinnamon extract was obtained and quality of cinnamon extract was evaluated by HPLC (High Performance Liquid Chromatography) analysis. In this study, we tested anti-tumor activity and elucidated action mechanism of cinnamon extract using various types of tumor cell lines including lymphoma, melanoma, cervix cancer and colorectal cancer in vitro and in vivo mouse melanoma model. Results Cinnamon extract strongly inhibited tumor cell proliferation in vitro and induced active cell death of tumor cells by up-regulating pro-apoptotic molecules while inhibiting NFκB and AP1 activity and their target genes such as Bcl-2, BcL-xL and survivin. Oral administration of cinnamon extract in melanoma transplantation model significantly inhibited tumor growth with the same mechanism of action observed in vitro. Conclusion Our study suggests that anti-tumor effect of cinnamon extracts is directly linked with enhanced pro-apoptotic activity and inhibition of NFκB and AP1 activities and their target genes in vitro and in vivo mouse melanoma model. Hence, further elucidation of active components of cinnamon extract could lead to development of potent anti-tumor agent or complementary and alternative medicine for the treatment of diverse cancers.

Atopic dermatitis (AD) is a complex inflammatory skin disease mediated by immune cells of both adaptive and innate types. Among them, CD4(+) Th cells are one of major players of AD pathogenesis. Although the pathogenic role of Th2 cells has been well characterized, Th17/Th22 cells are also implicated in the pathogenesis of AD. However, the molecular mechanisms underlying pathogenic immune responses in AD remain unclear. We sought to investigate how the defect in the AD susceptibility gene, Ets1, is involved in AD pathogenesis in human and mice and its clinical relevance in disease severity by identifying Ets1 target genes and binding partners. Consistent with the decrease in ETS1 levels in severe AD patients and the experimental AD-like skin inflammation model, T cell–specific Ets1-deficient mice (Ets1(ΔdLck)) developed severe AD-like symptoms with increased pathogenic Th cell responses. A T cell–intrinsic increase of gp130 expression upon Ets1 deficiency promotes the gp130-mediated IL-6 signaling pathway, thereby leading to the development of severe AD-like symptoms. Functional blocking of gp130 by selective inhibitor SC144 ameliorated the disease pathogenesis by reducing pathogenic Th cell responses. Our results reveal a protective role of Ets1 in restricting pathogenic Th cell responses and suggest a potential therapeutic target for AD treatment.

It is important to identify the reasons for implant failure when planning future dental treatment. No studies have distinguished the factors that affect implant failure by evaluating the appearance of failed fixtures.This study investigated these factors by evaluating the surface of implant fixtures using quantitative light-induced fluorescence-digital (QLF-D), which involves detecting the red fluorescence emitted by porphyrin metabolized by oral bacteria in a mature biofilm.The areas of red fluorescence in QLF-D images obtained from all aspects of the fixture surface were then analyzed using quantitative analysis software.Red fluorescence was evident on the surface of implants that failed after at least 2 years of occlusal loading and exhibited severe bone loss. Implants with no fluorescence exhibited a clean surface in blue-light images, and the bone loss could not be observed radiographically.The cases presented that failed dental implant surfaces caused by periimplantitis can be detected by the red fluorescence evident as QLF-D.

Introduction The ability to accurately detect tooth cracks and quantify their depth would allow the prediction of crack progression and treatment success. The aim of this in vitro study was to determine the capabilities of quantitative light-induced fluorescence (QLF) technology in the detection of enamel cracks. Methods Ninety-six extracted human teeth were selected for examining naturally existing or suspected cracked teeth surfaces using a photocuring unit. QLF performed with a digital camera (QLF-D) images were used to assess the ability to detect enamel cracks based on the maximum fluorescence loss value (ΔF max , %), which was then analyzed using the QLF-D software. A histologic evaluation was then performed in which the samples were sectioned and observed with the aid of a polarized light microscope. The relationship between ΔF max and the histology findings was assessed based on the Spearman rank correlation. The sensitivity and specificity were calculated to evaluate the validity of using QLF-D to analyze enamel inner-half cracks and cracks extending to the dentin-enamel junction. Results There was a strong correlation between the results of histologic evaluations of enamel cracks and the ΔF max value, with a correlation coefficient of 0.84. The diagnostic accuracy of QLF-D had a sensitivity of 0.87 and a specificity of 0.98 for enamel inner-half cracks and a sensitivity of 0.90 and a specificity of 1.0 for cracks extending to the dentin-enamel junction. Conclusions These results indicate that QLF technology would be a useful clinical tool for diagnosing enamel cracks, especially given that this is a nondestructive method.

Regulatory T (Treg) cells that express the transcription factor FoxP3 play a key role in self-tolerance and the control of inflammation. In mice and humans, there is a wide interindividual range in Treg frequency, but little is known about the underlying genetic or epigenetic mechanisms. We explored this issue in inbred strains of mice, with a special focus on the low proportion of Treg cells found in NZW mice. Mixed bone marrow chimera experiments showed this paucity to be intrinsic to NZW Treg cells, a dearth that could be tied to poor stability of the Treg pool and of FoxP3 expression. This instability was not a consequence of differential epigenetic marks, because Treg-specific CpG hypomethylation profiles at the Foxp3 locus were similar in all strains tested. It was also unrelated to the high expression of IFN signature genes in NZW, as shown by intercross to mice with an Ifnar1 knockout. NZW Tregs were less sensitive to limiting doses of trophic cytokines, IL-2 and -33, for population homeostasis and for maintenance of FoxP3 expression. Gene-expression profiles highlighted specific differences in the transcriptome of NZW Tregs compared with those of other strains, but no single defect could obviously account for the instability. Rather, NZW Tregs showed a general up-regulation of transcripts normally repressed in Treg cells, and we speculate that this network-level bias may account for NZW Treg instability.

Background The aim of this study was to evaluate validity and reliability of autofluorescence-based plaque quantification (APQ) method. Methods The facial surfaces of 600 sound anterior teeth of 50 subjects were examined. The subjects received dental plaque examination using Turesky modified Quigley Hein plaque index (QHI) and Silness & Loe plaque index (SLI). The autofluorescence images were taken before the plaque examination with Quantitative Light-induced Fluorescence-Digital, and plaque percent index (PPI) was calculated. Correlation between two existing plaque indices and the PPI of the APQ method was evaluated to find which level of plaque redness on tooth (Δ R ) by the APQ method shows the highest correlation. The area under the ROC curve (AUC) analysis and intra- and inter-examiner reliability tests were performed. Results The PPI Δ R 20 of the APQ method showed a moderate correlation with two existing plaque indices (rho of QHI = 0.48, SLI = 0.51). This methodology fell in the fair category and it had an excellent reliability. The APQ method also showed possibility to detect heavy plaque with fair validity. Conclusions The APQ method demonstrated excellent reliability, and fair validity, compared with 2 conventional indices. The plaque quantification described has the potential to be used in clinical evaluation of oral hygiene procedures.

Breast cancer is highly aggressive and is the leading cause of cancer-related mortality in women in developed countries. The ETS proto-oncogene 1 (Ets1) has versatile roles during the cellular processes of cancer development. It is often highly expressed in breast cancers and mediates migration and invasion of human breast cancer cells. However, underlying mechanisms of Ets1 gene expression is still ambiguous. Here, we identified a core-regulatory element (CRE) located in the Ets1 promoter region (−540/−80 bp from TSS) that contains elements responsible for associating with NFATs and NF-κBs. Compared with the less metastatic breast cancer cells, metastatic breast cancer cells (MDA-MB-231) show open chromatin configurations in the CRE, which facilitates direct binding of NFATc2 and/or NFKB1/RELA complex to trans-activate Ets1 transcription. Moreover, enhanced level of Nfatc2 and Nfkb1 positively correlated with Ets1 expression in the human breast cancer specimens. Deletion of the CRE region by CRISPR/Cas9 system resulted in significant reduction in Ets1 expression, which led to alterations of Ets1-mediated transcription programs including tumor invasiveness-related genes. Proper regulation of Ets1 gene expression by targeting the NFATc2 and NFKB1/RELA interaction could be a potential therapeutic target for Ets1-mediated metastatic breast cancer.

Background Some previous studies reported hearing ability can be reduced by impaired masticatory ability, but there has been little evidence reported of an association between hearing loss and unilateral mastication. Therefore, this study aimed to investigate the relationship between unilateral mastication (UM), estimated from individual functional tooth units (FTUs), and hearing loss in a representative sample of Korean adults. Methods The analyzed data were obtained from 1,773 adults aged 40-89 years who participated in Korean national survey. Hearing loss was defined as a pure-tone average of >25 dB at frequencies of 0.5, 1, 2, and 4 kHz in either ear. In each subject, UM was calculated as the difference in the sums of the FTU scores, which is an index of posterior tooth occlusion, on the two sides of the oral cavity. The scores were used to classify the UM into low, moderate, and high. The adjusted odds ratios (aORs) and their 95% confidence intervals (CIs) were calculated in multivariable logistic regression analyses. Results When controlling for sociodemographic factors, the aOR for hearing loss was 3.12 (95% CI, 1.21-8.03) for high UM relative to low UM. This association remained in a fully-adjusted model containing factors related to noise exposure (aOR 2.88; 95% CI, 1.12-7.46). Conclusion Adults with high UM as measured using FTUs showed a higher occurrence of hearing loss than those with low UM.

FoxP3+ regulatory T cells (Tregs) are a central element of immunological tolerance. FoxP3 is the key determining transcription factor of the Treg lineage, interacting with numerous cofactors and transcriptional targets to determine the many facets of Treg function. Its absence leads to devastating lymphoproliferation and autoimmunity in scurfy mutant mice and immunodysregulation polyendocrinopathy enteropathy X-linked (IPEX) patients. To finely map transcriptionally active regions of the protein, with respect to disease-causing variation, we performed a systematic alanine-scan mutagenesis of FoxP3, assessing mutational impacts on DNA binding and transcriptional activation or repression. The mutations affected transcriptional activation and repression in a variegated manner involving multiple regions of the protein and varying between different transcriptional targets of FoxP3. There appeared to be different modalities for target genes related to classic immunosuppressive function vs. those related to atypical or tissue-Treg functions. Relevance to in vivo Treg biology was established by introducing some of the subtle Foxp3 mutations into the mouse germline by CRISPR-based genome editing. The resulting mice showed Treg populations in normal numbers and exhibited no overt autoimmune manifestations. However, Treg functional defects were revealed upon competition or by system stress, manifest as a strikingly heightened susceptibility to provoked colitis, and conversely by greater resistance to tumors. These observations suggest that some of the missense mutations that segregate in human populations, but do not induce IPEX manifestations, may have unappreciated consequences in other diseases.

a b s t r a c t Objectives: Detection of approximal caries lesions can be difficult due to their anatomical position. This study aimed to assess the ability of the quantitative light-induced fluorescence-digital (QLF-D) in detecting approximal caries, and to compare the performance with those of the International Caries Detection and Assessment System II (ICDAS II) and digital radiography (DR). Methods: Extracted permanent teeth (n = 100) were selected and mounted in pairs. The simulation pairs were assessed by one calibrated dentist using each detection method. After all the examinations, the teeth (n = 95) were sectioned and examined histologically as gold standard. The modalities were compared in terms of sensitivity, specificity, areas under receiver operating characteristic curves (AUROC) for enamel (D1) and dentine (D3) levels. The intra-examiner reliability was assessed for all modalities. Results: At D1 threshold, the ICDAS II presented the highest sensitivity (0.80) while the DR showed the highest specificity (0.89); however, the methods with the greatest AUC values at D1 threshold were DR and QLF-D (0.80 and 0.80 respectively). At D3 threshold, the methods with the highest sensitivity were ICDAS II and QLF-D (0.64 and 0.64 respectively) while the method with the lowest sensitivity was DR (0.50). However, with regard to the AUC values at D3 threshold, the QLF-D presented the highest value (0.76). All modalities showed to have excellent intra-examiner reliability. Conclusions: The newly developed QLF-D was not only able to detect proximal caries, but also showed to have comparable performance to the visual inspection and radiography in detecting proximal caries. Clinical significance: QLF-D has the potential to be a useful detection method for proximal caries.

The aim of this study was to evaluate the desensitizing effects of a dentifrice containing nano-carbonate apatite (n-CAP) and Er,Cr:YSGG laser in the treatment of dentin hypersensitivity.Most studies of hypersensitivity treatment have been conducted with different methods of professional treatment and self-care in each study. Moreover, clinical studies that compare self-care and professional treatment have not yet been published.Eighty-two patients with dentin hypersensitivity were divided randomly into three groups: (1) a control group with strontium chloride dentifrice (SC), (2) n-CAP dentifrice (n-CAP), and (3) an Er,Cr:YSGG laser (laser) group. The study was conducted for 4 weeks: a treatment period of 2 weeks and a maintenance period of 2 subsequent weeks. The SC and n-CAP groups were instructed to brush their teeth twice a day for 1 min. The laser group visited twice at 1 week intervals for irradiation of the sensitive teeth. The desensitizing effect was evaluated by assessing the tactile sensitivity using the visual analogue scale (VAS), and evaporative air sensitivity was determined using an air blast score (ABS).The n-CAP group and the laser group showed significantly different desensitizing effects in VAS after 4 weeks (69% and 63%, respectively) and a 33% (p0.05) and 3% (p0.05) desensitizing effect, respectively, in VAS during the maintenance period.The n-CAP and the laser were effective in reducing dentin hypersensitivity. The laser had a superior desensitizing effect at the initial stage, whereas the n-CAP maintained its effect for a relatively longer time in clinical situations.

FoxP3 conditions the transcriptional signature and functional facets of regulatory T cells (Treg cells). Its mechanism of action, whether as an activator or a repressor, has remained unclear. Here, chromatin analysis showed that FoxP3 bound active enhancer elements, not repressed chromatin, around loci over- or under-expressed in Treg cells. We evaluated the impact of a panel of FoxP3 mutants on its transcriptional activity and interactions with DNA, transcriptional cofactors and chromatin. Computational integration, confirmed by biochemical interaction and size analyses, showed that FoxP3 existed in distinct multimolecular complexes. It was active and primarily an activator when complexed with the transcriptional factors RELA, IKZF2 and KAT5. In contrast, FoxP3 was inactive when complexed with the histone methyltransferase EZH2 and transcription factors YY1 and IKZF3. The latter complex partitioned to a peripheral region of the nucleus, as shown by super-resolution microscopy. Thus, FoxP3 acts in multimodal fashion to directly activate or repress transcription, in a context- and partner-dependent manner, to govern Treg cell phenotypes.

The aim of this study was to determine the occluding effects of a combination of dentifrice containing nano-carbonate apatite (n-CAP) and CO2 laser on dentinal tubules, and to evaluate the acid resistance of this combination after 4 days of treatment, by using an in situ model.The synergic effect of this combination was demonstrated in an in vitro study.This was a two period crossover, single-blind, randomized, four-treatment, split-mouth study. Ten healthy participants wore lower intraoral appliances during the treatment period. Specimens were divided into the following four groups: no treatment (control group), tooth-brushing using 20% n-CAP dentifrice (n-CAP group), CO2 laser irradiation (laser group), and laser irradiation after n-CAP application (combined group). Occluding effects were evaluated on 2 days (days 1 and 2), and then acid challenge was performed using grape juice on 2 days (days 3 and 4). All of the specimen surfaces were evaluated by a scanning electron microscope.The combined group showed a better occluding effect than control group compared with other treatment groups, and this effect was 20% higher than that in the n-CAP group. Also, the combined group had the smallest open dentinal tubular area among all of the treatment groups.The combined therapy is a promising method for ensuring a long-lasting effect of dentin hypersensitivity treatment in clinical practice.

Objective: A low-viscosity resin (infiltrant) was used to inhibit the progression of white spot lesions (WSLs) and resolve associated esthetic issues. An alternative pretreatment was explored to increase the pore volume of the surface layer of the WSLs. Also, the penetration effects of the infiltrant were evaluated for various pretreatments. Methods: Sixty two artificial lesions were fabricated on bovine teeth. As a positive control, 15% HCl gel was applied for 120 seconds. Further, 37% H3PO4 gel was applied for 30 seconds using three methods. The samples were divided as follows: H3PO4 only group, H3PO4 sponge group, and H3PO4 brush group. The acid was gently rubbed with the applicators (i.e., a sponge or brush) throughout the application time. To compare the effects of resin infiltration, twenty paired halves of specimens were treated with an infiltrant (ICON®). Results: Thicknesses of the removed surface layers and infiltrated areas were evaluated by confocal laser scanning microscope. The positive control and the 37% H3PO4 brush group failed to show significant differences in the removed thickness (p > 0.05); however, the mean percentage of the infiltrated area was higher in the 37% H3PO4 brush group (84.13 ± 7.58%) than the positive control (63.51 ± 7.62%, p < 0.001). Scanning electron microscope observations indicate higher pore volumes for the 37% H3PO4 brush group than for the positive control. Conclusions: Application of 37% H3PO4 with a brush for 30 seconds increased the pore volume of WSL surface layers and the percentage of infiltrated areas in comparison to the use of 15% HCl for 120 seconds.

Objectives The aims of this study were to investigate an alternative pre-treatment method for resin infiltration using 37% H3PO4 with a brush applicator and to evaluate the penetration effect of the infiltrant for masking the natural white spot lesions (WSLs) in human teeth. Methods Seventy extracted human molars and pre-molars with non-cavitated WSLs were collected. Thirty teeth met criteria of ICDAS code 2, and were sectioned, providing a total of sixty paired halves. For the control group, 15% HCl gel was applied for 120 s, and 37% H3PO4, gel was gently rubbed with a brush applicator for 30 s to the experimental group. Also, to evaluate the penetration effect of the infiltrant by pre-treatment, the specimens were treated with the infiltrant (Icon®). Thicknesses of the removed surfaces and percentages of the infiltrated areas (IA%) were evaluated by CLSM, and micro-morphological changes were observed by SEM. Results The mean thicknesses of removed surface layers were significantly different between the control group (36 ± 7.62 μm) and the experimental group (13 ± 2.76 μm) (p 0.05). In the SEM images, the prism cores were preferentially dissolved in the control group, while the prism peripheries were preferentially dissolved in the experimental group. Conclusions Applying 37% H3PO4 gel with an applicator brush for 30 s could increase the permeability and minimize removal of the surface layer of natural WSLs. Moreover, the effect of resin infiltration was similar to the control group which was pretreated 15% HCl gel for 120 s in vitro study. Clinical significance For resin infiltration, applying 37% H3PO4 gel with a brush applicator can preserve the protective surface layers of the WSLs with reduced application time.

Allergic contact hypersensitivity (CHS) is an inflammatory skin disease mediated by allergen specific T cells. In this study, we investigated the role of transcription factor NFAT1 in the pathogenesis of contact hypersensitivity. NFAT1 knock out (KO) mice spontaneously developed CHS-like skin inflammation in old age. Healthy young NFAT1 KO mice displayed enhanced susceptibility to hapten-induced CHS. Both CD4+ and CD8+ T cells from NFAT1 KO mice displayed hyper-activated properties and produced significantly enhanced levels of inflammatory T helper 1(Th1)/Th17 type cytokines. NFAT1 KO T cells were more resistant to activation induced cell death (AICD) and regulatory T cells derived from these mice showed a partial defect in their suppressor activity. NFAT1 KO T cells displayed a reduced expression of apoptosis associated BCL-2/BH3 family members. Ectopic expression of NFAT1 restored the AICD defect in NFAT1 KO T cells and increased AICD in normal T cells. Recipient Rag2−/− mice transferred with NFAT1 KO T cells showed more severe CHS sensitivity due to a defect in activation induced hapten-reactive T cell apoptosis. Collectively, our results suggest the NFAT1 plays a pivotal role as a genetic switch in CD4+/CD8+ T cell tolerance by regulating AICD process in the T cell mediated skin inflammation.

IL-10 is a multifunctional cytokine that plays a crucial role in immunity and tolerance. IL-10 is produced by diverse immune cell types, including B cells and subsets of T cells. Although Th1 produce IL-10, their expression levels are much lower than Th2 cells under conventional stimulation conditions. The potential role of E26 transformation-specific 1 (Ets-1) transcription factor as a negative regulator for Il10 gene expression in CD4+ T cells has been implicated previously. In this study, we investigated the underlying mechanism of Ets-1–mediated Il10 gene repression in Th1 cells. Compared with wild type Th1 cells, Ets-1 knockout Th1 cells expressed a significantly higher level of IL-10, which is comparable with that of wild type Th2 cells. Upregulation of IL-10 expression in Ets-1 knockout Th1 cells was accompanied by enhanced chromatin accessibility and increased recruitment of histone H3 acetylation at the Il10 regulatory regions. Reciprocally, Ets-1 deficiency significantly decreased histone deacetylase 1 (HDAC1) enrichment at the Il10 regulatory regions. Treatment with trichostatin A, an inhibitor of HDAC family, significantly increased Il10 gene expression by increasing histone H3 acetylation recruitment. We further demonstrated a physical interaction between Ets-1 and HDAC1. Coexpression of Ets-1 with HDAC1 synergistically repressed IL-10 transcription activity. In summary, our data suggest that an interaction of Ets-1 with HDAC1 represses the Il10 gene expression in Th1 cells.

We reported an innovative, targeted chemoimmuno drug-delivery system. Although chemoimmunotherapy, as an alternative to or in combination with conventional therapeutic systems, has been in the forefront of recent oncological research, as presently configured, such systems face several major obstacles for efficient clinical application. Here, we establish a novel nano-platform for effective chemoimmunotherapy designed to overcome the drawbacks of conventional cancer therapies, describing a delivery system based on a dendrimer and a single-strand DNA-A9 PSMA (prostate-specific membrane antigen) RNA aptamer hybrid. Employing these vehicles, we demonstrate the promising possibility of this chemoimmuno therapeutic system against prostate cancer in in vivo and in vitro models.

CD4(+) T cells play critical roles in the generation of protective immunity against a variety of pathogens. The main two types of effector CD4(+) T cells, Th1 and Th2 are characterized by their ability to produce signature cytokines. Among them, IL-10 is a multi-functional cytokine that plays a crucial role in maintaining the balance between immunity and tolerance. Although IL-10 is produced by both differentiated primary Th1 and Th2 cells, Th2 cells produce much higher levels of IL-10 upon stimulation. However, little information is available on the molecular mechanisms of IL-10 gene regulation at the transcriptional level. Interferon regulatory factor IRF4 is a member of the IRF family of transcription factors and plays critical roles in the development of CD4(+) T cells into Th2 cells. In this present study, we elucidate the underlying mechanism of IRF4 mediated IL-10 gene transcription in primary CD4(+) T cells. Th2 specific binding of IRF4 to the IRF4 responsive elements in IL-10 locus potentiated IL-10 expression in Th2 cells. Knockdown of IRF4 by siRNA decreased IL-10 expression level in Th2 cells. Nuclear translocation of IRF4 was much higher in Th2 cells upon stimulation, which contribute to maintain IL-10(high) phenotype of Th2 cells. Collectively, our results suggest that stimulation driven quantitative differences of IRF4 in the nucleus and its binding to IL-10 regulatory elements are crucial mechanisms to induce IL-10(high) gene expression in Th2 cells.

Forkhead transcription factor 3 (Foxp3) is critical for generating CD4(+)CD25(+) regulatory T cells. However, its role in microglia has not been identified. Here, we show that Foxp3 is expressed in microglia and is upregulated upon activation. In Foxp3 mutant mice (Foxp3(sf)), microglia release higher levels of inflammatory cytokines and mediators such as NO, MCP-1, CXCL10, and ROS upon liposaccharide treatment than the wild type, while TNF-alpha and IL-1 beta were not significantly different between wild and mutant microglial cells. In addition, Foxp3 silencing enhances inflammatory responses, suggesting that the major role of Foxp3 in microglia is that of a repressor of activation. Similarly, Foxp3 overexpression reduces inflammatory responses in microglia. We also demonstrate that Foxp3 interacts directly with NF-kappaB and modulates its transcriptional activities. These findings point to the importance of Foxp3 in NF-kappaB mediated inflammatory responses in microglia.

Occlusal discoloration due to staining frequently occurs on the pits and fissures of teeth. Noncariogenic discoloration (non-CD) refers to the attachment of staining chromogens to sound surfaces, whereas cariogenic discoloration (CD) represents the discoloration of porous structures due to bacterial metabolites and mineral loss from the enamel surface. This study evaluated whether it is possible to distinguish between non-CD and CD on stained occlusal surfaces with fluorescence assessed by the quantitative light-induced fluorescence (QLF) technology. Sixty-two extracted human permanent teeth with suspected discolorations on the pit and fissure were examined. The maximum values of fluorescence loss (ΔFmax) and red fluorescence gain (ΔRmax) were calculated using QLF images. Using histology as the gold standard, it was found that 12 teeth were sound (non-CD), while 50 teeth had enamel and dentine caries (CD). The validity tests at the enamel histological caries level, ΔRmax (ρ = 0.80) were strongly correlated with the histology (P

To evaluate immunomodulators from herbal medicines that affect the expression of IL-10 and TNF-α, we developed a cell-based high throughput screening system and tested 17 kinds of herbal medicine. We found that extracts of Coptidis Rhizoma can be used to stimulate TNF-α and IL-10 expression.