BAG6 inhibits influenza A virus replication by inducing viral polymerase subunit PB2 degradation and perturbing RdRp complex assembly.

The interaction between influenza A virus (IAV) and host proteins is an important process that greatly influences viral replication and pathogenicity. PB2 protein is a subunit of viral ribonucleoprotein (vRNP) complex playing distinct roles in viral transcription and replication. BAG6 (BCL2-associated athanogene 6) as a multifunctional host protein participates in physiological and pathological processes. Here, we identify BAG6 as a new restriction factor for IAV replication through targeting PB2. For both avian and human influenza viruses, overexpression of BAG6 reduced viral protein expression and virus titers, whereas deletion of BAG6 significantly enhanced virus replication. Moreover, BAG6-knockdown mice developed more severe clinical symptoms and higher viral loads upon IAV infection. Mechanistically, BAG6 restricted IAV transcription and replication by inhibiting the activity of viral RNA-dependent RNA polymerase (RdRp). The co-immunoprecipitation assays showed BAG6 specifically interacted with the N-terminus of PB2 and competed with PB1 for RdRp complex assembly. The ubiquitination assay indicated that BAG6 promoted PB2 ubiquitination at K189 residue and targeted PB2 for K48-linked ubiquitination degradation. The antiviral effect of BAG6 necessitated its N-terminal region containing a ubiquitin-like (UBL) domain (17-92aa) and a PB2-binding domain (124-186aa), which are synergistically responsible for viral polymerase subunit PB2 degradation and perturbing RdRp complex assembly. These findings unravel a novel antiviral mechanism via the interaction of viral PB2 and host protein BAG6 during avian or human influenza virus infection and highlight a potential application of BAG6 for antiviral drug development.

Relationship between 24-h activity behavior and body fat percentage in preschool children: based on compositional data and isotemporal substitution analysis.

OBJECTIVE: This study aims to elucidate the dose‒response relationship between 24-h activity behaviors and body fat percentage (BFP) in Chinese preschool children using a compositional isotemporal substitution model (ISM). METHODS: In a cross-sectional design, 881 children aged 3-6 from urban and rural areas of Jiangxi Province were sampled. Activity behaviors, including sedentary behavior (SB), low-intensity physical activity (LPA), and moderate- to high-intensity physical activity (MVPA), were measured using accelerometers. Sleep patterns were assessed through questionnaires, and BFP was determined by bioelectrical impedance analysis (BIA). The study employed compositional data analysis (CoDA) and ISM to estimate the impact of reallocating durations of different activity behaviors on BFP. RESULTS: Higher BFP was found in urban vs. rural children, decreasing with age. Overweight and obesity rates were 10.6% and 7.6%, respectively, above national averages. MVPA and LPA were negatively correlated with BFP, while SB was positively correlated. A 30-min MVPA reduction significantly increased zBFR, particularly in overweight children. Gender-specific nuances revealed that boys' MVPA negatively influenced zBFP (ß = -0.155), P < 0.05), while girls' SB positively impacted zBFP (ß = 0.636, P < 0.01). Isotemporal simulations emphasized amplified effects in overweight children, with boys' zBFR rising rapidly when MVPA was substituted and girls displaying a notable substitution effect between SB and LPA. CONCLUSION: BFP is closely linked to 24-h activity behaviors, notably in overweight and obese preschoolers. ISM identified MVPA as a critical influencer, with a 30-min reduction substantially increasing BFP. Gender disparities were evident, implicating MVPA in boys and LPA and SB in girls.

Alveolar bone morphology in patients with palatally-displaced maxillary lateral incisors before and after orthodontic treatment: A cone-beam computed tomography study.

INTRODUCTION: Palatal displacement of maxillary anterior teeth is common in clinical practice. Previous studies have reported that the labial bone around palatally-displaced incisors is thinner than that around normally-placed teeth. Therefore, it is necessary to elucidate alveolar bone changes after alignment to guide orthodontic treatment. In this study, we investigated the alveolar bone changes around palatally-displaced maxillary lateral incisors before and after treatment, and the effects of extraction and age using cone-beam computed tomography. METHODS: In this retrospective study, 55 patients with unilateral palatally-displaced maxillary lateral incisors were included. Three-dimensional alveolar bone changes were measured at three levels (25%, 50% and 75% of the root length) using cone-beam computed tomography. Group comparisons were made between displaced and control teeth, extraction and non-extraction groups, and adult and minor groups. RESULTS: After orthodontic treatment, labiopalatal and palatal alveolar bone widths decreased at all measured levels. Labial alveolar bone width increased significantly at P25, but decreased at P75. Concavity decreased, while tooth-axis angle, tooth length, B-CEJ and P-CEJ increased. Changes in LB and LP at P75, B-CEJ and P-CEJ were statistically significant. After treatment, the tooth-axis angle on the PD side increased by 9.46°. The change in tooth-axis angle on the PD side was significantly smaller, and LB and LP decreased more at P75, in the extraction group. CONCLUSIONS: Compared to the control teeth, alveolar bone thickness and height for the displaced teeth decreased more significantly after treatment. Tooth extraction and age also influenced alveolar bone changes.

Variations in the alveolar bone morphology in maxillary molar area: a retrospective CBCT study.

BACKGROUND: This study quantitatively analyzed the anatomic structure of the alveolar bone in the maxillary molar region at three potential locations for Temporary Anchorage Device (TAD) placement. Additionally, the study compared the variability in this region across different age groups, sagittal skeletal patterns, vertical facial types, and sexes. METHODS: In this retrospective cone-beam computed tomography study, the buccal alveolar bone was analyzed in the posterior molar area of 200 patients, the measurement items include buccal alveolar bone height, alveolar bone thickness, interradicular distance, and maxillary retromolar space. RESULTS: Buccal alveolar height was greatest in the U56 region. The interradicular space was largest in the U56 region and increased from the alveolar crest to the sinus floor. Buccal alveolar bone thickness was highest in the U67 region and generally increased from the alveolar crest to the sinus floor. The maxillary retromolar space gradually increased from the alveolar crest to the root apex. CONCLUSIONS: TADs are safest when placed in the buccal area between the maxillary second premolar and the first molar, particularly at the 9 mm plane. The U67 region is the optimal safe zone for TAD placement for maxillary dentition distalization. TADs placement in adolescents can be challenging. Maxillary third molar extraction can be considered for maxillary dentition distalization.

Nanofibers in Organelles: From Structure Design to Biomedical Applications.

Nanofibers are one of the most important morphologies of molecular self-assemblies, the formation of which relies on the diverse intermolecular interactions of fibrous-forming units. In the past decade, rapid advances have been made in the biomedical application of nanofibers, such as bioimaging and tumor treatment. An important topic to be focused on is not only the nanofiber formation mechanism but also where it forms, because different destinations could have different influences on cells and its formation could be triggered by unique stimuli in organelles. It is therefore necessary and timely to summarize the nanofibers assembled in organelles. This minireview discusses the formation mechanism, triggering strategies, and biomedical applications of nanofibers, which may facilitate the rational design of nanofibers, improve our understanding of the relationship between nanofiber properties and organelle characteristics, allow a comprehensive recognition of organelles affected by materials, and enhance the therapeutic efficiency of nanofibers.

Photo-Controlled Calcium Overload from Endogenous Sources for Tumor Therapy.

Designing reactive calcium-based nanogenerators to produce excess calcium ions (Ca2+ ) in tumor cells is an attractive tumor treatment method. However, nanogenerators that introduce exogenous Ca2+ are either overactive incapable of on-demand release, or excessively inert incapable of an overload of calcium rapidly. Herein, inspired by inherently diverse Ca2+ -regulating channels, a photo-controlled Ca2+ nanomodulator that fully utilizes endogenous Ca2+ from dual sources was designed to achieve Ca2+ overload in tumor cells. Specifically, mesoporous silica nanoparticles were used to co-load bifunctional indocyanine green as a photodynamic/photothermal agent and a thermal-sensitive nitric oxide (NO) donor (BNN-6). Thereafter, they were coated with hyaluronic acid, which served as a tumor cell-targeting unit and a gatekeeper. Under near-infrared light irradiation, the Ca2+ nanomodulator can generate reactive oxygen species that stimulate the transient receptor potential ankyrin subtype 1 channel to realize Ca2+ influx from extracellular environments. Simultaneously, the converted heat can induce BNN-6 decomposition to generate NO, which would open the ryanodine receptor channel in the endoplasmic reticulum and allow stored Ca2+ to leak. Both in vitro and in vivo experiments demonstrated that the combination of photo-controlled Ca2+ influx and release could enable Ca2+ overload in the cytoplasm and efficiently inhibit tumor growth.

The role of non-steroidal anti-inflammatory drugs as adjuncts to periodontal treatment and in periodontal regeneration.

Periodontitis is the sixth most prevalent chronic disease globally and places significant burdens on societies and economies worldwide. Behavioral modification, risk factor control, coupled with cause-related therapy have been the "gold standard" treatment for managing periodontitis. Given that host inflammatory and immunological responses play critical roles in the pathogenesis of periodontitis and impact treatment responses, several adjunctive strategies aimed at modulating host responses and improving the results of periodontal therapy and maintenance have been proposed. Of the many pharmacological host modulators, we focused on non-steroidal anti-inflammatory drugs (NSAIDs), due to their long history and extensive use in relieving inflammation and pain and reducing platelet aggregation. NSAIDs have been routinely indicated for treating rheumatic fever and osteoarthritis and utilized for the prevention of cardiovascular events. Although several efforts have been made to incorporate NSAIDs into the treatment of periodontitis, their effects on periodontal health remain poorly characterized, and concerns over the risk-benefit ratio were also raised. Moreover, there is emerging evidence highlighting the potential of NSAIDs, especially aspirin, for use in periodontal regeneration. This review summarizes and discusses the use of NSAIDs in various aspects of periodontal therapy and regeneration, demonstrating that the benefits of NSAIDs as adjuncts to conventional periodontal therapy remain controversial. More recent evidence suggests a promising role for NSAIDs in periodontal tissue engineering and regeneration.

A d-Methionine-Bonded Nanosized Heteropolyoxotungstate with Photocoloration.

A d-methionine-bonded nanosized arsenotungstate, Ba9K10H6[{As2W19O67(H2O)}2{AsW9O33}2{W3O6(H2O)(d-Met)}2{W2O4(OH)(d-Met)}]·60H2O [1; d-Met = d-methionine (C5H11NO2S)], is constructed without the use of lanthanide ions. The polyanion of 1 contains two {As2W19O67(H2O)}14- building blocks and two {B-ß-AsW9O33}9- subunits, integrated together with a {W2O4(OH)(d-Met)}2+ and two {W3O6(H2O)(d-Met)}5+ subclusters. Interestingly, 1 displays a highly reversible photocoloration property with a half-life (t1/2) time measured as about 0.793 min.

Pseudorabies Virus DNA Polymerase Processivity Factor UL42 Inhibits Type I IFN Response by Preventing ISGF3-ISRE Interaction.

Alphaherpesviruses are large dsDNA viruses with an ability to establish persistent infection in hosts, which rely partly on their ability to evade host innate immune responses, notably the type I IFN response. However, the relevant molecular mechanisms are not well understood. In this study, we report the UL42 proteins of alphaherpesvirus pseudorabies virus (PRV) and HSV type 1 (HSV1) as a potent antagonist of the IFN-I-induced JAK-STAT signaling pathway. We found that ectopic expression of UL42 in porcine macrophage CRL and human HeLa cells significantly suppresses IFN-α-mediated activation of the IFN-stimulated response element (ISRE), leading to a decreased transcription and expression of IFN-stimulated genes (ISGs). Mechanistically, UL42 directly interacts with ISRE and interferes with ISG factor 3 (ISGF3) from binding to ISRE for efficient gene transcription, and four conserved DNA-binding sites of UL42 are required for this interaction. The substitution of these DNA-binding sites with alanines results in reduced ISRE-binding ability of UL42 and impairs for PRV to evade the IFN response. Knockdown of UL42 in PRV remarkably attenuates the antagonism of virus to IFN in porcine kidney PK15 cells. Our results indicate that the UL42 protein of alphaherpesviruses possesses the ability to suppress IFN-I signaling by preventing the association of ISGF3 and ISRE, thereby contributing to immune evasion. This finding reveals UL42 as a potential antiviral target.

Quantitative prediction of ternary mixed gases based on an SnO2 sensor array and an SSA-BP neural network model.

This paper describes a tin oxide and copper doped tin oxide gas sensing material synthesized by a biological template method and simple hydrothermal reaction, which were used for the preparation of a gas sensor array. The sensor array is combined with the Sparrow Search Algorithm optimized BP neural network algorithm (SSA-BP) to predict and analyze the concentration of indoor toxic gases, including ammonia, xylene, and formaldehyde. Granular SnO2 was prepared by the biological template method and Cu/SnO2 doped with different copper ion concentrations was prepared by the hydrothermal method. The morphology of the synthesized nanomaterials was characterized by SEM, and the elemental composition and chemical state of the main elements were analyzed by XRD and XPS. The PL emission observed in the visible region is attributed to the defect level gap caused by oxygen. The optimal operating temperature, sensitivity, response/recovery time and the long-term stability of the sensor array have been studied. By combining the sensor array with the neural network algorithm in a simulated indoor environment at four humidity levels, the concentration information of the gas mixtures could be well predicted and the predicted concentration error was less than 0.84 ppm. Therefore, the sensor array prepared in this study combined with the SSA-BP algorithm achieved good results in predicting the concentrations of the three toxic mixtures.

Effect of socioeconomic status on the physical and mental health of the elderly: the mediating effect of social participation.

BACKGROUND: Previous studies have demonstrated the effect of socioeconomic status on the health status of the elderly. Nevertheless, the specific dimensions of the effect and the mechanism await further investigation. In this study, socioeconomic status was divided into three dimensions and we used social participation as the mediation variable to investigate the specific path of effect. METHODS: Using the 2018 Waves of Chinese Longitudinal Healthy Longevity Survey (CLHLS) dataset, a total of 10,197 effective samples of the elderly over 65 years old were screened out. Socioeconomic status included income, education level, and main occupation before retirement. The physical health and mental health of the elderly was measured by the Instrumental Activities of Daily Living Scale and the Minimum Mental State Examination, respectively. The social participation of the elderly was the mediation variable, including group exercise, organized social activities and interacting with friends. Omnibus mediation effect analysis was adopted to examine the mediation effect and mediation analysis was completed using the SPSS PROCESS program. RESULTS: First, the results showed that when the income gap between the elderly reached a certain level, there was a significant difference in health status. Significant differences existed in health status amongst with different education levels. There was no sufficient evidence to show that occupation has a significant effect on the physical health. But when the dependent variable was mental health, the effect was significant. Second, group exercise mediated 64.11% (aib = 0.24, 95% CI [0.17,0.3]) and up to 20.44% (aib = 0.12, 95% CI [0.07,0.17]) of the disparity in physical and mental health due to income gap, respectively. And it could mediate the effect up to 56.30% (aib = 0.62, 95% CI [0.52,0.73]) and 17.87% (aib = 0.50, 95% CI [0.4,0.61]) of education on physical and mental health status, respectively. The proportion of relative mediation effect of occupation was up to 28.74% (aib = 0.19, 95% CI [0.13,0.25]) on mental health. Interacting with friends mediated only on the path that the education affected the health status of the elderly. The proportion was up to 33.72% (aib = 0.29, 95% CI [0.16,0.44]). The relative mediation effect of organized social activities on the health gap caused by income or education level gap was significant at some levels. The proportion was up to 21.20% (aib = 0.33, 95% CI [0.26,0.4]). CONCLUSION: The SES of the elderly including relatively large income gap, different education levels and occupational categories could indeed have a significant effect on health status of the elderly, and the reason why this effect existed could be partly explained by the mediation effect of social participation. Policymakers should pay more attention to the social participation of the elderly.

Intratracheal exposure to polyhexamethylene guanidine phosphate disrupts coordinate regulation of FXR-SHP-mediated cholesterol and bile acid homeostasis in mouse liver.

A public health crisis in the form of a significant incidence of fatal pulmonary disease caused by repeated use of humidifier disinfectants containing polyhexamethylene guanidine phosphate (PHMG) recently arose in Korea. Although the mechanisms of pulmonary fibrosis following respiratory exposure to PHMG are well described, distant-organ effect has not been reported. In this study, we investigated whether intratracheal administration of PHMG affects liver pathophysiology and metabolism. Our PHMG mouse model showed a significant decrease in liver cholesterol level. An mRNA-seq analysis of liver samples revealed an alteration in the gene expression associated with cholesterol biosynthesis and metabolism to bile acids. The expression of genes involved in cholesterol synthesis was decreased in a real-time PCR analysis. To our surprise, we found that the coordinate regulation of cholesterol and bile acid homeostasis was completely disrupted. Despite the decreased cholesterol synthesis and low bile acid levels, the farnesoid X receptor/small heterodimer partner pathway, which controls negative feedback of bile acid synthesis, was activated in PHMG mice. As a consequence, gene expression of Cyp7a1 and Cyp7b1, the rate-limiting enzymes of the classical and alternative pathways of bile acid synthesis, was significantly downregulated. Notably, the changes in gene expression were corroborated by the hepatic concentrations of the bile acids. These results suggest that respiratory exposure to PHMG could cause cholestatic liver injury by disrupting the physiological regulation of hepatic cholesterol and bile acid homeostasis.

Progress of Nanomaterials-Based Photothermal Therapy for Oral Squamous Cell Carcinoma.

Oral squamous cell carcinoma (OSCC) is one of the top 15 most prevalent cancers worldwide. However, the current treatment models for OSCC (e.g., surgery, chemotherapy, radiotherapy, and combination therapy) present several limitations: damage to adjacent healthy tissue, possible recurrence, low efficiency, and severe side effects. In this context, nanomaterial-based photothermal therapy (PTT) has attracted extensive research attention. This paper reviews the latest progress in the application of biological nanomaterials for PTT in OSCC. We divide photothermal nanomaterials into four categories (noble metal nanomaterials, carbon-based nanomaterials, metal compounds, and organic nanomaterials) and introduce each category in detail. We also mention in detail the drug delivery systems for PTT of OSCC and briefly summarize the applications of hydrogels, liposomes, and micelles. Finally, we note the challenges faced by the clinical application of PTT nanomaterials and the possibility of further improvement, providing direction for the future research of PTT in OSCC treatment.

Information rates in Kerr nonlinearity limited optical fiber communication systems.

Achievable information rates of optical communication systems are inherently limited by nonlinear distortions due to the Kerr effect occurred in optical fibres. These nonlinear impairments become more significant for communication systems with larger transmission bandwidths, closer channel spacing and higher-order modulation formats. In this paper, the efficacy of nonlinearity compensation techniques, including both digital back-propagation and optical phase conjugation, for enhancing achievable information rates in lumped EDFA- and distributed Raman-amplified fully-loaded C -band systems is investigated considering practical transceiver limitations. The performance of multiple modulation formats, such as dual-polarisation quadrature phase shift keying (DP-QPSK), dual-polarisation 16 -ary quadrature amplitude modulation (DP-16QAM), DP-64QAM and DP-256QAM, has been studied in C -band systems with different transmission distances. It is found that the capabilities of both nonlinearity compensation techniques for enhancing achievable information rates strongly depend on signal modulation formats as well as target transmission distances.

Analysis of facial features and prediction of lip position in skeletal class III malocclusion adult patients undergoing surgical-orthodontic treatment.

OBJECTIVES: This study presents a retrospective study aimed to analyze the facial features at each stage of surgical-orthodontic treatment for skeletal class III malocclusion, and predict the changes in the lips after treatment. MATERIALS AND METHODS: There were 49 skeletal class III malocclusion patients treated with bimaxillary surgery and orthodontic treatment enrolled in this study. Lateral cephalograms were obtained before treatment (T0), 1 month before surgery (T1), 1 month after surgery (T2), and after debonding (T3) for cephalometric measurements. After the measurement of the required variables, paired t-test, Pearson's correlation analysis, and multiple linear regression were performed using SPSS 19.0. RESULTS: The main factors associated with changes in the upper lip included ΔUIE-V, ΔA-V, ΔU1A-V, and ΔL1A-V, and those associated with changes in the lower lip included ΔLIE-V, ΔL1A-V, ΔB-V, ΔPog-V, and Δfacial angle. The predicted regression equation for the horizontal change in the upper lip was represented as ΔUL-vertical reference line (VRL) = 9.430 + 0.779 (ΔUIE-VRL) - 0.542(VULT) (P < 0.05) with a mean error of 1.04 mm; the corresponding equation for the lower lip was ΔLL-VRL = -1.670 + 0.530 (ΔB-VRL) + 0.360 (Ls-E) + 0.393 (ΔLIE-VRL) (P < 0.05), with a mean error of 1.51 mm. CONCLUSIONS: This study explored the relationship between orthognathic surgery and changes in the lips and obtained the predictive equations of lip position after treatment by using multiple linear regression, which likely offers a reference for prediction of soft tissue changes before surgical-orthodontic treatment in patients with skeletal class III malocclusion. CLINICAL RELEVANCE: The findings can help dentists to rapidly predict the lip changes after surgical-orthodontic treatment in patients with skeletal class III malocclusion. The study has been registered with the Chinese Clinical Trial Registration (No: ChiCTR1800017694).

Investigation of Potting-Adhesive-Induced Thermal Stress in MEMS Pressure Sensor.

Thermal stress is one of the main sources of micro-electro-mechanical systems (MEMS) devices error. The Wheatstone bridge is the sensing structure of a typical piezoresistive MEMS pressure sensor. In this study, the thermal stress induced by potting adhesive in MEMS pressure sensor was investigated by experiments, calculated by analytics and analyzed by simulations. An experiment system was used to test the sensor at different air pressures and temperatures. The error becomes greater with the decrease in pressure. A set of novel formulas were proposed to calculate the stress-strain on Wheatstone bridge. The error increases with the temperature deviating from 25 °C. A full-scale geometric model was developed, and finite element simulations were performed, to analyze the effect of the stress on MEMS pressure sensor induced by different temperatures and thicknesses of potting adhesive. Simulation results agree well with the experiments, which indicated that there is a 3.48% to 6.50% output error in 0.35 mm potting adhesive at 150 °C. With the thickness of potting adhesive increasing, the variations of output error of the Wheatstone bridge present an N-shaped curve. The output error meets a maximum of 5.30% in the potting adhesive of 0.95 mm and can be reduced to 2.47%, by increasing the potting adhesive to 2.40 mm.

Investigation of Acoustic Injection on the MPU6050 Accelerometer.

Acoustic injection is one of the most dangerous ways of causing micro-electro-mechanical systems (MEMS) failures. In this paper, the failure mechanism of acoustic injection on the microprocessor unit 6050 (MPU6050) accelerometer is investigated by both experiment and simulation. A testing system was built to analyze the performance of the MPU6050 accelerometer under acoustic injection. A MEMS disassembly method was adopted and a MATLAB program was developed to establish the geometric model of MPU6050. Subsequently, a finite element model of MPU6050 was established. Then, the acoustic impacts on the sensor layer of MPU6050 were studied by acoustic-solid coupling simulations. The effects of sound frequencies, pressures and directions were analyzed. Simulation results are well agreed with the experiments which indicate that MPU6050 is most likely to fail under the sounds of 11,566 Hz. The failure mechanism of MPU6050 under acoustic injection is the relative shift of the capacitor flats caused by acoustic-solid resonance that make the sensor detect false signal and output error data. The stress is focused on the center linkage. MPU6050 can be reliable when the sound pressure is lower than 100 dB.

A novel IRF6 mutation causing non-syndromic cleft lip with or without cleft palate in a pedigree.

Non-syndromic cleft lip with or without cleft palate (NSCLP) is the most common congenital craniofacial malformation, and its harmful influence on affected individuals is apparent. Despite many studies, the causative genes and their mechanisms are not completely clear. We recruited a Han Chinese NSCLP family and explored the causative variant in this pedigree. We performed whole-exome sequencing on two patients. Bioinformatics screening and analysis were used to identify the mutation. We also performed species conservation analysis, mutation function predictions, and homology protein modelling to evaluate the influence of the mutation. We identified a rare mutation in interferon regulatory factor 6 (IRF6) (c.26G>A; p.Arg9Gln) as a candidate of causative mutation. This mutation was predicted to be deleterious. The codon is conserved in many species. The residue change caused by this mutation would affect the structure of IRF6 to a degree. Our study suggested that the rare IRF6 variant is probably the pathogenic mutation in this family. Our result adds evidence that IRF6 variants play a role in the aetiology of orofacial clefts.

A novel PTCH1 mutation underlies nonsyndromic cleft lip and/or palate in a Han Chinese family.

OBJECTIVES: Cleft lip and/or palate (CL/P) is the most common craniofacial congenital disease, and it has a complex aetiology. This study aimed to identify the causative gene mutation of a Han Chinese family with CL/P. SUBJECTS AND METHODS: Whole exome sequencing was conducted on the proband and her mother, who exhibited the same phenotype. A Mendelian dominant inheritance model, allele frequency, mutation regions, functional prediction and literature review were used to screen and filter the variants. The candidate was validated by Sanger sequencing. Conservation analysis and homology modelling were conducted. RESULTS: A heterozygous missense mutation c.1175C>T in the PTCH1 gene predicting p.Ala392Val was identified. This variant has not been reported and was predicted to be deleterious. Sanger sequencing verified the variant and the dominant inheritance model in the family. The missense alteration affects an amino acid that is evolutionarily conserved in the first extracellular loop of the PTCH1 protein. The local structure of the mutant protein was significantly altered according to homology modelling. CONCLUSIONS: Our findings suggest that c.1175C>T in PTCH1 (NM_000264) may be the causative mutation of this pedigree. Our results add to the evidence that PTCH1 variants play a role in the pathogenesis of orofacial clefts.

Vasoactive intestinal peptide downregulates proinflammatory TLRs while upregulating anti-inflammatory TLRs in the infected cornea.

TLRs recognize microbial pathogens and trigger an immune response, but their regulation by neuropeptides, such as vasoactive intestinal peptide (VIP), during Pseudomonas aeruginosa corneal infection remains unexplored. Therefore, C57BL/6 (B6) mice were injected i.p. with VIP, and mRNA, protein, and immunostaining assays were performed. After VIP treatment, PCR array and real-time RT-PCR demonstrated that proinflammatory TLRs (conserved helix-loop-helix ubiquitous kinase, IRAK1, TLR1, TLR4, TLR6, TLR8, TLR9, and TNFR-associated factor 6) were downregulated, whereas anti-inflammatory TLRs (single Ig IL-1-related receptor [SIGIRR] and ST2) were upregulated. ELISA showed that VIP modestly downregulated phosphorylated inhibitor of NF-κB kinase subunit α but upregulated ST2 ~2-fold. SIGIRR was also upregulated, whereas TLR4 immunostaining was reduced in cornea; all confirmed the mRNA data. To determine whether VIP effects were cAMP dependent, mice were injected with small interfering RNA for type 7 adenylate cyclase (AC7), with or without VIP treatment. After silencing AC7, changes in mRNA levels of TLR1, TNFR-associated factor 6, and ST2 were seen and unchanged with addition of VIP, indicating that their regulation was cAMP dependent. In contrast, changes were seen in mRNA levels of conserved helix-loop-helix ubiquitous kinase, IRAK1, 2, TLR4, 9 and SIGIRR following AC7 silencing alone; these were modified by VIP addition, indicating their cAMP independence. In vitro studies assessed the effects of VIP on TLR regulation in macrophages and Langerhans cells. VIP downregulated mRNA expression of proinflammatory TLRs while upregulating anti-inflammatory TLRs in both cell types. Collectively, the data provide evidence that VIP downregulates proinflammatory TLRs and upregulates anti-inflammatory TLRs and that this regulation is both cAMP dependent and independent and involves immune cell types found in the infected cornea.