Foot-and-mouth disease virus induces PERK-mediated autophagy to suppress the antiviral interferon response.

Foot-and-mouth disease virus (FMDV) is a picornavirus that causes contagious acute infection in cloven-hoofed animals. FMDV replication-associated viral protein expression induces endoplasmic reticulum (ER) stress and the unfolded protein response (UPR), in turn inducing autophagy to restore cellular homeostasis. We observed that inhibition of BiP (also known as HSPA5 and GRP78), a master regulator of ER stress and UPR, decreased FMDV infection confirming their involvement. Further, we show that the FMDV infection induces UPR mainly through the PKR-like ER kinase (PERK; also known as EIF2AK3)-mediated pathway. Knockdown of PERK and chemical inhibition of PERK activation resulted in decreased expression of FMDV proteins along with the reduction of autophagy marker protein LC3B-II [the lipidated form of LC3B (also known as MAP1LC3B)]. There are conflicting reports on the role of autophagy in FMDV multiplication. Our study systematically demonstrates that during FMDV infection, PERK-mediated UPR stimulated an increased level of endogenous LC3B-II and turnover of SQSTM1, thus confirming the activation of functional autophagy. Modulation of the UPR and autophagy by pharmacological and genetic approaches resulted in reduced numbers of viral progeny, by enhancing the antiviral interferon response. Taken together, this study underscores the prospect of exploring PERK-mediated autophagy as an antiviral target.

A new blocking ELISA for detection of foot-and-mouth disease non-structural protein (NSP) antibodies in a broad host range.

Large-scale monitoring of foot-and-mouth disease (FMD) in livestock is imperative in an FMD control program. Detection of antibodies against non-structural proteins (NSP) of FMD virus (FMDV) is one of the best tools to estimate the prevalence of past infection; availability of such a well-validated test is therefore essential. Using a FMDV 3B protein-specific monoclonal antibody, we have developed a new NSP antibody blocking ELISA (10H9 bELISA) and validated it on large panels of sera from different susceptible species. The diagnostic sensitivity of the ELISA was 95% with a specificity of 98%, similar to the values found using a commercial kit (PrioCHECK FMD NS test). The 10H9 bELISA can be used in a broad range of FMD susceptible species making it a very useful tool in monitoring the foot-and-mouth disease control programs by detection of virus circulation in the vaccinated populations. KEY POINTS: • A new ELISA for detection of foot and mouth disease (FMD) antibodies. • Diagnostic sensitivity of 95% and specificity of 98%. • Tested with panels of validated sera from broad host range.

Emergence of foot-and-mouth disease virus serotype Asia1 group IX in India.

Foot-and-mouth disease virus (FMDV) serotype Asia1 is prevalent in India and is responsible for a minor proportion of FMD outbreaks. Globally, serotype Asia1 is grouped into nine different groups (GI-IX) based on genetic analysis. In India, only Asia1/G-III and Asia1/G-VIII have been documented so far. Phylogenetic analysis of recent serotype Asia1 isolates from India revealed the emergence of Asia1/G-IX. The Asia1/G-IX lineage shares recent common ancestry with Asia1/G-VIII dating to 2016. The root state posterior probabilities of Asia1/G-VIII are inclusive and there may have been either an incursion of the virus from Bangladesh, where it was first identified, or in situ evolution of the virus within India, which is an intriguing possibility.

Reliability and Validity of a Nepalese Version of the Oral Health Impact Profile for Edentulous Subjects.

PURPOSE: To assess the reliability and to validate the translated Nepalese version of the Oral Health Impact Profile (OHIP-EDENT-N) in Nepalese edentulous subjects. MATERIALS AND METHODS: The international guidelines for translation and cross-cultural adaption of OHIP-EDENT were followed, and a Nepalese version of the questionnaire was adapted for this study. Eighty-eight completely edentulous subjects were then selected for the study and completed their responses for the questionnaire. The reliability of the OHIP-EDENT-N was evaluated using internal consistency. Validity was assessed as construct and convergent validity. Construct validity was determined using exploratory factor analysis (EFA). The correlation between OHIP-EDENT-N subscale scores and the global question was investigated to test the convergent validity. RESULTS: Cronbach's alpha for the total score of OHIP-EDENT-N was 0.78. Construct validity was assessed by factor analysis: 70.196% of the variance was accountable to five factors extracted from the factor analysis. Factor loadings above 0.40 were noted for all items. In terms of convergent validity, significant correlations could be established between OHIP-EDENT-N and global questions. CONCLUSIONS: This study has been able to establish the reliability and validity of the OHIP-EDENT-N, and OHIP-EDENT-N can be a considered a reliable tool to assess the oral health related quality of life in the Nepalese edentulous population.

Comparing bite force in untreated and orthodontically treated non- extraction cases with MBT appliance and self ligation technique using T-scan.

AIM: Aim of this cross sectional study was to compare the bite force and the individual tooth load among three groups of patients: cases finished with the Self ligation appliance (Damon-Q®, 0.022"X0.028" prescription) cases finished with the MBT (McLaughlin, Bennett and Trevisi) appliance system (0.022"X0.028"prescription) and orthodontically untreated cases using T-Scan® 9.0 version. MATERIAL & METHODS: A sample size of sixty non-extraction cases were divided equally into three groups - Group 1: patients treated with Self-ligation appliance (Damon Q®), Group 2: cases treated with the MBT prescription and Group 3: untreated subjects with Class I canine and molar relationship who were used as the control group. The bite force and distribution of force was measured on the right and left side as well as individual tooth loads were measured using the T-Scan® 9.0 device and inter-and intra-group comparisons made using Paired t-Test, ANOVA and Chi-square test. RESULT: The bite forces were balanced on both the sides in cases treated with Self-ligation and MBT appliance. The Self-ligation finishes were marginally better than the MBT group but the difference was statistically insignificant. CONCLUSION: Orthodontically treated individuals had a better (or more even) bite force distribution compared to untreated participants.

Assessment of fitness of foot-and-mouth disease virus A IND 27/2011 as candidate vaccine strain.

Antigenic profiling of recent field outbreak strains of foot-and-mouth disease virus (FMDV) serotype A in India has revealed considerable antigenic drift from the vaccine strain, A IND 40/2000, necessitating the selection of a new strain. The complete genome sequence of A IND 27/2011 was analysed. Vaccine quality attributes of the new candidate strain including potency as an inactivated vaccine in cattle were evaluated. The capsid coding region of A IND 27/2011 showed variation at eight antigenically critical amino acid positions from that of A IND 40/2000. The strain suited well with traits required by a vaccine in terms of its adaptability to adherent and suspension cell line, its immunogenicity, and potency as an inactivated vaccine formulation in cattle. Complete protection was observed upon homologous virus challenge at 4 weeks post-vaccination. Taken together, these data demonstrate the suitability of A IND 27/2011 as an effective vaccine strain of FMDV serotype A.

Differential expression of long non-coding RNAs under Peste des petits ruminants virus (PPRV) infection in goats.

Peste des petits ruminants (PPR) characterized by fever, sore mouth, conjunctivitis, gastroenteritis, and pneumonia, is an acute, highly contagious viral disease of sheep and goats. The role of long non-coding RNAs (lncRNAs) in PPRV infection has not been explored to date. In this study, the transcriptome profiles of virulent Peste des petits ruminants virus (PPRV) infected goat tissues - lung and spleen were analyzed to identify the role of lncRNAs in PPRV infection. A total of 13,928 lncRNA transcripts were identified, out of which 170 were known lncRNAs. Intergenic lncRNAs (7625) formed the major chunk of the novel lncRNA transcripts. Differential expression analysis revealed that 15 lncRNAs (11 downregulated and 4 upregulated) in the PPRV infected spleen samples and 16 lncRNAs (13 downregulated and 3 upregulated) in PPRV infected lung samples were differentially expressed as compared to control. The differentially expressed lncRNAs (DElncRNAs) possibly regulate various immunological processes related to natural killer cell activation, antigen processing and presentation, and B cell activity, by regulating the expression of mRNAs through the cis- or trans-regulatory mechanism. Functional enrichment analysis of differentially expressed mRNAs (DEmRNAs) revealed enrichment of immune pathways and biological processes in concordance with the pathways in which correlated lncRNA-neighboring genes were enriched. The results suggest that a coordinated immune response is raised in both lung and spleen tissues of the goat through mRNA-lncRNA crosstalk.

A single amino acid substitution in the VP2 protein of Indian foot-and-mouth disease virus serotype O vaccine strain confers thermostability and protective immunity in cattle.

Foot-and-mouth disease (FMD) is a significant threat to animal health globally. Prophylactic vaccination using inactivated FMD virus (FMDV) antigen is being practised for the control in endemic countries. A major limitation of the current vaccine is its susceptibility to high environmental temperature causing loss of immunogenicity, thus necessitating the cold chain for maintenance of its efficacy. Hence, the FMD vaccine with thermostable virus particles will be highly useful in sustaining the integrity of whole virus particle (146S) during storage at 4°C. In this study, 12 recombinant mutants of Indian vaccine strain of FMDV serotype O (O/IND/R2/1975) were generated through reverse genetics approach and evaluated for thermostability. One of the mutant viruses, VP2_Y98F was more thermostable than other mutants and the parent FMDV. The oil-adjuvanted vaccine formulated with the inactivated VP2_Y98F mutant FMDV was stable up to 8 months when stored at 4°C and induced protective antibody response till dpv 180 after primary vaccination. It is concluded that the VP2_Y98F mutant FMDV was thermostable and has the potential to replace the parent vaccine strain.

Genetic characterization of foot-and-mouth disease virus serotype O isolates collected during 2014-2018 revealed dominance of O/ME-SA/Ind2001e and the emergence of a novel lineage in India.

Foot-and-mouth disease (FMD) is endemic in India with a preponderance of outbreaks caused by FMD virus (FMDV) serotype O. Out of the 11 global topotypes of serotype O, only ME-SA topotype has been reported in the country so far. Lineage O/ME-SA/Ind2001 and O/ME-SA/PanAsia are documented as the most dominant ones in terms of the number of outbreaks caused by them. To understand the distribution of topotype/lineages in India and their antigenic behaviour during the year 2014-2018, a total of 286 FMDV serotype O viral isolates were sequence determined at the VP1 region, and 109 isolates were characterized antigenically. All the isolates grouped in the ME-SA topotype, being distributed in lineage O/ME-SA/Ind2001 (within sub-lineages O/ME-SA/Ind2001d and O/ME-SA/Ind2001e), and a new group designated here as O/ME-SA/2018 cluster. The sub-lineage O/ME-SA/Ind2001e reported for the first time in India during the year 2015, replaced sub-lineage O/ME-SA/Ind2001d gradually, which was dominating since 2008. During the years 2014-2018, the sub-lineage O/ME-SA/Ind2001e was found to be the most predominant one whose mean evolutionary rate was observed to be faster than that of the sub-lineage O/ME-SA/Ind2001d. The codon sites 45 and 85 of VP1 were found to be under diversifying selection in a large proportion of trees. The common ancestor predicted for sub-lineages O/ME-SA/Ind2001e and O/ME-SA/2018 dates back to 2012 and 2016, respectively. The sustenance and spread of the new O/ME-SA/2018 cluster need to be assessed by continued surveillance. The Indian vaccine strain O/INDR2/1975 was found to provide adequate antigenic coverage to the emerging and prevalent serotype O lineages. The trait association tests showed frequent virus exchange among different states, which could be an important confounder in the region-specific assessment of effectiveness of FMD control programme.

Differential antibody responses to the major antigenic sites of FMD virus serotype O after primo-vaccination, multiply-vaccination and after natural exposure.

Foot and mouth disease (FMD) virus serotype O is the predominant cause of FMD outbreaks in several regions of the world including India. Five independent neutralizing antigenic sites have been identified on the capsid surface of FMD virus serotype O. The relative importance of these neutralizing sites in eliciting antibody responses in the polyclonal sera collected from un-infected vaccinated (both primo and multiply-vaccinated) and naturally infected cattle populations were determined through a combination of reverse genetics and serology. The known critical amino acid residues present on the five antigenic sites of FMD virus serotype O Indian vaccine strain O IND R2/1975 were mutated through site-directed mutagenesis. The mutant viruses were rescued in cell-culture and analyzed through virus-neutralization assays along with parent virus using the polyclonal sera collected from three groups of cattle. In the polyclonal sera from primo-vaccinated cattle, significantly higher level of antibodies were directed towards antigenic site 2. In contrast, in polyclonal sera from multiply vaccinated animals, both antigenic sites 1 and 2 were equally important. In case of naturally infected animals, antibody responses were elicited against all the five antigenic sites. Although a drop in neutralization titres was observed for all the mutants, in one instance, increase in titre was noticed for a site 3 mutant. The findings from this study extend our knowledge on the antibody immunodominace following FMDV vaccination and infection, and may improve our strategies for vaccine strain selection and rational vaccine design.

Foot-and-Mouth Disease Virus: Immunobiology, Advances in Vaccines and Vaccination Strategies Addressing Vaccine Failures-An Indian Perspective.

A mass vaccination campaign in India seeks to control and eventually eradicate foot-and-mouth disease (FMD). Biosanitary measures along with FMD monitoring are being conducted along with vaccination. The implementation of the FMD control program has drastically reduced the incidence of FMD. However, cases are still reported, even in regions where vaccination is carried out regularly. Control of FMD outbreaks is difficult when the virus remains in circulation in the vaccinated population. Various FMD risk factors have been identified that are responsible for FMD in vaccinated areas. The factors are discussed along with strategies to address these challenges. The current chemically inactivated trivalent vaccine formulation containing strains of serotype O, A, and Asia 1 has limitations including thermolability and induction of only short-term immunity. Advantages and disadvantages of several new-generation alternate vaccine formulations are discussed. It is unfeasible to study every incidence of FMD in vaccinated animals/areas in such a big country as India with its huge livestock population. However, at the same time, it is absolutely necessary to identify the precise reason for vaccination failure. Failure to vaccinate is one reason for the occurrence of FMD in vaccinated areas. FMD epidemiology, emerging and re-emerging virus strains, and serological status over the past 10 years are discussed to understand the impact of vaccination and incidences of vaccination failure in India. Other factors that are important in vaccination failure that we discuss include disrupted herd immunity, health status of animals, FMD carrier status, and FMD prevalence in other species. Recommendations to boost the search of alternate vaccine formulation, strengthen the veterinary infrastructure, bolster the real-time monitoring of FMD, as well as a detailed investigation and documentation of every case of vaccination failure are provided with the goal of refining the control program.

Knowledge, Awareness, and Attitude Regarding Dental Implants among Dental Interns.

INTRODUCTION: Implant therapy is an elective dental procedure of dental rehabilitative treatment. Objective of this study was to assess the knowledge, awareness, and attitude of dental interns of Nepal toward dental implants and to see associations of the responses with gender and geographic location of dental college. METHODS: It was conducted from June 2016 to June 2017 after taking ethical clearance and approval from the research committee. The sample included the interns who were present at the time of the survey. Data collection was done through a cross-sectional questionnaire survey, during clinical postings of the interns at all the dental colleges of Nepal, supervised and monitored by the investigators themselves; coded and entered in Microsoft Excel 2013 and statistical analysis was done by SPSS 20 version. RESULTS: A majority of the interns said they are moderately well informed about dental implants 141 (50.36%); main advantage of dental implants is they are conservative 164 (58.6%); most important factor for implant success is case selection 143 (51.07%); the longevity of dental implants is 10-20yrs, 162 (57.85%), 188 (67.14%) agreed that they were not provided with sufficient information and 191 (68.21%) stated that they need more information; 180 (64.3%) felt the difficulty encountered while placing implants is average and 229 (81.78%) stated that they need more hygiene care than natural teeth. There were major differences in the mindset to some other questions. Significant associations were seen with the location of college for some responses. CONCLUSIONS: A majority of interns have knowledge regarding dental implants, but the percentage of those who gave unsatisfactory answers is also large. They have a positive attitude towards gaining more information through various means.

Preferred Source and Perceived Need of More Information about Dental Implants by the Undergraduate Dental Students of Nepal: All Nepal Survey.

OBJECTIVES: This study was conducted to know the preferred source and perceived need of more information about dental implants by the undergraduate students of Nepal and their association with academic levels and gender. MATERIALS AND METHODS: It was conducted in all the dental colleges of Nepal from June 2016 to June 2017 after taking ethical clearance and approval from the research committee of BPKIHS. It included all those who were present at the time of survey. Data collection was done through a cross-sectional questionnaire survey during the academic schedule of the colleges, supervised and monitored by the investigators themselves. The collected data were coded and entered in Microsoft excel 2013, and statistical analysis was done by SPSS 20 version. RESULT: A majority of the respondents agreed that they were not provided with sufficient information about implant treatment procedures during their BDS program (65.3%), would like more to be provided in the curriculum (95.1%), and would like to get additional reliable information from dental consultants and specialists (40.7%) and training on it from fellowship programs conducted by universities (39.2%). Significant association was seen between the responses and academic levels. CONCLUSION: Undergraduate dental students of Nepal want more information about dental implants through various means.

A comparative study to find out the relationship between the inner inter-canthal distance, interpupillary distance, inter-commissural width, inter-alar width, and the width of maxillary anterior teeth in Aryans and Mongoloids.

BACKGROUND: One of the most confusing and difficult aspects of complete denture prosthodontics is the selection of appropriately sized maxillary anterior denture teeth. Various guidelines have been suggested for determining the size of anterior teeth, but different opinions have been reported regarding their significance. In the study reported here, the relationships between facial measurements and the width of maxillary anterior teeth in two ethnic groups, namely Aryans and Mongoloids, were determined. OBJECTIVE: The aims of the study were to determine the inner inter-canthal distance (ICAD), inter-pupillary distance (IPD), inter-commissural width (ICOW), inter-alar width (IAW), and the combined width of maxillary anterior teeth (CW) in Aryans and Mongoloids and to determine the relationships between these measurements. MATERIALS AND METHODS: Impressions of the teeth of 170 dentulous subjects (85 males and 85 females) were made with alginate then Type IV gypsum product was poured in. Measurements of the cast maxillary anterior teeth at their widest dimension (contact areas) were made with the Boley gauge. ICAD, IPD, ICOW, and IAW distances were also measured with a Boley gauge. RESULTS: For all 170 subjects, 85 Aryans and 85 Mongoloids, Pearson's correlation coefficient (r) for IAW, IPD, ICOW, ICAD, and CW was calculated. In Aryans, highly significant (P<0.001) but weak correlations were found between CW and IAW, IPD, and ICOW. In Mongoloids, a highly significant (P<0.001) and weak correlation was found only between CW and IPD. CONCLUSION: Within the limitations of this study, the results suggest that the IAW, IPD, and ICOW for Aryans and IPD for Mongoloids can be used as a preliminary method for determining the width of the maxillary anterior teeth in edentulous patients.

Early treatment of Class III malocclusion by RME and modified Tandem appliance.

Patients with a skeletal Class III malocclusion and maxillary deficiency can be treated successfully using a combined protraction facemask and alternate rapid maxillary expansions and contractions (Alt-RAMEC). However, due to poor patient compliance during facemask therapy there has been growing interest in intraoral appliances for correcting Class III malocclusion. The tandem traction bow appliance (TTBA) is an intraoral appliance which has been used successfully for the treatment of growing Class III patients. This case report describes the management of a 10-year-old boy with a Class III malocclusion and maxillary deficiency treated with modified TTBA appliance.

Soft tissue airway dimensions and craniocervical posture in subjects with different growth patterns.

OBJECTIVE: To compare the dimensions of the nasopharynx and oropharynx of subjects with different growth patterns and to determine whether any correlation exists with their craniocervical posture. MATERIALS AND METHODS: Cephalometric radiograph of 60 subjects (16-25 years old), taken in natural head position, were divided into three groups according to the mandibular plane angle: hypodivergent (SN/MP <26°), normodivergent (SN/MP 26°-38°), and hyperdivergent (SN/MP <38°). Correlations were calculated between nasopharyngeal area, oropharyngeal area, and craniocervical posture. Continuous variables were compared by one-way analysis of variance, and the significance of mean difference between the groups was done by the Tukey post hoc test. A value of P < .05 was considered statistically significant. RESULTS: Patients in the hyperdivergent group were found to have significantly smaller nasopharyngeal and oropharyngeal areas than the other groups (P < .001 and P < .05, respectively). Similarly, the oropharyngeal area in the normodivergent group was significantly smaller than that in the hypodivergent group (P < .05). However, no significant differences were found in the nasopharyngeal area between the hypodivergent and normodivergent groups and between the hyperdivergent and normodivergent groups (P > .05). Reduced pharyngeal airways were typically seen in patients with a large craniocervical angle and a large mandibular inclination. CONCLUSIONS: Smaller nasopharyngeal and oropharyngeal airways were seen in connection with a large craniocervical and a large mandibular inclination. We therefore suggest that the vertical skeletal pattern may be one of the factors that contribute to nasopharyngeal and oropharyngeal obstruction.

Globalization and Livestock Biosecurity.

Globalization has resulted in enhanced trade in livestock and livestock products leading to increased risk of diseases to livestock and human beings. The emergence of highly contagious viral diseases of livestock and poultry such as foot-and-mouth disease, peste des petits ruminants, African swine fever (ASF), Newcastle disease, avian influenza and zoonotic diseases caused by viruses like Ebola, West Nile, Nipah, Hendra and swine influenza (H1N1) have necessitated the formulation of policies and regulatory frameworks for preventing ingress of exotic diseases and controlling dissemination of endemic diseases within the country. Biosecurity measures are important to maintain and improve animal health and reduce the risks. In this review, the risks of introduction of infections through livestock and livestock products, routes of transmission and general biosecurity measures to reduce these risks have been discussed.

Laser in dentistry: An innovative tool in modern dental practice.

The term LASER is an acronym for 'Light Amplification by the Stimulated Emission of Radiation'. As its first application in dentistry by Miaman, in 1960, the laser has seen various hard and soft tissue applications. In the last two decades, there has been an explosion of research studies in laser application. In hard tissue application, the laser is used for caries prevention, bleaching, restorative removal and curing, cavity preparation, dentinal hypersensitivity, growth modulation and for diagnostic purposes, whereas soft tissue application includes wound healing, removal of hyperplastic tissue to uncovering of impacted or partially erupted tooth, photodynamic therapy for malignancies, photostimulation of herpetic lesion. Use of the laser proved to be an effective tool to increase efficiency, specificity, ease, and cost and comfort of the dental treatment.