Assessment of the mineralisation stages of third molars and validation of Mincer et al.'s age estimation method: A retrospective, cross-sectional study in Western India population.

Context: Age estimation is one of the prime requisites in forensic human identification cases and the criminal justice system. There are several age estimation methods using dental parameters. A method proposed by Mincer et al. which uses the mineralization stages of third molars based on Demirjian's developmental stages is less tested in the Indian population. Aim: The present study aimed to assess the developmental status of the third molars and to apply and validate Mincer et al.'s method on the Western India population. Methods and Material: A total of 306 orthopantomograms (OPGs) from 128 males and 178 females with a mean age of 16.89 years ± 3.68 were analysed. Demirjian's A-H staging was applied to record the developmental stages of 1100 third molars. Mincer et al.'s mean age of attainment was applied based on the American Whites (Caucasian) population for males and females separately using stages of #18 and #38. Results: There was a slight overestimation of the chronological age (CA) in both #18 and #38. The females showed more accurate estimated age (EA) than males. Conclusions: Mincer et al.'s method is a convenient age estimation method using the third molar's developmental stage. The mean age of attainment mentioned in the original Mincer et al.'s study can be used in the Western Indian population, with a residual value ranging from 0.21 to 0.25 years.

Does third molar impaction affect age estimation in adolescent and young adults? A radiographic population based study.

Few studies consider the effects of impaction, resulting in inaccurate estimation of age using third molar. We aimed to assess the significance of impaction in estimating age using Modified Demirjian's method. 200 orthopantomograms of patients between 16-25 years were collected comprising 171 tests and 212 controls. Chronologic age (CA) and estimated age (EA) were calculated. Independent sample t-test, chi-square test, paired t-test and Kolmogrov-Smirnov test of normality were used. The mean difference between the CA and EA in both the tests and control samples were significant (mean difference test group = 2.6 ± 2.07, control group = 2.0 ± 1.8; p = 0.01). It provided a difference of 0.6 which is equal to 7.2 months. The results of our study indicate that impacted mandibular third molars are susceptible to undergo slower mineralization, leading to erroneous lower age estimation. These results in an Indian population are noteworthy. However further studies are needed.

Forensic genetics: Scope and application from forensic odontology perspective.

Forensic science corresponds to the employment of science to the law or legal matters. One of the major concerns of legal system is to deduce the recognition of an item or individual, involved in crime for which forensic expert plays a pivotal role. Forensic odontology, a budding branch in dentistry, involves the application of dentistry to the legal system. The dental characteristics are considered as one of the primary characteristics of identification as per Interpol DVI guidelines. Thus, establishing the identity of unknown human remains through dental features is considered as one of the core domains of forensic odontology. However, its reliability and its acceptability in the court of law are only secondary to the application of DNA technologies. Also, the acceptability of bite-mark analysis and its evidentiary role is debatable. However, the bite marks may also be a source of salivary DNA, to establish the linking of the perpetrator to the victim. The recent advancements in the DNA technologies and the use of teeth and saliva as sources of DNA are the added advantages in the application of DNA as person identifiers especially in badly mutilated, decomposed and charred bodies and in linking the perpetrator to the crime. With this background, we present here a review on the application of forensic genetics from a forensic odontology point of view.

The association of fingerprint patterns with oral potentially malignant disorders and oral cancer: A dermatoglyphic study.

Background: Dermatoglyphics play an important role in the identification of a person as well as identifying a person with a genetic abnormality. Hence, the present study was conducted with an aim to explore the association between the fingerprint patterns and oral potentially malignant disorders, oral squamous cell carcinoma (OSCC), individuals with habit and without lesion and the control group. Materials and Methods: A cross-sectional study comprising of 100 individuals divided into 5 groups with 20 individuals in each group based on their habit and the presence of oral lesions. Group 1 included individuals with oral leukoplakia (OL), Group 2 included individuals with oral submucous fibrosis (OSMF), Group 3 included individuals who had OSCC, Group 4 individuals had tobacco habits but without lesion and Group 5 comprised of control individuals. The fingerprints of all the study subjects were recorded using a standard ink method. They were analyzed qualitatively and quantitively. Results: The present study showed a predominance of that loop patterns in all the groups. The frequency of the whorled pattern was higher in subjects with OSMF as compared to OL and OSCC. The arch pattern was comparatively lesser in subjects with OL than OSMF subjects. The right-left comparison of the fingerprint patterns in all the five groups, except Group 4, showed a significant association and correlation. The Group 4 individuals were showing the least correlation between sides. Conclusion: The study concluded that dermatoglyphics can be used as a potential marker to identify the subjects with risk for potentially malignant oral disorders and oral cancers.

Inter-observer agreement in the radiographic interpretation of Demirjian's developmental stages in the mandibular second and third molars - A comparative study.

Background: The developmental stages of the teeth in the radiographs are graded on an ordinal scale. The present study was conducted using 123 digital orthopantomograms from individuals in the age group of 5 to 22 years and to analyze and evaluate the inter-observer agreement in grading the developmental stages of second and third molars. Four observers with different levels of practical experience in age estimation participated in the study. The development stages of both the molars in the lower left quadrant (3rd quadrant) were assigned based on the Demirjian's 10 stage chart. The percentage agreement and Kappa statistics were used to test the agreement between the observers. The data of the observer 1 were used as the standard for the comparison. RESULTS: There was 70.0%-75.6% agreement among the observers in staging second molar and 52%-68.3% for the third molars. There was an excellent agreement (k > 080) between observer 1 and observer 2 and a good agreement (k = 060-0.79) between observer 1 and the other two observers for both the molars. The Fleiss Kappa revealed moderate to a good overall agreement for both the molars (k = 0.51-0.66). The Freidman's test revealed a significant difference in the grading of third molars between all the raters (χ2= 25.02, df 3, P < 0.001) and for the second molar the difference was not significant (χ2= 3.89, df 3, P > 0.05). The stage-wise overall agreement was fair for Stage 3 in the second molar and Stage 9 and Stage 4 in the third molar. Conclusions: In conclusion, proper training in the radiographic interpretation of developmental stages may minimize the errors during the age estimation methods.

Does the Surgeon-reported Outcome Correlate with Patient-reported Outcome after Total Knee Arthroplasty? A Cohort Study.

BACKGROUND: Total knee arthroplasty (TKA) has been proved to be a successful and cost-effective treatment for improving pain and function in patients with knee arthritis. Total knee arthroplasty (TKA) is one of the most common orthopaedic surgeries performed worldwide and advancement in surgical techniques and prosthetic designs have improved the patient outcomes. However, concerns and priorities of patients and surgeons relating to joint replacement may differ. MATERIALS AND METHODS: 306 TKAs in 223 patients were evaluated for functional outcome using surgeon reported American Knee Society Knee Score (KS)/Functional Score (FS) and patient-reported Oxford Knee Score (OKS). We have also assessed the correlation between FS and OKS at midterm follow up. RESULTS: The mean preoperative KS, FS, and OKS in 223 patients were 42.76, 42.4, and 38.84 and the midterm mean KS, FS, and OKS were 84.29, 73.40, and 30.26, respectively. There was a statistically significant improvement in the KS, FS, and OKS at midterm follow up in Category A (CAT A) (bilateral TKA or unilateral with asymptomatic contralateral knee), CAT B (unilateral TKA with symptomatic other knee) and CAT C (inflammatory arthritis). Overall, the correlation between the midterm FS and OKS was fair. However, in CAT A and CAT B, there was no significant correlation between FS and OKS, but CAT C had a strong correlation. There was a statistically significant improvement in the KS, FS, and OKS when midterm follow up scores were compared with preoperative scores. However, no significant correlation between the American knee society FS and OKS in osteoarthritic patients at midterm follow up signifies acceptable outcome may vary between patients and physicians. CONCLUSION: All patients should be counseled preoperatively to assess their expectations and sensitize them to information regarding the expected functional outcome following TKA in their cultural context.

Determination of normal maxillary transverse dimension by using intercanine width and interpalatal first molar width.

BACKGROUND: Malocclusion in the vertical and sagittal planes is easy to observe whereas it is difficult to diagnose malocclusion in the transverse plane. There are yet no criteria defining a normal transverse occlusion. AIM: The aim of this study was to identify a ratio that defines a normal transverse occlusion. STUDY DESIGN: This was a retrospective study. MATERIALS AND METHODS: A total sample of 151 maxillary dental models (M = 55, F = 96) in the age range of 12-16 years were randomly selected, with 73 models in control group and 78 in the study group. All the models were mixed up and given to an operator for the measurement of intercanine width (ICW) and inter-palatal molar width (IPMW). STATISTICAL ANALYSIS USED: The data were statistically analyzed using SPSS software version 16.0. Chi-square test was performed to test the statistical significance difference between the groups at p ≤ 0.05. RESULTS: Nearly 98.60% of normal arches were found to have an IPMW to ICW ratio of 1:1 ± 0.05. On further analysis, it was found that when IPMW ≥37.45 mm, then the case may be considered to have normal arches, and if the ratio between IPMW to ICW is 1.15:1 ± 0.05 or more and IPMW is <34.92 mm, then the case may be identified as deficient arch. CONCLUSION: The IPMW-to-ICW ratio can be used to differentiate deficient and normal arches. This hypothesis (Banker's hypothesis) is an easy and noninvasive method, using only two parameters, developed to help clinicians in identifying normal transverse dimension.

Efficacy of exogenous hormone (GnRHa) for induced breeding of climbing perch Anabas testudineus (Bloch, 1792) and influence of operational sex ratio on spawning success.

The climbing perch, Anabas testudineus, is an air-breathing fish having great consumer preference as a food fish and is considered a prime candidate species for aquaculture. Spawning success is an important issue while using hormones for captive induced breeding. In the first experiment, a trial was conducted to assess the efficacy of a synthetic Gonadotropin Releasing Hormone analog (sGnRHa) on the spawning success of climbing perch. Female fish were administered six different doses each with a single intramuscular injection of sGnRHa hormone at 0.002 (TOD1), 0.005 (TOD2), 0.01 (TOD3), 0.015 (TOD4), 0.02 (TOD5), 0.03 (TOD6) µg/g body weight. Similarly, males were administered half of the hormone dose of females in all the respective treatment groups. The greatest (P<0.05) relative fecundity (715.13±15.0 eggs/g female body weight) and fertilization percentage rates (93.1±8.0%) occurred when female fish were treated at the 0.015µg/g body weight dose. There was a reduction in relative fecundity and hatching rate in female fish injected with the largest dose (1.5µL/g body weight) of sGnRHa. A second experiment was conducted to assess the effect of a different male-female ratio on optimum spawning success in climbing perch. For this study a different female to male ratio (1:1, 1:2, 1:3 and 1:4) and male to female ratio (1:1, 1:2 and 1:3) were used. There were a greater (P<0.05) relative fecundity (886.62±17.9 eggs/g female body weight), fertilization (98±6.7%) and hatching (99±5.4%) rates with the female to male ratio of 1:2. This indicated that the hormone dose of 0.015µg/g body weight and a female-male ratio of 1:2 are optimal for enhanced spawning success in the climbing perch.

Carbon sequestration capacity of sediments, algae, and zooplankton from fresh water aquaculture ponds.

The contribution of aquaculture and allied activities to the emission of green house gases and consequently to global warming is an emerging concern among environmentalists in the recent past. However, there exists ample scope for aquaculture activities to sequester carbon and thus compensate for the carbon emissions linked to aquaculture. This article attempts to elucidate the carbon sequestration capacity of sediments, algae, and zooplankton from fresh water aquaculture ponds. The percent organic carbon in the pond sediments ranged from 0.39 to 1.31 with an average value of 0.912 ± 0.321 whereas the carbon sequestration capacity ranged from 0.442 to 1.882 MgC/ha (1 Mg = 10(6) g) with an average value of 1.018 ± 0.447 MgC/ha. In the case of zooplankton and algae from pond, the percent organic carbon was 7.688 ± 0.196 and 2.354 ± 0.047, respectively, whereas the total estimated carbon burial rate was 0.009 ± 0.005 and 0.150 ± 0.003 MgC/ha, respectively. These findings are discussed with the previous reports available at present and are found to be in comparable ranges.

Detection of goldfish haematopoietic necrosis herpes virus (Cyprinid herpesvirus-2) with multi-drug resistant Aeromonas hydrophila infection in goldfish: First evidence of any viral disease outbreak in ornamental freshwater aquaculture farms in India.

This outbreak report details of a mortality event where Cyprinid herpes virus-2 (CyHV-2) was detected in association with multidrug-resistant Aeromonas hydrophila infection in goldfish, Carassius auratus, from commercial farms. The goldfish exhibited large scale haemorrhages on the body, fins and gills, lepidorthosis, necrosed gills, protruded anus and shrunken eyes. White nodular necrotic foci in spleen and kidneys were noticed, along with necrosis and fusion of gill lamellae. Transmission electron microscopy of affected tissues revealed the presence of mature virus particles. Involvement of CyHV-2 was confirmed by PCR, sequencing and observed cytopathic effect in koi carp fin cell line along with experimental infection study. A bacterium isolated from the internal organs of affected fish was found to be pathogenic Aeromonas hydrophila having resistance to more than 10 classes of antibiotics. We postulate that CyHV-2 was the primary etiological agent responsible for this outbreak with secondary infection by A. hydrophila. The experimental infection trials in Labeo rohita and koi carp by intraperitoneal challenge with CyHV-2 tissue homogenates failed to reproduce the disease in those co-cultured fish species. This is the first report of a viral disease outbreak in organised earthen ornamental fish farms in India and bears further investigation.

Development of Antimicrobial Peptide Prediction Tool for Aquaculture Industries.

Microbial diseases in fish, plant, animal and human are rising constantly; thus, discovery of their antidote is imperative. The use of antibiotic in aquaculture further compounds the problem by development of resistance and consequent consumer health risk by bio-magnification. Antimicrobial peptides (AMPs) have been highly promising as natural alternative to chemical antibiotics. Though AMPs are molecules of innate immune defense of all advance eukaryotic organisms, fish being heavily dependent on their innate immune defense has been a good source of AMPs with much wider applicability. Machine learning-based prediction method using wet laboratory-validated fish AMP can accelerate the AMP discovery using available fish genomic and proteomic data. Earlier AMP prediction servers are based on multi-phyla/species data, and we report here the world's first AMP prediction server in fishes. It is freely accessible at http://webapp.cabgrid.res.in/fishamp/ . A total of 151 AMPs related to fish collected from various databases and published literature were taken for this study. For model development and prediction, N-terminus residues, C-terminus residues and full sequences were considered. Best models were with kernels polynomial-2, linear and radial basis function with accuracy of 97, 99 and 97 %, respectively. We found that performance of support vector machine-based models is superior to artificial neural network. This in silico approach can drastically reduce the time and cost of AMP discovery. This accelerated discovery of lead AMP molecules having potential wider applications in diverse area like fish and human health as substitute of antibiotics, immunomodulator, antitumor, vaccine adjuvant and inactivator, and also for packaged food can be of much importance for industries.

Complete mitochondrial genome sequence of Heteropneustes fossilis obtained by paired end next generation sequencing.

In the present study, the complete mitochondrial genome sequence of Heteropneustes fossilis is reported using massive parallel sequence technology. The complete mitogenome of H. fossilis is obtained by de novo assembly of paired end Illumina sequences using CLC Genomics Workbench version 7.0.4, which is 16,489 bp in length. It comprised of 13 protein- coding genes, 22 tRNAs, 2 rRNA genes and a putative control region along with the gene order and organization, being similar to most of vertebrates. The mitogenome in the present study has 99% similarity to the complete mitogneome sequence of H. Fossilis, as reported earlier. Phylogenetic analysis of Siluriformes depicted that Heteropneustids were closer to Clariids. The mitogenome sequence of H. fossilis contributes better understanding of population genetics, phylogenetics and evolution of Indian catfish species.

Complete mitochondrial genome sequence of Labeo fimbriatus (Bloch, 1975).

The complete mitochondrial genome of Labeo fimbriatus, the fringe-lipped carp, was determined for the first time. The genome was 16,614 bp in length and consisted of 13 protein-coding genes, 22 tRNA genes, 2 rRNA genes and one control region. Organization of genes and their order are in accordance with other vertebrates. The overall base composition on the plus strand was A: 32.35%, G: 15.46%, C: 27.54%, T: 24.647%, and the A + T content 57%. The control region contains a microsatellite, (TA)12, a putative termination-associated sequence and three conserved sequence blocks. This mitogenome sequence data would play an important role in population genetics and phylogenetics of cultivable carps of India.

Expression of Mx Gene in Cirrhinus mrigala (Hamilton, 1822) to OmpC Protein of Aeromonas hydrophila and Bacterial Infection.

The aims of this study were to identify alternative myxovirus (Mx) stimulatory compounds in Cirrhinus mrigala and to characterize the kinetics and intensity of their stimulated responses by semi-quantitative RT-PCR. Mx transcripts were measured in C. mrigala injected with Aeromonas OmpC (outer membrane protein) at a dose 0.4 mg/fish. At day 1, day 2, day 3, day 5, day 10, day 20 and day 30, samples were collected from kidney, spleen, liver, heart brain, gill, intestine and muscle for the study of Mx transcript and housekeeping gene ß-actin. Similarly, Mx gene expression was also studied in Aeromonas hydrophila-infected fish for a period of 10 days. Mx/ß-actin ratio was constitutively expressed from all the organs of OmpC-vaccinated fish. The expression was significantly highest (P ≤ 0.05) in spleen, followed by liver, kidney, intestine, gill, heart, muscle and brain. The expression was highest in day 2 except spleen (on day 3) and subsequently reduced up to day 30. Control fish also showed Mx expression. Similarly, A. hydrophila-infected fish showed Mx/ß-actin ratio upregulated significantly in the spleen and kidney on day 5, liver on day 2 and intestine on day 3. This study revealed that OmpC of A. hydrophila and its infection could stimulate the antiviral Mx gene of C. mrigala.

Low-depth shotgun sequencing resolves complete mitochondrial genome sequence of Labeo rohita.

Labeo rohita, popularly known as rohu, is a widely cultured species in whole Indian subcontinent. In the present study, we used in-silico approach to resolve complete mitochondrial genome of rohu. Low-depth shotgun sequencing using Roche 454 GS FLX (Branford, Connecticut, USA) followed by de novo assembly in CLC Genomics Workbench version 7.0.4 (Aarhus, Denmark) revealed the complete mitogenome of L. rohita to be 16 606 bp long (accession No. KR185963). It comprised of 13 protein-coding genes, 22 tRNAs, 2 rRNAs and 1 putative control region. The gene order and organization are similar to most vertebrates. The mitogenome in the present investigation has 99% similarity with that of previously reported mitogenomes of rohu and this is also evident from the phylogenetic study using maximum-likelihood (ML) tree method. This study was done to determine the feasibility, accuracy and reliability of low-depth sequence data obtained from NGS platform as compared to the Sanger sequencing. Thus, NGS technology has proven to be competent and a rapid in-silico alternative to resolve the complete mitochondrial genome sequence, thereby reducing labors and time.

Complete mitochondrial genome sequence of E. suratensis revealed by next generation sequencing.

The complete mitochondrial genome of Etroplus suratensis, the Green chromide cichlid, was determined for the first time through NGS method. The genome is 16,467 bp (Accession no. KU301747) in length and consisted of 13 protein-coding genes, 22 tRNA genes, 2 rRNA genes and one control region. Organization of genes and their order are in accordance with other vertebrates. The overall base composition on plus strand was A: 28.3%, G: 15.2%, C: 30.9%, T: 25.6%, and the A + T content 53.9%. The control region contains a putative termination-associated sequence and three conserved sequence blocks. This mitogenome sequence data would play an important role in population genetics and phylogenetics of cichlid fish of India.

Assembly and variation analyses of Clarias batrachus mitogenome retrieved from WGS data and its phylogenetic relationship with other catfishes.

Whole genome sequencing (WGS) using next generation sequencing technologies paves the way to sequence the mitochondrial genomes with greater ease and lesser time. Here, we used the WGS data of Clarias batrachus, generated from Roche 454 and Ion Torrent sequencing platforms, to assemble the complete mitogenome using both de novo and reference based approaches. Both the methods yielded almost similar results and the best assembled mitogenome was of 16,510 bp size (GenBank Acc. No. KM259918). The mitogenome annotation resulted in 13 coding genes, 22 tRNA genes, 2 rRNA genes and one control region, and the gene order was found to be identical with other catfishes. Variation analyses between assembled and the reference (GenBank Acc. No. NC_023923) mitogenome revealed 51 variations. The phylogenetic analysis of coding DNA sequences and tRNA supports the monophyly of catfishes. Two SSRs were identified in C. batrachus mitogenome, out of which one was unique to this species. Based on the relative rate of gene evolution, protein coding mitochondrial genes were found to evolve at a much faster pace than the d-loop, which in turn are followed by the rRNAs; the tRNAs showed wide variability in the rate of sequence evolution, and on average evolve the slowest. Among the coding genes, ND2 evolves most rapidly. The variations present in the coding regions of the mitogenome and their comparative analyses with other catfish species may be useful in species conservation and management programs.

Preliminary genetic linkage map of Indian major carp, Labeo rohita (Hamilton 1822) based on microsatellite markers.

Linkage map with wide marker coverage is an essential resource for genetic improvement study for any species. Sex-averaged genetic linkage map of Labeo rohita, popularly known as 'rohu', widely cultured in the Indian subcontinent, was developed by placing 68 microsatellite markers generated by a simplified method. The parents and their F1 progeny (92 individuals) were used as segregating populations. The genetic linkage map spans a sex-averaged total length of 1462.2 cM, in 25 linkage groups. The genome length of rohu was estimated to be 3087.9 cM. This genetic linkage map may facilitate systematic searches of the genome to identify genes associated with commercially important characters and marker-assisted selection programmes of this species.

In-silico analysis of heat shock protein 47 for identifying the novel therapeutic agents in the management of oral submucous fibrosis.

BACKGROUND: Heat shock proteins-47 (HSP47) is a collagen specific molecular chaperone, involved in the processing and/or secretion of procollagen. It seems to be regularly upregulated in various fibrotic or collagen disorders. Hence, this protein can be a potential target for the treatment of various fibrotic diseases including oral submucous fibrosis (OSF), which is a collagen metabolic disorder of oral cavity and whose etiopathogeneic mechanism and therapeutic protocols are still not well documented. AIM: The aim of this study is to identify the novel therapeutic agents using in-silico methods for the management of OSF. OBJECTIVES: The objectives of this study are to identify the binding sites of HSP47 on the collagen molecule and to identify the lead compound with anti-HSP47 activity from the library of natural compounds, using in-silico methodology. MATERIALS AND METHODS: The web-based and tool based in-silico analysis of the HSP47 and collagen molecules are used in this study. The crystal structure of collagen and HSP47 were retrieved from Protein Data Bank website. The binding site identification and the docking studies are done using Molegro Virtual Docker offline tool. RESULTS: Out of the 104 Natural compounds, six ligands are found to possess best binding affinity to the binding amino acid residues. Silymarin binds with the 4AU2A receptor and the energy value are found to be -178.193 with four Hbonds. The other best five natural compounds are hesperidin, ginkgolides, withanolides, resveratrol, and gingerol. Our findings provide the basis for the in-vitro validation of the above specified compounds, which can possibly act as "lead" molecules in designing the drugs for OSF. CONCLUSION: HSP47 can be a potential candidate to target, in order to control the production of abundance collagen in OSF. Hence, the binding sites of HSP47 with collagen are identified and some natural compounds with a potential to bind with these binding receptors are also recognized. These natural compounds might act as anti-HSP47 lead molecules in designing novel therapeutic agents for OSF, which are so far unavailable.