Genome Sequence Analysis of Calcifying Bacteria Bacillus paranthracis CT5 and Its Biomineralization Efficacy to Improve the Strength and Durability Properties of Civil Structures.

Bacteria producing urea amidohydrolases (UA) and carbonic anhydrases (CA) are of great importance in civil engineering as these enzymes are responsible for microbially induced calcium carbonate precipitation (MICCP). In this investigation, genomic insights of Bacillus paranthracis CT5 and the expression of genes underlying in MICCP were studied. B. paranthracis produced a maximum level of UA (669.3 U/ml) and CA (125 U/ml) on 5th day of incubation and precipitated 197 mg/100 ml CaCO3 after 7 days of incubation. After 28 days of curing, compressive strength of bacterial admixed and bacterial cured (B-B) specimens was 13.7% higher compared to water-mixed and water-cured (W-W) specimens. A significant decrease in water absorption was observed in bacterial-cured specimens compared to water-cured specimens after 28 days of curing. For genome analysis, reads were assembled de novo producing 5,402,771 bp assembly with N50 of 273,050 bp. RAST annotation detected six amidohydrolase and three carbonic anhydrase genes. Among 5700 coding sequences found in genome, COG gene annotation grouped 4360 genes into COG categories with highest number of genes to transcription (435 genes), amino acid transport and metabolism (362 genes) along with cell wall/membrane/envelope biogenesis and ion transport and metabolism. KEGG functional classification predicted 223 pathways consisting of 1,960 genes and the highest number of genes belongs to two-component system (101 genes) and ABC transporter pathways (98 genes) enabling bacteria to sense and respond to environmental signals and actively transport various minerals and organic molecules, which facilitate the active transport of molecules required for MICCP.

Isolation and characterization of thermostable and alkali-tolerant cellulase from litter endophytic fungus Bartalinia pondoensis.

Endophytic fungi in plant tissues produce a wide range of secondary metabolites and enzymes, which exhibit a variety of biological activities. In the present study, litter endophytic fungi were isolated from a fire-prone forest and screened for thermostable cellulases. Among nine endophytic fungi tested, two isolates, Bartalinia pondoensis and Phoma sp., showed the maximum cellulase activity. Bartalinia pondoensis was further selected for its cellulase production and characterization. Among the carbon and nitrogen sources tested, maximum cellulase production was observed with maltose and yeast extract, and the eucalyptus leaves and rice bran served as the best natural substrates. The cellulase activity increased with increasing temperature, with maximum activity recorded at 100 °C. The maximum CMCase activity was observed between pH 6.0 and 7.0 and retained 80% of its activity in the pH range of 8-10. Partially purified cellulase of B. pondoensis retained 50% of its activity after 2 h of incubation at 60 °C, 80 °C and 100 °C. These results suggest that litter endophytic fungus B. pondoensis is a potential source for the production of thermostable and alkali-tolerant cellulase.

Evaluation of immunohistochemical expression of epithelial- mesenchymal transition markers E-cadherin, Twist and Snail in oral submucous fibrosis and their possible association with malignant transformation.

BACKGROUND: Oral submucous fibrosis (OSMF) is a chronic, potentially malignant condition of the oral mucosa, predominantly seen in people of Asian descent. The reported malignant transformation rate of OSMF is 7%-13%. In the context of the understanding progression of OSMF, the study of prime molecular expressions is essential. Various markers have received more attention, one of them is E-cadherin. Various factors which promote epithelial-mesenchymal transition (EMT) and inhibit E-Cadherin include Snail1, Snail2, Twist and EF1/ZEB1. The intended study was undertaken to evaluate the possible role of E-cadherin and its regulatory markers Twist1 and Snail1 in OSMF. AIMS AND OBJECTIVES: To evaluate the expression of E-cadherin, Twist1 and Snail1 in OSMFTo evaluate their possible association with malignant transformation of OSMF. MATERIALS AND METHODS: Histologically proven 20 cases of each OSMF with and without dysplasia were taken as the study group, 20 normal mucosa as control group and were subjected for immunohistochemical (IHC) expression with E-cadherin, Twist1 and snail1. RESULTS: Immunohistochemical expression of all the three markers showed statistically significant expression of all the three markers. Intensity and percentage of staining between the groups were statistically significant for E-cadherin between normal oral mucosa (NOM) and OSMF with dysplasia (OSMFD), no significance was found between NOM and OSMF, whereas Snail1 and Twsit1 were statistically significant between NOM and OSMF and also between NOM and OSMFD. However, no significance was found for all the three markers when compared between the groups OSMF and OSMFD. CONCLUSION: The increased expression of Snail1 and Twist1 with concomitant loss of E -cadherin in OSMF suggests the role of the EMT.

"SKILL TO KILL" - Oral cancer and potentially premalignant oral epithelial lesions (PPOELs): A survey approach. Emerging of a new system and professionals.

BACKGROUND: Oral cancer is said to be the 6th most common cancer in men and 12th in women. Potentially malignant disorders/potentially premalignant oral epithelial lesion (PMDs/PPOELs) have shown an increased risk of progressing to cancer. In this regard, lack of awareness about identification of oral PMDs among healthcare providers in general and oral pathologists in particular is said to be responsible for the diagnostic delay. Oral cancer is said to be the most common cancer in men and 3rd most common in women in the Indian subcontinent. PMDs have shown an increased risk of progressing to cancer. The various rates of conversion include 14%-51% for erythroplakia, 60%-100% for proliferative verrucous leukoplakia and 7%-26% for oral submucous fibrosis (OSMF). In this regard, early detection at PPOEL level will lead to prevention of malignant transformation. AIMS AND OBJECTIVES: The purpose of this survey was: 1. To determine if awareness among professionals is essential/key to challenge the progression of PMDs of the oral cavity. 2. To examine and/ordiscern if oral pathologists felt enough was being done to address the issue of early detection and prevention and how the scenario could be improved further. MATERIALS AND METHODS: A survey was designed to assess the interest, zeal, knowledge and skill of healthcare providers in general and oral pathologists in particular in careful examination of the oral cavity and early detection of PMDs. A questionnaire with 18 questions was designed to address these issues/points and distributed among post-graduate students and practicing oral pathologists on online platforms. RESULTS: The results of the esurvey were collected, analyzed and the results discussed question wise. The validity and reliability of the questionnaire was assessed and confirmed with Aiken's index for validity and Cronbach's alpha for reliability. Inferential statistical analysis was performed using Chi square test with P = 0.05 being statistically significant. CONCLUSION: On evaluation of the survey, we found that 85% of the surveyees are on agreement that PPOELs should be registered in a standard format and should be included in the list of recognizable diseases. 89.2% and 87.8% of the respondents would like to be part of an active body for early detection and diagnosis and for Cancer screening in our country respectively. In conclusion, the oral pathology fraternity is eager to work for and tackle these PPOELs head on, provided the right opportunities and training are meted out to them.

Investigation of hub genes and their nonsynonymous single nucleotide polymorphism analysis in Plasmodium falciparum for designing therapeutic methodologies using next-generation sequencing approach.

BACKGROUND: Incidences of resistance to current drugs by Plasmodium is increasing, hence, it is necessary to investigate and explore new drug targets to combat malarial disease. OBJECTIVE: Analysis of the transcriptome sequence information to characterize hub genes and their nonsynonymous single nucleotide polymorphisms (nsSNPs) to derive therapeutic objectives for Plasmodium falciparum. MATERIALS AND METHODS: Differentially expressed genes between Ring and other stages of P. falciparum were identified using Cufflinks tool. Using DAVID and KAAS programs, the gene ontology and pathway analysis were performed. The networks of protein-protein interaction (PPI) were developed by Search Tool for the Retrieval of Interacting Genes/Proteins and Cytoscape, and the node degree in the network was calculated by using Network Analyzer, and MCODE plugins of Cytoscape. SIFT, PROVEAN, and PredictSNP programs were used to study the genetic variations, which affect protein functions. RESULTS: A list of 4196 nonredundant genes was used for functional annotation cluster analysis, and 8 significant hub genes have been picked from the PPI network using MCODE plugins of Cytoscape. Various nsSNPs were identified in these 8 hub genes and were investigated both for its native and mutant stage for solvent accessibility and alteration in secondary structure protein residues. CONCLUSION: Hub genes identified in this study serve as potential targets to develop therapy to suppress the pathogenic action of P. falciprum through experimental techniques.

The synergy of tobacco and alcohol and glutathione S-transferase θ 1 gene deletion and oral squamous cell carcinoma.

BACKGROUND: Oral squamous cell carcinoma (OSCC) is the leading cancer among males in India. It is related to tobacco habits and alcohol consumption as well as the individual susceptibility for xenobiotic metabolizing enzyme polymorphisms. Glutathione S-transferase θ 1 (GSTT1) is a Phase II metabolic enzyme which is directly involved in catalyzing chemicals to mutagenic intermediates. This gene is characterized by genetic polymorphism resulting in complete gene deletion and subsequent absence of the enzyme, which ultimately dictates the risk of cancer development. Scraping buccal mucosa to obtain DNA from the cells is a simple, readily acceptable and rapid method to detect and assess the gene. AIM: To assess GSTT1 gene deletion in individuals giving a history of tobacco smoking and/or chewing and alcohol consumption and absence of clinically detectable lesions; and in OSCC cases to gauge if GSTT1 gene deletion confers protection to an individual and whether it can be used as a "single" marker to arrive at this conclusion. To validate the use of buccal scrape for determining the genotype of an individual by assessing the polymorphism at GSTT1 gene locus (22q11.2). MATERIALS AND METHODS: Fifty-two cases were evaluated using buccal mucosal scrapes of tobacco habituates for 8 or more years, without clinically evident lesion (Group I) and from mucosa of tobacco habituates with clinically evident and histopathologically confirmed OSCC (Group II). DNA extraction and genotype at GSTT1 gene locus was determined by polymerase chain reaction assay. STATISTICAL ANALYSIS: The results were statistically analyzed using Chi-square test. RESULTS: 90.66% of subjects had GSTT1 null genotype in Group I subjects. In Group II, subjects with both clinically and histopathologically diagnosed oral cancer, about 76.96% had GSTT1 null genotype. CONCLUSION: GSTT1 null genotype confers protection to individuals with tobacco habits and alcohol consumption, predominantly to those who used chewable form of tobacco and especially among female population. However, the influence of many other environmental, genetic and epigenetic factors should be considered for the genesis/occurrence of cancer.

Quantification of Porphyromonas gingivalis in chronic periodontitis patients associated with diabetes mellitus using real-time polymerase chain reaction.

INTRODUCTION: Periodontal diseases, if left untreated, can lead to tooth loss and affect at least one tooth in 80% of adults worldwide, with the main cause being a bacterial plaque. Among subgingival plaque bacterial species, Porphyromonas gingivalis has been implicated as a major etiological agent causing tooth loss. Diabetics and smokers are two patient groups at high risk for periodontal disease. The increase in the number of this organism with the coexistence of other pathogenic microbes leads to rapid destruction of the periodontium, premature loss of teeth and also because of its virulence has implications in systemic pathology. Our aim was to observe the involvement of P. gingivalis in diabetes mellitus (DM) patients associated with periodontitis with and without tobacco-associated habits and to compare them with periodontitis patients having no other systemic pathologies. MATERIALS AND METHODS: Subgingival plaque samples from a total of seventy subjects were included in the study. DNA was isolated from the collected sample and was quantified using spectrophotometer for standardizing the polymerase chain reaction. The quantity of the isolated DNA was checked in a ultraviolet-visible spectrophotomer. STATISTICS: One-way ANOVA and Tukey's multiple post hoc procedures were carried out. RESULTS: The maximum score of P. gingivalis was seen in periodontitis patients having DM, whereas the least score was seen in periodontitis patients having DM with tobacco smoking habit compared to the other groups. CONCLUSION: P. gingivalis count is significantly reduced in periodontitis patients having DM with smoking habit; it is concluded that P. gingivalis might not be a key causative organism responsible for the periodontal destruction in case of smokers despite the DM condition. The decrease in counts may be attributed to change in the local environment like chemical (tobacco nitrosamines) and physical changes preventing the growth of P. gingivalis.

Adjunctive role of dental restorations in personal identification of burnt victims.

BACKGROUND: Fire remains one of the major causes of morbidity and mortality throughout the world and identification of a body from the fatal fire remains a daunting task. Several forensic cases involve interpretation of burnt human bodies from airline and automobile accidents, bombings and unlawful cremation. Fire is also involved in homicides, suicides, accidental death, arson and in attempts to destroy forensic evidence in criminal cases. Soft tissue destruction from fire can be so extensive that conventional methods of identification may be impossible. However, teeth survive even high temperatures due to their resistant composition and so, obviously, the restorative material housed in the teeth are even more secure and can yield valuable information in personal identification. AIM: To assess the usefulness of most common restorations in personal identification in burnt cases. MATERIALS AND METHODS: The study was conducted on 40 extracted teeth which were divided into four groups (Group 1 - Unrestored teeth, Group 2 - Amalgam restored, Group 3 - Glass ionomer restored and Group 4 - Composite resin restored teeth. The effect of incineration at 200°C, 400°C, 600°C, 800°C, 1000°C for 15 min at each target temperature followed by subsequent cooling was studied. RESULTS: Amalgam restoration was resistant and intact even at 1000°C, whereas GIC and composite restoration are identifiable till 600°C, the residual cavity preparation leaves a clue for narrowing down the spectrum of identification.

Antioxidant and cytotoxic activity of carotenes produced by Dunaliella salina under stress.

Context Dunaliella salina Teodoresco (Dunaliellaceae) is one of the promising microalgae consumed as food and medicine for many years. Objective Dunaliella salina was grown under different stress conditions for enhancing carotene production. The carotene enriched extract was evaluated for antioxidant and cytotoxic activity. Materials and methods Carotene content was calculated under salinity, nitrogen and temperature stress conditions. Antioxidant activity was determined through DPPH assay by incubating the samples for 45 min with 250 µg/mL of extract and reducing power assay was performed with 50, 100, 150 and 200 µg/mL of extract. Cytotoxicity was determined by incubating ∼2 × 10(4) MCF-7 (breast cancer) cells with 250 µg of extract in each well for 72 h by MTT assay. Result Carotene content was significantly increased to 9.8 (3.5 M NaCl), 13.9 (37 °C), 8.2 (250 mM KNO3) and 10.6 µg/mL (nitrogen-depleted medium) as compared with 3.2 µg/mL in normal conditions (1.7 M NaCl, 0.75 mM KNO3 and 28 °C). Free radical scavenging activity increased at 3.0 and 3.5 M NaCl (27.8 and 57.5%, respectively), 37 °C (31.4%) and in nitrogen-depleted medium (41.9%) compared with normal (15%) conditions. Carotene content and scavenging activity were positively correlated under salinity (r = 0.97), temperature (r = 0.85) and nitrogen (r = 0.7) stress conditions. Cytotoxicity against MCF-7 cell lines increased due to increase in carotene content suggesting that cytotoxicity may be associated with carotene accumulation. Discussion and conclusions Carotene content enhanced by D. salina under stress conditions increased the antioxidant and cytotoxic activity.

A phase contrast cytomorphometric study of squames of normal oral mucosa and oral leukoplakia: Original study.

UNLABELLED: Oral leukoplakia represents the most common potentially malignant oral disorder, representing 85% of such lesions. The worldwide prevalence of leukoplakia is 1.5- 4.3%. Leukoplakia is often associated with carcinogenic exposures, such as from use of tobacco, alcohol or betel nut. The level of risk for malignant transformation of leukoplakia is associated with lesion histology. The overall malignant transformation rates for dysplastic lesions range from 11% to 36%, depending on the length of follow-up. Exfoliative cytology is a simple and minimally invasive method. Phase contrast microscope, an essential tool in the field of biology and medical research provides improved discrimination of cellular details. AIMS: To study and compare the cytomorphological and cytomorphometric features of squames obtained from the mucosa of normal individuals, tobacco habituates with and without clinically evident leukoplakia. To assess the role of phase contrast microscopy as an alternative and easy method of cytological evaluation of wet and unstained smears. MATERIALS AND METHODS: Fifty cases from each group were taken. Fixed, unstained smears were viewed under phase contrast microscope and were evaluated morphologically and morphometrically for nuclear and cellular diameters. RESULTS: The study showed a significant increase in the mean nuclear diameter and decrease in the mean cellular diameter. CONCLUSION: Cytomorphometric changes could be the earliest indicators of cellular alterations. This indicates that there could be a cause-effect relationship between tobacco and quantitative alterations.

Expression of Ki-67 in normal oral epithelium, leukoplakic oral epithelium and oral squamous cell carcinoma.

AIMS AND OBJECTIVE: To demonstrate the presence, location and pattern of cell proliferation in different histological grades of oral epithelial dysplasia (OED), oral squamous cell carcinoma (OSCC) and normal oral epithelium (NOE) using an antibody directed against the Ki-67 antigen and its intensity of staining evaluated respectively. MATERIALS AND METHODS: A total number of 100 archival paraffin embedded blocks obtained from Department of Oral and Maxillofacial Pathology were studied. The case details were retrieved which consisted of histopathologically diagnosed cases of OSCC (n = 20), low risk OED (n = 30), high risk OED (n = 30) and normal appearing mucosa (n = 20) were taken as standard for comparison. Ki-67 immunostaining was detected. Ki-67 positive cells were counted in the five random high power fields in each case. RESULTS: Ki-67 labeling Index (LI) was restricted to the basal and parabasal layers of the normal oral epithelium irrespective of age, sex and site whereas it was seen in the basal, suprabasal and spinous layers in OED. Ki-67 LI is increased in high risk cases than the low risk cases of OED. Ki-67 positive cells in OSCC were located in the periphery of the tumor nests than the center, where frequent mitoses were observed. CONCLUSION: The architectural alteration evaluated by Ki-67 antibody in proliferating cell distribution in the layers of epithelial dysplasias may provide useful information to evaluate the grading of OED. Ki-67 LI increased in high risk cases than low risk cases of OED. This study showed that over expression of Ki-67 antigen between well-differentiated and poorly differentiated OSCC was in accordance with histologic grade of malignancy but not in accordance with moderately differentiated OSCC.

Chondromyxoid fibroma of zygoma: A rare case report.

Chondromyxoid fibroma (CMF) is a rare benign mesenchymal tumor of the bone. Clinically, it is characterized by a lobular growth pattern and histologically by chondroid and myxoid differentiation. The tumor is rare in the craniofacial bones with only 2% of all reported cases. Extragnathic location in the facial skeleton is extremely rare. Most of the cases reported either originate from gnathic sites or in the cranium. A case of CMF in a 3½-year-old male is presented here, which arose from the root of zygomatic arch. A detailed clinical history and histopathological picture of one more case is added to the literature. It is important to document such cases so that better light can be shed on future reviews and conclusions. This shall facilitate better treatment approaches and prognosis. This case is the first reported case of involvement of the zygomatic arch in a pediatric patient.

Leiomyoma: A rare tumor in the head and neck and oral cavity: Report of 3 cases with review.

Leiomyomas are benign tumors arising from smooth muscle, most commonly seen in uterine myometrium, gastrointestinal tract, skin and lower extremities of middle-aged women. Leiomyomas are uncommon in the oral cavity with reported incidence of 0.065%, which accounts for 0.42% of all soft-tissue neoplasms in the oral cavity. Leiomyomas of head and neck region account for less than 1% of all leiomyomas. The most common site of leiomyoma in the head and neck region is the lips (27.46%) followed by tongue (18.30%), cheeks and palate (15.49%), gingiva (8.45%) and mandible (5.63%). The purpose of this article is to present three cases of leiomyoma comprising of an intraoral vascular leiomyoma and two solid leiomyomas in the head and neck region. The clinical features, etiology, differential diagnosis and treatment of leiomyoma are discussed with review of the literature.

Primary melanoma of oral mucosa: A case report and review of literature.

Primary oral melanoma is a rare neoplasm of melanocytic origin, accounting for 0.5% of all oral malignancies. The "chameleonic" presentation of a mainly asymptomatic condition, rarity of this lesion, poor prognosis, and the necessity of a highly specialized treatment are factors that should be seriously considered by the involved health provider. Here is a case report presenting a malignant melanoma of oral mucosa in 48-year-old male patient on maxillary gingiva. The lesion was removed by partial maxillectomy and patient is disease free after 11 months of regular followup. This case provides an example of how dental clinicians play a major role in the identification of pigmented lesions of oral cavity and also emphasize on the fact that any pigmented lesion detected in the oral cavity may exhibit potential growth and should be submitted to biopsy to exclude malignancy.

Clear cell unicystic ameloblastoma.

Clear cell differentiation in unicystic ameloblastoma with inclusion of many other histologic variants in the same tumor is a very rare occurrence. Here, we report a case of a well-circumscribed large mandibular swelling in a 22 - year old female. The lesion was histopathologically diagnosed as unicystic ameloblastoma which showed multiple histologic patterns and clear cell differentiation. The tumor was treated with surgical enucleation and chemical cauterization. A follow up of 20 months has shown no recurrence after initial surgery.