Wnt Signaling Pathway Collapse upon ß-Catenin Destruction by a Novel Antimicrobial Peptide SKACP003: Unveiling the Molecular Mechanism and Genetic Activities Using Breast Cancer Cell Lines.

Despite progress in breast cancer treatment, the survival rate for patients with metastatic breast cancer remains low due to chemotherapeutic agent resistance and the lack of specificity of the current generation of cancer drugs. Our previous findings indicated that the antimicrobial peptide SKACP003 exhibited anticancer properties, particularly against the MCF-7, MDA-MB-231, and MDA-MB-453 breast cancer cell lines. However, the mechanism of SKACP003-induced cancer cell death is unknown. Here, we investigated the molecular mechanism by which SKACP003 inhibits the cell cycle, cell proliferation, and angiogenesis in breast cancer cell lines. The results revealed that all the breast cancer cell lines treated at their IC50 values significantly inhibited the replicative phase of the cell cycle. The SKACP003-induced growth inhibition induced apoptosis, as evidenced by a decrease in BCL-2 and an increase in BAX and caspase gene (Cas-3, Cas-8, and Cas-9) expression. Reduced expression of the ß-Catenin signaling pathway was associated with the SKACP003-induced apoptosis. SKACP003-treated breast cancer cells showed decreased expression of Wnt/ß-Catenin targeting genes such as C-Myc, P68, and COX-2 and significant downregulation of CDK-4 and CDK-6 genes. Furthermore, cytoplasmic ß-catenin protein levels in SKACP003-treated cell lines were significantly lower than in control cell lines. The results of the current study suggest that the newly identified antimicrobial peptide SKACP003 has great potential as a candidate for specifically targeting the ß-catenin and thus significantly reducing the progression and prognosis of breast cancer cell lines.

Knockdown of sirtuin6 positively regulates acetylation of DNMT1 to inhibit NOTCH signaling pathway in non-small cell lung cancer cell lines.

BACKGROUND AND AIM: Sirtuin proteins (1-7) are nicotinamide adenine dinucleotide (NAD)-dependent deacetylases and ADP-ribosyl transferases (class III histone deacetylase enzymes (HDAC)) mainly involved in the removal of the acetyl group from histone proteins. SIRT6, one of the sirtuins, plays a major role in cancer progression in many types of cancer conditions. We recently reported that SIRT6 acts as an oncogene in NSCLC; thus, silencing of SIRT6 inhibits cell proliferation and induces apoptosis in NSCLC cell lines. NOTCH signaling has been reported to be involved in cell survival and regulates cell proliferation and differentiation. However, recent studies from different groups have converged on the notion that NOTCH1 may be an important oncogene in NSCLC. The abnormal expression of NOTCH signaling pathway members is a relatively frequent event in patients with NSCLC. SIRT6 and the NOTCH signaling pathway might play a critical role in tumorigenesis since they are highly expressed in NSCLC. This study has been performed to explore the exact mechanism by which SIRT6 inhibits cell proliferation and induces the apoptosis of NSCLC cell lines and its correlation with NOTCH signaling. MAIN METHODS: In vitro experiments with human NSCLC cells have been performed. Immunocytochemistry study was used to analyze the expression of NOTCH1 and DNMT1 in A549 and NCI-H460 cell lines. RT-qPCR, Western Blot, Methylated DNA specific PCR, and Co-Immunoprecipitation were performed to explore the key events in the regulation of NOTCH signaling by silencing SIRT6 in NSCLC cell lines. KEY FINDINGS: The findings of this study suggest that silencing of SIRT6 significantly promotes the acetylation status of DNMT1 and stabilizes it. Consequently, acetylated DNMT1 translocates into the nucleus and methylates the NOTCH1 promoter region, resulting in the hindering of NOTCH1-mediated NOTCH signaling.

Bactericidal and non-cytotoxic activity of bacteriocin produced by Lacticaseibacillus paracasei F9-02 and evaluation of its tolerance to various physico-chemical conditions.

This study aims to explore novel lactic acid bacteria (LAB) from breast-fed infants' faeces towards characterizing their antimicrobial compound, bacteriocin. The LAB, Lacticaseibacillus paracasei F9-02 showed strong antimicrobial activity against clinical pathogens. Their proteinaceous nature was confirmed as the activity was completely abolished when treated with proteinaceous enzymes and retained during neutral pH and catalase treatment. The purified bacteriocin showed antimicrobial activity at the minimum inhibitory concentration (MIC) value of 7.56 µg/ml against vancomycin-resistant Enterococcus sp. [vancomycin-resistant enterococcal (VRE)], and methicillin-resistant Staphylococcus aureus (MRSA), 15.13 µg/ml against Klebsiella pneumoniae, Pseudomonas aeruginosa, Salmonella enterica subsp. enterica serotype typhi and 30.25 µg/ml against Shigella flexneri. Present study also proved the bactericidal, non-cytotoxic and non-hemolytic nature of bacteriocin. Additionally, bacteriocin retained their stability under various physico-chemical conditions, broad range of pH (2-10), temperature (40-121°C), enzymes (amylase, lipase and lysozyme), surfactants [Tween-20, 80, 100 and sodium dodecyl sulfate (SDS)], metal ions (CaCl2 , FeSO4 , ZnSO4 , MgSO4 , MnSO4 , CuCl2 ) and NaCl (2%-8%). The molecular weight of bacteriocin (~28 kDa) was determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), functional and active groups were assessed by Fourier Transform-Infrared (FT-IR). To our knowledge, this is the first study reporting L. paracasei from breast-fed infants' faeces and assessing their antimicrobial compound, bacteriocin. The study results furnish the essential features to confirm the therapeutic potential of L. paracasei F9-02 bacteriocin.

Phenotypic Assessment of Probiotic and Bacteriocinogenic Efficacy of Indigenous LAB Strains from Human Breast Milk.

Breast milk is the combination of bioactive compounds and microflora that promote newborn's proper growth, gut flora, and immunity. Thus, it is always considered the perfect food for newborns. Amongst their bioactives, probiotic communities-especially lactic acid bacteria (LAB)-are characterized from breast milk over the first month of parturition. In this study, seven LAB were characterized phenotypically and genotypically as Levilactobacillus brevis BDUMBT08 (MT673657), L. gastricus BDUMBT09 (MT774596), L. paracasei BDUMBT10 (MT775430), L. brevis BDUMBT11 (MW785062), L. casei BDUMBT12 (MW785063), L. casei BDUMBT13 (MW785178), and Brevibacillus brevis M2403 (MK371781) from human breast milk. Their tolerance to lysozyme, acid, bile, gastric juice, pancreatic juice, and NaCl and potential for mucoadhesion, auto-aggregation, and co-aggregation with pathogens are of great prominence in forecasting their gut colonizing ability. They proved their safety aspects as they were negative for virulence determinants such as hemolysis and biofilm production. Antibiogram of LAB showed their sensitivity to more than 90% of the antibiotics tested. Amongst seven LAB, three isolates (L. brevis BDUMBT08 and BDUMBT11, and L. gatricus BDUMBT09) proved their bacteriocin producing propensity. Although the seven LAB isolates differed in their behavior, their substantial probiotic properties with safety could be taken as promising probiotics for further studies to prove their in vivo effects, such as health benefits, in humans.

Production of eco-friendly PHB-based bioplastics by Pseudomonas aeruginosa CWS2020 isolate using poultry (chicken feather) waste.

Nowadays, the accumulation of non-degradable plastics and other disposed wastes leads to environmental pollution across the world. The production of eco-friendly and cost-effective poly-ß-hydroxybutyrate (PHB) could be a better alternative to conventional petroleum-based plastics and prevent environmental pollution. Besides, the area in and around Namakkal, Tamil Nadu, India is well known for poultries, currently facing the number of environmental issues due to the accumulation of chicken feather waste. This study focused on the production of eco-friendly PHB by recycling poultry (chicken feather) waste as the substrate. The native PHB producers were screened from the chicken waste disposal site in Namakkal by Sudan black B staining method. Further, the potent bacterial isolate was identified as Pseudomonas aeruginosa (NCBI accession MF18889) by phenotypic and genotypic characteristics. The PHB production media with chicken feather waste was statistically optimized by response surface methodology. The dry weight of PHB produced under optimized condition (15.96 g/L chicken feather waste, 37 °C temperature, 19.8 g/L glucose and 6.85 pH) was found to be 4.8 g/L. Besides, PHB was characterized and confirmed by thin-layer chromatography, Fourier-transform infrared spectroscopy and Gas chromatography-mass spectrometry analysis. Thus, this study concludes that poultry waste could be a complex nitrogen source for improving the growth of PHB producers and substantially increasing the yield of PHB, and it will be an eco-friendly and low-cost production in bioprocess technology.

Knowledge and Awareness of Oral Manifestations of Diabetes Mellitus and Oral Health Assessment among Diabetes Mellitus Patients- A Cross Sectional Study.

BACKGROUND AND OBJECTIVES: Diabetes mellitus has increased rapidly throughout the world. The objectives of our study were to assess the knowledge and awareness about oral manifestations of diabetes, among type 2 diabetes mellitus patients, their risk for developing oral diseases due to complications associated with diabetes mellitus, and at same time, to perform an oral examination to detect these oral symptoms, if present any, along with the recording of Decayed Missing Filled Teeth Index (DMFT) and Community Periodontal Index (CPI) index. METHODOLOGY: Structured questionnaires consisting of 12 different statements on the knowledge base of oral manifestations of diabetes mellitus were distributed to 447 Type 2 diabetes mellitus patients. Following this oral examination, brushing and dental visit history were noted, and CPI index and DMFT indices were recorded in all the patients. RESULTS: Results showed that the knowledge about oral manifestations of diabetes mellitus was poor with a mean value of 4.92 out of a possible score of 12. Among the study subjects, the average score of men was 4.42 while that of females, was 5.41. These scores, when subjected to statistical analysis, were highly significant. (P value- 0.005) Subjects also showed significantly high DMFT (P value <0.001) and CPI scores (P value- 0.270). CONCLUSION: Our study concluded that there is a significant lack of knowledge about oral manifestations of diabetes mellitus among patients and hence steps have to be taken to increase their awareness through various outreach programs. All health professionals need to work together for promoting better oral health so that oral complications of diabetes can be brought under control.

Effectiveness of Theobromine on Enamel Remineralization: A Comparative In-vitro Study.

Background Remineralizing agents demonstrate potential to reverse early carious lesions. Theobromine containing dentifrices claim to remineralize enamel lesions effectively. The aim of this in-vitro study was to evaluate and compare the remineralization potential of dentifrices containing theobromine, 0.21% sodium fluoride (NaF) with functionalized tricalcium phosphate (f-TCP) and amine fluoride on artificial enamel caries. Materials and methods Sound extracted human premolars were demineralized to produce deep artificial carious lesions. The teeth were sectioned longitudinally and allocated to three treatment groups with nine specimens in each group: Group A (NaF + f-TCP), Group B (amine fluoride), and Group C (theobromine). The specimens were then subjected to pH cycling for seven days. Confocal laser scanning microscopy (CLSM) was utilized to record the patterns of demineralization and remineralization. One-way ANOVA and paired t-test were used to analyze changes in lesion depth. The level of significance was set at p<0.05. Results All three dentifrices effectively remineralized artificial carious lesions (paired t-test, p<0.001). Of the groups, Group A (54.97%) reported the highest percentage change in lesion depth values followed closely by Group B (51.51%) and Group C (31.71%), respectively. Conclusion Within this in-vitro study, theobromine containing dentifrice was effective in remineralizing lesions of enamel. However, theobromine demonstrated less remineralization potential in comparison to dentifrices containing NaF + f-(TCP) and amine fluoride.

A Comparative Data Analysis of 1835 Road Traffic Accident Victims.

AIMS AND OBJECTIVES: The purpose of the study was to analyze and compare the incidence of road traffic accidents (RTA) with head injuries and maxillofacial injuries in two arterial roads, Old Mahabalipuram road (OMR) and East Coast road (ECR), connecting with Chennai city and outlining the need of safety precautions to be followed to reduce the incidence of morbidity. MATERIALS AND METHODS: This study involved the medical records of about 1835 trauma victims who reported to Chettinad Health city, kelambakkam, between August 2008 and June 2013. The data analyzed were, age of trauma victims, gender, type of trauma, type of vehicle, accident time, accident zone, presence of head injury, maxillofacial injury and history of alcohol consumption. RESULTS: Trauma victims were predominantly male (84.3%), with majority of individuals in the age group of 21-40 yrs (56%). About 42% of the reported accidents occurred in OMR and 18.3% of accidents occurred in ECR. About 51.2% of the reported road traffic accidents occurred in the busy traffic hours, between 7am-10am and 5pm-9pm. About 66.4% of RTAs were due to two wheeler vehicles and 21.6% were due to four wheeler vehicles. The incidence of head injury was 47.5% and about 1417 (77.2%) patients reported with maxillofacial injuries. CONCLUSION: RTAs are more common in OMR than in ECR, involving mostly male victims and two wheeler vehicles, during the peak traffic hours. Rash driving and over speeding of vehicles are the preventable causative factors. Wearing of Helmets by the two wheeler riders and seat belts by the four wheeler riders are essential to prevent morbidity. We stress the need of separate lane for Ambulance on the roads for faster transport of accident victims to nearby Hospital and trauma care centers.

Toluidine Blue Staining in Identification of a Biopsy Site in Potentially Malignant Lesions: A Case-control Study.

OBJECTIVE: Oral cancer is a significant threat to public health all over the world, especially in Southeast Asia. At the present time, screening of oral cancer, its premalignant stages as well as its early detection, is still largely based on visual examination of the mouth. Visual examination is highly subjective and hence lacks the specificity and sensitivity. The objective of this study was to determine the usefulness of toluidine blue in marking a biopsy site in potentially malignant disorders. METHODS: In this study, a total of 500 patients were screened. The study was a case-control study which included 17 lesion cases and 23 normal controls. Toluidine blue staining was taken into consideration to identify clinically doubtful oral potentially malignant lesions and to compare the clinical evaluation with toluidine blue stain followed by a punch biopsy and histological evaluation. SPSS Statistics version 16.0 and Chi-square test were used for statistical analyses. RESULTS: The most common site for potentially malignant lesions was found to be the buccal mucosa. The sensitivity of toluidine blue was found to be 88.89%, while specificity was found to be 74.19%. The positive predictive and negative predictive values were 50% and 97.83%, respectively. P = 0.000672 was considered statistically significant. CONCLUSIONS: The results seem to be promising, but many such studies have to be done at larger scales to exactly help us in identifying the capability of toluidine blue in the long run.