Saliva Based Diagnostic Prediction of Oral Squamous Cell Carcinoma using FTIR Spectroscopy.

Oral cancer ranks as the sixth most prevalent form of cancer worldwide, presenting a significant public health concern. According to the World Health Organization (WHO), within a 5-year period following diagnosis, the mortality rate among oral cancer patients of all stages stands at 45%. In this study, a total of 60 patients divided into 2 groups were recruited. Group A included 30 histo-pathologically confirmed OSCC patients and Group B included 30 healthy controls. A standardized procedure was followed to collect saliva samples. FTIR spectroscopy was done for all the saliva samples collected from both Group A and B. An IR Prestige-21 (Shimadzu Corp, Japan) spectrometer was used to record IR spectra in the 40-4000 cm-1 range SVM classifier was applied in the classification of disease state from normal subjects using FTIR data. The peaks were identified at wave no 1180 cm-1, 1230 cm-1, 1340 cm-1, 1360 cm-1, 1420 cm-1, 1460 cm-1, 1500 cm-1, 1540 cm-1, 1560 cm-1, and 1637 cm-1. The observed results of SVM demonstrated the accuracy of 91.66% in the classification of Cancer tissues from the normal subjects with sensitivity of 83.33% while specificity and precision of 100.0%. The development of oral cancer leads to noticeable alterations in the secondary structure of proteins. These findings emphasize the promising use of ATR-FTIR platforms in conjunction with machine learning as a reliable, non-invasive, reagent-free, and highly sensitive method for screening and monitoring individuals with oral cancer.

Salivary transforming growth factor beta in oral submucous fibrosis: A diagnostic and predictive marker.

CONTEXT: Growth factors and cytokines like transforming growth factor beta (TGF-ß) play a key role in the pathogenesis of oral submucous fibrosis. AIMS: To elucidate the role of Salivary TGF-ß isoforms as a predictive and diagnostic marker for oral submucous fibrosis. SETTINGS AND DESIGN: A total of 30 OSMF and 10 control patients were included in this study, and their clinic-epidemiological data was recorded. METHODOLOGY: The expression of TGF-ß genes-TGF-ß1, TGF-ß2, TGF-ß3-was studied by a real-time polymerase chain reaction in tissue and saliva. Patients were given medicinal intervention for 12 weeks along with jaw-opening exercises. Expression of salivary TGF-ß genes was studied at 12 weeks. STATISTICAL ANALYSIS USED: SPSS software version 20. RESULT: Expression of salivary TGF beta isoforms in OSMF was more than in the control group. There was an increase in salivary TGF-ß1, ß2, ß3 expressions with increasing clinical grades of OSMF and advancing the stage of the disease. Expression of all the TGF beta isoforms was decreased after treatment with statistically significant results. Statistically significant correlations were found between the mean difference of TGF-ß1 and the mean difference between mouth opening and tongue protrusion. CONCLUSION: Salivary TGF-ß isoforms may be used in diagnosis, risk assessment, and screening of the entire population at risk of OSMF after its clinical validation. However, adequate sample size and segmental assessment of the expression of TGF-ß isoforms are needed for further evaluation.

Efficacy of photobiomodulation in the management of oral Lichen Planus in comparison to topical corticosteroids: Systematic review, meta-analysis, and GRADE-based assessment of certainty of evidence.

BACKGROUND: There is an urgent need to update the evidence available on the efficacy of photobiomodulation (PBM) in comparison to topical corticosteroids in the management of Oral Lichen Planus (OLP). METHODS: Cochrane Oral Health's Trials Register, Cochrane Central Register of Controlled Trials in the Cochrane Library, MEDLINE Pubmed, SCOPUS, Lilacs and Google Scholar were searched. Other sources included US National Institutes of Health Ongoing Trials Register, World Health Organization International Clinical Trials Registry Platform and Clinical Trial Registry - India were searched using variations of the keywords "Laser" and "Oral Lichen Planus". Randomized controlled trials comparing PBM and topical corticosteroids in the resolution of pain among OLP patients were included. The studies were assessed using the Cochrane Risk-of-Bias assessment tool. Publication bias was assessed using a funnel plot, and the certainty of evidence was evaluated according to the GRADE guidelines. RESULTS: Ten studies were included for qualitative assessment and of these eight were included in the meta-analysis. The included studies used laser parameters of varying strengths and duration. Meta-analysis favoured PBM (n = 274, MD =-0.48, CI -0.66- -0.30) for pain score. No adverse effects were reported for laser therapy. There was high heterogeneity and moderate certainty of evidence, and most studies had a high risk of bias. CONCLUSION: There is improvement in the clinical parameters of OLP when treated with PBM in comparison to topical corticosteroids. However, the strength of the evidence for these findings in limited. It is recommended to conduct better long-term trials with large sample size.

Colchicine in the Management of Oral Submucous Fibrosis: A Systematic Review, Meta-analysis, and GRADE-Based Assessment of Certainty of Evidence.

Purpose: To study the efficacy of Colchicine in the management of Oral Sub mucous Fibrosis (OSMF) through systematic review and meta-analysis. Methods:An extensive literature search was conducted on databases such as PubMed, Cochrane Library database, LILACS, Google Scholar, CTRI, and Google search engine. The search comprises all articles published from 2013 to 2022. A total of 10 randomized control trial studies involving colchicine as one of the interventions were included. [CRD42022377674] Results:10 randomized control comprising 456 subjects were included. 3 studies are included in the meta-analysis comprising 90 subjects. All three studies show that there are significant differences between colchicine and other intervention regarding mouth opening and burning sensation. It indicates that Colchicine is significantly more effective in the reduction of burning sensation than other interventions used as a control in OSMF. Conclusion: The evidence generated through this systemic review and meta-analysis suggest that the use of an oral form of colchicine is an effective measure in the management of OSMF, especially the subjective symptom such as burning sensation where as it is not found to be much effective in case of mouth opening. If colchicine is used as co drug along with conventional injection therapy better will the result. However there is marked diversity among the studies reviewed; therefore, the results of this review should be interpreted very carefully.

Oxidative Stress Enzyme as Markers in Oral Potentially Malignant Disorders and Oral Squamous Cell Carcinoma Patients.

Oxidative stress is a recognized factor that poses a significant risk for the development of Oral Squamous Cell Carcinoma (OSCC) and Oral Submucous Fibrosis (OSMF), as it leads to the generation of Reactive Oxygen Species. In recent years, there has been significant research on the enzymes MDA (malondialdehyde) and SOD (superoxide dismutase), investigating their potential role in the development of OPMD and OSCC. These enzymes have emerged as promising biomarkers due to their ability to provide a less invasive, cost-effective, and objective diagnostic method. Furthermore, they can be used to monitor disease progression and assess the effectiveness of therapy. The aim of this study was to assess the levels of MDA and SOD in the serum of patients diagnosed with OSCC and OSMF. Study group comprised of 60 patients, out of which 20 cases of clinically diagnosed OSCC patients and 20 cases of OSMF and 20 cases pf control comprising of healthy patients were recruited. Estimation MDA and SOD was done by ELISA. The statistical analysis was done using SPSS analysis. When comparing the levels of MDA and SOD between the OSCC and OSMF groups and the control group, statistically significant findings indicated elevated levels of malondialdehyde and reduced levels of superoxide dismutase in both the OSCC and OSMF groups. In this study, the assessment of lipid peroxidation through MDA levels revealed elevated concentrations in both the OSCC and OSMF groups when compared to the control group. Specifically, the order of MDA levels was observed as OSCC > OSMF > Control. Conversely, antioxidant enzyme levels, such as SOD, exhibited decreased concentrations in the OSCC and OSMF groups compared to the control group, with the order of SOD levels being Control > OSMF > OSCC. Consequently, the findings suggest that MDA and SOD can be considered potential biomarkers for identifying and monitoring OSCC and OSMF diseases.

Phytonutrient Inhibitors of SARS-CoV-2/NSP5-Encoded Main Protease (Mpro) Autocleavage Enzyme Critical for COVID-19 Pathogenesis.

The genomic reshuffling, mutagenicity, and high transmission rate of the SARS-CoV-2 pathogen highlights an urgent need for effective antiviral interventions for COVID-19 control. Targeting the highly conserved viral genes and/or gene-encoded viral proteins such as main proteinase (Mpro), RNA-dependent RNA polymerase (RdRp) and helicases are plausible antiviral approaches to prevent replication and propagation of the SARS-CoV-2 infection. Coronaviruses (CoVs) are prone to extensive mutagenesis; however, any genetic alteration to its highly conserved Mpro enzyme is often detrimental to the viral pathogen. Therefore, inhibitors that target the Mpro enzyme could reduce the risk of mutation-mediated drug resistance and provide effective antiviral protection. Several existing antiviral drugs and dietary bioactives are currently repurposed to treat COVID-19. Dietary bioactives from three ayurvedic medicinal herbs, 18 ß-glycyrrhetinic acid (ΔG = 8.86 kcal/mol), Solanocapsine (ΔG = 8.59 kcal/mol), and Vasicoline (ΔG = 7.34 kcal/mol), showed high-affinity binding to Mpro enzyme than the native N3 inhibitor (ΔG = 5.41 kcal/mol). Flavonoids strongly inhibited SARS-CoV-2 Mpro with comparable or higher potency than the antiviral drug, remdesivir. Several tannin hydrolysates avidly bound to the receptor-binding domain and catalytic dyad (His41 and Cys145) of SARS-CoV-2 Mpro through H-bonding forces. Quercetin binding to Mpro altered the thermostability of the viral protein through redox-based mechanism and inhibited the viral enzymatic activity. Interaction of quercetin-derivatives with the Mpro seem to be influenced by the 7-OH group and the acetoxylation of sugar moiety on the ligand molecule. Based on pharmacokinetic and ADMET profiles, several phytonutrients could serve as a promising redox nutraceutical for COVID-19 management.

Targeting COVID-19 (SARS-CoV-2) main protease through active phytochemicals of ayurvedic medicinal plants - Withania somnifera (Ashwagandha), Tinospora cordifolia (Giloy) and Ocimum sanctum (Tulsi) - a molecular docking study.

COVID-19 (Coronavirus disease 2019) is a transmissible disease initiated and propagated through a new virus strain SARS-CoV-2 (Severe Acute Respiratory Syndrome Coronavirus-2) since 31st December 2019 in Wuhan city of China and the infection has outspread globally influencing millions of people. Here, an attempt was made to recognize natural phytochemicals from medicinal plants, in order to reutilize them against COVID-19 by the virtue of molecular docking and molecular dynamics (MD) simulation study. Molecular docking study showed six probable inhibitors against SARS-CoV-2 Mpro (Main protease), two from Withania somnifera (Ashwagandha) (Withanoside V [10.32 kcal/mol] and Somniferine [9.62 kcal/mol]), one from Tinospora cordifolia (Giloy) (Tinocordiside [8.10 kcal/mol]) and three from Ocimum sanctum (Tulsi) (Vicenin [8.97 kcal/mol], Isorientin 4'-O-glucoside 2″-O-p-hydroxybenzoagte [8.55 kcal/mol] and Ursolic acid [8.52 kcal/mol]). ADMET profile prediction showed that the best docked phytochemicals from present work were safe and possesses drug-like properties. Further MD simulation study was performed to assess the constancy of docked complexes and found stable. Hence from present study it could be suggested that active phytochemicals from medicinal plants could potentially inhibit Mpro of SARS-CoV-2 and further equip the management strategy against COVID-19-a global contagion. HighlightsHolistic approach of Ayurvedic medicinal plants to avenge against COVID-19 pandemic.Active phytoconstituents of Ayurvedic medicinal plants Withania somnifera (Ashwagandha), Tinospora cordifolia (Giloy) and Ocimum sanctum (Tulsi) predicted to significantly hinder main protease (Mpro or 3Clpro) of SARS-CoV-2.Through molecular docking and molecular dynamic simulation study, Withanoside V, Somniferine, Tinocordiside, Vicenin, Ursolic acid and Isorientin 4'-O-glucoside 2″-O-p-hydroxybenzoagte were anticipated to impede the activity of SARS-CoV-2 Mpro.Drug-likeness and ADMET profile prediction of best docked compounds from present study were predicted to be safe, drug-like compounds with no toxicity.Communicated by Ramaswamy H. Sarma.

In silico screening of Pueraria tuberosa (PTY-2) for targeting COVID-19 by countering dual targets Mpro and TMPRSS2.

COVID-19 pandemic was started in Wuhan city of China in December 2019; immensely affected global population. Herein, an effort was made to identify potential inhibitors from active phytochemicals of Pueraria tuberosa (PTY-2) via molecular docking study. Our study showed five potential inhibitors (Robinin, Genistin, Daidzin, Hydroxytuberosone, Tuberostan) against Mpro and five inhibitors (Robinin, Anhydrotuberosin, Daidzin, Hydroxytuberosone, Stigmasterol) against TMPRSS2. Out of these, Robinin, Daidzin and Hydroxytuberosone were common inhibitors for Mpro and TMPRSS2. Among these, Robinin showed the highest binding affinity, therefore, tested for MD simulation runs and found stable. ADMET analysis revealed the best-docked compounds are safe and follow the Lipinski Rule of Five. Thus, it could be suggested that phytochemicals of PTY-2 could serve as potential inhibitors for COVID-19 targets.Communicated by Ramaswamy H. Sarma.

Incretin hormones receptor signaling plays the key role in antidiabetic potential of PTY-2 against STZ-induced pancreatitis.

AIMS: Incretin therapy is one of the most potential approaches in the treatment of diabetes. In contrast to markedly available drugs, the herbal incretin modulators have lesser side effects with low economic cost. The main aim of this work was to analyze the potential of previously reported DPPIV inhibitor, aqueous extract of Pueraria tuberosa tubers (PTY-2) as incretin hormones receptor agonist against streptozotocin (STZ)-induced diabetes. METHODS: Chronic diabetes was induced with STZ (65mg/kg bw) in rats for 60days and grouped into diabetic control and PTY-2. Expression of genes was assessed by PCR, IHC, and ELISA. Morphological analysis of tissue was observed using H & E stain. In silico molecular docking approach has been used to see the interaction of active phytochemicals of PTY-2 on the basis of their binding energy [kcal/mol] and dissociation constant [pM] using YASARA software. Interactive visualization was done using Discovery studio 3.0. RESULTS: In comparison to diabetic control, the size and number of islet cells along with the plasma level of GLP-1, GIP, and pancreatic expressions of GLP-1R, GIP-R, Bcl2, and insulin were enhanced significantly after PTY-2 treatment. Through in silico molecular docking, tuberostan showed the best interaction for GLP-1R with binding energy at 8.15kcal/mol and dissociation constant at 1061624.125 pM. Puererone showed the best interaction for GIP-R with binding energy at 8.31kcal/mol and dissociation constant at 810381 pM. CONCLUSIONS: In addition to previously studied DPPIV inhibitor, PTY-2 also acts as incretin receptors agonist and protects against STZ-induced diabetes by down regulating ß cells apoptosis.

Active phytochemicals of Pueraria tuberosa for DPP-IV inhibition: in silico and experimental approach.

BACKGROUND: We had earlier reported that the extract of Pueraria tuberosa significantly inhibits DPP-IV enzyme, resulting in glucose tolerance response in rats. In this study, we have explored the active phytochemicals responsible for this potential. The results have been validated in both fasting and postprandial states in the plasma of normal rats and also in fasting blood and intestinal homogenates of diabetic models. METHODS: Pueraria tuberosa water extract (PTWE) was administered to normal Charles Foster rats for 35 days and to diabetic model (65 mg/kg bw) for 10 days. After treatments, oral glucose tolerance test (OGTT) and insulin was done for 90 min, and the changes in the levels of GLP-1, GIP, and DPP-IV activities were monitored in fasting and postprandial states. In the case of the diabetic model, DPP-IV activity was measured in intestinal homogenate and basal insulin in plasma. The components of PTWE were analyzed via HPLC-MS based on their chemical formula, molecular mass, and retention time. Using the molecular docking study, we have selected the top five components having strong binding energy with DPP-IV. RESULTS: The increase in secretion of GLP-1 and GIP was significantly higher in the postprandial state when compared to fasting condition. GLP-1 plasma concentration increased by 5.8 and 2.9 folds and GIP increased by 8.7 and 2.4 folds in PTWE and control rats, respectively. In contrast, the postprandial decrease in DPP-IV specific activities was recorded at 2.3 and 1.4 folds. The response in OGTT and insulin was also consistent with these changes. In comparison to diabetic controls, PTWE-administered rats showed decreased DPP-IV activity in the intestine, leading to enhanced basal insulin concentration. Through molecular docking, we found Puerarone and Robinin to be the most potential phytochemicals of PTWE for DPP-IV inhibition. Binding energy (kcal/mol) and dissociation constant (pM) of Robinin with DPP-IV protein were found to be 7.543 and 2,957,383.75, respectively. For Puerarone, it was 7.376 and 3,920,309, respectively. CONCLUSIONS: Thus, this study provides the novel active components that contribute to the DPP-IV inhibitory property of PTWE.

Epilithonimonas ginsengisoli sp. nov., isolated from soil of a ginseng field.

A novel Gram-staining-negative, rod-shaped bacterium, designated DCY78(T), was isolated from soil of a ginseng field in Yeon-cheon province (38° 04' 00″ N 126° 57' 00″ E), Republic of Korea. The phylogenetic analysis based on 16S rRNA gene sequences showed that strain DCY78(T) belonged to the genus Epilithonimonas and was most closely related to Epilithonimonas lactis DSM 19921(T) (98.5 % sequence similarity) and Epilithonimonas tenax DSM 16811(T) (97.8 %). Growth occurred at 10-30 °C with an optimum temperature of 28 °C. The pH range for growth was pH 5.5-8.0. The major polar lipids were found to be phosphatidylethanolamine three unidentified amino lipids and one unidentified polar lipid. The only predominant quinone was MK-6. The major polyamines were sym-homospermidine and spermidine. The major fatty acids were summed feature 3 (comprising C16 : 1ω6c and/or C16 : 1ω7c), iso-C15 : 0 and iso-C17 : 0 3-OH. The DNA G+C content was 37.9 mol%. On the basis of the phenotypic and genotypic analysis, the isolate is classified as representative of a novel species in the genus Epilithonimonas, for which the name Epilithonimonas ginsengisoli is proposed. The type strain is DCY78(T) ( = KCTC 32174(T) = JCM 19896(T)).

Amelioration of insulin resistance by Rk1 + Rg5 complex under endoplasmic reticulum stress conditions.

BACKGROUND: Diabetes mellitus is a metabolic syndrome exaggerated by stress conditions. Endoplasmic reticulum stress (ERS) impairs the insulin signaling pathway making the diabetic conditions worsen. Pharmacological agents are supplied externally to overcome this malfunction. Ginsenosides from Panax ginseng C.A Meyer possesses many pharmacological properties and are used for the treatment of diabetes. OBJECTIVE: To investigate the effects of the Rk1 +Rg5 complex on the amelioration of insulin resistance in 3T3-L1 cells under endoplasmic reticulum stress conditions. MATERIALS AND METHODS: Heat-processed ginseng extracts are found to contain many pharmacologically active ginsenosides. Among them Rk1 +Rg5 is found to be present in higher concentrations than the other minor ginsenosides. The Rk1 +Rg5 complex was tested for its effect in the 3T3-L1 insulin-resistant model and subjected to the MTT assay, glucose oxidase assay and gene expression studies using RT-PCR and real-time PCR under endoplasmic reticulum stress conditions. RESULTS: Rk1 +Rg5 treatment is found to increase the glucose uptake into the cells when compared to that of a positive control (tunicamycin treatment group, TM). Further we have analyzed the role at gene expression level. The Rk1 +Rg5 complex was found to show an effect on the IGF 2R receptor, CHOP-10, and C/EBP gene at a particular treated concentration (50 µM). Moreover, stress condition (about 50% decreases) was overcome by the ginsenoside treatments at 50 µM. CONCLUSION: The present results showed that under endoplasmic reticulum stress conditions Rk1 +Rg5 complex exhibits a potential protective role in insulin-resistant 3T3-L1 cells.

Phenethyl isothiocyanate inhibits 12-O-tetradecanoylphorbol-13-acetate-induced inflammatory responses in mouse skin.

Phenethyl isothiocyanate (PITC) is the hydrolysis product of the glucosinolate gluconasturtiin in cruciferous vegetables. This study was conducted to determine whether PITC inhibits 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced inflammation in the mouse ear. Topical application of 5 nmol of TPA to mouse ears markedly increased the ear weight, expression of the inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 protein, and phosphorylation of the inhibitor of κB (IκB) α, AKT, and extracellular signal-regulated protein kinase (ERK) 1/2 and reduced IκBα protein levels. Pretreatment with PITC (150-450 nmol) significantly suppressed these TPA-induced inflammatory responses. We also determined whether low concentrations of PITC (0.5-5 µmol/L) inhibited lipopolysaccharide (LPS)-stimulated inflammatory responses in Raw264.7 cells. PITC dose-dependently reduced the LPS-induced secretion of nitric oxide, prostaglandin E(2), interleukin (IL)-1ß, IL-6, and tumor necrosis factor (TNF)-α, as well as COX-2 and iNOS protein expression. PITC also attenuated LPS-induced increases in iNOS, COX-2, IL- 6, IL-1ß, and TNF-α mRNA levels, as well as the promoter-dependent transcriptional activation of the genes for iNOS and COX-2. PITC inhibited LPS-induced IκBα phosphorylation and degradation and subsequently reduced LPS-induced p65 nuclear translocation and the transcriptional activity of nuclear factor-κB (NF-κB), which was accompanied by a reduction in ERK1/2 and AKT phosphorylation. The results of this study demonstrated that PITC effectively inhibits inflammatory responses in vivo and in vitro, which may be mediated via the inhibition of AKT and ERK1/2 activation, leading to subsequent inhibition of the transcriptional activity of NF-κB.