Exploring the neuropharmacological properties of scopoletin-rich Evolvulus alsinoides extract using in-silico and in-vitro methods.

OBJECTIVES: This study investigates the neuropharmacologic properties of Scopoletin, a bioactive compound in Evolvulus alsinoides (EA) extract, for managing cognitive impairment using in-vitro, in-silico, and zebrafish embryo toxicity assays. METHODS: The study estimates Scopoletin concentration in EA extract using HPTLC, assesses antioxidant properties using 2,2-Diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing ability of plasma (FRAP) assays, and uses bioinformatic tools for scopoletin targets. Zebrafish embryo toxicity (ZET) is used to assess its toxicological profile. RESULTS: 0.0076% w/w Scopoletin in the samples was quantified using HPTLC, further studies on the DPPH (0.5 mM) and FRAP gave EC50 at 440.0 µg/ml and 84.29 µg/ml respectively. Twelve common targets associated with cognitive impairment (CI) were identified, along with possible pathways and molecular interactions. Our results indicate significant binding affinities of Scopoletin with ERAP1, SCN3A, and COMT. Molecular dynamics simulations further confirm the stability of these interactions. ZET assessment demonstrated mortality after 450 µg/ml concentration of EA extract. CONCLUSION: The study verifies the presence of Scopoletin in EA, along with their targets playing a crucial role in neurogenesis and neuroplasticity. The ZET demonstrated concentration-dependent effects, emphasizing the importance of dosage considerations in developing new formulations or therapeutics. This comprehensive study contributes valuable insight into the therapeutic potential of Scopoletin from EA for cognitive impairment, paving the way for further research.

Impact of Heartfulness meditation practice on anxiety, perceived stress, well-being, and telomere length.

Objective: Exhaustion, stress, and burnout have all been found to be reduced using techniques like yoga and meditation. This study was carried out to check the effectiveness of Heartfulness practice (a form of meditation) on certain psychological and genetic variables. Methods: A total of 100 healthy individuals (aged 18-24) were recruited and randomized into two groups-Heartfulness intervention and control group. The intervention was carried out for 03 months. Participants from both groups were analysed for their cortisol levels and telomere length before and after the intervention. Psychometric measures of anxiety, perceived stress, well-being and mindfulness were carried out using Beck Anxiety Inventory (BAI), Perceived Stress Scale (PSS), WHO-Well-being Index (WHO-WBI) and Five Facet Mindfulness Questionnaire (FFMQ). Results: The cortisol levels in the meditators group significantly decreased (p < 0.001) after the intervention as compared to the non-meditators group, whereas, the telomere length increased in the mediators group. This increase was not significant (p > 0.05). Anxiety and perceived stress also decreased post intervention, and well-being as well as mindfulness increased, as assessed by the questionnaire tools, although the decrease in perceived stress was statistically insignificant (p > 0.05). A negative correlation was observed between telomere length and cortisol (stress biomarker), whereas a positive correlation was found between telomere length and well-being. Conclusion: Our data provide evidence that Heartfulness meditation practice can improve our mental health. Additionally, telomere length is shown to be affected by cortisol levels, and this meditation practice can also help to increase telomere length, and thereby slow down cellular aging. However, future studies with larger sample size are required to confirm our observations.

Aldehyde dehydrogenase 1: Its key role in cell physiology and oral carcinogenesis.

Aldehyde dehydrogenase (ALDH), an aldehyde-metabolizing enzyme, is a cytosolic antioxidant. It performs many important physiological catalytic and non-catalytic functions in mammalian cells. Apart from physiological functions, like the biosynthesis of vital molecules, this NAD(P)+ substrate-dependent enzyme superfamily is primarily involved in catalyzing the oxidation of highly reactive exogenous and endogenous aldehydes to their respective carboxylic acids. Among ALDH isoenzymes, ALDH1 has gained much attention as a prominent stem cell marker, as it is associated with the maintenance of stemness and the differentiation of normal stem cells, in addition to involvement in oncogenic functions, like cell proliferation, anti-apoptosis and the reduction of oxidative stress in cancer stem cells (CSCs). In this context, the authors review the physiological functions of ALDH1 in normal cells, normal stem cells and CSCs, along with the discussion of the putative role of ALDH1 in oral carcinogenesis by commenting on its expression in normal oral mucosa cells, oral potentially malignant disorders (OPMDs), like leukoplakia and dysplastic lesions, and oral squamous cell carcinoma (OSCC).

Altered immunoexpression of SOX2, OCT4 and Nanog in the normal-appearing oral mucosa of tobacco users.

BACKGROUND: Tobacco use is causatively associated with various human cancers, including oral carcinoma. A number of pathways have been delineated to describe its etiopathological link with oral carcinogenesis, including alterations in the expression of stem cell markers. Embryonic stem cell markers, such as sex-determining region Y-box 2 (SOX2), octamer-binding protein 4 (OCT4) and homeobox protein Nanog, which are mainly involved in the maintenance of stemness and pluripotency, have been positively associated with the pathogenesis of oral potentially malignant disorders and oral cancers. In this context, we attempted to explore the subcellular impact of tobacco through examining the expression of these stem cell markers in normal and normal-appearing oral mucosa in non-tobacco users and tobacco users. OBJECTIVES: The aim of the study was to analyze the immunoexpression of SOX2, OCT4 and Nanog in the normal-appearing oral mucosa (NAOM) of tobacco users as compared to the normal oral mucosa (NOM) of non-tobacco users. MATERIAL AND METHODS: The tissue samples of tobacco users and non-tobacco users (n = 50 per group) were immunohistochemically stained to assess the expression of SOX2, OCT4 and Nanog. RESULTS: In the oral mucosa of non-tobacco users, a peculiar parabasal expression pattern of SOX2 and OCT4 was observed, whereas Nanog was non-reactive. The grade of inflammation was found to be a predictive variable influencing the expression of the 2 markers. In tobacco users, variables such as male gender, mixed habit and basilar hyperplasia significantly controlled the basilar and suprabasilar expression of SOX2, OCT4 and Nanog. The expression of SOX2 and OCT4 was higher in tobacco users; in particular, OCT4 positivity was significantly increased (p < 0.001) in comparison with non-tobacco users. CONCLUSIONS: The altered expression of the examined stem cell markers could be an indication of early molecular changes in NAOM under the influence of tobacco.

Identification of genetic instability in peripheral blood lymphocyte of oral squamous cell carcinoma patients assess by comet assay.

Context: Studies established that human cancer is principally a genetic disease; it arises as accumulation of a set of genetic changes. In the pathogenesis of cancer, genetic instability is the sequential event to a carcinogenic stimulus resulting in various genomic changes including DNA damage. Aims: To assess genetic instability, as susceptibility to DNA damage, we used single-cell gel electrophoresis (comet assay) to study double strand breaks in associated with the risk of oral squamous cell carcinoma (OSCC). Materials and Methods: We used comet assay to measure double strand break in individual peripheral blood lymphocytes from 50 individuals with OSCC and 30 healthy control subjects. All personal information was gathered from subjects including tobacco history. DNA damage was visualized as comet assay and quantified by movement of damaged strands as length of tail. Results: Study results of OSCC patients were observed in relation to clinical staging and histological grading of carcinoma. On the basis of clinical observation, cases were grouped in to Stage I, Stage II, Stage III and Stage IV. No stage I cases were in study sample. The mean DNA damage migration length was observed 4.600 ± 0.4613 µm in stage II, whereas in Stage III and Stage IV, it was observed to be 4.961 ± 0.5620 µm and 4.883 ± 0.410 µm, respectively. The DNA damage length in histological grades of squamous cell carcinoma patients in Grade I was 4.6437 ± 0.3061 µm and Grade II was 5.3533 ± 0.3831 µm. In comparison with control group and squamous cell carcinoma group, it was observed in the range of 0.02-0.36 µm and varied from 4.04 to 5.84 µm range, respectively. Thus, the results were statistically significant with the histological grading of OSCC. Statistical Analysis: Unpaired' test and "ANOVA" test are used for statistics. Statistical Analysis: Unpaired' test and "ANOVA" test are used for statistics. Conclusion: The amount of DNA strand breaks in peripheral lymphocytes are measured by comet assay which is associated with relative risk of OSCC.

Green Synthesis to Develop Iron-Nano Formulations and Its Toxicity Assays.

OBJECTIVES: In the past few years, herbal medicines have gained popularity over synthetic drugs because of their natural source and minimal side effects which has led to a tremendous growth of phytopharmaceuticals usage. With the development of nanotechnology, it provides alternative approaches to overcome several limitations using nano-formulations. In spite of considerable quantity of antianemic preparations with different iron forms available, currently additives are used and represented in modern pharmaceutical market. Iron deficiency anemia is a major global public health problem which particularly affects pregnant women, children and elderly persons. The situation is complicated because of disadvantages and drug side effects from existing antianemic medicines. There is a great demand for the development of new antianemic preparations. Green synthesis of iron oxide nanoparticles, possess high potential in this field. METHODS: Our study focuses on developing green synthesis of iron oxide nanoparticles (IONPs) of 10-50 nm with spherical shape where different dosages were used -1 mg/kg, 10 mg/kg and 100 mg/kg for exposure in Wistar albino female rats for 28 days. The toxicity was assessed using various parameters such as measurements of the rat body and organ mass, hematology, biochemical evaluation and histopathological examinations. RESULTS: No significant differences were observed in body and organ weights. Hematological indices also indicated no significant differences whereas biochemical factors showed increase in levels of direct bilirubin and globulin of medium as well as high dose and SGPT levels were increased only in high dose. The major organs (heart, kidney and liver) showed histopathological alterations in 10 and 100 mg/kg whereas brain showed only in 100 mg/kg. CONCLUSION: The toxicity of IONPs was found to be more significant when the concentration was increased; however, low doses can be used for further investigation as an antianemic preparation.

SOX2, OCT4 and NANOG: The core embryonic stem cell pluripotency regulators in oral carcinogenesis.

Embryonic stem cells provide their major contribution to embryogenesis through formation of germ layers as they have pluripotency potential and capacity for self-renewal. Retention of pluripotency of these stem cells depends on expression/level of transcription factors, i.e., SOX2, OCT4 and NANOG. During organogenesis, the altered expression of the molecules also influences these stem cells to lose their pluripotency and turn toward the lineage selection. As the differentiation progresses, the maintenance of the somatic cells including the oral squamous cells also depends on differential expression of the transcription factors to some extent. Recently, many experimental and observational studies documented the significant contribution in carcinogenesis of various human cancers. In this review, we have attempted to summarize the evidences indicating about the putative role of these master pluripotency regulators in various phases of oral carcinogenesis i.e. initiation , progression and prognosis of oral squamous cell carcinoma.

Embryonic Zebrafish Model - A Well-Established Method for Rapidly Assessing the Toxicity of Homeopathic Drugs: - Toxicity Evaluation of Homeopathic Drugs Using Zebrafish Embryo Model.

OBJECTIVES: Advancements in nanotechnology have led to nanoparticle (NP) use in various fields of medicine. Although the potential of NPs is promising, the lack of documented evidence on the toxicological effects of NPs is concerning. A few studies have documented that homeopathy uses NPs. Unfortunately, very few sound scientific studies have explored the toxic effects of homeopathic drugs. Citing this lack of high-quality scientific evidence, regulatory agencies have been reluctant to endorse homeopathic treatment as an alternative or adjunct treatment. This study aimed to enhance our insight into the impact of commercially-available homeopathic drugs, to study the presence of NPs in those drugs and any deleterious effects they might have, and to determine the distribution pattern of NPs in zebrafish embryos (Danio rerio). METHODS: Homeopathic dilutions were studied using high-resolution transmission electron microscopy with selected area electron diffraction (SAED). For the toxicity assessment on Zebrafish, embryos were exposed to a test solution from 4 - 6 hours post-fertilization, and embryos/larvae were assessed up to 5 days post-fertilization (dpf) for viability and morphology. Toxicity was recorded in terms of mortality, hatching delay, phenotypic defects and metal accumulation. Around 5 dpf was found to be the optimum developmental stage for evaluation. RESULTS: The present study aimed to conclusively prove the presence of NPs in all high dilutions of homeopathic drugs. Embryonic zebrafish were exposed to three homeopathic drugs with two potencies (30CH, 200CH) during early embryogenesis. The resulting morphological and cellular responses were observed. Exposure to these potencies produced no visibly significant malformations, pericardial edema, and mortality and no necrotic and apoptotic cellular death. CONCLUSION: Our findings clearly demonstrate that no toxic effects were observed for these three homeopathic drugs at the potencies and exposure times used in this study. The embryonic zebrafish model is recommended as a well-established method for rapidly assessing the toxicity of homeopathic drugs.

Physicochemical characterization and antibacterial activity of Rajata Bhasma and silver nanoparticle.

INTRODUCTION: Bhasma, an Ayurvedic metallo-mineral preparation, is claimed to be biologically produced nanoparticles. Rajata (silver) is a noble metal known for its antimicrobial activity. Rajata Bhasma (RB) is expected to be composed of nanoparticles. With all these facts in place, this study was conducted to evaluate RB for the presence of silver nanoparticle (SNP) and its antimicrobial effect. AIM: The aim of this study is to analyze the physicochemical characterization, antibacterial activity of RB, and SNP. MATERIALS AND METHODS: RB was commercially ordered and SNP was prepared by Turkevich method. Characterization of RB was carried out by scanning electron microscopy (SEM) and inductively coupled plasma-atomic emission spectroscopy (ICP-AES). SNP was characterized by ultraviolet (UV)-visible spectroscopy, transmission electron microscopy (TEM), and ICP-AES. Antibacterial activity of RB and SNP was carried out by well-diffusion method. RESULTS: Analysis of RB by SEM revealed particles in range from 10 to 60 nm. UV-visible spectrum of the aqueous medium containing SNPs showed absorption peak at around 423 nm. The TEM analysis showed that SNP was spherical in the range of 5-50 nm and uniformly distributed without significant agglomeration. The content of silver in RB measured with ICP-AES was found to be 70.56% whereas in case of SNP was 65.23%. Staphylococcus aureus was found to be sensitive to both RB and SNP. Escherichia coli, Pseudomonas aeruginosa, and Enterococcus faecalis were found to be resistant to RB as well as SNP. CONCLUSION: The current study shows that RB does have silver particles in the size of nanometers and also has mild antibacterial activity.

Histopathological and ultra structural effects of nanoparticles on rat testis following 90 days (Chronic study) of repeated oral administration.

BACKGROUND: Nanoparticles (Ag NPs) have recently received much attention for their possible applications in biotechnology and biomedical. However, little is known about the toxicity in reproductive organs of animal model following exposure to nanoparticles. OBJECTIVE: This study therefore, tried to examine the effects of nanoparticles with a diameter range of 5-20 nm on the histology of the testis of wistar rats and correlate it with Transmission Electron Microscopy results. MATERIALS AND METHODS: Sixteen wistar rats were randomly divided into two groups of 8 rats each. Each group received the following via gavage technique for 90 days: Control Group (Group-1)-tap water; Experimental group (Group 2) - nanoparticles (20ug/kg/day). After ninety days (chronic study), rats were sacrificed and testis tissues was processed for histology and transmission electron microscopic study. RESULTS: There was significant difference between the observations of group-1 and group 2. The changes observed in the testis were disarray of the spermatogenic cells and disorientation of the testis. These changes were observed to have been disappearing from normal histological features. Detailed structural damages were observed with TEM analysis, such as depletion of germ cells, germinal cells necrosis, especially in spermatogonia and Leydig cells had an abnormal fibroblast-like appearance, abnormal space between neighboring sertoli cells, mitochondria, lost cristae and vacuolated (none energized) with those animals exposed to nanoparticles. CONCLUSION: It seems that nanoparticles have acute and significant effects on spermatogenesis and number of spermatogenic cells. More experimental investigations are necessary to elucidate better conclusion regarding the safety of nanoparticles on male reproduction system.