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  • Open access
  • 28 Reads
Polyherbal Oral Spray for Instant Mouth Refreshing

In today’s busy lifestyle, people are experiencing constant shortage of time for their personal care. Focus on personal health and hygiene has reduced to the significant extent. In such situation, health and hygiene of the people are continuously downgrading due to lack of attention towards it. As the solution of such situations, market is full of many instantly acting products containing harmful chemicals and ingredients. Mouth health and hygiene is of prime importance as it is the main gateway of food and also for verbal communication. Tackling the increasing problems related to the health and hygiene of oral cavity and also emerging issues due to excessive use of instant acting products for it is the need of the hour. Herbal formulations are proven treatment options that can deal with such situations without any untoward side effects. We had studied many traditionally used natural items to screen a few of the potent ingredients to formulate an oral spray. Developed polyherbal spray formulation using Clove oil, Peppermint oil, Fennel oil, Piper Betel oil and Cardamom oil was evaluated for various basic parameters. As all of the above herbal ingredients are already proven their activities for maintaining and improving oral hygiene and health, the final product was not evaluated for specific activities. Moreover it don’t require additional facilities like In the nutshell, developed polyherbal spray was found to be a probable alternative for instant mouth refreshning product dealing with majority of oral health and hygiene issues, especially foul smell.

  • Open access
  • 25 Reads
Antipsychotic drugs counteract autophagy and mitophagy in multiple sclerosis

Background: Multiple sclerosis (MS) is a neuroinflammatory and neurodegenerative disease characterized by myelin damage followed by axonal and ultimately neuronal loss. The etiology and physiopathology of MS are still elusive, and no fully effective therapy is yet available

Objectives: We investigated the role in MS of autophagy (physiologically, a controlled intracellular pathway regulating the degradation of cellular components) and of mitophagy (a specific form of autophagy that removes dysfunctional mitochondria).

Methods: Our study has been performed by using in vitro, ex vivo and in vivo model of MS. Furthermore, experiments have been also conducted in human biofluids obtained from healthy and MS-affected individuals

Results: Three main findings emerge from the present work. First, autophagy and its selective forms occur in MS patients and in experimental models of MS; second, these phenomena play a causal role in MS be-cause their inhibition prevents myelin loss; third, two clinically used drugs can inhibit autophagy, prevent demyelination, induce remyelination, and revert MS behavioral deficits

Conclusion: Our findings suggest to repurpose Food and Drug Administration–approved drugs for the treatment of MS, at least in patients with MS variants that are more closely modeled by CPZ, like type III and IV. Such compounds may accelerate recovery from a demyelinating attack and prevent relapses.

  • Open access
  • 171 Reads
GC-MS based Metabolite profiling, and anti-inflammatory activity of Aqueous extract of Myrica esculenta through In vitro and In-Silico approach.

The present study was to determine the anti-inflammatory activity of aqueous extract of bark and root of Myrica esculenta and their active phytoconstituents through In-Vitro and In-Silico studies. The bioactive phytoconstituent of Myrica esculenta determined by GC-MS spectroscopy techniques. After that total phenolic and flavonoid content of both bark and root extract was determined. Furthermore, In-vitro anti-inflammatory activity was determined in both extracts. The molecular docking analysis determined the binding affinity of bioactive compounds against inflammatory proteins COX-1, COX-2, IL-10, and TNF-α. The present study revealed bark extract of Myrica esculenta has the highest total phenolic and flavonoid content compared with root extract (553.44±18.38mg GAE/g equivalent and 336.02±8.04mg quercetin/g equivalent respectively). Similarly, the bark extract showed good inhibitory activity with 5-LOX and HYA assay (IC50 11.26±3.93 and 21.61±8.27 µg/mL respectively), but in 15-Lox inhibitory assay root extract showed the highest inhibitory activity, IC50 16.95±5.92 µg/mL. The Docking result showed that myrecitin, Arjunolic acid, and myricanone have the highest binding affinity with all inflammatory proteins in respective order: myrecitin>arjunolic acid>celecoxib>myricanone>myricitrin>3-epi-ursonic acid. The MD simulation of COX-1 and myrecitin showed the highest stability and low deviation at 310K through RMSD values (1.07-2.3 Å) as compared with COX-1 and myricitrin (0.193-1.885 Å) and TNF-α and myricanone (1.377 to 3.457Å) respectively when analyzed at 100 ns time frame. Extract and their active constituents showed good anti-inflammatory activity. Further study is essential to define their mechanism of action.

  • Open access
  • 78 Reads
Signature garlic phytochemical as a potential anti-candidal candidate targeting virulence factors in Candida albicans

Anti-microbial resistance to presently available antifungal medications, as well as an increase in their toxicity, is becoming a severe concern throughout the world. It is necessary to investigate innovative, more effective medications especially derived from medicinally active plants with lesser chances of side effects. Allyl methyl sulfide (AMS) is an organosulfur derived from garlic oil which is responsible for typical garlic odour is explored for its therapeutic potential against Candida albicans. The effect of AMS on major virulence factors has been studied in Candida albicans reference strains, with MIC and MFC values reported to be 200 µg/ml and 400 µg/ml, respectively. Fungal growth in C. albicans was entirely inhibited at their respective MIC values, as demonstrated by growth curve and time kill kinetics experiments. In C. albicans, AMS treatment also reduces attachment to buccal cell epithelial tissues as measured microscopically. After treatment with AMS, C. albicans' release of extracellular proteinases, phospholipases, and biofilm formation were significantly inhibited. AMS exhibited little toxicity to human red blood cells (RBCs), indicating that it might be a feasible alternative to commonly used antifungal medications. More research is needed to determine its mechanism of action and precise target areas. The current work should be supported by molecular in silico and in vivo research.

  • Open access
  • 38 Reads
Silybin, the main active component of Silybum marianum, affects blood coagulation: an in vitro pilot study

The health-promoting properties of Silybum marianum have been acknowledged since antiquity. This plant is credited with substantial hepatoprotective properties and is also protective in cardiovascular diseases, diabetes mellitus, and neurodegeneration, mainly for its anti-inflammatory and antioxidant effects. Only a few experimental studies have described the impact of Silybum marianum extract on the blood coagulation process; furthermore, these data are unsatisfactorily fragmented and need to be supplemented to understand the plant’s properties better. The predominant biologically active flavonolignan extracted from Silybum marianum is silybin, a mixture of two diastereomers, silybin A and silybin B, in approximately equimolar ratio. This study investigated the effect of silybin on the fundamental laboratory parameter for blood coagulation, namely prothrombin time (PT), an assay used to assess the extrinsic and common coagulation pathways.
To evaluate the effect of silybin on PT, we prepared three solutions of silybin (Silybin (A + B mixture), PhytoLab GmbH & Co. KG, Vestenbergsgreuth, Germany) in 0.1% dimethylsulfoxide (DMSO, Sigma-Aldrich, Co., St. Louis, MO, USA): 10 μM, 50 μM, and 100 μM. PT was measured on a Coag 4D coagulometer (DIAGON Kft., Budapest, Hungary) using rabbit calcium thromboplastin (Dia-PT, DIAGON Kft., Budapest, Hungary) and control plasma which is pooled plasma obtained from healthy donors (Dia-CONT, DIAGON Kft., Budapest, Hungary). 10 µl of silybin solution was added to 40 µl of plasma, the sample was incubated for two minutes at 37 °C, and then 100 µl of thromboplastin, pre-warmed to 37 °C, was added to the mixture. The coagulometer automatically gives the PT result in seconds (s). At the same time, PT was measured in the control plasma both without additional solutions and with the addition of tris-buffered saline (TBS) and 0.1% DMSO (10 µl of TBS or DMSO + 40 µl of plasma). Each measurement was made eight times. Student’s t-test and the Friedman test with post-hoc analysis were used in the statistical analysis (Statistica 13, TIBCO Software Inc. Palo Alto, CA, USA).
In the first step of our study, we tested how the dilution of the plasma sample affected PT. We did not observe statistically significant differences in PT between the control plasma and the control plasma supplemented with TBS (mean ± standard deviation 14.00±0.77 s vs. 13.88±0.38 s, p=0.606). We also found no statistically significant differences in PT between the control plasma and the control plasma with the addition of 0.1% DMSO (mean ± standard deviation 14.00±0.77 s vs. 14.10±0.26 s, p=0.728); therefore, we further analyzed the effect of silybin on PT using DMSO at this level (0.1%). The addition of silybin solutions to the control plasma resulted in a statistically significant PT shortener (p<0.001). Post-hoc analysis revealed a substantial shortening of PT under the influence of 50 μM (median 13.55 s) and 100 μM solution (median 13.40 s) of silybin compared to plasma with the addition of 0.1% DMSO alone (median 14.10 s) and plasma with the addition of the lowest, 10 μM, level of silybin (median 14.20 s). At the same time, PT in the plasma with the addition of a 50 μM and 100 μM solution of silybin did not significantly differ statistically.
Our in vitro analysis characterized the possible effect of Silybum marianum on the blood coagulation process. These results require further investigation to validate their validity and clinical utility.

  • Open access
  • 34 Reads
HPLC screening of phytoestrogens from soybeans in conjunction with chemometric data analysis: a tool for selecting the best raw materials for producing dietary supplements for menopausal health

Soybeans are widely grown crops that have numerous uses, including as a food source, animal feed, and biofuel, as well as for the production of various chemicals. They are a rich source of plant-based protein and are also high in fiber, vitamins, and minerals. Due to their nutritional value and health benefits, there is an increasing concern on breeding soybeans with improved protein, fatty acid and isoflavone content. Soybeans are currently one of the primary dietary sources of isoflavones; these phytoestrogens have numerous health benefits, particularly for women's health, being increasingly used in the production of dietary supplements in the last decade. Isoflavones have been shown to have positive effects on human health, such as: alleviating the menopausal symptoms, reducing the risk of certain types of cancer (breast, prostate, and colon cancer), lowering the cholesterol levels, improving heart health by decreasing inflammation and increasing the blood flow, protective effect against osteoporosis, reducing the risk of depression and anxiety, even a positive effect on cognitive function and mental health. In this context, the goal of this research is to provide a combined approach of high performance liquid chromatography (HPLC) and chemometric techniques, able to highlight in a fast and convenient way the isoflavones’ content of soybean seeds belonging to different genotypes. A fast and sensitive HPLC method have been developed, optimized and validated for the analysis of the target analytes: isoflavone analysis was accomplished using a Flexar UHPLC system with UV detection, enabling the separation of genistein, glycitein, daidzein, daidzin, glycitin and genistin in less than 9 minutes. The resulted chromatographic data were further preprocessed (autoscalled), then subjected to principal component analysis (PCA) using Matlab. In this study, soybean seeds originating from genotypes harvested at the Research & Development Station for Agriculture, Turda - Romania were used. The analyzed genotypes showed particular isoflavones profiles, depending on genetic factors, the major isoflavones being the glycosides daidzin and genistin (more than 100 mg/ 100 g). The resulted PCA model revealed both the genotypes with the best quality attributes and similarities between the studied ones. Overall, the proposed approach can be considered not only for quality control assessment purposes, but also for assisting breeding programs targeted to develop new genotypes with the desired isoflavones’ content.

  • Open access
  • 50 Reads
EXPERIMENTAL IDENTIFICATION OF ABERRANLY EXPRESSED LONG NON-CODING RNAS IN BREAST CANCER

Relevance. Breast cancer (BC) remains one of the leading causes of cancer deaths among women worldwide. However, so far very little is known about the exact mechanisms of the occurrence of this type of cancer and the regulation of carcinogenesis, which complicates the diagnosis, prognosis and therapy. Recently, studies of long non-coding RNAs (lncRNAs) involved in the regulation of various signaling pathways in cells have become increasingly important, since they demonstrate great prognostic potential in cancer.

The aim of our work was to identify new aberrantly expressed long non-coding RNAs in breast cancer.

Materials and methods. Total RNA was isolated from paired breast cancer samples according to the standard method. Analysis of the expression of 84 lncRNAs in tumor and histologically normal adjacent breast tissue was performed using RT² lncRNA PCR Arrays (LAHS-002Z, QIAGEN). Additionally, data on lncRNAs expression in breast cancer from the GEPIA database (http://gepia.cancer-pku.cn/) were analyzed.

Results. We have identified 30 aberrantly expressed lncRNAs in breast cancer. For most lncRNAs, a decrease in the expression level by 2.34–13.2 times (p<0.05) was noted, and only for lncRNA TERC, an increase in the expression level by 2.24 times (p=0.034) was noted. Of greatest interest are the data obtained for lncRNAs ADAMTS9-AS2, EMX2OS, HOTAIRM1 and MEG3, as they are consistent with the data of bioinformatics analysis. However, further studies of larger group of samples are needed to evaluate the biomarker potential of the identified lncRNAs in breast cancer.

Conclusions. Experimental and bioinformatic analysis of changes in lncRNA expression in breast cancer revealed lncRNAs TERC, ADAMTS9-AS2, EMX2OS, HOTAIRM1, and MEG3 with biomarker potential.

This work was supported by the Russian Science Foundation grant no. 22-75-00132.

  • Open access
  • 72 Reads
The development of medical shampoo with plant-based pharmaceutical substance for treatment of seborrheic dermatitis

Dermatological diseases of the scalp significantly affect the life quality of people. According tostatistical data, seborrheic dermatitis (SD) is the most common dermatological disease in morethan 50% of the population all over the world. There are many possible causes of SD includingsebum accumulation, less frequent shampooing, and sensitivity to hair care products.Maintaining personal cleanliness and reducing unwanted effects on scalp can be achieved byapplication of special shampoo. Most commercial antidandruff shampoos employ syntheticsubstances as active ingredients, but their regular use changes a biodiversity of scalp microfloraand determines subsequent antimicrobial resistance. Therefore, the aim of study was thedevelopment of medical shampoo with plant-based pharmaceutical substance for treatment ofSD. The inhibitory activity against S. epidermidis, S. aureus, C. albicans of the plant-basedsubstance based on M. alternifolia essential oil, 1,8-cineol (eucalyptol) and α-(-)-bisabolol andshampoo containing the substance were investigated by micro-broth dilution and broth dilutionmethod, respectively. The medical shampoo with 0.75% substance possessed high antibacterialactivity against S. epidermidis and S. aureus with log CFU>1.0. Interestingly, the antifungalactivity against C. albicans (model of Malassezia spp.) was compatible to that of ketoconazole,climbazole and piroctone olamine. Shampoo with developed combination of surfactants as laurylglucoside, sodium coco-sulfate, coco-glucoside and cocamidopropyl betaine had the bestformulation in terms of microbiology and pH-stability, density foam characteristics, hairsmoothness, soothing and scalp cleansing. Therefore, this medical shampoo with natural originingredients was suggested for use in treatment of SD and dandruff.

  • Open access
  • 23 Reads
Modulatory Action of Phenolic-enriched Combretum paniculatum Vent Ethanolic Extract on Oxidoinflammatory Anomalies in Experimental Animals

The use of medicinal plants with favourable therapeutic effects has gained interest over conventional drugs in the treatment of oxidative stress and inflammatory-mediated diseases. The antioxidant and anti-inflammatory activities of Combretum paniculatum ethanolic extract (CPEE) were investigated in this study using in vitro and in vivo analysis. The results of phytochemical screening recorded in mg/100 g revealed that CPEE contains phenols (2711.02 ± 60.66), tannins (21.12 ± 0.41), flavonoids (49.00 ± 6.74), alkaloids (605.83 ± 10.10), steroids (0.64 ± 0.06), terpenoids (12.17 ± 0.55), reducing sugar (57.03 ± 0.12), and glycosides (2.59 ± 0.82). Our in vitro analysis revealed that CPEE inhibited nitric oxide, phospholipase A2, and thiobarbituric acid reactive substance activities with half-maximal inhibitory concentration (IC50) values of 6.55, 361.1, and 2.28 µg/mL, respectively. Furthermore, the in vivo study showed that implantation of cotton pellets elicited an increase in granuloma tissue formation, levels of malondialdehyde (MDA), and C-reactive protein (CRP) while decreasing the activities of superoxide dismutase (SOD), catalase (CAT), and reduced glutathione (GSH) in the untreated groups compared to normal rats. Interestingly, the groups treated with 100 and 200 mg/kg body weight of CPEE had decreased granuloma tissue, MDA, and CRP with an increase in the activities of SOD, CAT, and GSH. These findings suggest that CPEE ameliorated chronic inflammatory-induced oxidative stress in the experimental animals. Thus, it could be applied as an effective remedy for developing antioxidant and anti-inflammatory drugs.

  • Open access
  • 18 Reads
Nepeta nuda L. plant extract preserves the morphologhy of red blood cells subjected to oxidative stress

Medicinal plant extracts are widely explored for their beneficial effects on human health. A number of previous reports highlight the protective effect of plant extracts against oxidative damage, including on erythrocytes (e.g. Luqman et al. Pharmaceutical Biology, 2009; 47(6): 483–490). The catmint Nepeta nuda L. (Lamiaceae) was reported to exhibit antioxidant effects due to its phenolic compounds and iridoids, however to the best of our knowledge, no studies have been performed so far on red blood cells (RBC).

In this work, we evaluate the effect of water extract from catmint flowers prepared at 60 °С on RBC resistance to hemolysis and oxidative stress-induced deformation. Control studies are performed with 0.1 M Trolox antioxidant that is a water-soluble analogue of the vitamin E. First we show that the utilized extract is not hemotoxic since RBC (1 mg haemoglobin/ml) treatment with 0.01 – 1 mg/ml extract for 1 h does not induce cell hemolysis and does not alter significantly the RBC shape. In vitro induced oxidative stress by 0.8 M H2O2 leads to significant increase in lipid peroxidation (evaluated by malondialdehyde level). This is accompanied by reduction in the number of biconcave RBC and increase in the amount of echinocytes. Pre-treatment of RBC with catmint extract and Trolox does not reduce the lipid peroxidation level, however it results in partial restoration of the relative abundance of biconcave cells and respective reduction in echinocytes number.

In conclusion, our data show that water catmint extract exhibits protective effect on RBC morphology in the conditions of H2O2-induced oxidative stress.

Acknowledgment: Financial supported was obtained by the Bulgarian National Science Fund (BNSF), Grant No. KP-06-N56/9/12.11.2021. Equipment of the Distributed Scientific Infrastructure INFRAMAT (National Roadmap of Bulgaria for Scientific Infrastructure), financially supported by the Ministry of Education and Science was utilized.

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