Inhibition Of Hemolysis Research Articles

Antioxidant, anti-inflammatory, antiulcerogenic, and haemolytic properties of hydroethanolic and aqueous extracts from Tabernaemontana crassa Benth. fruit

Anti-inflammatory, antioxidant, antiulcer, and cytotoxic properties of hydroethanolic and aqueous extracts from the fruits of Tabernaemontana crassa were investigated. Four phenolic compounds, including three phenolic acids (gallic, caffeic, and chlorogenic acids) and one flavonol (rutin), were detected by HPLC-PDA. The hydroethanol extract (HE) exhibited high antioxidant activity and inhibition of haemolysis against red blood cells. Administration of HE (200 mg kg−1) significantly decreased the paw edoema induced by carrageenan compared to diclofenac sodium (20 mg kg−1). At the same dose, HE exhibited appreciable gastric cytoprotective capacity compared to Lanzoprazole (20 mg kg−1). These results were further supported by the histological analysis. Crude fruit extracts demonstrated a low haemolytic effect against rat red blood cells. Our findings support the use of T. crassa fruit in pharmacological and therapeutic fields.

Development of a Topical Cream from the Ethanolic of Agave sisalana Residues with Anti-Inflammatory and Analgesic Properties

Brazil is the largest producer in the world of the species Agave sisalana, sisal. The residue of the sisal, which is the result of the extraction of fibers from its leaves, represents 95% of its weight. Considering that sisal leaves have high concentrations of sapogenins and aiming at a future phytotherapeutic, in this study, the alcoholic fraction of sisal, AFS, was developed, and the sapogenins were characterized. In vitro, the cytotoxicity (MTT) and the anti-inflammatory effect of AFS (phagocytosis and hemolysis inhibition) were evaluated. In vivo, the analgesic (formalin test—FT) and anti-inflammatory (paw edema test—PET) activities of AFS, orally, and the cream containing AFS, topical, were analyzed. The results demonstrated that AFS contains hecogenin and tigogenin and is not cytotoxic. In vitro, 0.5, 1, and 2 mg/mL of AFS showed anti-inflammatory activity similar to the positive control (PC). In the FT, the dose of 25 mg/kg did not differ from the PC in the neurogenic phase (p > 0.05). In the PET, 25 and 50 mg/kg of AFS differed from the negative control (NC) (p < 0.05), and the cream with AFS (5 mg/g) showed activity similar to the PC. The therapeutic activities of AFS probably result from sapogenins. In the future, we expect to develop an anti-inflammatory from the thousands of tons of sisal waste discarded in Brazil.

Antisickling, hemolytic activities and toxicological evaluation of three plants species from the Nigerien Pharmacopoeia: Annona senegalensis Linné. Boscia senegalensis (Pers) Lam. and Ampelocissus afr

The aim of this study consits to evaluate the antisickling and antihemolytic activities and to determine the subacute toxicity of three plants Annona senegalensis L.; Boscia senegalensis (Pers) Lam.; and Ampelocissus africana (Lour) Merr.. The plant material contains leaves and bark of Annona senegalensis roots, leaves and bark root of Boscia senegalensis and the roots of Ampelocissus africana. A single method (80% ethanol maceration) is used to extract the five samples. The antisickleemic and antihemolytic effects are established with sickle cells disease (SCD) in the presence of sodium metabisulfite. Subacute toxicity is tested on Wistar strain rats. All samples showed an antisickling effect but leaves of Annona senegalensis (LAS) showed the highest percentage of antisickling activity (89.55%) at 50mg/ml. The anti-hemolytic effect was demonstrated in all samples by inhibition of hemolysis in the presence of a hemolysant. The toxicity results showed that the aqueous extracts of Annona senegalensis leaves and Boscia senegalensis root bark have non-toxic effects to wistar strain rats. However, some manifestations including fatigue, stretching and slight dyspenea were observed at doses of 150mg/mL and 300mg/mL. This work can contribute as a starting point towards the development of an improved traditional medicine (ITM) method which can help to treat the sickle cell disease. Keywords: Annona senegalensis L., Boscia senegalensis (pers) Lam and Ampelocissus Africana (Lour) Merr. Antisickness activity, Toxicity.

Glycyrrhetinic acid reduces lung inflammation caused by pneumococcal infection by reducing the toxicity of pneumolysin

ObjectiveIn this study, to provide new methods for the treatment of Streptococcus pneumoniae infection, we aimed to describe the anti-inflammatory and antibacterial value of glycyrrhetinic acid on the basis of its inhibitory effect on bacterial growth (without killing the bacteria) and its reduction of the toxicity of S. pneumoniae. MethodsA mouse model was established via intranasal administration of Streptococcus pneumoniae D39, and glycyrrhetinic acid was subcutaneously injected for treatment. The wet‒dry ratio, bacterial flora content and inflammatory factor levels in the mouse lungs were determined. Cell experiments were used to evaluate glycyrrhetinic acid-mediated inhibition of PLY hemolysis and A549 cell death, and WB was used to measure glycyrrhetinic acid-mediated inhibition of PLY oligomerization. ResultsGlycyrrhetinic acid reduced the levels of inflammatory factors, the dry‒wet ratio, the abundance of S. pneumoniae in the lungs of infected mice, pneumolysin-mediated A549 cell death, erythrocyte hemolysis and PLY oligoplasia. ConclusionGlycyrrhetinic acid can reduce the virulence of S. pneumoniae by preventing the oligomerization of PLY.

Antioxidant and Biological Activity of Mexican Madroño Fruit (Arbutus arizonica).

The fruit of the Mexican madroño (Arbutus arizonica) has been consumed since pre-Columbian times by North American tribes and native groups in Mexico. Despite this, reports on its chemical composition and biological activity are limited. This work aims to determine the antioxidant, antiproliferative, and digestive enzyme inhibition activities of the methanol amberlite-retained extract of Mexican madroño. Results showed that madroño fruit is rich in antioxidants: DPPH (EC50 = 0.89 ± 0.03 mg/mL), TEAC (1078 ± 4.9 μM/g), and hemolysis inhibition (IC50 = 358.07 μg/mL), with high phenolic and flavonoid content at 15.92 ± 3.2 mg GAE/g and 4.33 ± 0.3 mg CA/g, respectively. Using analytical chromatography, gallic acid, vanillic acid, chlorogenic acid, ferulic acid, quercetin, and rutin were quantified. The extract also showed α-glucosidase inhibition (IC50 = 3.1 ± 0.17 mg/mL), but no inhibition against α-amylase and lipase (>5 mg/mL), while showing antiproliferative activity against HeLa, HT-29, and MCF-7 cancer cell lines. These results point towards an interesting potential for the fruit of the A. arizonica as chemopreventive and hold potential for elaborating functional foods.

Biological activities of gaba-producing probiotics from fermented panchagavya

Abstract Panchagavya, a venerable elixir from the Ayurvedic tradition, merits scholarly inspection. It denotes the sacred offerings of cow’s milk, urine, dung, ghee and curd, holding medicinal import within Ayurveda’s ancient practices. This study aims to illuminate differences between probiotic variants V2 and V7, through findings on their pharmacological applications and biochemical concentrations. It is proposed that fermented panchagavya’s psychobiotic strains may exhibit favorable reactions worth examining. Chiefly, the organic synthesis of neurotransmitters by beneficial bacteria is of interest. The regulatory neurotransmitter gamma-aminobutyric acid (GABA) could potentially be generated by a panchagavya-derived probiotic. Through serial dilution culture of bacteria obtained from fermented panchagavya, their growth and GABA formation, as evaluated by thin-layer chromatography and ultraviolet-visible spectrometric analysis, are observed. Strain V2 demonstrates promising pharmacological properties including antioxidant, anti-inflammatory, anti-diabetic and anti-obesity effects, as well as inhibition of albumin denaturation and heat-induced hemolysis. It presents a higher concentration of such activities than V7. Biochemical tests further indicate V2’s larger percentage of GABA synthesis, supporting prospective therapeutic applications. After sequencing selected strains to explore psychobiotic functioning, their isolation holds medical import as GABA-producing beneficial bacteria from fermented panchagavya.

Quercetin disrupts biofilm formation and attenuates virulence of Aeromonas hydrophila.

Aeromonas hydrophila poses significant health and economic challenges in aquaculture owing to its pathogenicity and prevalence. Overuse of antibiotics has led to multidrug resistance and environmental pollution, necessitating alternative strategies. This study investigated the antibacterial and antibiofilm potentials of quercetin against A. hydrophila. Efficacy was assessed using various assays, including antibacterial activity, biofilm inhibition, specific growth time, hemolysis inhibition, autoaggregation, and microscopic evaluation. Additionally, docking analysis was performed to explore potential interactions between quercetin and virulence proteins of A. hydrophila, including proaerolysin, chaperone needle-subunit complex of the type III secretion system, and alpha-pore forming toxin (PDB ID: 1PRE, 2Q1K, 6GRK). Quercetin exhibited potent antibacterial activity with 21.1 ± 1.1 mm zone of inhibition at 1.5 mg mL-1. It also demonstrated significant antibiofilm activity, reducing biofilm formation by 46.3 ± 1.3% at the MIC and attenuating autoaggregation by 55.9 ± 1.5%. Hemolysis was inhibited by 41 ± 1.8%. Microscopic analysis revealed the disintegration of the A. hydrophila biofilm matrix. Docking studies indicated active hydrogen bond interactions between quercetin and the targeted virulence proteins with the binding energy -3.2, -5.6, and -5.1 kcal mol⁻1, respectively. These results suggest that quercetin is an excellent alternative to antibiotics for combating A. hydrophila infection in aquaculture. The multifaceted efficacy of quercetin in inhibiting bacterial growth, biofilm formation, virulence factors, and autoaggregation highlights the potential for aquaculture health and sustainability. Future research should delve into the precise mechanisms of action and explore synergistic combinations with other compounds for enhanced efficacy and targeted interventions.

Insights into the antioxidant, anti-inflammatory and anti-microbial potential of Nigella sativa essential oil against oral pathogens

Oral disorders can exert systemic ramifications beyond their localized effects on dental tissues, implicating a wide array of physiological conditions. The utilization of essential oils (EOs) for protection of oral health represents a longstanding practice. Consequently, in this investigation, essential oil derived from Nigella sativa seeds (NSEO) underwent isolation via the hydro-distillation process, followed by a comprehensive evaluation of its antioxidant, anti-inflammatory, anti-fungal, antibacterial activities, and cytocompatibility. The isolated NSEO manifested as a pale-yellow substance and was found to harbor a diverse spectrum of bioactive constituents, including steroids, triterpenoids, flavonoids, phenols, proteins, alkaloids, tannin, sesquiterpenoid hydrocarbons, monoterpenoid alcohol, and monoterpenoid ketone (thymoquinone). Notably, the total phenolic content (TPC) and total flavonoid content (TFC) of NSEO were quantified at 641.23 μg GAE/gm and 442.25 μg QE/g, respectively. Furthermore, NSEO exhibited concentration-dependent inhibition of protein denaturation, HRBC membrane stabilization, and hemolysis inhibition. Comparative analysis revealed that NSEO and chlorhexidine (CHX) 0.2% displayed substantial inhibition of hemolysis compared to aspirin. While NSEO and CHX 0.2% demonstrated analogous antibacterial activity against Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa, NSEO showcased heightened efficacy against Lactobacillus acidophilus and Candida albicans. Additionally, NSEO exhibited pronounced effects against periodontal pathogens such as Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, Tannerella forsythia, and Prevotella intermedia. Importantly, no cytotoxicity was observed on human gingival fibroblast cell lines. These findings underscore the potential of NSEO as a potent antibacterial and antifungal agent in the management of oral microbial pathogens, thereby offering avenues for the development of innovative therapies targeting diverse oral inflammatory conditions. Nevertheless, further investigations are imperative to unlock its full therapeutic repertoire.

INVESTIGATION OF POTENTIAL ACTIVITIES OF PEGANUM HARMALA SEEDS: IN SILICO AND IN VITRO ANALYSES

Seeds of Peganum harmala L. have been traditionally used in Algerian medicine. This study investigates whether the antioxidant, antihemolytic, and anti-inflammatory activities of Peganum harmala extracts (PHE) are attributed to polyphenolic compounds, which are abundant in methanol, chloroform, and ethyl acetate extracts. Extraction and fractionation of polyphenols involved solvents with different polarities, resulting in a crude extract (CrE), a chloroform extract (CHE), and an ethyl acetate extract (EAE). The antioxidant potential of CrE and its fractions was assessed using the ferrothiocyanate (FTC) and thiobarbituric acid (TBA) assays. The inhibition of mice erythrocyte hemolysis was evaluated for methanol, chloroform, and ethyl acetate extracts in the presence of the oxidant (AAPH). PMA-induced mouse ear edema was used as an in vivo model for inflammation. The FTC assay demonstrated the strong antioxidant effect of CrE (87.64 ± 0.003%). EAE showed potent antioxidant activity with low MDA absorption levels. Assessment of antihemolytic effects against AAPH-induced oxidative hemolysis revealed significant protective effects of CrE and EAE, with EAE showing the most pronounced effect. In the in vivo model, CrE (100 mg/kg) exhibited substantial anti-inflammatory activity, inhibiting the ear edema. Computational analyses using molecular docking simulations showed that chlorogenic acid, hesperetin, and rutin have promising potential as inhibitors of COX-2 protein, which is a key component in inflammatory pathways. This study highlights the potent antioxidant properties of P. harmala, particularly in CrE, and its anti-inflammatory effects. The bioactive compounds, such as chlorogenic acid, hesperetin, and rutin, exhibit potential as anti-inflammatory agents. P. harmala could be a valuable natural source for potential medical applications, suggesting the need for further exploration in medical treatments.

Hesperidin - loaded PVA/alginate hydrogel: targeting NFκB/iNOS/COX-2/TNF-α inflammatory signaling pathway.

Skin injuries represent a prevalent form of physical trauma, necessitating effective therapeutic strategies to expedite the wound healing process. Hesperidin, a bioflavonoid naturally occurring in citrus fruits, exhibits a range of pharmacological attributes, including antimicrobial, antioxidant, anti-inflammatory, anticoagulant, and analgesic properties. The main objective of the study was to formulate a hydrogel with the intention of addressing skin conditions, particularly wound healing. This research introduces a methodology for the fabrication of a membrane composed of a Polyvinyl alcohol - Sodium Alginate (PVA/A) blend, along with the inclusion of an anti-inflammatory agent, Hesperidin (H), which exhibits promising wound healing capabilities. A uniform layer of a homogeneous solution comprising PVA/A was cast. The process of crosslinking and the enhancement of hydrogel characteristics were achieved through the application of gamma irradiation at a dosage of 30 kGy. The membrane was immersed in a Hesperidin (H) solution, facilitating the permeation and absorption of the drug. The resultant system is designed to deliver H in a controlled and sustained manner, which is crucial for promoting efficient wound healing. The obtained PVA/AH hydrogel was evaluated for cytotoxicity, antioxidant and free radical scavenging activities, anti-inflammatory and membrane stability effect. In addition, its action on oxidative stress, and inflammatory markers was evaluated on BJ-1 human normal skin cell line. We determined the effect of radical scavenging activity PVA/A (49 %) and PVA/AH (87%), the inhibition of Human red blood cell membrane hemolysis by PVA/AH (81.97 and 84.34 %), hypotonicity (83.68 and 76.48 %) and protein denaturation (83.17 and 85.8 %) as compared to 250 μg/ml diclofenac (Dic.) and aspirin (Asp.), respectively. Furthermore, gene expression analysis revealed an increased expression of genes associated with anti-oxidant and anti-inflammatory properties and downregulated TNFα, NFκB, iNOS, and COX2 by 67, 52, 58 and 60%, respectively, by PVA/AH hydrogel compared to LPS-stimulated BJ-1 cells. The advantages associated with Hesperidin can be ascribed to its antioxidant and anti-inflammatory attributes. The incorporation of Hesperidin into hydrogels offers promise for the development of a novel, secure, and efficient strategy for wound healing. This innovative approach holds potential as a solution for wound healing, capitalizing on the collaborative qualities of PVA/AH and gamma irradiation, which can be combined to establish a drug delivery platform for Hesperidin.

Assessment of Sourdough Fermentation Impact on the Antioxidant and Anti-Inflammatory Potential of Pearl Millet from Burkina Faso.

Millet, a gluten-free cereal, has received attention for its environmental friendliness and higher protein content than other grains. It represents a staple food in many African countries, where fermentation is traditionally used for preserving food products and preparing different cereal-based products. This study aimed to assess the impact of sourdough fermentation on bioactive compounds and antioxidant and anti-inflammatory properties of pearl millet from Burkina Faso. Phenolic compounds were investigated spectrophotometrically and by HPLC-DAD. The antioxidant activity of unfermented (MF) and fermented (FeMF) millet was evaluated in vitro by spectrophotometric and fluorometric assays and ex vivo on oxidized human erythrocytes for hemolysis inhibition. Finally, the potential anti-inflammatory effect of FeMF and MF was evaluated on human adenocarcinoma cell line (HT-29) exposed to TNF-α inflammatory stimulus. Results revealed significantly higher levels of polyphenols, flavonoids, and in vitro antioxidant activity following millet fermentation. Notable differences in phenolic composition between FeMF and MF are observed, with fermentation facilitating the release of bioactive compounds such as gallic acid, quercetin, and rutin. A dose-dependent protection against oxidative hemolysis was observed in both FeMF- and MF-pretreated erythrocytes. Similarly, pretreatment with FeMF significantly reduced the levels of inflammatory markers in TNF-α-treated cells, with effects comparable to those of MF. Fermentation with sourdough represents a simple and low-cost method to improve the bioactive compounds content and in vitro antioxidant activity of millet flour with promising nutraceutical potential.

Proximate composition, bioactive constituents, and therapeutic potentials of pasteurized Noni juice derived from <em>Morinda citrifolia</em> (L.) growing in Sri Lanka

Morinda citrifolia (L.), also known as Noni or Ahu in Sri Lanka, is infrequently consumed as a dietary supplement within the country due to its association with myths and perceived toxicity. Nonetheless, in Sri Lanka, limited scientific research has been undertaken in the past to explore the therapeutic potential of noni, with public awareness primarily focused on its exportdriven cultivation. The study focused on evaluating the proximate composition, bioactive constituents, and therapeutic potential of hydro-methanolic extracts from pasteurized noni fruit juice, employing an 80% methanol extraction method, filtration, and assessment through standard assays to elucidate its potentia health benefits. The juice showed noteworthy antioxidant potential through DPPH scavenging activity (57.90 ± 0.40 μg/mL) and ABTS scavenging activity (42.95 ± 1.33 μg/mL), as well as high nitric oxide inhibition activity (164.56 ± 3.22 μg/mL), suggesting its anti-inflammatory properties. The juice also displayed potential as an anti-diabetic agent by inhibiting alpha-amylase (27.68 ± 0.36 μg/mL) and alpha-glucosidase (25.67 ± 1.03 μg/mL). Furthermore, it exhibited potential in protecting against certain pathological processes through heat-induced hemolysis inhibition (36.08 ± 1.08 μg/mL), protein denaturation inhibition (56.14 ± 1.11 μg/mL), and proteinase inhibitory activity (38.04 ± 0.56 μg/mL). These findings support the potential use of pasteurized fruit juice as a functional food ingredient, offering benefits to both the food industry and consumers. Further research can be directed towards the development of Noni fruit-incorporated food products that possess desirable sensory attributes to mitigate any potential aversion associated with its taste and odor profiles.

Development and characterization of a hemolysis inhibition assay to determine functionality of anti-Streptolysin O antibodies in human sera

The high burden of disease and the long-lasting sequelae following Streptococcus pyogenes (Strep A) infections make the development of an effective vaccine a global health priority. Streptolysin O (SLO), is a key toxin in the complex pathogenesis of Strep A infection. Antibodies are elicited against SLO after natural exposure and represent a key target for vaccine-induced immunity. Here we present the setup and characterization of a hemolysis assay to measure functionality of anti-SLO antibodies in human sera. Assay specificity, precision, linearity, reproducibility, and repeatability were determined. The assay was demonstrated to be highly sensitive, specific, reproducible, linear and performed well in assessing functionality of anti-SLO antibodies induced by exposed individuals. Moreover, different sources of critical reagents, in particular red- blood cells, have been compared and had minimal impact on assay performance. The assay presented here has throughput suitable for evaluating sera in vaccine clinical trials and sero-epidemiological studies to gain further insights into the functionality of infection- and vaccine-induced antibodies.

Watercress oil loaded with gel: evaluation of hemolysis inhibition, antioxidant, antimicrobial, and healing properties.

Plant-derived compounds are renowned for their remarkable pharmacological properties, holding immense promise for therapeutic interventions in human health. In this study, we aimed to assess the antimicrobial, anti-hemolytic, antioxidant, and wound healing attributes of watercress oil incorporated into Vaseline gel (OLG) compared to watercress oil alone. OLG was formulated through a meticulous process involving the addition of Vaseline gel to the oil under agitation conditions. High-performance liquid chromatography analysis of watercress oil unveiled a rich array of phenolic compounds, including gallic acid (10.18μg/mL), daidzein (3.46μg/mL), and hesperetin (3.28μg/mL). The inhibitory zones caused by watercress oil alone against a spectrum of pathogens, including Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Enterococcus faecalis, and Candida albicans, were measured at 25 ± 0.3, 26 ± 0.1, 22 ± 0.2, 25 ± 0.2, and 24 ± 0.1mm, respectively. Notably, OLG exhibited slightly larger zones of inhibition (27 ± 0.2, 30 ± 0.2, 24 ± 0.1, 28 ± 0.1, and 25 ± 0.3mm) against the same microbial strains. Furthermore, the minimum inhibitory concentration (MIC) of OLG against E. coli and E. faecalis was lower compared to watercress oil alone, indicating enhanced efficacy. Similarly, the minimum bactericidal concentration (MBC) of OLG was notably lower across all tested bacteria compared to watercress oil alone. Inhibition of bacterial hemolysis, particularly K. pneumoniae, was significantly enhanced with OLG treatment, showcasing reductions of 19.4%, 11.6%, and 6.8% at 25%, 50%, and 75% MIC concentrations, respectively, compared to watercress oil alone. The antioxidant activity of both oil and OLG was quantified with IC50 values of 2.56 and 3.02μg/mL, respectively. Moreover, OLG demonstrated remarkable efficacy in wound healing assays, with notable enhancements in migration rate, wound closure, and area difference compared to control cells. In light of the observed antibacterial, antifungal, anti-hemolytic, and wound healing properties of OLG, this formulation holds therapeutic potential in treating microbial infections and promoting wound healing.

Chemical composition's effect on Solanum nigrum Linn.'s antioxidant capacity and erythrocyte protection: Bioactive components and molecular docking analysis.

Oxidative stress has been widely believed to be the mechanism responsible for developing diseases such as arthritis, asthma, dementia, and aging. Solanum nigrum Linn. is a common edible medicinal herb that belongs to the family Solanaceae which has more than 180 chemical components that have so far been discovered. The main bioactive components of these are steroidal saponins, alkaloids, phenols, and polysaccharides. This article presents comparative phytochemical profiling including total phenolic, total flavonoid, alkaloid, proanthocyanidins, tannin, and vitamin C contents of three Algerian S. nigrum samples collected from three different locations in the Algerian desert. Additionally, the potential antioxidant activity of the three samples was assessed by 2,2-diphenyl-1-picrylhydrazyl, ferric reducing antioxidant power, and oxidative hemolysis inhibition assay. Moreover, the correlation between the major phenolic phytoconstituents previously reported and isolated from the plant and antioxidant activity has also been done by in silico molecular docking. Ten bioactive compounds were docked with selected proteins, arachidonate-5-lipoxygenase (PDB: 6n2w) and cytochrome c peroxidase (PDB: 2x08), to check their affinity with binding sites of these proteins for the possible mechanism of action. The docking scores suggest that S. nigrum's quercetin and kaempferol may play a significant role in its antioxidant action.

Evaluation of the Effect of Rosa Damascena and Jasminum Sambac on Dental Plaque Regrowth Inhibition

Objective: To evaluate the effect of Rosa damascena and Jasminum Sambac on dental plaque regrowth by the Plaque Glycolysis and Regrowth Method (PGRM). Methods: Ethanolic extracts of fresh flowers of Rosa damascena and Jasminum Sambac were prepared and subject to evaluation of antimicrobial property against Streptococcus mutans and also evaluated for their antioxidant property, anti-inflammatory property and their effect on dental plaque regrowth by the PGRM. Results: Both Rosa damascena and Jasminum Sambac showed inhibition of Growth of S. mutans, and MIC for both was found to be 30mg/ml. The antioxidant activity of both showed a dose-dependent increase, with the highest activity of 90.70% at 100 μg/mL concentration for Rosa damascena and 82.93 % at 100 μg/mL concentration for Jasminum Sambac. Both showed some inhibition of RBC haemolysis in the Membrane Protection assay, indicating anti-inflammatory activity, at the concentration of 300 μg/ml. The Percentage RBC Protection for Rosa damascena and Jasminum Sambac was 11.92% and 9.27%. The PGRM assay established that at concentration of 30mg/ml, Extracts of Rosa damascena and Jasminum Sambac showed inhibition of biofilm formation, with highest inhibition observed between 4 hours and 6 hours of incubation, and all were statistically significant. Conclusion: Study concludes that Rosa damascena and Jasminum Sambac have an inhibitory effect on regrowth of dental plaque and has antioxidant, antibacterial and inflammatory properties.

Phytochemical characterization and phospholipase A2 inhibitory effect of Vitex negundo L. root extracts

Ethnopharmacological relevanceSnake bites are a critical health issue in many parts of the world particularly in Asian countries lacking efficient health facilities in rural areas. Cobra is the most common snake type in Asia and is responsible for a large number of mortalities particularly in rural areas. Plants are usually considered the most effective and easy-to-approach treatment for snake bites in rural areas of various countries. Vitex negundo L. is an important medicinal plant traditionally used to treat snake bite envenomation in many countries of Asia. Aim of the studyFrom literature survey of plants traditionally used in the treatment of snake bites in Asian countries including India, Pakistan and Sri Lanka, roots of V. negundo were selected for the present study. Anti-snake venom potential of its roots was assessed through various in vitro assays targeting the phospholipase A2 enzyme. Materials and methodsV. negundo roots were sequentially extracted in different organic solvents to get fractions and in methanol to get total extract. The extracts were evaluated for phospholipase A2 (PLA2) inhibitory potential through inhibition of venom-induced hemolysis, ADP-induced platelet aggregation, PLA2-induced fatty acid hydrolysis and anticoagulant effect of cobra venom. Antioxidant power was determined using DPPH and superoxide radical scavenging assays. GC-MS and HPLC analysis was performed for the total methanol extract. ResultsStrong PLA2 inhibitory effect was observed for all the extracts. The ethyl acetate, acetone and methanol fractions significantly inhibited toxic effects of cobra venom under in vitro conditions. Radical scavenging potential of these fractions was also significantly high as compared to non-polar fractions in both DPPH and superoxide scavenging assays. Phytochemical analysis indicated high phenolic and flavonoid contents in these fractions. GC-MS and HPLC analysis of total methanol extract confirmed the presence of bis(2-ethylhexyl) phthalate, phenol, o-Guaiacol, palmitic acid-methyl ester, methyl stearate, quercetin and kaempferol in the plant. ConclusionThe study concluded that the roots of V. negundo, particularly their polar extracts, have strong PLA2 inhibitory effect against cobra venom confirming their traditional use to manage snake bites. The roots of this plant can be further studied for isolation of plant-based antisera.

Fermentation, Identification, and Antioxidant Activity of Saponins Produced by a Wild Ginseng Endophytic Fungus Umbelopsis dimorpha Strain NSJG

Background: Endophytes from ginseng plants may produce saponins through in vitro fermentation, and some saponins possess antioxidant activity. This study aimed to enhance the total saponin yield of an endophytic fungus isolated from wild ginseng, determine saponin types, and explore whether saponin extracts from optimized fermentation protocols have antioxidant activity. Methods: Umbelopsis dimorpha strain NSJG was fermented in liquid under different conditions. The types of saponins were analyzed by Liquid Chromatography/Mass Spectrometry (LC/MS). The antioxidant activity of saponin extracts was evaluated by measuring their hemolytic inhibition rate on erythrocytes. Results: The endophytic fungus strain NSJG produced 1.236 mg/mL total saponins through liquid potato dextrose (PD) fermentation. The total saponin concentrations of optimal protocols G-ED-8 (3.107 mg/mL) and G-DP-8 (2.045 mg/mL) were three times and twice that of G-PD. Saponin extracts from G-ED-8 and G-DP-8 contained seven and eight types of ginsenosides and had higher hemolysis inhibition rates on erythrocytes than Vitamin C (VC), which were 36.05% and 46.69%. Conclusions: The research highlights the potential implications of wild ginseng endophytic fungus strain NSJG for the in vitro production of ginsenosides and the development of novel antioxidants.

SPECTROPHOTO AND DIGITAL COLORIMETRIC DETERMINATION INDICATOR OF BETA-ADRENOREACTIVITY OF ERYTHROCYTE MEMBRANES OF PATIENTS WITH BRONCHIAL ASTHMA

The change in the adrenoreactive properties of membrane and cellular structures under the influence of catecholamines makes it possible to consider the beta-adrenergic reactivity of the erythrocyte membrane (-ARM) as a systemic indicator of the organism's adrenoreactivity. To assess adrenoreactivity, a technique is used based on a quantitative assessment of the degree of inhibition of hypoosmotic hemolysis of erythrocytes in vitro in the presence of the beta-blocker propranolol. The article presents data on the comparison of spectrophoto- and colorimetric determination of erythrocyte membrane beta-adrenoreactivity based on the registration of erythrocyte hemolysis with digital signal processing. A smartphone was used to register the images of the analyzed samples. Photography was carried out in a special box under standard lighting conditions. The chromaticity of the obtained images was evaluated using the Photoshop program within the RGB color model intended for computer color rendering. The obtained RGB values were used for quantitative determinations. The study was performed on 20 blood samples of patients suffering from bronchial asthma. The discrepancy between the results of determining -ARM using the colorimetric and spetrophotometric methods was no more than 5.6%, which indicates the possibility of replacing the spectrophotometer with image registration in digital format using software.

Utilizing Extraepitopic Amino Acid Substitutions to Define Changes in the Accessibility of Conformational Epitopes of the Bacillus cereus HlyII C-Terminal Domain.

Hemolysin II (HlyII)-one of the pathogenic factors of Bacillus cereus, a pore-forming β-barrel toxin-possesses a C-terminal extension of 94 amino acid residues, designated as the C-terminal domain of HlyII (HlyIICTD), which plays an important role in the functioning of the toxin. Our previous work described a monoclonal antibody (HlyIIC-20), capable of strain-specific inhibition of hemolysis caused by HlyII, and demonstrated the dependence of the efficiency of hemolysis on the presence of proline at position 324 in HlyII outside the conformational antigenic determinant. In this work, we studied 16 mutant forms of HlyIICTD. Each of the mutations, obtained via multiple site-directed mutagenesis leading to the replacement of amino acid residues lying on the surface of the 3D structure of HlyIICTD, led to a decrease in the interaction of HlyIIC-20 with the mutant form of the protein. Changes in epitope structure confirm the high conformational mobility of HlyIICTD required for the functioning of HlyII. Comparison of the effect of the introduced mutations on the effectiveness of interactions between HlyIICTD and HlyIIC-20 and a control antibody recognizing a non-overlapping epitope enabled the identification of the amino acid residues N339 and K340, included in the conformational antigenic determinant recognized by HlyIIC-20.