Inhibition Of Hemolysis Research Articles

Aesculus hippocastanum L.: A Simple Ornamental Plant or a Source of Compelling Molecules for Industry?

Aesculus hippocastanum L., also known as horse chestnut, is an ornamental tree whose seeds are mostly discarded in landfills in the regions where they are grown. However, recent studies have shown that these seeds can be a source of interesting compounds for several industries. This work aimed to chemically characterize horse chestnut seeds at the level of compounds recognized for their wide bioactivity, i.e., organic acids, including phenolic compounds, using chromatographic methodologies (UFLC-DAD and LC-DAD-ESI/MSn). In addition, the bioactivity of these seeds was evaluated by in vitro methodologies, seeking to relate the respective (bio)activity to the compounds present in the endocarp (husk), seed coat (skin), and peeled seed (pulp). The antioxidant activity (lipid peroxidation inhibition and oxidative haemolysis inhibition), antibacterial potential (against Gram-positive and Gram-negative bacteria) and cytotoxicity (in human tumour cell lines and porcine liver primary cells) were evaluated. Kaempferol-O-pentoside-O-hexoside-O-hexoside was the main phenolic identified in the pulp. At the same time, (-)-epicatechin and β-type (epi)catechin dimer were the major phenolics present in husk and skin, respectively. In general, A. hippocastanum extracts presented antioxidant and antibacterial potential, without toxicity up to the maximal tested dose. Overall, these findings anticipate potential applications of A. hippocastanum seeds in food- or pharmaceutical-related uses.

Effects of simulated gastrointestinal digestion on chickpea protein, and its hydrolysate physicochemical properties, erythrocyte haemolysis inhibition, and chemical antioxidant activity

Chickpea protein and its hydrolysates have good antioxidant activity. Proteins and peptides are degraded and modified in the gastrointestinal tract before being digested and absorbed, a process that changes their physicochemical and biological properties. Changes in structure and antioxidant capacity of the chickpea protein and its hydrolysates after simulated gastrointestinal digestion were studied. The secondary structure, amino acid composition, and chemical antioxidant activity of chickpea protein and its hydrolysates were determined, and their protective effects on AAPH-induced erythrocyte oxidative damage were studied. Results showed that chickpea protein hydrolysate was easier to be digested by the gastrointestinal tract than chickpea protein. After digestion, the chemical antioxidant capacity of chickpea protein and its hydrolysates increased in a dose-dependent manner, and showed a protective effect against erythrocytes AAPH-induced oxidative damage. Chickpea protein and its hydrolysates can be used as natural antioxidants to promote healthy digestion.

Pharmacognostic standards for Uvaria chamae P. Beauv. leaves (Annonaceae)

Background : Standardisation as a tool in quality control ensures purity, safety and efficacy of herbal medicines. Uvaria chamae from the family Annonaceae has wide medicinal application which includes its use as a febrifuge and as an anti-inflammatory.Objective: The study was aimed at developing some Pharmacognostic standards for the leaves of U. chamae.Material and Methods: Uvaria chamae plant was collected, identified and authenticated. Fresh leaves were examined macroscopically and microscopically based on standard procedures. Fluorescence behavior was assessed by adding various reagents and viewing at daylight, 254 nm and 365 nm. Chemomicroscopy and physicochemical analysis of powdered samples were done using standard procedures. Phytochemical and Thin layer chromatographic analysis of the extracts were also carried out.Results: Leaves of U. chamae are alternately arranged and elliptic in shape. The abaxial surface has straight polygonal epidermal cells with paracytic stomata. Vascular bundles are arranged in fan shape from the transverse section of the leaf. Chemomicroscopic evaluation revealed the presence of lignin, starch and oil. Alcohol and water soluble extractive values were 4.00 ± 0.00 and 5.6 ± 0.67, respectively. The membrane stabilization activity of the extracts at 0-1 mg exceeded that of the standard drug diclofenac. The ethyl acetate fraction had the highest percentage of inhibition of haemolysis at 0.4 mg.Conclusion: The features established in this study may contribute in the identification and quality control of U. chamae.

The Application of Cinnamon Twig Extract as an Inhibitor of Listeriolysin O against Listeria monocytogenes Infection

As a major virulence factor of Listeria monocytogenes (L. monocytogenes), listeriolysin O (LLO) can assist in the immune escape of L. monocytogenes, which is critical for the pathogen to evade host immune recognition, leading to various infectious diseases. Cinnamon twig (CT), as a traditional medicine, has been widely used in clinics for multiple functions and it has exhibited excellent safety, efficacy and stability. There are few reports on the effects of the extracts of traditional medicine on bacterial virulence factors. CT has not been reported to be effective in the treatment of L. monocytogenes infection. Therefore, this study aims to explore the preventive effect of CT against L. monocytogenes infection in vivo and in vitro by targeting LLO. Firstly, a hemolysis assay and a cell viability determination are used to detect the effect of CT extract on the inhibition of the cytolytic activity of LLO. The potential mechanism through which CT extract inhibits LLO activity is predicted through network pharmacology, molecular docking assay, real-time polymerase chain reaction (RT-PCR), Western blotting and circular dichroism (CD) analysis. The experimental therapeutic effect of CT extract is examined in a mouse model infected with L. monocytogenes. Then, the ingredients are identified through a high-performance liquid chromatography (HPLC) and thin layer chromatography (TLC) analysis. Here we find that CT extract, containing mainly cinnamic acid, cinnamaldehyde, β-sitosterol, taxifolin, catechin and epicatechin, shows a potential inhibition of LLO-mediated hemolysis without any antimicrobial activity. The results of the mechanism research show that CT extract treatment can simultaneously inhibit LLO expression and oligomerization. Furthermore, the addition of CT extract led to a remarkable alleviation of LLO-induced cytotoxicity. After treatment with CT extract, the mortality, bacterial load, pathological damage and inflammatory responses of infected mice are significantly reduced when compared with the untreated group. This study suggests that CT extract can be a novel and multicomponent inhibitor of LLO with multiple strategies against L. monocytogenes infection, which could be further developed into a novel treatment for infections caused by L. monocytogenes.

Valorization of rambutan (Nephelium lappaceum L.) peel: Chemical composition, biological activity, and optimized recovery of anthocyanins

Chemical constituents and bioactive properties of rambutan (Nephelium lappaceum L.) peel were characterized and heat-/ultrasound-assisted extractions (HAE/UAE) of anthocyanins were optimized by response surface methodology. Five organic acids, the α-, γ-, and δ-tocopherol isoforms, and twenty-five fatty acids (36.8 % oleic acid) were identified, as well as a phenolic profile composed of ellagitannin derivatives, geraniin isomers, ellagic acid, and delphinidin-O derivatives. The extract showed antioxidant activity via lipid peroxidation (IC50 = 2.79 ± 0.03 µg/mL) and oxidative hemolysis (IC50 = 72 ± 2 µg/mL) inhibition, and displayed antibacterial and antifungal properties (MIC ≤ 1 mg/mL). On the other hand, no cytotoxicity was observed in tumor and non-tumor cell lines up to 400 µg/mL. The recovery of anthocyanins was more effective using HAE than UAE, allowing greater yields (16.2 mg/g extract) in just 3 min and using lower ethanol proportions. Overall, rambutan peel could be upcycled into bioactive ingredients and natural colorants for industrial applications.

Antioxidant, Anti-inflammatory, Antimicrobial and Thrombolytic Activities of Eclipta alba L. Growing in Bangladesh

Eclipta alba L. (Family: Asteraceae) is an evergreen herb of ethnomedicinal significance in the Indian subcontinent. The present study was carried out to determine the in vitro antioxidant, anti-inflammatory, antimicrobial and thrombolytic activities of n-hexane, chloroform and aqueous soluble fractions of ethanol crude extract of E. alba whole plant. Preliminary phytochemical screening was performed by qualitative tests which revealed the presence of alkaloids, steroids, glycosides, tannins, flavonoids, saponins and amides in E. alba. During the DPPH (2,2-diphenyl-1-picrylhydrazyl) scavenging assay, the aqueous soluble fraction of E. alba at a dose of 100 μg/ml showed maximum 87.5% inhibition in scavenging the DPPH radical with IC50 value of 7.86 μg/ml. Anti-inflammatory activity was evaluated by egg albumin denaturation technique and RBC membrane stabilizing method. In both experiments, all the test samples of E. alba showed a concentration-dependent anti-inflammatory potential while the aqueous soluble fraction at the dose of 500 μg/ml demonstrated the maximum 90.12% inhibition of egg albumin denaturation and 78.42% inhibition of RBC membrane hemolysis against hypotonic solution. Antimicrobial sensitivity was evaluated by disc diffusion assay against several bacterial and fungal species. All the test samples at a concentration of 500 μg/disc exhibited noticeable antibacterial (zone of inhibition = 11-23 mm) and antifungal (zone of inhibition = 10-21 mm) activities when compared to the respective standard drug. The aqueous soluble fraction showed maximum activity against Lactobacillus casai (zone of inhibition = 23 mm). During antifungal test, the maximum activity was exhibited by the n-hexane fraction against Pityrosporum ovale (zone of inhibition = 21 mm). During in vitro thrombolytic assay, the aqueous soluble fraction revealed 56.84% lysis of blood clot, as compared to the standard streptokinase (84.60%). The results of our study suggest that E. alba can be considered for further investigation in order to discover the pharmacologically active natural products. Bangladesh Pharmaceutical Journal 26(1): 20-27, 2023 (January)

Comparative analysis of antioxidant and anti-inflammatory activities of red, blue, and black tea for health benefits

The current COVID-19 predicament necessitates a greater emphasis on developing immunity. Herbal teas are abundant in antioxidants which are important for strengthening the immune system. Hot water decoction of Hibiscus rosa sinensis flowers (red tea), Clitoria ternatea flowers (blue tea) and commercially available black tea were evaluated by comparing for in vitro antioxidant and anti-inflammatory properties. Anthocyanin pigment in red, blue, and black tea demonstrated Rf values of 0.52, 0.86 and 0.78 respectively. Blue and black teas exhibited dominance of polyphenols, flavonoids, tannins, glycosides, terpenoids, saponins as compared to red tea. The highest total phenolic (12.25 ± 0.245 mg GAE/gm extract-1) and flavonoid (15.84 ± 0.268mg QE/gm extract-1) content were observed with black and blue tea respectively. Blue tea, and black tea extracts exhibited FRPA values of 1.81 ± 0.413 mg and 1.93 ± 0.178 mg AAE per gram extract-1 respectively. Black tea exhibits the highest antioxidant capacity in reducing molybdate ions (1.94 ± 0.354 mg AAE per gram extract-1) followed by blue tea (1.56 ± 0.199 mg AAE per gram extract-1). Blue tea extract at a very low concentration showed highest percentage hemolytic inhibition (57.14 ± 0.567%). According to the study, blue tea is a rich source of antioxidants with significant anti-inflammatory properties. The research may offer a valuable supplementary strategyfor its therapeutic applications.

The In Vitro Simulated Gastrointestinal Digestion Affects the Bioaccessibility and Bioactivity of Beta vulgaris Constituents.

Beetroot (Beta vulgaris L.) is an important root vegetable crop and a valuable food source of micronutrients and bioactive constituents. In this study, the bioaccessibility of minerals, organic acids, and betacyanins in beetroot powder during simulated gastrointestinal digestion was investigated, as well as the antioxidant activity of the final fractions of each phase of the process. Mineral elements were analyzed by inductively coupled plasma optical emission spectroscopy (ICP-OES), organic acids by ultra-fast liquid chromatography with photodiode array detection (UFLC-PDA), and betacyanins by liquid chromatography with diode-array detection and mass spectrometry (HPLC-DAD-ESI/MSn). The oxidative hemolysis inhibition assay was used to assess the ex vivo antioxidant activity. The bioaccessibility of minerals at the end of gastrointestinal digestion ranged from 43 to 65%, depending on the mineral element. Among these, Mg was the most bioaccessible, while Ca and Fe had the lowest bioaccessibility. For organic acids, a major release during digestion was observed for quinic acid. It was also found that betanin (the major betalain in beetroot) was highly unstable during the digestion process, probably due to its hydrophilic nature, which agreed with the significant (p < 0.05) decrease in antioxidant/antihemolytic activity. These results suggest that beetroot antioxidant compounds are unstable under gastrointestinal conditions, and could be useful for future development of novel and more stable beetroot food formulations.

Evaluation of Senna tora (L.) Roxb. leaves as source of bioactive molecules with antioxidant, anti-inflammatory and antibacterial potential

Senna tora (L.) Roxb. is an ethno-medicinal herb used by rural and tribal people of the Satpura region of Madhya Pradesh in India and the Phatthalung Province of Thailand for treating rheumatism, bronchitis, ringworm, itches, leprosy, dyspepsia, liver disorders and heart disorders. It is also used in Chinese and Ayurvedic medicine. This study was conducted to investigate the potential of Senna tora (L.) Roxb. as a source of drug candidates against oxidants, inflammation, and bacterial infection. Preliminary phytochemical screening (PPS) and GC-MS were performed to identify the phytochemicals in the ethyl acetate extract of Senna tora (L.) Roxb. leaves (EAESTL). The in vitro antioxidant activity was assessed by 2,2-diphenyl-1-picrylhydrazyl (DPPH)- and H2O2-scavenging tests; the in vitro anti-inflammatory activity was determined by bovine serum albumin (BSA) denaturation and red blood cell (RBC) hemolysis inhibition; and the antibacterial activity was evaluated by agar-well diffusion methods. Cytotoxicity was estimated by Artemia salina larvae lethality, while acute toxicity was evaluated by oral delivery of the extract to mice. In silico antioxidant, anti-inflammatory, and antibacterial activities were predicted by the Prediction of Activity Spectra for Substances (PASS) program. The pharmacokinetics related to ADME and toxicity tests were determined by the admetSAR2 and ADMETlab2 web servers, and drug-able properties were assessed by the SwissADME server. GC-MS detected fifty-nine phytochemicals that support the types of compounds (phenols, flavonoids, tannins, terpenoids, saponins, steroids, alkaloids, glycosides and reducing sugar) identified by phytochemical screening. EAESTL exhibited dose-dependent antioxidant, anti-inflammatory, and antibacterial activities without any adverse effects or fluctuations in body weight. The PASS program predicted that the identified phytochemicals have antioxidant, anti-inflammatory and antibacterial activities. Among 51 phytochemicals, 16 showed good ADME, and 8 fulfilled drug-able properties without toxicity. Altogether, four phytochemicals, viz., benzyl alcohol, 3-(hydroxy-phenyl-methyl)-2,3-dimethyl-octan-4-one, phenylethyl alcohol and 2,6,6-trimethylbicyclo [3.1.1] heptane-3-ol, showed good pharmacokinetics and drug-able properties without toxicity, along with antioxidant, anti-inflammatory, and antibacterial activities. The obtained results suggest that Senna tora (L.) Roxb. leaves contain bioactive phytochemicals that have the potential to fight against oxidants, inflammation, and bacterial infection as potential drug candidates.

Antisickling and Antihemolytic Mechanism of Spirulina platensis (Oscillatoriaceae): A Nutraceutical Commonly Used in Cameroon

Sickle cell anaemia (SCA) is a widespread genetic disease in Africa, associated with chronic hemolytic anaemia and vaso‐occlusive and infectious complications. The most commonly used means of management and treatment such as blood transfusions and allografting are expensive and predispose patients to the risk of infections. This research study aimed at evaluating the antisickling and antihemolytic activities of aqueous extracts of Spirulina platensis from Cameroon for optimising the management of this disease using natural substances. The Spirulina platensis harvested in Nomayos‐Yaounde was dried, crushed, and macerated for 24 h in distilled water and the filtrate was freeze dried. The determination of the inhibition rates of falciformation induced by 2% sodium metabisulfite (MBS) and the sickling reversibility rate was carried out at different concentrations (100, 200, 400, 800, and 1600 μg·mL−1) of the spirulina extract at different time intervals (2 h, 4 h, and 24 h). Aspirin, hypotonic solution, Triton X‐100, and hydrogen peroxide were used as hemolytic inducers and the antihemolytic activity of the extract was studied at 800 μg·mL−1 and 1600 μg·mL−1 using the colorimetric method. The extraction yield was 14.015%. The maximum duration of induced falciformation was 2 h 30 min and the percentage of falciformation increased from 27.99 ± 3.15% (at the initial time) to 91.44 ± 3.70%, giving a falciformation induction rate of 69.3%. The falciformation inhibition rate after 2 h 30 min ranges from 15.10 ± 0.60% to 66.09 ± 4.69% for the concentration of 100 μg·mL−1 to 1600 μg·mL−1 of the spirulina extract. This rate of inhibition of falciformation was found to be dose‐dependent. The best concentration of the extract was 1600 μg·mL−1. The reversibility rate of falciformation at 800 μg·mL−1 and 1600 μg·mL−1 varied from 37.54 ± 6.35% to 82.34 ± 5.63% as a function of time. 1600 μg·mL−1 was the most active concentration after 24 h. In addition, the extract improved the Fe2+/Fe3+ ratio with an increase in the rates of 69.78 ± 8.81 and 69.78 ± 13.82 at 800 μg·mL−1 and 1600 μg·mL−1, respectively. According to each inducer at 800 μg·mL−1 and 1600 μg·mL−1, respectively, of the spirulina extract, the following rates of inhibition of hemolysis were found: 53.03 ± 9.46% and 96.67 ± 5.77% (aspirin); 80 ± 8.66% and 71.25% (hypotonic solution); 36.56 ± 9.53% and 45.67 ± 22.55% (Triton X‐100); 24.26 ± 9.55% and 36.76 ± 1.27% (hydrogen peroxide). At the end of this study, the best antickling activities were obtained at the concentrations 800 μg·mL−1 and 1600 μg·mL−1. It also has antihemolytic properties on various hemolysis inducers at concentrations 800 μg·mL−1 and 1600 μg·mL−1 with inhibition rates varying from 36% to 96%.

Chemical Composition and Biological Activity of Commelina erecta: An Edible Wild Plant Consumed in Brazil.

In recent years, the interest in products of natural origin has boosted the exploitation and use of plants as food and sources of bioactive compounds, especially wild plants widely used in different cultures for several purposes. Commelina erecta is a wild edible plant (WEP) traditionally used as food and medicine, about which few studies exist. Thus, this study aimed at enhancing the knowledge about its nutritional, chemical and bioactive profile, considering different plant parts and development stages, in order to increase its inclusion in the diet of South American communities. The nutritional profile was found to be similar to other WEP frequently consumed in Brazil. Thirteen phenolic compounds (HPLC-DAD-ESI/MS) were tentatively identified, with apigenin, luteolin and quercetin derivatives being the most abundant. Fructose and oxalic acid were the major sugar and organic acid, respectively, in the aerial parts of C. erecta, and four isoforms of tocopherols were also identified. Regarding the plant's antioxidant activity, the EC50 values varied between 18.4 and 1060 µg/mL in the inhibition of lipid peroxidation assay (TBARS) and between 53 and 115 µg/mL in the oxidative haemolysis inhibition (OxHLIA) assay. The hydroethanolic extract obtained from stems at the flowering stage also presented anti-inflammatory activity. In general, all the extracts evidenced promising antimicrobial activity. Altogether, these results reinforce the traditional use of this plant species as food and medicine to support the diet of needier populations and also promote food sovereignty and sustainability.

Effects of Carboxymethyl Modification on the Acidic Polysaccharides from Calocybe indica: Physicochemical Properties, Antioxidant, Antitumor and Anticoagulant Activities.

An acidic polysaccharide fraction was obtained from Calocybe indica (CIP3a) after subjecting it to hot water extraction followed by purification through DEAE-cellulose 52 and Sepaharose 6B column chromatography. The CIP3a was further modified using chloroacetic acid to yield carboxymethylated derivatives (CMCIP3a). The modified polysaccharide was characterized using various spectroscopic methods. In addition, further antioxidant, antitumor and anticoagulant activities were also investigated. The polysaccharides CIP3a and CMCIP3a were heterogeneous in nature and composed of various molar percentages of glucose, arabinose and mannose with molecular weights of 1.456 × 103 and 4.023 × 103 Da, respectively. The NMR and FT-IR data demonstrated that the carboxymethylation on the polysaccharide was successful. In comparison to CIP3a polysaccharides, the modified derivatives had lower sugar and protein contents, and higher levels of uronic acid. The in vitro antioxidant activity showed that CMCIP3a with higher molecular weight displayed an elevated ability in scavenging the DPPH radical, ABTS, superoxide, hydroxyl radical, ferric reducing power, cupric reducing power and erythrocyte hemolysis inhibition with an EC50 value of 2.49, 2.66, 4.10, 1.60, 3.48, 1.41 and 2.30 mg/mL, respectively. The MTT assay results revealed that CMCIP3a displayed a dose-dependent inhibition on five cancer cells (HT29, PC3, HeLa, Jurkat and HepG-2) in the range of 10-320 μg/mL. The APTT, PT and TT were significantly extended by CMCIP3a in relation to dosage, indicating that the anticoagulant effect of CIP was both extrinsic and intrinsic, along with a common coagulation pathway. These findings demonstrated that carboxymethylation might effectively improve the biological potential of the derivatives and offer a theoretical framework for the creation of novel natural antioxidants, low-toxicity antitumor and antithrombotic drugs.

Protective effects of Brenania brieyi (De Wild) E.M.A.Petit root bark fractions against inflammatory-mediated hemolysis and dyslipidemia in rats

Background and Aims: The inflammatory response, though protective, is the major cause of debilitating diseases when provoked excessively or if left unresolved. Brenania brieyi (De Wild) E.M.A.Petit is widely used in folk medicine for the treatment of inflammatory-related diseases. This study investigated the protective effects of methanol and chloroform root bark fractions of Brenania brieyi on inflammation-induced hemolysis and dyslipidemia. Methods: Anti-inflammatory activity was investigated by inserting 20 mg of autoclaved cotton pellets into forty-five rats randomly distributed into nine groups (n=5), this excluded group 1 (baseline). The extent of hemolysis and dyslipidemia in the inflamed rats was ascertained from hematological parameters, lipid profile, and lipidemic index, while the possible underlying mechanisms of inflammation were determined using standard procedures. Results: Treatment with varying doses of the root bark fractions of B. brieyi elicited a significant (p< 0.05) decrease in granuloma tissue and an increase (p<0.05) in hemoglobin, red and white blood cell count, packed cell volume, and platelets compared with the untreated group 2. A significant (p<0.05) decrease in cholesterol, triacylglycerols, and low-density lipoprotein, and a non-significant (p>0.05) increase in high-density lipoprotein were observed in almost all the test groups compared with group 2. There was a significant restoration of atherogenic and dyslipidemia indices and inhibition of acetic acid-induced vascular permeability, membrane hemolysis, and platelet aggregation in the fraction-treated groups compared with the control. Conclusion: The findings from this study suggest that B. brieyi inhibits exudation and proliferation of granuloma-forming cells and also has the potential to restore the hematological parameters and lipid anomalies to their physiologic state under chronic inflammation. The possible mechanisms of its action could be inhibition of vascular permeability, stabilization of the membrane, or inhibition of platelet aggregation. This justifies the use of the plant in traditional medicine and also demonstrates its potential as a target for the discovery of new anti-inflammatory agents.

THE EFFECT OF AUXILIARY DRUGS ON PHOTODYNAMIC CYTOLYSIS IN PHOTODYNAMIC THERAPY

The effect of pyridine-containing and phenolic compounds (Mexidol, Emoxypine, Vitamin В6 , Paracetamol, Propofol, Coenzyme Q10) and imidazole-containing Dexdor on photoinduced hemolysis in PDT was studied in an experiment (on erythrocytes of healthy people’s blood). It was found that Mexidol and Emoxypine showed themselves not as antioxidants, but as powerful agents to enhance the hemolytic effect, while in the area of small doses they showed a membraneprotective antihemolytic effect. Therefore, the possibility of their use as concomitant drugs in PDT is problematic and additional clinical studies are required. The anesthetic Dexdor (dexmedetomidine) did not have a noticeable effect on the rate of hemolysis, which makes it possible to recommend it as a priority drug for drug support of PDT. Sedative Propofol (2,4-diisopropylphenol) and Paracetamol (p-acetoaminophenol) as most phenolic compounds caused significant inhibition of hemolysis, which makes their use in PDT problematic. Coenzyme Q10 (its reduced form – ubiquinol) showed a pronounced inhibitory effect on the hemolysis process, however, it can be recommended as an effective membrane-protective drug to protect the membranes of healthy cells

In vitro Comparative Studies of the Leaf Extracts of Four Different Types of Calamus Species

The leaf extracts of Calamus tenius, Calamus viminalis, Calamus guruba and Calamus erectus possessed various phytochemicals including alkaloids, carbohydrates, saponins, phenols, tannins, flavonoids, glycosides, fixed oils and steroids. The point of the present investigation was to compare the antioxidant profile of different leaf extracts of these species. In the present study, methanol, ethanol and chloroform extract of leaf of C. tenius, C. viminalis, C. guruba and C. erectus were explored for in-vitro thrombolytic activity, membrane stabilizing activity by using hypotonic solution-induced & heat-induced hemolysis and antioxidant profile. Ethanol extracts of all plant species demonstrated the highest percentage of clot lysis. During hypotonic solution-induced hemolysis, methanol extract of C. erectus inhibited 59.74% whereas during heat induced condition ethanol extracts of all plant species demonstrated the highest percentage inhibition of hemolysis of RBCs. The screenings of the plants revealed that leaf extracts contain phenolic and flavonoid contents in great amount with the high level of antioxidant capacity. The chloroform extract of C. tenius (2.45 μg/ml) and methanol extract of C. erectus (1.74 μg/ml) leaf showed potent DPPH and H2O2 scavenging activity, respectively. The nitric oxide scavenging activity was not significantly different for methanol, ethanol and chloroform extracts. Methanol, ethanol and chloroform extracts of these four plants showed medium to moderate reducing power and cupric reducing capacity in comparison to standard. A strong relationship was found between the antioxidant activity of the extracts with their total phenolic and flavonoid contents. Dhaka Univ. J. Pharm. Sci. 21(2): 173-181, 2022 (December)

EFFECT OF POLYHERBAL FORMULATION OF Boerhavia diffusa, Sesbania grandiflora, Moringa olifera, AND Solanum nigrum AGAINST GASTROENTERITIS AND ITS VALIDATION USING In silico APPROACH

Boerhavia diffusa, Sesbania grandiflora, Moringa olifera, and Solanum nigrum are indigenous plants that have been used in traditional medicine for treating various gastroenteritis. The present study aimed to demonstrate the Acid-neutralizing capacity, invitro anti-urease, anti-oxidant, inhibition of lipid peroxidation ability, anti- Inflammatory, and H+/K+ ATPase inhibition activity of hydroalcoholic extract of the polyherbal formulation. The phytochemicals present in the polyherbal formulation (PHF) were identified by GC – MS analysis. The total Phenolic Content and total Flavonoid content of phytochemicals were quantified. Phytochemical interactions with the active sites of urease were supported by Molecular docking analysis. The results revealed that 0.02 mg of phenolic content and 0.013 mg of flavonoids per gram of plant extract were observed, PHF had acid- neutralizing capacity better than the standard antacid tablet, 38% of anti-urease activity towards urease enzyme, 64% of inhibition of lipid peroxidation against oxidation of lipid bilayer, 59% of inhibition of protein denaturation, 37% of haemolysis inhibition, 80% of H+/K+ ATPase inhibition activity in the goat parietal cells. There were 14 functional compounds tend to interact with the enzyme urease. But two compounds directly interacted with the active site of ureases.

Effectiveness of Withania frutescens root extract on testicular damage induced by lead acetate in adult albino rats

Withania frutescens was used previously in traditherapy against poisoning, gastric ulceration, and dysentery treatments. Because no previous studies reporting on its therapeutic effects on male reproductive system and fertility disorders, this study aims to examine its effect on lead induced testicular damages as well as sperm count and hormonal status in rats. The present study is performed to determine their phytochemical compositions using GC-MS analysis, their antioxidant and anti-inflammatory activities in-vitro using spectrophotometry and then to estimate testosterone levels, sperm count, histopathological features, as well as spermatogenesis (TDI) and spermiogenesis (SPI) indices. The experiment is conducted for three months using four groups (Group A: control rats; Group B: exposed rats to lead-acetate; Group C: exposed rats to lead-acetate and 200 mg/kg of W. frutescens extract; Group D: treated rats with 200 mg/kg of W. frutescens extract). The obtained results show a total of 10 identified components from GC-MS analysis. Whereas a total phenolic content of 63.23 ± 3.82 GAE/g of extract, 25.16 ± 1.21 µg/mL of anti-free radical activity, and reducing power of 163.19 ± 6.01 µg/mL. A high anti-inflammatory activity is determined by hemolysis inhibition (IC50 =12.71 ± 1.06 µg/mL) and protein denaturation inhibition (IC50 =6.8 ± 1.23 µg/mL). Besides, lead exposure causes histological alterations in testis and decreases serum testosterone level, sperm count, and TDI and SPI indices. W. frutescens treated and co-treated animals showed no toxic effects throughout the experiment. However, it is found to improve testosterone level, increase sperm count, attenuate the testicular histopathological effect of lead, and increase TDI and SPI. These findings . these findings suggest that W. frutescens is a better source of bioactive compounds, which play an effective role against lead testicular damages. Furthermore, this natural extract can be utilized potentially in pharmaceutical and medicinal applications.

Extraction of bird blood nuclei and its effect on inhibition of hemolysis

تمت دراسة دور أنوية كريات الدم الحمر للدجاج في تثبيط تحلل كريات الدم الحمر وأختبار الهشاشة التنافذية لدم بعض الاشخاص غير المصابين بمرض ذو علاقة بتحلل كريات الدم الحمر. تم أستخدام السابونين وبيروكسيد الهيدروجين (H2O2) كمسببات لتحلل كريات الدم الحمر خارج الجسم Invitro وملاحظة التحلل الحاصل . ثم تم أضافة مستخلص أنوية كريات الدم الحمر للدجاج وملاحظة دورها في تثبيط التحلل الحاصل وكانت هذه الاختبارات بخمسة طرق وهي : أولاً: أستخدام محلول كلوريد الصوديوم (0.5%) . ثانياً: أستعمال محلول كلوريد الصوديوم مع محلول بيروكسيد الهيدروجين (0.6%). ثالثاً: أستخدام محلول كلوريد الصوديوم مع محلول السابونين (0.12%). رابعاً: أستعمال محلول كلوريد الصوديوم مع محلول بيروكسيد الهيدروجين ومستخلص أنوية كريات الدم الحمر للدجاج. خامساً: أستخدام محلول كلوريد الصوديوم مع محلول السابونين ومستخلص أنوية كريات الدم الحمر للدجاج. أوضحت الدراسة ان نتائج الطريقة الرابعة والخامسة متوافقة مع نتائج الطريقة الاولى من حيث بداية التحلل ومن ثم أكتماله.

Preparation, Characterization and Pharmacological evaluation of Bauhinia variegata Lauha Bhasma

The study was carried out to prepare, characterize, and investigates the pharmacological evaluation of Bauhinia variegata lauha bhasma (BVLB). The Lauha bhasma (LB) was prepared according to the reported standard method that involved the destruction of metallic state followed by conversion of crude product into bhasma state. The purification was done using Triphala extract prepared in cow-urine. The destruction of the metallic test was carried out using process juice of BV. At the last, the preparation of BVLB is subjected to a closed crucible system to get the final product. The characterization of BVLB was done by some preliminary examination followed by specific methods viz. X-ray diffractometry (XRD), Infrared (IR) spectroscopy, and UV-visible spectrophotometry. The In-vitro anti-inflammatory, anti-arthritic and antioxidant activity was carried out for BVLB at different concentrations of 50, 100, 200, and 250μg/mL. The said In-vitro activities were screened using percent inhibition of heat-induced hemolysis, the percent inhibition of protein denaturation, proteinase inhibition, and DPPH free radical scavenging activity. BVLB preliminary investigation showed 1.53 % aqueous and 3.95 % HCL solubility whereas specific characterization methods showed the formation of micro and nanocrystals of F2CO3. The BVLB showed % inhibition of hemolysis of in between 5.26 ± 0.95 to 31.20±4.12 whereas percent inhibition of protein denaturation in between 12 ± 1.22 to 52.22 ± 4.69. The percent proteinase inhibition of BVLB at a different concentration from 50, 100, 200, and 250μg/mL was estimated and found in between 3.26 ± 0.86 to 51.22 ± 3.12. The free radical scavenging activity of BVLB was found from 10.11 1 ± 0.89 to 52.05 ± 2.05. From the current investigation, it can be conducted that BVLB has significant In-vitro anti-inflammatory, anti-arthritic and antioxidant activity. The formed nanocrystals and presence of BV active constituents would be the underlying intermediaries for significant activity.

Sustained Complement C1s Inhibition with Sutimlimab in Patients with Cold Agglutinin Disease Results in Continued Efficacy during Part B of the Randomized Placebo-Controlled Phase 3 Cadenza Study (NCT03347422)

Background: Cold agglutinin disease (CAD) is a rare chronic autoimmune hemolytic anemia characterized by classical complement pathway (CP)-mediated hemolysis. Sutimlimab (SUT) is a first-in-class humanized monoclonal antibody that prevents CP activation by selectively inhibiting C1s; both the alternative and lectin pathways remain intact. In the randomized, double-blind, placebo-controlled Part A (26 weeks) of Phase 3 CADENZA, SUT treatment resulted in rapid and sustained improvements in hemoglobin (Hb), hemolytic markers and quality of life (QoL). Aims: To report the long-term safety and efficacy of SUT treatment in patients with CAD from the open-label Part B extension of CADENZA. Methods: In Part A, eligible patients with CAD without a history of recent blood transfusion (≤1 during previous 12 months; 0 during last 6 months), received SUT or placebo (PBO) through intravenous infusions on Days 0 and 7, then biweekly. All patients completing Part A were eligible to enter Part B and receive biweekly SUT for ≥1 year after the last patient completed Part A. Efficacy data up to Week 79 as well as the last available patients' values are reported. Efficacy endpoints included change from baseline (BL) in Hb, hemolytic and pharmacodynamic markers, Functional Assessment of Chronic Illness Therapy (FACIT)-Fatigue, and blood transfusions. Safety data was recorded throughout the study; treatment emergent adverse event (TEAE) and serious TEAE (TESAE). Results: Of the 42 patients enrolled in Part A, 39 completed Part A and entered Part B, with 32 (82.1%) completing Part B; 7 (17.9%) patients discontinued the study prematurely, due to TESAE (n=1), lack of efficacy (n=3), withdrawal of consent (n=2) and withdrawal by sponsor (n=1). SUT treatment rapidly improved Hb levels; mean (SE) Hb at the end of Part A was 11.51 (0.40) g/dL and 9.43 (0.40) g/dL for SUT and PBO groups, respectively. In Part B, improvements in Hb were sustained in the SUT group and the ex-PBO group saw rapid and comparable increases in Hb upon initiation of SUT; at Week 79 mean (SE) Hb levels were 11.86 (0.54) g/dL and 11.76 (0.58) g/dL in the SUT only and ex-PBO groups, respectively (Panel A). Mean total bilirubin was normalized with SUT treatment in Part A and sustained in Part B; similar decreases were observed for patients in the ex-PBO group when they started receiving SUT in Part B (Panel B). Improvements in mean [SE] FACIT-Fatigue scores (BL: 32.96 [1.79]) observed in Part A were sustained in Part B in the SUT only group (44.31 [2.19] at Week 87); the mean FACIT-Fatigue score increased to comparable levels in the ex-PBO group (41.40 [2.71] at Week 87). Improvements in Hb, bilirubin, and FACIT-Fatigue correlated with normalization of C4 and near-complete inhibition of CP activity, that was maintained through the end of treatment. Reductions in mean absolute reticulocyte count and increases in haptoglobin levels observed with SUT treatment in Part A were maintained in Part B, and upon initiation of SUT in Part B, the ex-PBO group reached comparable levels. In Part B, 9 (23.1%) patients received a ≥1 transfusion. Thirty-six (92.3%) patients experienced ≥1 TEAE in Part B. Seven (17.9%) patients experienced a total of 11 TESAEs; one TESAE of hypertension was assessed as related to SUT by the investigator. Thromboembolic events (transient ischemic attack [n=1]; deep vein thrombosis [n=1]) were observed in 2 (5.1%) patients with underlying risk factors for thromboembolism. Both events were assessed as non-serious and unrelated to SUT by the Investigator. One TESAE of infection (urinary tract infection Grade ≥3) was reported. No meningococcal infections, serious events of hypersensitivity, anaphylaxis or systemic lupus erythematosus were reported. One patient with a history of tobacco use (received PBO in Part A) experienced a TESAE of squamous cell carcinoma of the lung with fatal outcome. SUT was withdrawn due to this TESAE prior to the patient's death. Conclusion: Long-term treatment with SUT, an anti-C1s CP inhibitor, maintained mean Hb levels >11 g/dL, achieved sustained normalization of bilirubin and led to clinically meaningful improvements of FACIT-Fatigue scores, while maintaining a favourable safety profile. Improvements in anemia, inhibition of hemolysis and favourable effects on QoL were rapidly achieved and sustained to a similar extent in those patients administered SUT throughout the study and those who switched to SUT treatment in Part B. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal