Evaluation of in-vitro Anti-Inflammatory Activity of Gallic Acid

Gallic acid's anti-inflammatory effect was studied at various concentrations, including 50,100,150,200, and 250 ug/ml. Gallic acid's anti-inflammatory effect was assessed using two in-vitro assays: proteinase inhibition and albumin denaturation. The greatest proteinase inhibition activity of 52.83 percent was achieved at a concentration of 250ug/ml, according to the results. It also revealed that at 250ug/ml, the maximum percentage inhibition in albumin denaturation was 74.79 percent. Gallic acid's antiproteinase and albumin denaturation activities both increase with increasing concentrations, according to this research. Keywords: Gallic acid, Anti-Inflammatory, In-Vitro Assays, Albumin Denaturation, Proteinase activity.

Unlocking the Real Potential of Black Soldier Fly (Hermetia illucens) Larvae Protein Derivatives in Pet Diets

Black soldier fly larvae (BSFL)-derived proteins are gaining popularity as sustainable pet food ingredients. According to the literature, these ingredients have strong antioxidant and antimicrobial activities. Due to the ability of BSFL protein derivatives to donate hydrogen atoms and/or electrons to counterpoise unstable molecules, they could possibly help in the prevention of osteoarthritis. During this study, the antiarthritic potential of BSFL protein derivatives was evaluated using the following assays: (1) proteinase inhibition, (2) erythrocyte membrane stability, (3) reactive oxygen species (ROS) production by activated macrophages, (4) ROS production by monocytes, and (5) cellular toxicity. Additionally, the glucosamine content of these ingredients was also evaluated. Chicken meal is commonly used in pet food formulations and was used as an industrial benchmark. The results obtained during this study demonstrated the strong antiarthritic potential of BSFL protein derivatives. We found that BSFL protein derivatives are not only useful in preventing the development of arthritis but could also help to cure it due to the presence of glucosamine. We also found that chicken meal could contribute to the development of arthritis by increasing ROS production by monocytes.

Serpins in cartilage and osteoarthritis: what do we know?

Serpins (serine proteinase inhibitors) are an ancient superfamily of structurally similar proteins, the majority of which use an elegant suicide inhibition mechanism to target serine proteinases. Despite likely evolving from a single common ancestor, the 36 human serpins have established roles regulating diverse biological processes, such as blood coagulation, embryonic development and extracellular matrix (ECM) turnover. Genetic mutations in serpin genes underpin a host of monogenic disorders — collectively termed the ‘serpinopathies’ — but serpin dysregulation has also been shown to drive pathological mechanisms in many common diseases. Osteoarthritis is a degenerative joint disorder, characterised by the progressive destruction of articular cartilage. This breakdown of the cartilage is driven by the metalloproteinases, and it has long been established that an imbalance of metalloproteinases to their inhibitors is of critical importance. More recently, a role for serine proteinases in cartilage destruction is emerging; including the activation of latent matrix metalloproteinases and cell-surface receptors, or direct proteolysis of the ECM. Serpins likely regulate these processes, as well as having roles beyond serine proteinase inhibition. Indeed, serpins are routinely observed to be highly modulated in osteoarthritic tissues and fluids by ‘omic analysis, but despite this, they are largely ignored. Confusing nomenclature and an underappreciation for the role of serine proteinases in osteoarthritis (OA) being the likely causes. In this narrative review, serpin structure, biochemistry and nomenclature are introduced, and for the first time, their putative importance in maintaining joint tissues — as well as their dysregulation in OA — are explored.

THE EFFECT OF VARYING LEVELS OF RAW LIMA BEAN (Phasealus lunatus) ON THE RAT'S PANCREATIC AND INTESTINAL TRYPSIN (EC 3.4.21.4) AND CHYMOTRYPSIN (EC 3.4.21.1) ACTIVITIES

Two series of assays involving a total of 120 growing rats were carried out to investigate the effect of varying levels of raw lima bean (RLB) on pancreatic and intestinal trypsin EC 3.4.21.4) and chymotrypsin (EC 3.4.21.1) activities. Experiment one indicates significant (P<0.01) inhibtion of both pancreatic trypsin and chymotrypsin due to RLB feeding. Enzyme activities in both the small and large intestine were also significantly (P<0.01) depressed while enzyme values in the caecum were not. Age x Diet interaction was non significant with respect to these enzymes. Pancreatic trypsin and chymotrypsin correlated significantly (P<0.01; P<0.05) with RLB with respective R2, coefficient of multiple determination, of 0.94 and 0.67. Trypsin activity in both the small and large intestine was also significantly (P<0.01) correlated with respective R2 of 0.78 and 0.96. The second study suggests a less than 10% replacement of cooked lima bean by the raw to avert significant pancreatic proteinase inhibition, and a less than 15% replacement of the cooked lima bean by the raw to aver t significantinhibition of the proteinases especially in the small intestine.

Studies on In-vitro Anti-inflammatory and Antioxidant Potentials of Annona muricata Leaf Extracts

Inflammation has stimulated significant worldwide scientific interest because of its implication in many human diseases. Most inflammations are caused by reactive oxygen species or free radicals. Annona muricataleaf extracts were investigated for their in-vitroantioxidant and anti-inflammatory potentials. Annona muricataleavesweredried at room temperature, blended using a mill.and extracted with solvents of varying degree of polarities. The solventsused were hexane, ethyl acetate,and ethanol. After sequential extraction, the crude extracts were examined for their in-vitroanti-inflammatory activities on lipoxygenase inhibition, proteinase inhibition, albumin denaturation inhibition,and red blood cell membrane stabilization assays,while the antioxidant activities were examined using DPPH, ABTS and hydrogen peroxide assays. The results showed that the ethanol extract had significantlyhigher albumin denaturation inhibition activity at 500 μg/mL (p &lt; 0.01). The activity of all the extracts on proteinase inhibition decreased with the increase in concentration of the extracts. Indomethacin (standard), ethanol extract,and ethyl acetate extract exhibited a dose dependent increase in lipoxygenase activity. The ethanol extract showed highred blood cell membrane stabilization activity at 500 μg/mL and the activity was comparable with that of the standard (diclofenac). Hydrogen peroxide scavenging activity of the extracts and standard (Vitamin C) were comparable at 20 –100 μg/mL. The ethanol extract showed significantly higher(p &lt; 0.01) DPPH radical scavenging activity compared with other extracts. A similar trend was also observed for ABTS radical scavenging activity. Generally,the ethanol extract exhibited higher anti-inflammatory and antioxidant activities in most of the assays, this could be attributed to the polar compounds present in the extract.

Anti-Snake Venom, Anti-Arthritic and Cytotoxic Activities of Tectona grandis L. f. Stem Bark (Lamiaceae)

Tectona grandis L. f. (Lamiaceae) is famous for timber production and has been used in traditional medicine for treating bronchitis, liver-related roubles, urinary discharge amongst other diseases. Methanol extract of the stem bark was investigated for in vivo antiophidian assay against Bitis arietans and Naja nigricollis. Also, the extract and chromatographic fractions were subjected to cytotoxicity tests using tadpole model and antiarthritic assay by proteinase inhibition. Chromatography of crude methanol extract afforded three highly polar vacuum liquid chromatography (VLC) fractions (BVLC-1, BVLC-2 and BVLC-3). BVLC-2 further gave four semi-pure uncharacterized isolates (I, II, III, IV). Only BVLC-2 and BVLC-3 were cytotoxic at 10 - 80 mg/ml, with BVLC-3 being most potent (100% mortality, LC50 40 mg/ml). Concentration-dependent proteinase enzyme inhibition (24 - 71%) at 200 - 1000 μg/ml of BVLC-2 was observed, and this was less (IC50 659.24 μg/ml) than the activity of the standard drug, acetyl salicylic acid. However, neutralization of B. arietans and N. nigricollis snake venoms using methanol extract was not dose-dependent, but the extract atthe least dose, 50 mg/kg offered better protection (75%) on Naja nigricollis envenomed-mice in 48 h. Its activity was comparable to that of the positive antivenin tested at 0.2 mg/kg. These findings justify the folkloric use of T. grandis in the treatment of snake bites, arthritic conditions, and oxidative stress-induced diseases. Keywords: Tectona grandis, methanol extract, anti-snake venom activity, tadpole cytotoxicity, anti-arthritic activity

Comparative Proximate Nutraceutical Study of Poor Man’s Pulse, Horsegram [Macrotyloma uniflorum] with the Other Common Legume Crops: A Review

Horsegram is an underutilized drought hardy crop and mainly neglected by the farmers in Northern region of India. However, the present study reveals the hidden comparative analysis of nutraceutical use with well-known legumes like Phaseolus vulgaris, Vigna mungo, Cicer arietinum, Vicia faba, Cajanus cajan, Vigna radiata, Pisum sativum and Lens culinaris. This pulse crop is an excellent source of carbohydrate, protein and dietary fiber. This present study shows that amount of energy in horsegram falls in the range of 376.12-377.21 Kcal/100 g which is maximum than the other legumes. The ash, protein, dietary fibres, carbohydrates, fat and starch content of horse gram falls in the range (2.24% to 5.16%), (18.15% to 28.8%), (5% to 16.3%), (50% to 63.4%), (1.10 to 1.9%) and (31.86% to 47.5%) respectively. Horsegram is found to be less fat and more dietary food fibers than the most common legumes. Hence, it is an excellent source of food for diabetic patients and useful in weight management. The unique anti-uroliathiatic activity of horsegram is well known against calcium oxalate crystals, calcium phosphate crystals and uric acid crystals. Anticholelithiatic, Anti-histaminic, Hemolytic, Larvicidal, Proteinase inhibition and Anti-HIV are among other unique medicinal properties of horsegram which are not reported in any other legumes.

Biologically valuable components, antioxidant activity and proteinase inhibition activity of leaf and callus extracts of Salvia sp.

Sage is medicinal plant, known for its antioxidant and anti-inflammatory effects. Eight extract samples were tested in this study: extract from Salvia officinalis L. varieties from two different geographical localities (Jaslovské Bohunice and Pobedim, Slovakia), Salvia officinalis L., variety “bicolor”, Salvia officinalis L., variety “purpurescens”, Salvia apiana, Salvia divinorum, and two callus cultures of Salvia sclarea L. and Salvia aethiopis L. The highest values for composite parameters were observed for extract from Salvia apiana. It can be concluded that prepared sage extract samples are rich on polyphenolic acids (2 950±265 μg.mL−1 GAeq.) and amines (197±5.50 μg.mL−1 TRPeq.). HPLC analysis confirmed the dominant content of rosmarinic acid in the extracts; the highest content was detected in the Salvia apiana extract (1 120±15 μg.mL−1). Extract from Salvia apiana expressed too the highest antioxidant activity (1 710 – 4 669 μg.mL−1TEAC). Similarly, the highest inhibition activity was observed for this extract on thrombin (57±3.3 %) and on other proteinases (over 80 %). Spearman correlation analysis and PCA analyses revealed a coherence between antioxidant activity of samples and their content of rosmarinic acid as well as inhibitory activity towards particular proteases, and revealed the significance of thiol based secondary metabolites. Cluster analysis demonstrates the differences of Salvia apiana extract from extracts of S. officinalis L., the group of S. divinorum extract and from callus cultures.

TIMP-3 as a therapeutic target for cancer

Tissue inhibitor of metalloproteinase-3 (TIMP-3), a secreted glycoprotein, plays an important role in carcinogenesis. It can bind to many proteinases to suppress their activity and thus protect the extracellular matrix from degradation. TIMP-3 may have many anticancer properties, including apoptosis induction and antiproliferative, antiangiogenic, and antimetastatic activities. This review summarizes the structure, proteinase inhibition ability, genetic and epigenetic regulation, cancer therapy potential, and contribution to cancer development of TIMP-3. Furthermore, in this review we discuss its potential as a biomarker for predicting cancer progression and the current state of drugs that target TIMP-3, either alone or in combination with clinical treatment. In conclusion, TIMP-3 can be a biomarker of cancer and a potential target for cancer therapy. This review article can serve as a basis to understand how to modulate TIMP-3 levels as a drug target of cancers.

IN VITRO ANTI-INFLAMMATORY ACTIVITY OF SYRINGIC ACID

Objective: The present study was carried out to investigate the in vitro anti-inflammatory activity of syringic acid (SA). Methods: SA was tested for it's in vitro anti-inflammatory activity at different concentrations in protein denaturation, proteinase inhibition and human red blood cell (HRBC) membrane stabilization assay. The reference drugs used were aspirin and diclofenac sodium. Results: SA showed concentration-dependent inhibition of protein denaturation and proteinase activity with a half-maximal inhibitory concentration (IC50) value of 49.38±0.56 µg/ml and 53.73±0.27 µg/ml respectively. Heat-induced haemolysis was inhibited by SA with an IC50 value of 57.13±0.24 µg/ml. SA also inhibited the hypotonicity-induced haemolysis (IC50 value of 53.87±0.72 µg/ml). Conclusion: From the present study, we can conclude that SA possesses appreciable anti-inflammatory effect against denaturation of proteins, proteinase activity, and human red blood membrane stabilization assays. Further studies are required for determining the possible mechanisms behind its anti-inflammatory action.