Enzyme Inhibitory Properties Research Articles

Tropolones and Thailandepsin B as Lead-like Natural Compounds in the Development of Potent and Selective Histone Deacetylase Inhibitors.

Tropolone and thailandepsin B are naturally occurring substances that are primarily isolated from fungi and plants, although they can also be found in certain bacteria. Tropolones belong to an important class of aromatic compounds with a seven-membered nonbenzenoid ring structure. Thailandepsins are a group of natural products that were initially discovered in the culture broth of the Gram-negative bacterium Burkholderia thailandensis. Tropolonebased structures have been identified in over 200 natural compounds, ranging from simple tropolone derivatives to complex multicyclic systems like pycnidione and pyrerubrine A. These natural compounds exhibit a diverse range of pharmacological effects, including antibacterial, antifungal, insecticidal, phytotoxic, anti-inflammatory, antimitotic, anti-diabetic, enzyme inhibitory, anticancer, cytoprotective, and ROS scavenging properties. It is worth noting that thujaplicane, a compound similar to tropolone, displays all of the listed biological activities except for antimitotic action, which has only been observed in one natural tropolone compound, colchicine. Tropolone can be synthesized from commercially available seven-membered rings or derived through various cyclization and cycloaddition reactions. Thailandepsin B, on the other hand, can be synthesized by macro-lactonization of the corresponding secoacid, followed by the formation of internal disulfide bonds. It is important to mention that thailandepsin B exhibits different selective inhibition profiles compared to FK228. We investigated the HDAC inhibitory activity of the Tropolones and Thailandepsin B and discussed the biosynthesis of the naturally occurring compounds and their synthetic scheme. It has been observed that Tropolone derivatives act as isoenzyme-selective inhibitors of proven anticancer drug targets, histone deacetylases (HDACs). Some monosubstituted tropolones show remarkable levels of selectivity for HDAC2 and strongly inhibit the growth of T-lymphocyte cell lines. And Thailandepsins have different selective inhibition profiles than FK228. They exhibit comparable inhibitory activities to FK228 against human HDAC1, HDAC2, HDAC3, HDAC6, HDAC7, and HDAC9, but less potent inhibitory activities than FK228 toward HDAC4 and HDAC8, the latter of which may be useful. Thailandepsins possess potent cytotoxic activities toward some types of cell lines.

Antimicrobial, antifibrinolytic, enzyme inhibitory and wound healing properties of zinc borate

Boron containing compounds (BCGs) have recently been used for pharmaceutical applications. Zinc, an essential element, is known to be one of the most promising biodegradable metals. The present study was conducted to determine the wound healing properties of zinc borate with its antimicrobial, antifibrinolytic and enzyme inhibitory characteristics. In vitro scratch wound healing assay revealed that zinc borate at 0.01 µg/mL concentration stimulated the proliferation of 3T3 fibroblast cells after 24 h of scar formation. The highest enzyme inhibition was observed against collagenase at 1 mg/mL (81.5%). Minimum inhibition concentration (MIC) values were determined as 1 mg/mL and 0.5 mg/mL against Candida albicans and Staphylococcus aureus, respectively. Zinc borate did not have any fibrinolytic activity at 1, 0.5 and 0.1 mg/mL concentrations. It can be suggested that zinc borate can be used effectively to improve the wound healing process and to prevent the possible wound infections.

Potential and limitations of rabbit meat in maintaining food security in Ukraine

The aim of the present review was to assess whether rabbit meat production can help to improve food security in Ukraine and whether it can make a meaningful contribution to the national economy. Rabbit meat is a nutrient-rich and affordable food with a lower environmental impact than red meat from cattle or sheep. Rabbit meat is considered healthy, as it is lower in fat, cholesterol and sodium than other types of meat, and rich in protein. In addition, rabbit meat proteins can serve as a source of bioactive peptides with the angiotensin converting enzyme (ACE) inhibition properties. The health-promoting properties of rabbit meat and role of associated by-products in providing nutritious and safe food for consumers, and in reducing waste in the meat sector, should be promoted. In Ukraine, rabbit meat products are already recommended as part of a healthy diet for children, pregnant women and the elderly, and the market could be broadened by introducing more rabbit-based products for the whole population. Rabbit production in Ukraine is a promising area of economic activity and with a broader market this segment could increase further and become a significant part of the economy. However, the rabbit farming sector in Ukraine needs comprehensive improvement to address existing issues, including the introduction of sustainable production methods in accordance with European animal welfare and meat quality standards.

Piperazin incorporated Schiff Base derivatives: Assessment of in vitro biological activities, metabolic enzyme inhibition properties, and molecular docking calculations.

The cytotoxic activities of the compounds were determined by the 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) method in human breast cancer (MCF-7), human cervical cancer (HeLa), and mouse fibroblast (L929) cell lines. The compounds MAAS-5 and four modified the supercoiled tertiary structure of pBR322 plasmid DNA. MAAS-5 showed the highest cytotoxic activity in HeLa, MCF-7, and L929 cells with IC50 values of 16.76 ± 3.22, 28.83 ± 5.61, and 2.18 ± 1.22 µM, respectively. MAAS-3 was found to have almost the lowest cytotoxic activities with the IC50 values of 93.17 ± 9.28, 181.07 ± 11.54, and 16.86 ± 6.42 µM in HeLa, MCF-7, and L929 cells respectively at 24 h. Moreover, the antiepileptic potentials of these compounds were investigated in this study. To this end, the effect of newly synthesized Schiff base derivatives on the enzyme activities of carbonic anhydrase I and II isozymes (human carbonic anhydrase [hCA] I and hCA II) was evaluated spectrophotometrically. The target compounds demonstrated high inhibitory activities compared with standard inhibitors with Ki values in the range of 4.54 ± 0.86-15.46 ± 8.65 nM for hCA I (Ki value for standard inhibitor = 12.08 ± 2.00 nM), 1.09 ± 0.32-29.94 ± 0.82 nM for hCA II (Ki value for standard inhibitor = 18.22 ± 4.90 nM). Finally, the activities of the compounds were compared with the Gaussian programme in the B3lyp, HF, M062X base sets with 6-31++G (d,p) levels. In addition, the activities of five compounds against various breast cancer proteins and hCA I and II were compared with molecular docking calculations. Also, absorption, distribution, metabolism, excretion, and toxicity analysis was performed to investigate the possibility of using five compounds as drug candidates.

Chemical Characterization of Different Extracts from Artemisia annua and Their Antioxidant, Enzyme Inhibitory and Anti-Inflammatory Properties.

Artemisia annua L. (Asteraceae Family) is an important plant in Asia that has been used for treating different diseases, including fever due to malaria, wounds, tubercolisis, scabues, pain, convulsions, diabetes, and inflammation. In this study we aimed to evaluate the effects of different polarity extracts (hexane, dichloromethane, ethyl acetate, ethanol, ethanol/water (70 %) and water) from A. annua against the burden of inflammation and oxidative stress occurring in colon tissue exposed to LPS. In parallel, chemical composition, antiradical, and enzyme inhibition effects against α-amylase, α-glucosidase, tyrosinase, and cholinesterases were evaluated. The water extract contained the highest content of the total phenolic with 34.59 mg gallic acid equivalent (GAE)/g extract, while the hexane had the highest content of the total flavonoid (20.06 mg rutin equivalent (RE)/g extract). In antioxidant assays, the polar extracts (ethanol, ethanol/water and water) exhibited stronger radical scavenging and reducing power abilities when compared to non-polar extracts. The hexane extract showed the best AChE, tyrosinase and glucosidase inhibitory effects. All extracts revealed effective anti-inflammatory agents, as demonstrated by the blunting effects on COX-2 and TNFα gene expression. These effects seemed to be not related to the only phenolic content. However, it is worthy of interest to highlight how the higher potency against LPS-induced gene expression was shown by the water extract ; thus suggesting a potential phytotherapy application in the management of clinical symptoms related to inflammatory colon diseases, although future in vivo studies are needed to confirm such in vitro and ex vivo observations.

Okra seeds proteins: Amino acid profile, free radical scavenging activities and inhibition of diabetes and hypertensive converting enzymes indices

The study was aimed to determine bioactivity of okra seed proteins against oxidative stress, diabetes, and hypertension. Whole okra seed flour (OKPF), protein isolate (OKPI) and hydrolysate (OKPH, using proteolytic Lactic Acid Bacteria) were produced by standard methods. Samples were evaluated for protein contents, amino acids, in-vitro antioxidant, and enzymes (pancreatic lipase, α-amylase, α-glucosidase, and Angiotensin-1 converting) inhibitory properties. Protein content (g/100 g) of OKPH (97.45) was more than OKPI (80.09) and OKPF (26.42). In-vitro protein digestibility indicated that OKPH (87.17%) had the highest value followed by OKPI (77.63%) and OKPF (66.06%), respectively. Amino acid composition (mg/100 g protein), i.e., hydrophobic (32.36), aromatic (7.56), antioxidant (19.15), and branch chain (13.0) in OKPH were higher than in OKPF and OKPI. Functional properties of OKPH showed better attributes than in OKPI and OKPF, respectively. Protein hydrolysate (OKPH) exhibited highest antioxidant and free radical scavenging activity than in OKPI and OKPF. Sample OKPH exhibited highest percentage inhibition against carbohydrate-hydrolyzing and anti-hypertensive enzyme inhibition indices than in OKPF and OKPI. This study established that OKPH exhibited highest antioxidant, pancreatic lipase, angiotensin-1 converting, α-amylase and α-glucosidase enzyme activities. Hence, it may be suitable as nutraceutical agent for antidiabetic and antihypertension.

Chemical composition, antioxidant and multi-enzymatic inhibitory potential of licorice harvested from wild populations in Iraq

The current work focuses on comparing the chemical composition, the antioxidant activities and the enzyme inhibitory properties of licorice collected from ecologically distinct regions of Iraq, namely: Wasit and Al-Diwaniyah. The chemical compositions of the aerial parts volatile oils, extracted via hydrodistillation, were characterized by gas chromatography-flame ionization detector (GC-FID) and gas chromatography-mass spectrometry (GC–MS) techniques. Furthermore, the evaluation of enzyme inhibitory properties of both samples against acetylcholinesterase, butyrylcholinesterase, tyrosinase, α-amylase, and α-glucosidase were performed. Subsequently, the antioxidant properties were assessed using DPPH and ABTS radical scavenging methods, cupric and Ferric reducing power (CUPRAC and FRAP), metal chelating ability (MCA) and phosphomolybdenum (PBD) assay. Thus, the chemical analysis showed the richness of Glycyrrhiza glabra Linn. volatile oils in oxygenated aliphatic hydrocarbons, with Palmitic acid identified as the major constituent (41.1%-51.6%). Conclusively and as a result of antioxidant assays, licorice from Al-Diwaniyah region was more active than same specie located in Wasit. Both volatile oils exhibited significant cupric ion reducing antioxidant capacity with values ranging between (36.66 and 41.68 mg TE/g). Moreover, G. glabra presented a significant anti-tyrosinase activity, with inhibition values of 58.33 and 52.53 mg KAE/g for Wasit and AL-Diwaniyah populations respectively.

Synthesis and enhanced biological activities of methyl 5-amino-β-resorcylate capped silver nanoparticles

The biomedical potential of silver nanoparticles loaded and conjugated with a drug exhibited diverse effects. Compared to free molecules, these nanoconjugates demonstrated enhanced biological activities, requiring a reduced quantity of the active ingredient for effectiveness. To achieve this, silver nanomaterials (AgMRA) with a size of 19 nm were prepared and modified with methyl-5-amino-β-resorcylate (MRA) through the reduction of an aqueous solution of AgNO3. The capping of AgMRA was analyzed using TEM, FT-IR, and UV–vis techniques. The stability of the silver nanomaterials (AgMRA) was investigated in relation to salt concentration, pH, and temperature. The resulting nanoconjugates were soluble in water, and their suspension remained stable at a salt concentration of 4 mol/L, pH range of 2–13, and elevated temperatures. Multiple biological activities, including enzyme inhibition, antioxidant effects, and antimicrobial properties, were identified for the silver nanomaterials (AgMRA) and compared to the pure MRA and commercially available drugs. The urease inhibition and antioxidant properties of AgMRA were comparable to or even superior to those of MRA and the standard drugs. Notably, the synthesized nanoconjugates contained 25% of silver nanomaterials conjugated with MRA by weight, allowing for a four-fold reduction in the required dose of MRA for achieving the desired biological activities.

Assessment of antioxidant and enzyme inhibition properties of Myrtus communis L. leaves

This study investigated the antioxidant and enzyme inhibitory properties of Myrtus communis leaves. Three different solvents including methanol, 50%-methanol, and water were used to extract of M. communis. In addition, total bioactive compounds were evaluated by using total phenolic and total flavonoid content assays. In antioxidant assays, water extract displayed the highest antioxidant potential. The MeOH extract demonstrated the highest inhibitory effect against AChE (4.38 mg GALAE/g), BChE (1.58 mg GALAE/g), α-amylase (0.56 mmol ACE/g), and tyrosinase (132.20 mg KAE/g). The M. communis leaves extract could be used as a promising raw material source in food and medicine industries.

Evaluation of black seed (Nigella sativa L.) cake and its protein in muffins as a valuable potential functional source for obesity control

Background:Due to the use of high-calorie foods and reduced physical activity caused by the modern lifestyle, obesity is on the rise. Obesity occurs by the increase of fat accumulation in the body, causing different complications such as diabetes, cardiovascular diseases, cancer, respiratory disorders, blood pressure, etc., which has become a global problem. Objectives:The production of functional food that has medicinal properties for controlling or inhibiting diseases is increasingly popular. With the expanding concerns about obesity and its related diseases, it is very important to provide natural controlling or inhibiting compounds. In this study, the functional properties of black seed cake (BSC) and its protein (BSCP) were investigated for obesity control through the inhibition of pancreatic lipase, α-glucosidase, and α-amylase enzymes as well as its antioxidant properties. Methods:The textural properties and sensory evaluation of muffins, enzyme inhibition properties (inhibiting the effect of enzymes in the obesity process), and antioxidant properties (control or inhibition of side effects caused by body fat oxidation) of BSC and BSCP were investigated.Results:The highest and lowest inhibitory levels of pancreatic lipase are related to black seed (Nigella sativa)cake protein (BSCP) (59.21%) and black seed cake (BSC) (32.90%), respectively. The highest and lowest levels of α-amylase inhibition are related to BSCP (32.51%) and black seed cake in ethanol 75% (BSC-E75%) (3.62%), respectively. The highest and lowest α-glucosidase inhibitory levels are related to BSC-E30% (71%) and BSCP (10.10%), respectively. BSC and BSCP showed high antioxidant activity. The use of BSC and BSCP in the production of muffins (a widely consumed and popular product) to replace flour and oil in the formulation of muffins caused a decrease in L*, b*, but an increase in a*. Hardness, chewiness, springiness, and cohesiveness decreased with the increase of BSC and BSCP content. Samples BSM 3%, BSM 6%, PBM 1%, and PBM 2% were similar to control muffins in sensory evaluation.Conclusions :From the result of this study, it can be concluded that black seed cake and its protein can be considered a potential efficient source for the production of functional muffins and possibly other functional foods that aim to manage and control obesity, blood glucose, and blood fat. However, more research is needed to fully determine the effectiveness of black seed cake and its protein.Keywords: Obesity, Black seed cake, Muffins, Diabetes, Blood pressure, Blood fat

Orbitrap Mass Spectrometry-Based Profiling of Secondary Metabolites in Two Unexplored Eminium Species and Bioactivity Potential.

The study aimed at the metabolite profiling and evaluation of antioxidant and enzyme inhibitory properties of methanol extracts from flowers, leaves, and tubers of unexplored Eminium intortum (Banks & Sol.) Kuntze and E. spiculatum (Blume) Schott (Araceae). A total of 83 metabolites, including 19 phenolic acids, 46 flavonoids, 11 amino, and 7 fatty acids were identified by UHPLC-HRMS in the studied extracts for the first time. E. intortum flower and leaf extracts had the highest total phenolic and flavonoid contents (50.82 ± 0.71 mg GAE/g and 65.08 ± 0.38 RE/g, respectively). Significant radical scavenging activity (32.20 ± 1.26 and 54.34 ± 0.53 mg TE/g for DPPH and ABTS) and reducing power (88.27 ± 1.49 and 33.13 ± 0.68 mg TE/g for CUPRAC and FRAP) were observed in leaf extracts. E. intortum flowers showed the maximum anticholinesterase activity (2.72 ± 0.03 mg GALAE/g). E. spiculatum leaves and tubers exhibited the highest inhibition towards α-glucosidase (0.99 ± 0.02 ACAE/g) and tirosinase (50.73 ± 2.29 mg KAE/g), respectively. A multivariate analysis revealed that O-hydroxycinnamoylglycosyl-C-flavonoid glycosides mostly accounted for the discrimination of both species. Thus, E. intortum and E. spiculatum can be considered as potential candidates for designing functional ingredients in the pharmaceutical and nutraceutical industries.

Blood glucose lowering and anti-oxidant potential of erythritol: An in vitro and in vivo study.

Diabetes is a major cause of death worldwide and Nigeria is not an exception. The quest to lower sugar levels has become a major factor in the management of diabetes; this has occasioned the use of substitutes for refined sugar in beverages. Erythritol is a natural sweetener gaining immense interest in recent times. Like metformin, erythritol has shown hydroxyl radical scavenging ability and has metabolic profile suitable for diabetics. Therefore, the blood glucose-lowering and anti-oxidant properties of erythritol under in vitro and in vivo systems were accessed. Radical scavenging assay (ABTS and DPPH) and inhibition of carbohydrate digestive enzymes (alpha-amylase and alpha-glucosidase) were employed to determine in vitro anti-oxidant and glucose regulatory function of erythritol respectively. Molecular docking studies were performed between 3D structures of human pancreatic alpha-amylase and alpha-glucosidase, isomaltase from saccharomyces cerevisiae with erythritol. The drug-like activity of erythritol was also assessed.Thereafter, we investigated the effect of erythritol on blood glucose and antioxidant status of normal and streptozocin- nicotinamide-induced diabetes rats which were grouped into five (n = 5); Normal, Ery (normal and administered erythritol), Db (diabetic control), Db + Ery (diabetic and administered erythritol), and Db + Met (diabetic and administered metformin). Erythritol showed a considerable radical scavenging activity and an ability to inhibit alpha-amylase and alpha-glucosidase in vitro. Also, a significant reduction in glucose intolerance, blood glucose and hemoglobin A1c levels and improved antioxidant level was seen in erythritol-treated diabetic rats. Erythritol showed anti-oxidant activity, alpha amylase and glucosidase enzyme inhibition property, improved antioxidant status and ameliorated blood glucose, HbA1c, and glucose intolerance following diabetes.

Evaluation of the multifunctional dipeptidyl-peptidase IV and angiotensin converting enzyme inhibitory properties of a casein hydrolysate using cell-free and cell-based assays.

The objective of the study was the evaluation of the potential pleiotropic effect of a commercial casein hydrolysate (CH). After an analysis of the composition, the BIOPEP-UWM database suggested that these peptides contained numerous sequences with potential inhibitory activities on angiotensin converting enzyme (ACE) and dipeptidyl-peptidase IV (DPP-IV). The anti-diabetic and anti-hypertensive effects of these peptides were thus assessed using either cell-free or cell-based assays. In the cell-free system, CH displayed inhibitory properties against DPP-IV (IC50 value equal to 0.38 ± 0.01 mg/mL) and ACE (IC50 value equal to 0.39 ± 0.01 mg/mL). Further, CH reduced the DPP-IV and ACE activities expressed by human intestinal Caco-2 cells by 61.10 ± 1.70% and 76.90 ± 4.47%, respectively, versus untreated cells, after 6 h of treatment at the concentration of 5 mg/mL. This first demonstration of the multifunctional behavior of this material suggests that it may become an anti-diabetic and/or anti-hypertensive ingredient to be included in the formulation of different functional food or nutraceutics.

Chemical composition, antioxidant, and enzyme inhibitory properties of Rauwolfia vomitoria extract.

Rauwolfia vomitoria is one ethno-botanicals in Nigeria used by traditional health practitioners in managing several human diseases. However, necessary information regarding its effect on enzymes implicated in the development and progression of erectile dysfunction is missing in the literature. Thus, this study investigated the antioxidant property and impact of Rauwolfia vomitoria extract on erectile dysfunction-related enzymes invitro. High performance liquid chromatography was used to identify and quantify Rauwolfia vomitoria's phenolic components. Then, utilizing common antioxidant assays, the extract's antioxidant properties were evaluated and finally the effect of the extract on some enzymes (AChE, arginase and ACE) implicated in erectile dysfunction was investigated invitro. The results showed that the extract inhibited AChE(IC50=388.72 μg/mL), arginase (IC50=40.06 μg/mL) and ACE (IC50=108.64 μg/mL) activities. In addition, phenolic rich extract of Rauvolfia vomitoria scavenged radicals and chelated Fe2+ in concentration dependent manner. Furthermore, rutin, chlorogenic acid, gallic acid, and kaempferol were found in large quantities by HPLC analysis. Therefore, one of the potential reasons driving Rauwolfia vomitoria's use in folk medicine for the treatment of erectile dysfunction could be its antioxidant and inhibitory activities on several enzymes linked to erectile dysfunction invitro.

The natural products discovered in marine sponge-associated microorganisms: structures, activities, and mining strategy

It is well known that microorganisms are abundant in sponges, which make up 50%-60% of the host biomass. Moreover, mounting evidence suggests that bacteria, fungi, and cyanobacteria, all associated with sponges, are the true creators of the bioactive compounds identified from sponges. The discovery of 774 structurally active compounds from 1998 to 2017 presents a good overview of natural product resources in sponge-associated microorganisms. During the last 5 years, many new molecules, including peptides, polyketides, alkaloids, and terpenes, have been identified from sponge-associated microorganisms through various mining strategies, exhibiting a wide range of biological activities, such as anti-microbial, anti-cancer, enzyme inhibition, and antioxidant properties. In this paper, 140 compounds produced by sponge-associated microorganisms from 2017 to 2022 are systematically discussed in terms of their structures, biological activities, and strain sources, as well as the mining strategies, which not only further updates the natural product library of sponge-associated microorganisms but also provides a new guideline for exploring the “dark matter” in sponges.

Palynological, chemical, antimicrobial, and enzyme inhibition properties of Cannabis sativa L. propolis

Palynological, chemical, antimicrobial, and enzyme inhibition properties of Cannabis sativa L. propolis

Production and Characterization of ACE Inhibitory and Anti-Diabetic Peptides from Buffalo and Camel Milk Fermented with Lactobacillus and Yeast: A Comparative Analysis with In Vitro, In Silico, and Molecular Interaction Study.

The investigation aimed at assessing a comparative study on the production and characterization of ACE inhibitory, anti-diabetic, and anti-inflammatory activities, along with the production of ACE inhibitory and anti-diabetic peptides through the fermentation of buffalo and camel milk by Limosilactobacillus fermentum (KGL4) and Saccharomyces cerevisiae (WBS2A). The angiotensin-converting enzyme (ACE) inhibitory and anti-diabetic properties were evaluated at particular time intervals (12, 24, 36, and 48 h) at 37 °C, and we discovered maximum activity at 37 °C after 48 h of incubation. The maximum ACE inhibitory, lipase inhibitory activities, alpha-glucosidase inhibitory, and alpha-amylase inhibitory activities were found in the fermented camel milk (77.96 ± 2.61, 73.85 ± 1.19, 85.37 ± 2.15, and 70.86 ± 1.02), as compared to the fermented buffalo milk (FBM) (75.25 ± 1.72, 61.79 ± 2.14, 80.09 ± 0.51, and 67.29 ± 1.75). Proteolytic activity was measured with different inoculation rates (1.5%, 2.0%, and 2.5%) and incubation times (12, 24, 36, and 48 h) to optimize the growth conditions. Maximum proteolysis was found at a 2.5% inoculation rate and at a 48 h incubation period in both fermented buffalo (9.14 ± 0.06) and camel milk (9.10 ± 0.17). SDS-PAGE and 2D gel electrophoresis were conducted for protein purification. The camel and buffalo milk that had not been fermented revealed protein bands ranging from 10 to 100 kDa and 10 to 75 kDa, respectively, whereas all the fermented samples showed bands ranging from 10 to 75 kDa. There were no visible protein bands in the permeates on SDS-PAGE. When fermented buffalo and camel milk were electrophoresed in 2D gel, 15 and 20 protein spots were detected, respectively. The protein spots in the 2D gel electrophoresis ranged in size from 20 to 75 kDa. To distinguish between different peptide fractions, water-soluble extract (WSE) fractions of ultrafiltration (3 and 10 kDa retentate and permeate) of fermented camel and buffalo milk were employed in RP-HPLC (reversed-phase high-performance liquid chromatography). The impact of fermented buffalo and camel milk on inflammation induced by LPS (lipopolysaccharide) was also investigated in the RAW 264.7 cell line. Novel peptide sequences with ACE inhibitory and anti-diabetic properties were also analyzed on the anti-hypertensive database (AHTDB) and bioactive peptide (BIOPEP) database. We found the sequences SCQAQPTTMTR, EMPFPK, TTMPLW, HPHPHLSFMAIPPK, FFNDKIAK, ALPMHIR, IPAVFK, LDQWLCEK, and AVPYPQR from the fermented buffalo milk and the sequences TDVMPQWW, EKTFLLYSCPHR, SSHPYLEQLY, IDSGLYLGSNYITAIR, and FDEFLSQSCAPGSDPR from the fermented camel milk.

Investigation of NADPH-Oxidase’s Binding Subunit(s) for Catechin Compounds Induce Inhibition

Catechins are natural polyphenolic compounds with ability to minimize excess free radicals through different mechanisms including inhibition of NADPH oxidase (NOX) activity. NOX is a complex enzyme made-up of several subunits, where molecules including catechins binds to exert their effect. Hence, the attempt to probe the NOX enzyme’s binding subunit of catechins-induce effect. Several in-silico techniques were deployed in probing the NOX enzyme’s binding subunit of the catechins. The catechins were downloaded from PubChem database in SDF files. The five NOX subunits with PDB ID: 3A1F, 1OV3, 1HH4, 1OEY, and 7CFZ were downloaded from the protein databank. Drug-likeness properties and biological activities were predicted using ADMETMESH software. Catechin-NOX subunits’ interactions was performed via molecular docking, and the docked conformations were analyzed using Protein-plus software. The results of the study predicted the catechin compounds; epicatechin (E), epicatechin gallate (EG) and epigallocatehin gallate (EGG) are drug-like in nature and possess enzymes inhibitory properties. Docking result predicted catechins are capable of interacting with the various NADPH oxidase (NOX) subunits but in a varied degree. Their (catechins) strongest affinities was predicted on p40phox and p67phox PB1 subunit (PBD: 1OEY) with binding energies in the ranges of -8.3 to -9.9kcal/mol in this order; Apocynin>EGG>EG>E. While a weak affinity was predicted between the catechin compounds and gp91 (phox) subunit (PDB: 3A1F) with binding energies (-4.9 to -6.5 kcal/mol) in this sequence; E<apocynin<EGG<EG. In conclusion, the study predicted catechin compounds possess drug-likeness properties and has affinities for interaction with NADPH-oxidase subunits particularly the p40phox and p67phox PB1 probably to exert their antioxidant effects. Therefore, in vitro and in vivo study is recommended to verify this claim.

Glutamate, Humic Acids and Their Combination Modulate the Phenolic Profile, Antioxidant Traits, and Enzyme-Inhibition Properties in Lettuce.

Lettuce (Lactuca sativa L., Asteraceae) is a popular vegetable leafy crop playing a relevant role in human nutrition. Nowadays, novel strategies are required to sustainably support plant growth and elicit the biosynthesis of bioactive molecules with functional roles in crops including lettuce. In this work, the polyphenolic profile of lettuce treated with glutamic acid (GA), humic acid (HA), and their combination (GA + HA) was investigated using an untargeted metabolomics phenolic profiling approach based on high-resolution mass spectrometry. Both aerial and root organ parts were considered, and a broad and diverse phenolic profile could be highlighted. The phenolic profile included flavonoids (anthocyanins, flavones, flavanols, and flavonols), phenolic acids (both hydroxycinnamics and hydroxybenzoics), low molecular weight phenolics (tyrosol equivalents), lignans and stilbenes. Overall, GA and HA treatments significantly modulated the biosynthesis of flavanols, lignans, low molecular weight phenolics, phenolic acids, and stilbene. Thereafter, antioxidant capacity was evaluated in vitro with 2,2-diphenyln-1-picrylhydrazyl (DPPH), 2,2'-Azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS), ferric reducing antioxidant power (FRAP), and cupric ion reducing antioxidant capacity (CUPRAC) assays. In addition, this study examined the inhibitory properties of enzymes, including acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), tyrosinase, alpha-amylase, and alpha-glucosidase. Compared to individual treatments, the combination of GA + HA showed stronger antioxidant abilities in free radical scavenging and reducing power assays in root samples. Moreover, this combination positively influenced the inhibitory effects of root samples on AChE and BChE and the tyrosinase inhibitory effect of leaf samples. Concerning Pearson's correlations, antioxidant and enzyme inhibition activities were related to phenolic compounds, and lignans in particular correlated with radical scavenging activities. Overall, the tested elicitors could offer promising insights for enhancing the functional properties of lettuce in agricultural treatments.

Density Functional Theory Calculations and Molecular Docking Analyses of Flavonoids for Their Possible Application against the Acetylcholinesterase and Triose-Phosphate Isomerase Proteins of Rhipicephalus microplus.

Ticks and tick-borne diseases constitute a substantial hazard to the livestock industry. The rising costs and lack of availability of synthetic chemical acaricides for farmers with limited resources, tick resistance to current acaricides, and residual issues in meat and milk consumed by humans further aggravate the situation. Developing innovative, eco-friendly tick management techniques, such as natural products and commodities, is vital. Similarly, searching for effective and feasible treatments for tick-borne diseases is essential. Flavonoids are a class of natural chemicals with multiple bioactivities, including the inhibition of enzymes. We selected eighty flavonoids having enzyme inhibitory, insecticide, and pesticide properties. Flavonoids' inhibitory effects on the acetylcholinesterase (AChE1) and triose-phosphate isomerase (TIM) proteins of Rhipicephalus microplus were examined utilizing a molecular docking approach. Our research demonstrated that flavonoids interact with the active areas of proteins. Seven flavonoids (methylenebisphloridzin, thearubigin, fortunellin, quercetagetin-7-O-(6-O-caffeoyl-β-d-glucopyranoside), quercetagetin-7-O-(6-O-p-coumaroyl-β-glucopyranoside), rutin, and kaempferol 3-neohesperidoside) were the most potent AChE1 inhibitors, while the other three flavonoids (quercetagetin-7-O-(6-O-caffeoyl-β-d-glucopyranoside), isorhamnetin, and liquiritin) were the potent inhibitors of TIM. These computationally-driven discoveries are beneficial and can be utilized in assessing drug bioavailability in both in vitro and in vivo settings. This knowledge can create new strategies for managing ticks and tick-borne diseases.