Angiotensin Converting enzyme-I Research Articles

In silico development of novel angiotensin-converting-enzyme-I inhibitors by Monte Carlo optimization based QSAR modeling, molecular docking studies and ADMET predictions

Within the realm of pharmacological strategies for cardiovascular diseases (CVD) like hypertension, stroke, and heart failure, targeting the angiotensin-converting enzyme I (ACE-I) stands out as a significant treatment approach. This study employs QSAR modeling using Monte Carlo optimization techniques to investigate a range of compounds known for their ACE-I inhibiting properties. The modeling process involved leveraging local molecular graph invariants and SMILES notation as descriptors to develop conformation-independent QSAR models. The dataset was segmented into distinct sets for training, calibration, and testing to ensure model accuracy. Through the application of various statistical analyses, the efficacy, reliability, and predictive capability of the models were evaluated, showcasing promising outcomes. Additionally, molecular fragments derived from SMILES notation descriptors were identified to elucidate the activity changes observed in the compounds. The validation of the QSAR model and designed inhibitors was carried out via molecular docking, aligning well with the QSAR results. To ascertain the drug-worthiness of the designed molecules, their physicochemical properties were computed, aiding in the prediction of ADME parameters, pharmacokinetic attributes, drug-likeness, and medicinal chemistry compatibility.

In vitro evaluation of the anti-hypertensive potential of albumin and globulin hydrolysates from cocoa bean (Theobroma cacao L. var. Forastero)

According to reports from the World Health Organization (WHO), hypertension is considered the most common cardiovascular risk factor, affecting 25 to 30% of the world's population. One of the primary causes of hypertension is angiotensin-converting enzyme-I (ACE), which plays a crucial role in regulating blood pressure through the modulation of the renin-angiotensin-aldosterone system. Consequently, there is a wide range of drugs available that can inhibit ACE. However, these drugs can cause adverse side effects such as cough, headache, skin rash, altered taste, angioedema, nausea, and allergic reactions. For this reason, there is considerable interest in obtaining ACE inhibitors from natural sources that reduce side effects and dependence on drugs. Specifically, bioactive peptides present in enzymatic protein hydrolysates have shown anti-hypertensive properties by inhibiting ACE, making them a promising alternative in the prevention and treatment of hypertension. The objective of this study was to evaluate the ACE inhibitory activity (in vitro) of enzymatic hydrolysates obtained from cocoa bean proteins (albumin and globulin), with the purpose of considering them as a natural source of potential anti-hypertensive agents. The results demonstrated that the enzymatic treatment (with alcalase) of albumin and globulin from cocoa beans is a viable bioprocess for producing hydrolysates with ACE inhibitory activity. In particular, hydrolysates and peptide fractions (< 3000 Da) with greater ACE inhibitory capacity were generated from globulin, making them a suitable choice for designing functional foods.

Bio-functional and prebiotics properties of products based on whole grain sorghum fermented with lactic acid bacteria.

Products fermented with lactic acid bacteria based on whole grain flours of red or white sorghum (Sorghum bicolor (L.) Moench) added with malted sorghum flour, or with skim milk (SM) were developed. Composition, protein amino acid profile, total acidity, pH, prebiotic potential, and bio-functional properties after simulation of gastrointestinal digestion were evaluated. In all cases, a pH of 4.5 was obtained in approximately 4.5 h. The products added with SM presented higher acidity. Products made only with sorghum presented higher total dietary fiber, but lower protein content than products with added SM, the last ones having higher lysine content. All products exhibited prebiotic potential, white sorghum being a better ingredient to promote the growth of probiotic bacteria. The addition of malted sorghum or SM significantly increased the bio-functional properties of the products: the sorghum fermented products added with SM presented the highest antioxidant (ABTS•+ inhibition, 4.7 ± 0.2 mM Trolox), antihypertensive (Angiotensin converting enzyme-I inhibition, 57.3 ± 0.5%) and antidiabetogenic (dipeptidyl-peptidase IV inhibition, 31.3 ± 2.1%) activities, while the products added with malted sorghum presented the highest antioxidant (reducing power, 1.6 ± 0.1 mg ascorbic acid equivalent/mL) and antidiabetogenic (α-amylase inhibition, 38.1 ± 0.9%) activities. The fermented whole grain sorghum-based products could be commercially exploited by the food industry to expand the offer of the three high-growth markets: gluten-free products, plant-based products (products without SM), and functional foods. © 2023 Society of Chemical Industry.

Bioactivity and nutritional quality of nutgall (Rhus semialata Murray), an underutilized fruit of Manipur.

Underutilized fruits plays a significant role in socio economic, cultural, nutritional and ethnomedicinal status of tribal people. However, scientific studies on the nutritional and other pharmaceuticals/biological activities of these fruits are meagre. Hence, the present study dealt with the quantification of nutritional quality and deciphering the bioactivity of nutgall (Rhus semialata Murray syn. Rhus chinensis Mill.), an underutilized fruit crop mainly found in foothill tracks of Eastern Himalaya, India, China, Japan, Korea and other South East Asian countries. The Rhus semialata Murray fruits were collected from five different locations in Purul sub-division, Senapati district, Manipur, India. The nutritional composition of the fruit pulp was analysed. Further the fruit pulp was extracted in methanol and water. The methanol and water extracts were studied for bioactivity properties such as antioxidant, antihyperglycemic, antihypertensive, antihyperuricemia, anti-tyrosinase, and antimicrobial activity. The fruit was rich in essential fatty acids. The presence of linoleic and oleic acids, along with traces of docosahexaenoic acid and eicosapantaenoic acid, revealed the potential food value of the fruit. 59.18% of the total amino acid composition of the protein present was constituted by essential amino acids. The IC50 value of methanolic extract (MExt) and Water extract (WExt) of the fruit were recorded as 4.05 ± 0.22 and 4.45 ± 0.16 μg/mL, respectively, in the DPPH assay and 5.43 ± 0.37 and 11.36 ± 2.9 μg/mL, respectively, in the ABTS assay as compared to Ascorbic acid (3 and 5.4 μg/mL in DPPH and ABTS assay, respectively). The CUPRAC assay also showed a high antioxidant potential of MExt and WExt (1143.84 ± 88.34 and 456.53 ± 30.02 mg Ascorbic Acid Equivalent/g, respectively). MExt and WExt of the fruit were more active against α-glucosidase (IC50 of 1.61 ± 0.34 and 7.74 ± 0.54 μg/ mL, respectively) than α-amylase enzyme (IC50 14.15 ± 0.57 and 123.33 ± 14.7 μg/mL, respectively). In addition, the methanolic fruit extract showed low to moderate pharmacological potential in terms of antihypertensive (Angiotensin converting enzyme-I inhibition), antihyperuricemia (xanthine oxidase inhibition), anti-tyrosinase, and antimicrobial activity. The IC50 values of angiotensin-converting enzyme I inhibition, xanthine oxidase inhibition and tyrosinase inhibition were recorded as 13.35 ± 1.21 mg/mL, 93.16 ± 4.65 mg/mL, and 862.7 ± 12.62 μg/mL, respectively. The study evidently indicates that nutgall fruit is a potential source of phytonutrients, bestowed with commercially exploitable, multifaceted health benefits.

Fermented quinoa flour: Implications of fungal solid-state bioprocessing and drying on nutritional and antioxidant properties

Quinoa (Chenopodium quinoa Willd) is a pseudocereal rich in protein and excellent source of vitamins, antioxidants, and minerals. Developing new quinoa-based products would be a strategy to extend its consumption. The aim of this study is to evaluate the impact of solid-state fermentation with Pleurotus ostreatus and drying (hot-air at 50, 60 and 70 °C and lyophilization) on the nutritional and functional properties of fermented white and black quinoa flours. Changes in proximal composition revealed that white variety was more suitable for solid-state fermentation (SSF), according to the higher biomass production, higher increase of protein (2.11%) and insoluble fiber (7.82%) content. SSF was highly effective in phytic acid reduction (90%) regardless the variety. However, after fermentation, quinoa seeds presented reduced total phenolic content (TPC) and antioxidant capacity, especially on black variety. Nonetheless, hot-air drying improved the antioxidant properties (increased TPC and antioxidant capacity) and promoted changes in the phenolic profile. These changes were characterized by a release of gallic acid together with a notable reduction of vanillic, caffeic and ferulic acids, quercetin-3-glucoside and/or quercetin. A notable increase of the angiotensin converting enzyme-I inhibitory activity was found on the fermented flours mainly due to SSF.

Neurologic Complications in Adult and Pediatric Patients with SARS-CoV-2 Infection

SARS-CoV-2 has an impact on the nervous system as a result of pathological cellular and molecular events at the level of vascular and neural tissue. Severe neurologic manifestations including stroke, ataxia, seizure, and depressed level of consciousness are prevalent in patients with SARS-CoV-2 infection. Although the mechanism is still unclear, SARS-CoV-2 has been associated with the pathogenesis of intravascular coagulation and angiotensin-converting enzyme-I, both exacerbating systemic inflammation and contributing to hypercoagulation or blood–brain barrier leakage, resulting in ischemic or hemorrhagic stroke. On the other hand, the SARS-CoV-2 spike protein in neural tissue and within the cerebrospinal fluid may induce neural dysfunction, resulting in neuroinflammation, which is exacerbated by peripheral and neural hypercytokinemia that can lead to neuronal damage and subsequent neuroinflammation. A deeper understanding of the fundamental biological mechanisms of neurologic manifestations in SARS-CoV-2 infection can pave the way to identifying a single biomarker or network of biomarkers to help target neuroprotective therapy in patients at risk for developing neurological complications.

Anti-hypertensive Peptide Predictor: A Machine Learning-Empowered Web Server for Prediction of Food-Derived Peptides with Potential Angiotensin-Converting Enzyme-I Inhibitory Activity.

Angiotensin converting enzyme-I (ACE-I) is a key therapeutic target of the renin-angiotensin-aldosterone system (RAAS), the central pathway of blood pressure regulation. Food-derived peptides with ACE-I inhibitory activities are receiving significant research attention. However, identification of ACE-I inhibitory peptides from different food proteins is a labor-intensive, lengthy, and expensive process. For successful identification of potential ACE-I inhibitory peptides from food sources, a machine learning and structural bioinformatics-based web server has been developed and reported in this study. The web server can take input in the FASTA format or through UniProt ID to perform the in silico gastrointestinal digestion and then screen the resulting peptides for ACE-I inhibitory activity. This unique platform provides elaborated structural and functional features of the active peptides and their interaction with ACE-I. Thus, it can potentially enhance the efficacy and reduce the time and cost in identifying and characterizing novel ACE-I inhibitory peptides from food proteins. URL: http://hazralab.iitr.ac.in/ahpp/index.php.

Conoidecyclics A-C from marine macroalga Turbinaria conoides: Newly described natural macrolides with prospective bioactive properties

Intertidal marine brown alga Turbinaria conoides (J.Agardh) Kützing (family Sargassaceae) is considered as one of the largely abundant species, available in the coastal zones of the Indian subcontinent. Bioactivity-guided chromatographic fractionation of the organic extract of T. conoides resulted in three previously undescribed macrocyclic lactone homologues, named as conoidecyclics A-C. Conoidecyclic A displayed greater attenuation potential against cyclooxygenase-2 (IC50 1.75 mM) and 5-lipoxygenase (IC50 4.24 mM) in comparison with other analogues. Conoidecyclic A exhibited higher attenuation potential against 5-lipoxygenase than that displayed by an anti-inflammatory agent, ibuprofen (IC50 4.51 mM). The higher selectivity index of conoidecyclic A (1.79) recognized its selective attenuation potential against the inducible cyclooxygenase-2 enzyme. Inhibition potential of conoidecyclic A against angiotensin converting enzyme-I (IC50 1.23 mM) and protein tyrosine phosphatase-1B (IC50 1.39 mM) were non-competitive, as deduced by kinetic studies. In-silico molecular modeling study of conoidecyclic A with the allosteric sites of the targeted enzymes exhibited least binding energy of −14.51 to −11.27 kcal mol−1 compared to those exhibited by other studied macrolide homologues. Reaction kinetic studies of conoidecyclic A coupled with lesser apparent Vmax inferred that it could efficiently bind with the allosteric site of targeted enzymes in a non-competitive manner to diminish the reaction velocity resulting in enzyme inhibition. Drug-likeness and predictive pharmacokinetic parameters of conoidecyclic A exhibited an acceptable oral bioavailability. These reports inferred that conoidecyclic A encompassing pentacosa macrocyclic moiety could be a promising therapeutic lead to inhibit the enzymes related to the development and progression of pathological conditions leading to inflammation, hypertension and type-2 diabetes.

Functional and Bioactive Properties of Protein Extracts Generated from Spirulina platensis and Isochrysis galbana T-Iso

There is growing consumer demand for food products derived from microalgae, driven largely by the perceived health benefits associated with them. The functional and bioactive potential of proteins isolated from two microalgae—Spirulina sp. and Isochrysis galbana T-Iso—were determined. The results obtained show the potential of microalgal protein extracts for use in the beverage industry, based on solubility values and other functional characteristics, including water and oil holding capacities, foaming, emulsifying activities and stabilities, water activities, solubility and pH. The solubility of algal proteins was pH-dependent, and they were largely insoluble at pH values between 2 and 11. However, the proteins were increasingly soluble at a pH of 12, and they have potential use in formulating foods with higher viscosities or gels, where they could act as fillers to strengthen networks. Compared with whey and flaxseed proteins, the Spirulina sp. protein extract had a superior oil-holding capacity (OHC). The OHC is important in developing texture in food products such as meats. Overall, better foam stability was observed for both Spirulina sp. and Isochrysis sp. soluble protein extracts, compared with flaxseed protein at pH values from 2 to 10 over a period of 120 min. The foam capacity and stability increase the physical properties of foods. However, the emulsion activity and stability values for soluble algal protein extracts were less than the values observed for flaxseed and whey proteins. Algal proteins would not be suitable for use in creaming and food processing involving flocculation. In addition, algal protein extracts inhibited Angiotensin-converting enzyme-I (ACE-I) and renin, and they have potential for use in functional food ingredient applications to maintain heart health and also to act as meat substitutes.

Multifunctional in vitro bioactive properties: Antioxidant, antidiabetic, and antihypertensive of protein hydrolyzates from tarwi (Lupinus mutabilis Sweet) obtained by enzymatic biotransformation

AbstractBackground and objectivesTarwi is a legume that grows in the Andean region of Peru. This grain is characterized by having a high protein content (~42%). In order to generate added‐value products from this resource, the possibility of obtaining tarwi protein hydrolyzates (TPH) with in vitro bioactive properties was evaluated using Alcalase, Neutrase, and Flavourzyme enzymes, acting alone or in combination for up to 240 min. The in vitro evaluated properties were as follows: antioxidant (ABTS assay), antidiabetic (α‐amylase, α‐glucosidase and dipeptidyl peptidase IV (DPP‐IV) inhibition), and antihypertensive (angiotensin‐converting enzyme‐I (ACE) inhibition).FindingsThe TPH obtained from the different enzyme treatments exhibited important antioxidant, antidiabetic (specially for DPP‐IV inhibition), and antihypertensive properties. One‐stage treatment using Alcalase for 240 min and two sequential stage treatments with Alcalase–Neutrase for 180 min presented the highest ABTS antioxidant activity and lowest IC50 values for DPP‐IV and ACE corresponding to 2.0–1.95 µmol TE/mg, 2.14–2.61 and 0.16–0.11 mg/ml, respectively.ConclusionsThe results of this research demonstrate that TPH exhibit in vitro multifunctional bioactive properties.Significance and noveltyTPH could potentially be used in the prevention or management of chronic diseases related to the development of oxidative processes, diabetes, and hypertension.

Characterization of Proteolytic Activity of Artichoke (Cynara scolymus L.) Flower Extracts on Bovine Casein to Obtain Bioactive Peptides.

Simple SummaryRecently, dairy proteins, in addition to their basic nutritional role in the diet, were recognized as a source of bioactive peptides. Such peptides are encoded within the primary structure of the protein and can be released by enzymatic hydrolysis. The growing interest in the development of functional foods for the benefit of consumer health led to a recent increase in research on the production of bioactive peptides from different matrices and production methods. The use of aspartic proteases from stigmas of mature artichoke (Cynara scolymus L.) flowers to obtain hydrolytic enzymes (cinarases) in the production of bioactive peptides would involve the utilization of an agricultural residue of a plant species of great socio-economic importance. In the present study, the characterization of the optimal hydrolysis conditions of artichoke flower extracts was carried out for the production of peptides from bovine casein. Furthermore, the angiotensin-converting enzyme-I inhibitory activity and the antioxidant capacity against 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2’-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid (ABTS) free radicals in vitro were determined for the obtained hydrolysates. The results revealed that the water-soluble extract of artichoke flower could be suitable for the production of bioactive peptides from whole bovine casein.The aim of this work is to establish the most suitable proteolysis conditions to obtain bovine casein hydrolysates containing peptides with antioxidant and antihypertensive capacity. To this end, the proteolytic activity of Cynara scolymus L. flower extracts was characterized on whole bovine casein, evaluating the effect of several factors (pH, temperature, substrate concentration, enzyme concentration, and hydrolysis time). The optimal conditions to carry out the hydrolysis with the C. scolymus L. extract were as follows: pH 6.2, 50 °C, and 0.023 mg·mL−1 of extract-protein concentration. A Michaelis constant (Km) value of 5.66 mg·mL−1 and a maximum rate of reaction (Vmax) of 8.47 mUAbs∙min−1 were observed. The optimal hydrolysis time was 17 h. The casein hydrolysates obtained with these conditions contained peptides with antioxidant activity (1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging capacity: 30.89%; Trolox equivalent antioxidant capacity (TEAC) against 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) free radical (ABTS●+): 4.43 mM Trolox equivalent·mg−1 peptide) and antihypertensive activity, showing 55.05% angiotensin-converting enzyme-I inhibition in vitro.

Pharmacological Properties of Seaweeds against Progressive Lifestyle Diseases

ABSTRACT Seaweeds are consumed as staple food items in the cuisines of many Asian countries and are considered as essential components of a healthy diet. The pharmacological potential of ethyl acetate-methanol (EtOAc-MeOH) extract of 10 species of seaweeds belonging to the subclasses Dictyotophycidae, Rhodymeniophycidae, and Corallinophycidae against progressive lifestyle diseases was determined by various in vitro models. Among the studied species, the organic extracts of Gracilaria salicornia and Padina tetrastromatica exhibited potential dipeptidyl-peptidase-4 inhibiting activity (IC50 ~ 0.03 mg mL−1) and angiotensin converting enzyme-I inhibitory property (ACE-I) (IC50 0.12 mg mL−1) compared to those displayed by other studied species. The organic extracts of G. salicornia and P. tetrastromatica also displayed significantly greater attenuation properties against pro-inflammatory cyclooxygenase-2 and 5-lipoxygenase (IC50 0.98–1.34 mg mL−1, P < .05) compared to those derived from other species (IC50 1.99–5.02 mg mL−1). The nuclear magnetic resonance-guided dereplication of organic extracts of the studied seaweed species recognized the co-linearity between the bioactive potential and downfield electronegative functionalities. These results recognized the significance of the organic extracts of G. salicornia and P. tetrastromatica to isolate compounds for functional food applications against oxidative stress and induced diseases, such as diabetes, hypertension, and inflammation.

Functional Properties of Ethyl Acetate-methanol Extract of Commonly Edible Molluscs

ABSTRACTMarine molluscs are consumed as culinary delicacies and represent a relatively untapped source of natural functional food ingredients. The bioactive potential of ethyl acetate-methanol (C2H5OAc/CH3OH) extract of molluscs Uroteuthis duvaucelii, Amphioctopus marginatus, Crassostrea madrasensis, and Sepiella inermis were demonstrated using different in vitro systems. The C2H5OAc/CH3OH extract of A. marginatus displayed greater 2, 2-diphenyl-1-picryl-hydrazil (IC90 1.76 mg mL−1) radical quenching capacity along with potential hydroxymethylglutaryl coenzyme-A reductase (HMGCR)-inhibiting activity (IC90 1.21 mg mL−1) and angiotensin converting enzyme-I inhibitory property (ACE-I) (IC90 0.87 mg mL−1) compared to those displayed by other molluscs. The organic extract of S. inermis set forth significantly higher pro-inflammatory 5-lipoxygenase inhibitory property (IC90 1.96 mg mL−1) along with potential antidiabetic activity as determined by dipeptidyl peptidase-4 (DPP-4) antagonistic activities (IC90 2.62 mg mL−1, P < .05). Significant correlations were observed between bioactive properties and electronegative groups of organic extracts of a mollusc. The free radical quenching property of the extracts exhibited a positive correlation towards other bioactivities, implicating the role of antioxidant property of the compounds in the extract to inhibit lifestyle ailments. The present study indicated that S. inermis and A. marginatus could be utilized as functional food ingredients to combat hypertension, hyperlipidemia, and inflammation-related risks.

Food-derived bioactive peptides and their role in ameliorating hypertension and associated cardiovascular diseases.

Non-communicable diseases including cardiovascular diseases (CVDs) and associated metabolic disorders are responsible for nearly 40 million deaths globally per year. Hypertension or high blood pressure (BP) is one of the primary reasons for the development of CVDs. A healthy nutritional strategy complementing with physical activity can substantially reduce high BP and prevent the occurrence of CVD-associated morbidity and mortality. Bioactive peptides currently are the next wave of the promising bench to clinic options for potential targeting chronic and acute health issues including hypertension. Peptides demonstrating anti-inflammatory, anti-oxidant, and angiotensin-converting enzyme-I inhibitory activity are widely studied for the amelioration of hypertension and associated CVDs. Isolating these potent bioactive peptides from different food sources is a promising endeavor toward nutraceutical based dietary management and prevention of hypertension. Understanding the pathophysiology of hypertension and the action mechanisms of the bioactive peptides would complement in designing and characterizing more potent peptides and suitable comprehensive dietary plans for the prevention of hypertension and associated CVDs.

Characterization of nanoliposomes containing bioactive peptides obtained from sheep whey hydrolysates

Whey is recognized by the excellent nutritional and functional properties of its proteins, which can be hydrolyzed to peptides with an array of biological activities. In this work, sheep whey was treated with a novel proteinase from Bacillus sp. P7, resulting in hydrolysates with antioxidant and angiotensin-converting enzyme I (ACE-I) inhibitory activity. The hydrolysates were purified by gel filtration chromatography on Sephadex G-25 and the active fractions were subjected to ultrafiltration through 10 kDa cut-off membranes. The fraction enriched in bioactive peptides was encapsulated into phosphatidylcholine liposomes, which were characterized and evaluated by their ability to maintain biological activities. The physicochemical parameters of the liposomes showed constant values during 30 days, with mean diameter around 166 nm and zeta potential −17 mV. Both antioxidant and ACE-I-inhibitory activities were detected in the liposome preparations. In addition, the residual antioxidant activity was 87% after 30 days storage, suggesting the bioactive peptides are stable in this formulation. Liposome encapsulated peptides may offer the possibility of application in areas such as food science and technology, in order to increase the nutritional value and shelf life of food products, as well as for the development of functional foods.

Pharmacological properties of marine macroalgae-associated heterotrophic bacteria.

The rich diversity of marine macroalgae and their associated bacterial flora represent a potential reservoir of bioactive compounds with valuable biotechnological and pharmaceutical use. Heterotrophic bacteria associated with the intertidal macroalgae were isolated and evaluated for their pharmacological properties using various in vitro models. Among 148 cultivable isolates, more than 50% were dominated by γ-Proteobacteria and Firmicutes, wherein 53 of them showed consistent antibacterial activity against a broad spectrum of clinically significant pathogens. The bacteria were characterized by extensive microbiological, molecular and chemical identification tools. The heterotrophs Bacillus amyloliquefaciens MTCC 12716 and Shewanella algae MTCC 12715 isolated from a red marine macroalga Hypnea valentiae exhibited potential anti-infective properties against multidrug-resistant pathogens, such as methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococcus faecalis (minimum inhibitory concentration of 6.25-12.5µg/mL). The organic extract of B. amyloliquefaciens displayed significantly greater antioxidative properties (IC90 < 1mg/mL) and the activities showed considerable positive correlation (r2 > 0.8, P < 0.05) with the inhibitory activities against angiotensin converting enzyme-I, pro-inflammatory cyclooxygenases and 5-lipoxygenase, dipeptidyl peptidase-4 and hydroxymethylglutaryl coenzyme A reductase, which were associated with hypertension, inflammation, diabetes, and hypercholesterolemia, respectively. The applications of nuclear magnetic resonance-based fingerprinting to analyze the characteristic signals in the solvent extracts and to correlate them with the pharmaceutical properties were underlined. The heterotrophic bacterium B. amyloliquefaciens MTCC 12716 might, therefore, serve as a potential therapeutic candidate to develop products with wide pharmaceutical applications.

Optimization of fast and simple real-time PCR-based method for genotyping the angiotensin converting enzyme-I I/D polymorphism

A inserção ou deleção de 288 pb no íntron 16 do gene que codifica a enzima conversora de angiotensina (ECA) foi a primeira variante genética associada com o desempenho físico e uma das mais estudadas nos últimos 15 anos. Carreadores da deleção em pelo menos um dos alelos, podem apresentar maior atividade enzimática, resultando em uma maior resposta vasoconstritora. Estes indivíduos podem também possuir uma melhor resposta ao treinamento de força e potência, assim como carreadores da inserção podem possuir uma melhor predisposição ao treinamento de endurance. Tradicionalmente, para determinar o genótipo do indivíduo (I/I, I/D ou D/D), o método utilizado é o PCR convencional. Este método envolve dois estágios; primeiramente a reação da PCR é realizada e depois o gel de agarose contendo o produto da PCR é submetido à eletroforese tornando possível visualizar por luz UV as bandas de DNA indicando o genótipo. Para o uso desta metodologia em larga escala, como no caso de estudos de associação, utilizados para avaliar a influência da genética no esporte, este duplo processo consumo muito tempo. Este artigo tem como objetivo apresentar um protocolo rápido e eficiente para a genotipagem deste polimorfismo por meio da PCR em tempo real, utilizando DNA genômico coletado de células bucais. O protocolo discutido no texto foi inicialmente proposto em 2001, contudo a sua configuração original apresenta limitações e utiliza uma quantidade grande de material. Variáveis do protocolo, tais como: concentração do primer, volume de reação e resolução da curva de dissociação que indica o genótipo foram ajustadas. Após este ajustamento, o protocolo permaneceu efetivo com uma quantidade reduzida de custo, adequado para o uso em estudos em larga-escala envolvendo genética e esporte.

Characterization and in vitro evaluation of seaweed species as potential functional ingredients to ameliorate metabolic syndrome

This study aimed at identifying seaweed species with optimal characteristics to develop multifunctional foods for metabolic syndrome (MetS) management. Mass spectrometry chemical characterization and bioactive profile evaluation of methanolic extracts from seven commonly consumed seaweeds were compared. Monomeric sugars namely mannitol, fucitol, xylitol and their sulphated analogs as well as lipids (phosphatidic acid, octadecenoic acid, and prostaglandin 2α) were detected in seaweeds. Himanthalia elongata showed the highest phenolic content (24.04 µmol GAE/g), and antioxidant activity. This species was the only one showing angiotensin converting enzyme-I inhibitory activity (IC50 = 65 µg/mL). U. pinnatifida and H. elongata extracts were notably more effective reducing pro-inflammatory molecules in lipopolysaccharide-induced RAW 264.7 macrophages. Finally, Ulva spp., Palmaria palmata, U. pinnatifida and H. elongata significantly inhibited triglyceride accumulation in mature 3T3-L1 adipocytes (43–52% inhibition). Among seaweeds, H. elongata showed the highest potential to be used as ingredient in the development of new functional foods for MetS management.

Novel furanyl derivatives from the red seaweed Gracilaria opuntia with pharmacological activities using different in vitro models

Two previously undescribed furanyl compounds, characterized as 5-(7-(5-ethyl-3,4-dimethoxycyclooctyl) benzofuran-6-yl)-7-methyl-3,4,7,8-tetrahydro-2H-oxocin-2-one (compound 1) and 2-(3-ethyl-9-(2-methoxyethoxy)-1-oxo-2,3,4,9-tetrahydro-1H-xanthen-2-yl) ethyl-5-hydroxy-9-methoxy-7,8-dimethyl-8-(5-methylfuran-2-yl) nona-3,6-dienoate (compound 2) were derived from the ethyl acetate–methanol (EtOAc:MeOH) crude extract of red seaweed Gracilaria opuntia. The isolated compounds are the first furanyl natural products featuring methoxycyclooctyl benzofuran with tetrahydro-2H-oxocin framework and tetrahydro-1H-xanthenyl methoxy methylfuran skeletons. These compounds were assessed for anti-inflammatory activities against pro-inflammatory cyclooxygenase-2/5-lipoxygenase (COX-1, 2, and 5-LOX) and antioxidative effects in various in vitro models. The methylfuran derivative exhibited comparable inhibitory activities towards 5 LOX (IC50 0.209 × 10−2 M) with synthetic non-steroidal anti-inflammatory drugs (NSAID) ibuprofen (IC50 0.451 × 10−2 M, P 0.146 × 10−2 M), and were similar to those displayed by the synthetic antioxidants (butylated hydroxytoluene (BHT)/ butylated hydroxyanisole (BHA) (IC50 ~0.144–0.189 × 10−2 M, P < 0.05). The anti-inflammatory selectivity indices of the isolated compounds recorded significantly greater values (SI: anti-COX-1IC50/anti-COX-2IC50 ~1.08–1.09) than NSAIDs (aspirin, and ibuprofen, SI: 0.02 and 0.44, respectively, P < 0.05), and consequently, appeared to be safer. The isolated compounds showed significant anti-diabetic properties as determined by α-amylase/α-glucosidase (IC50 < 0.052 × 10−2 M) and dipeptidyl peptidase-4 (DPP-4, IC50 < 0.002 × 10−2 M) inhibitory activities. The angiotensin converting enzyme-I (ACE-I) inhibitory activity of the compounds (IC50 0.023–0.024 × 10−2 M) was found to be comparable with that recorded by commercial ACE inhibitor, captopril (IC50 0.037 × 10−2 M).

Pharmacological potential of sulfated polygalactopyranosyl-fucopyranan from the brown seaweed Sargassum wightii

A sulfated polygalactopyranosyl-fucopyranan characterized as ∙∙∙∙ → 1)-α-Fucp-(2SO3−)-(3 → 1)-α-Fucp-(2SO3−)-(4 → 1)-β-Galp-(4 → 1)-β-Galp-(4 → ∙∙∙∙ was isolated from the brown seaweed Sargassum wightii and evaluated for pharmacological properties with reference to antioxidant, anti-inflammatory, antidiabetic, and antihypertensive activities using different in vitro models. The sulfated polygalactopyranosyl-fucopyranan displayed potential di(phenyl)-(2,4,6-trinitrophenyl) iminoazanium (DPPH), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS+) radical scavenging, and Fe2+ ion chelating activities (IC90 ~ 1 mg mL−1). The studied polysaccharide displayed higher anti-inflammatory selectivity towards inducive pro-inflammatory enzyme cyclooxygenase-2 (COX-2, IC90 1.13 mg mL−1) than constitutive cyclooxygenase-1 (COX-1, IC90 > 1.20 mg mL−1) resulting in greater selectivity index (IC90 COX-2/COX-1, 0.93) than synthetic non-steroidal anti-inflammatory drug aspirin (0.88) and also showed potent lipoxygenase-5 inhibition (LOX-5, IC90 1.02 mg mL−1). The studied polysaccharide displayed significantly higher (P < 0.05) antidiabetic properties compared to the antidiabetic agents acarbose and diprotein-A in terms of α-amylase (IC90 0.93 mg mL−1), α-glucosidase (IC90 1.48 mg mL−1), and dipeptidyl peptidase-4 (IC90 0.11 mg mL−1) enzyme inhibition potentials. The sulfated polygalactopyranosyl-fucopyranan also displayed potential antihypertensive activity with reference to angiotensin-converting enzyme-I inhibitory activity (IC90 0.2 mg mL−1). Extensive spectroscopic experiments in conjugation with monosaccharide compositional analysis attributed (1 → 3)-linked α-fucopyranose units in the polygalactofucan chain with C-2 sulfation and C-4 substituents as O-acetyl/O-methyl/(1 → 4)-linked β-galactopyranose. The previously undescribed sulfated polygalactopyranosyl-fucopyranan could function as a potential pharmacophore lead against inflammation, type 2 diabetics, hypertension and utilization as natural antioxidant.