Fraction F1 Research Articles

Antimicrobial and antioxidant activity of proteins isolated from Melipona beecheii honey

Proteins from Melipona beecheii honey were purified by concanavalin A (conA) affinity chromatography and eluted with a stepwise glucose gradient into fractions named F2-F5. The conA-unbound fraction (F1) was further separated by molecular exclusion into fractions named MbF1-1,2 and MbF1-3. All fractions were evaluated for antibacterial activity against foodborne pathogens and antioxidant capacity. F1 fraction possessed highest antimicrobial activity against S. aureus, L. monocytogenes, S. Typhimurium, E. coli and P. aeruginosa with MIC’s 1.4 ± 0.2, 15 ± 1, 39 ± 2, 1 ± 0.1, and 75 ± 2 µg/mL, respectively. F1, MbF1-1,2 and MbF1-3 had bactericidal effect except against P. aeruginosa. When the antioxidant capacity of the fractions was determined, F2 had the highest antioxidant activity measured by DPPH radical scavenging activity (IC50 = 2.4 ± 0.4 µg/µL) and reducing power of Fe(III) (IC50 = 1.8 ± 0.2 µg/µL). We provide evidence that M. beecheii honey proteins possess broad spectrum antibacterial and antioxidant activity, the latter probably through their reducing agent and free radical scavenger properties.

Remediation of As and Cd contaminated sediment by biochars: Accompanied with the change of microbial community

As and Cd pollution in sediment has posed a potential threat to ecosystem owing to their high toxicity. Fixing heavy metals and transforming their speciation into stable forms can be a sustainable method. In this study, nine biomass feedstocks were pyrolyzed into biochars for immobilization of As and Cd in sediment, and the accompanied changes in the microbial community were analyzed to evaluate their effect on bioavailability. The heavy metal immobilization capacity of biochars in sediment were assessed by measuring the change of As and Cd concentrations in aqueous phases (overlying water (COW) and pore water (CPW)) and the leachate of Toxicity Characteristic Leaching Procedure (TCLP), and metal speciation after biochar treatment. The maximum reduction of As and Cd concentrations could reach 95% and 78% (COW), 73% and 59% (CPW), 81% and 37% (leachate), respectively, and the F1, F2, F3 fraction (bioavailable form) of As and Cd significantly decreased (up to 18.0%) in the biochar-treated sediments. The ACE (Abundance-based Coverage Estimator) and Shannon index results revealed that biochars efficiently enriched the microbial biodiversity and population in the sediment. Water hyacinths biochar (WHB) exhibited the better performance among all biochar samples mainly due to the abundant oxygen-containing functional groups on the surface and the higher O/C. The overall results highlighted the inhibition effects of biochar on heavy metal mobility and bioavailability, and helped to provide guidance for optimization of biochars for sediment remediation.

Unravelling the Anti-Inflammatory and Antioxidant Potential of the Marine Sponge Cliona celata from the Portuguese Coastline.

Inflammation is a double-edged sword, as it can have both protective effects and harmful consequences, which, combined with oxidative stress (OS), can lead to the development of deathly chronic inflammatory conditions. Over the years, research has evidenced the potential of marine sponges as a source of effective anti-inflammatory therapeutic agents. Within this framework, the purpose of this study was to evaluate the antioxidant and the anti-inflammatory potential of the marine sponge Cliona celata. For this purpose, their organic extracts (C1–C5) and fractions were evaluated concerning their radical scavenging activity through 2,2-diphenyl-1-picrylhydrazyl radical (DPPH), ferric reducing antioxidant power (FRAP), oxygen radical absorbance capacity (ORAC), and anti-inflammatory activity through a (lipopolysaccharides (LPS)-induced inflammation on RAW 264.7 cells) model. Compounds present in the two most active fractions (F5 and F13) of C4 were tentatively identified by gas chromatography coupled to mass spectrometry (GC-MS). Even though samples displayed low antioxidant activity, they presented a high anti-inflammatory capacity in the studied cellular inflammatory model when compared to the anti-inflammatory standard, dexamethasone. GC-MS analysis led to the identification of n-hexadecanoic acid, cis-9-hexadecenal, and 13-octadecenal in fraction F5, while two major compounds, octadecanoic acid and cholesterol, were identified in fraction F13. The developed studies demonstrated the high anti-inflammatory activity of the marine sponge C. celata extracts and fractions, highlighting its potential for further therapeutic applications.

Metabolic Profile and Mycoherbicidal Activity of Three Alternaria alternata Isolates for the Control of Convolvulus arvensis, Sonchus oleraceus, and Xanthium strumarium.

Alternaria alternata isolates C1, S1, and X3 were isolated respectively from the weeds Convolvulus arvensis, Sonchus oleraceus, and Xanthium strumarium in Algiers during 2016 and identified by morphological and molecular analyses. The aim of this investigation was to chemically characterize the exometabolome of these fungi and to evaluate the myco-herbicidal potential of their culture filtrates, crude extracts, or fractions towards target weeds. Results revealed a great heterogeneity in the biochemical profiles of the exometabolome with the remarkable presence of two compounds: tenuazonic acid (TeA) and triprenyl phenol-7 (SMTP-7). To the best of our knowledge, SMTP-7—found in all the isolates—as well as 12-methoxycitromycin detected in the culture filtrate of isolate C1, have never been reported to be produced by A. alternata. Some fractions of isolates C1 and S1 showed symptoms (necrosis and chlorosis) on the detached leaves of C. arvensis and S. oleraceus, respectively with up to 100% phytotoxic effect at low concentration. In conclusion, biochemical characterization revealed great difference of C1, S1, and X3 exometabolome that is likely to explain the difference in their phytotoxic activity. Some fractions (d1, e1, h1, i1, a2, and f2) of isolates C1 and S1 of A. alternata caused severe necrosis and chlorosis on the injured detached leaves of C. arvensis and S. oleraceus, respectively.

Mallotus Mollissimus and Solanum Erianthum Exhibit Antikinase, Antiphosphatase and Anti-Cancer Properties

Cancer is a leading cause of death worldwide and caused by dysregulated signal transduction from kinase and phosphatases. Inhibitors of kinase and phosphatase have demonstrated anticancer properties. Therefore, this study aimed to investigate the antikinase, antiphosphatase and cytotoxic properties of Mallotus mollissimus (M. mollissimus) and Solanum erianthum (S. erianthum). Toxic activities against PP1, MKK1 and MSG5 assays were demonstrated by S. erianthum methanol extract. Bioassay-guided fractionation of the methanolic extracts showed that chloroform fraction (CE) of M. mollissimus exhibited toxic activity against PP1. Meanwhile, CE of S. erianthum showed positive activity on PP1 assay. Column chromatography separation of the CE has revealed that fractions F1 and F2 of M. mollissimus are toxic against PP1. Meanwhile, F1 and F2 CE fractions of S. erianthum were positive against PP1 and F9 fraction showed toxic activity in PP1 assay. Chloroform extracts of both plants exhibit cytotoxicity activity against HeLa, CaOV3 and MCF7 cell lines. This study demonstrated the potential of M. mollissimus and S. erianthum extracts in antikinase, antiphosphatase and anti-cancer activities which warrant further purification and identification.

Production and Evaluation of Fractionated Tamarind Seed Oil Methyl Esters as a New Source of Biodiesel

Biodiesel has attracted considerable interest as an alternative biofuel due to its many advantages over conventional petroleum diesel such as inherent lubricity, low toxicity, renewable raw materials, biodegradability, superior flash point, and low carbon footprint. However, high production costs, poor low temperature operability, variability of fuel quality from different feedstocks, and low storage stability negatively impact more widespread adoption. In order to reduce production costs, inexpensive inedible oilseed alternatives are needed for biodiesel production. This study utilized inedible tamarind (Tamarind indica) seed oil as an alternative biodiesel feedstock, which contained linoleic (31.8%), oleic (17.1%), and lauric (12.0%) acids as the primary fatty acids. A simple and cost-effective high vacuum fractional distillation (HVFD) methodology was used to separate the oil into three fractions (F1, F2, and F3). Subsequent transesterification utilizing basic, acidic, and enzymatic catalysis produced biodiesel of consistent quality and overcame the problem of low temperature biodiesel performance. The most desirable biodiesel with regard to low temperature operability was produced from fractions F2 and F3, which were enriched in unsaturated fatty acids relative to tamarind seed oil. Other properties such as density and cetane number were within the limits specified in the American and European biodiesel standards.

Phenolic Compound Composition and Biological Activities of Fractionated Soybean Pod Extract

The objective of this study was to determine the potential of dry extracts from soybean pods as a source of bioactive compounds. The phenolic compound composition of the extract (E) and fraction (F1-F3) as well as their respective lipophilicity were analyzed by UPLC-ESI-QTOF-MS. The biological activity as measured by antioxidant and cytotoxic activity was also evaluated. The in vitro anticancer potential of the ethanolic extract and fraction of soybean pods was measured through MTT assay using human colorectal carcinoma (HTC-116) and prostate cancer (PC-3) cell lines. The F2 fraction, with medium lipophilicity, produced the most pronounced cytotoxic effect in PC-3 cells. Analysis of Pearson’s correlation demonstrated a high relationship between the content of phenolic compounds, the antioxidant activity of the extract and fractions, and cytotoxic activity towards PC-3 cells. UPLC-ESI-QTOF-MS analysis of soybean pod extract resulted in putative identification of 50 polyphenols belonging to three different chemical families (phenolic acids, flavonoids, and other polyphenols), the first time such an analysis has been performed. Among the fractions with variable lipophilicity, fraction F2 was the most abundant in phenolic compounds. This was confirmed in the quantitative analysis, as a result of which the highest total content of phenolic compounds was found in the extract (E) and fraction F2. Soybean pods might be useful material for obtaining preparations with anti-radical and anti-cancer properties. They can also be an active food additive or a component in dietary supplements.

Inhibitory effects of Chinese quince fruit proanthocyanidins with different polymerisation degrees on the formation of heterocyclic aromatic amines in chemical model systems

SummaryProanthocyanidins with different polymerisation degrees were extracted from Chinese quince fruits by subcritical ethanol/water into four fractions (F1‐F4), which were then analysed using UPLC‐MS/MS, GPC and thiolysis combined with RP‐HPLC MS and MALDI‐TOF MS (matrix‐assisted laser desorption/ionisation time‐of‐flight mass spectrometry). The inhibitory effects of these proanthocyanidins on the formation of heterocyclic aromatic amines (HCAs), such as norharman, harman and PhIP, in chemical model systems were determined. The results reflected that the mean degree of polymerisation (DP) of proanthocyanidins in fractions F1, F2, F3 and F4 were 8.0, 12.7, 27.5 and 64.9, respectively. All four fractions prominently and dose‐dependently reduced the formation of HCAs at concentrations of 0.5, 1 and 2 mg mL−1 in chemical models. F1, the fraction richest in oligomers, had the most effective inhibition of harman, norharman and PhIP, with the levels reduced by 65.71%, 37.15% and 72.99%, respectively, when used at a concentration of 2 mg mL−1. The mean DP of proanthocyanidins showed negative correlation with the inhibition of HCA formation (R2= −0.697 for norharman, −0.799 for PhIP and −0.758 for harman). These findings suggest that proanthocyanidins from Chinese quince fruit have the potential to be used as additives to reduce the levels of heterocyclic amines in foods.

Trends and environmental factors of arsenic in sediments from the five lake ecoregions, China.

The behavior and risk of arsenic (As) closely relate to its geochemical fractionation and environmental factors in sediments. The soluble (F1), reducible (F2), oxidizable (F3), and residual fraction (F4) of As were extracted in the sediments from Lake Hulun, Wuliangsuhai, and Dalinor of Inner Mongolia Plateau. Coupled with lakes from Eastern and Northeast Plain, Yunnan-Guizhou and Qinghai-Tibetan Plateau, the responses of As fractions to environmental conditions were investigated according to the spatial distribution of As fractionations in five lake ecoregions at a national scale of China. Generally, F1 was more sensitive to environmental changes, and the pH presented significantly negative effects on the amount of soluble As, while water depth played an important role in regulating the distribution of the fraction F2 and F4. The As pools in surface lake sediments presented a latitudinal zonation due to the gradient effects of climate and anthropogenic activities on nutrient decomposition, and their influence on the capacity of sediments holding As. This work indicated that nutrients played a coordinating role in regulating the impacts of climate and environmental factors on As fractionation in aquatic environments.

Investigation of S.limacinum microalgae digestibility and production of antioxidant bioactive peptides

Schizochytrium limacinum (S. limacinum) has potential as a high protein food ingredient. The digestibility of this protein was performed in the conditions of simulated stomach phase (with pepsin enzyme) and simulated small intestine phase (with α-chymotrypsin and trypsin). The whole hydrolysate was fractionated into various ranges of molecular weight cutoff (MWCO) by using ultrafiltration (UF) technique. Antioxidant activities of hydrolysate fractions were evaluated using direct free radical scavenging activity such as 2,2-diphenyl-1-picrylhydrazyl Radical (DPPH•) scavenging, activity, 2,2-azino-bis(3-ethylbenzthiazoline)-6-sulfonic acidradical cation (ABTS•+) sscavengin activity and reducing power activity. In addition, the protective effect of purified peptide against oxidation of low-density lipoprotein (LDL) was determined. The microalgae proteins were completely hydrolyzed by digestive enzymes and the enzymatic hydrolysis of proteins resulted in increased antioxidant activity of peptides. The peptide fraction F3 (MWCO = 5–10 kDa) presented the highest antioxidant activity out of all the methods described. The protein hydrolysates showed higher reducing power, radical scavenging capacity and significantly inhibited LDL oxidation. These results suggest that the enzymatic hydrolysates from S. limacinum can act as novel antioxidants in functional foods.

Chemical Evaluation, Antioxidant, Antiproliferative, Anti-Inflammatory and Antibacterial Activities of Organic Extract and Semi-Purified Fractions of the Adriatic Sea Fan, Eunicella cavolini.

Due to sedentary lifestyle and harsh environmental conditions, gorgonian coral extracts are recognized as a rich source of novel compounds with various biological activities, of interest to the pharmaceutical and cosmetic industries. The presented study aimed to perform chemical screening of organic extracts and semi-purified fractions obtained from the common Adriatic gorgonian, sea fan, Eunicella cavolini (Koch, 1887) and explore its abilities to exert different biological effects in vitro. Qualitative chemical evaluation revealed the presence of several classes of secondary metabolites extended with mass spectrometry analysis and tentative dereplication by using Global Natural Product Social Molecular Networking online platform (GNPS). Furthermore, fractions F4 and F3 showed the highest phenolic (3.28 ± 0.04 mg GAE/g sample) and carotene (23.11 ± 2.48 mg β-CA/g sample) content, respectively. The fraction F3 inhibited 50% of DPPH (2,2-diphenyl-1-picryl-hydrazyl-hydrate) and ABTS (2,2′-azino-bis (3-ethylbenzthiazolin-6-yl) sulfonic acid) radicals at the concentrations of 767.09 ± 11.57 and 157.16 ± 10.83 µg/mL, respectively. The highest anti-inflammatory potential was exhibited by F2 (IC50 = 198.70 ± 28.77 µg/mL) regarding the inhibition of albumin denaturation and F1 (IC50 = 254.49 ± 49.17 µg/mL) in terms of soybean lipoxygenase inhibition. In addition, the most pronounced antiproliferative effects were observed for all samples (IC50 ranging from 0.82 ± 0.14–231.18 ± 46.13 µg/mL) against several carcinoma cell lines, but also towards non-transformed human fibroblasts pointing to a generally cytotoxic effect. In addition, the antibacterial activity was tested by broth microdilution assay against three human pathogenic bacteria: Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus. The latter was the most affected by fractions F2 and F3. Finally, further purification, isolation and characterization of pure compounds from the most active fractions are under investigation.

Changes in potentially toxic element concentration and potential ecological risk in topsoil caused by sewage sludge application on forestland: A 3-year field trial

Sewage sludge (SS) application on forest plantation soils as a fertilizer/soil amendment is increasingly becoming a forestry management measure. However, the potential risk of SS-derived potentially toxic elements (PTEs) is still a cause for concern. This research was carried out to evaluate PTEs behavior in SS applied as an amendment for forestland application purposes. Speciation and transformation perspectives on residual effects during a 3-year field experiment were evaluated. Sewage sludge was applied to soils under Eucalyptus urophylla S.T. Blake, Schima superba Gardn. et Champ, and Pinus elliottii Engelm plantations at 30 Mg·ha−1 (dry weight). We investigated the total concentrations of Cu, Zn, Pb, Cd, and Ni, as well as their concentrations of five fractions: exchangeable (F1), bound to carbonates (F2), Fe–Mn oxides (F3), organic matter (F4), and residual (F5) along with the 0–10 cm of a surface A horizon (topsoil) one day, one, two, and three years after SS application, respectively. Sewage sludge caused significant increase in PTEs total concentrations and proportion of fraction F1 or F2. The mobility index and potential ecological risk confirmed that the risk was linked to the presence of these PTEs, especially for Cd. The total concentration and corresponding ecological risks of PTEs in most cases decreased, suggesting a potential loss of PTEs from the topsoil system. The percentage of all PTEs in fractions F1–F4 decreased and that in F5 increased with time. These results illustrate that the residual and mobility of PTEs in the topsoil during SS utilization was lower than expected and more attention should be paid to the risk of PTEs transferring to other environments.

Purification, identification and in silico studies of antioxidant, antidiabetogenic and antibacterial peptides obtained from sorghum spent grain hydrolysate

The identification of antioxidant, antidiabetogenic and antimicrobial peptides obtained from sorghum spent grain (SSG) protein hydrolysate was performed. Antioxidant activity (AOA) was evaluated by ABTS+ radical cation scavenging, antidiabetogenic activity (ADA) by dipeptidyl peptidase IV (DPP-IV) inhibition, and antimicrobial activity (AMA) through the minimum inhibitory concentration (MIC) method against 2 Gram-negative and 3 Gram-positive bacteria. SSG peptides were fractionated using a Sephadex G-25 molecular exclusion column, and fractions collected (F1–F6) were evaluated through AOA, ADA and AMA assays. The chromatographic process allowed increasing 20% the AOA and 35% the ADA in F2, and 90% the AMA in fractions F5 and F6. The most active fractions (F2, F5 and F6) were evaluated using LC-ESI-Q-TOF tandem mass spectrometry, which allowed identifying six new peptides with high bioactivity. Moreover, the docking refinement protocol showed that identified peptides could interact with the DPP-IV enzyme, two of them in a competitive inhibition mode. Sorghum spent grain (SSG) could be an important source of bioactive peptides. Additionally, this is the first report indicating the presence of antibacterial peptides encrypted in sorghum proteins.

Bioactivity of a Linoleic Acid–Rich Fraction of Ricinus communis L. (Euphorbiaceae) Leaves against the Yellow Sugarcane Aphid, Sipha flava (Hemiptera: Aphididae)

The insecticidal activity of a Ricinus communis leaf hexane extract and its fractions against adult yellow sugarcane aphids (Sipha flava) was evaluated using a contact bioassay after fumigation. The n-hexane extract at 10,000 ppm achieved the highest mortality (80%); the positive control had 100% mortality and the negative control had only 4% mortality over the 72-h experiment time. Chemical fractionation of the hexane extract of R. communis leaves produced multiple fractions, and 10,000 ppm of the F4 fraction resulted in 92% aphid mortality at 72 h. Gas chromatography-mass spectrometry of the F4 fraction revealed linoleic acid as the major compound (84.5%). The R. communis n-hexane extract and linoleic acid could be used for integrated pest control as an ecologically friendly alternative to synthetic chemical insecticides.

Evaluation of Anti-Diabetic and Anti-Hyperlipidemic Activity of Isolated Bioactive Compounds of Leaves of Annona reticulata Linn.

Background: The leaves of Annona reticulata Linn. have been traditionally used by the tribes of Assam as a source of medicine to mitigate a range of health ailments, including diabetes and obesity. Objectives: The current study aimed to evaluate the anti-diabetic and anti-hyperlipidemic potential of bioactive fractions isolated from the methanolic extract of Annona reticulata Linn. leaves using Nicotinamide + Streptozotocin (60 mg/kg, i.p.) induced diabetic rats. Methods: The partially purified bioactive fractions, namely F1, F2, F3 and F4 were administered to diabetic rats with a dose of 200 mg/kg, per oral (p.o.) and the effect of the fractions on serum glucose was studied up to 21 days. The potent fractions were further subjected to spectral analysis for the identification of the isolated active compounds. Results: The in-vivo anti-diabetic activity of the isolated fractions F2 and F3 was found significant controlling blood glucose level, alike glibenclamide. Interestingly, F2 and F3 treated animals were found significant in restoring the lipid and liver enzymes profile in streptozotocin challenge rats. Further, spectral analysis revealed that F2 and F3 were comprised of Quercetin and Gallic acid, respectively. Conclusion: The outcome of finding demonstrates the anti-diabetic and anti-hyperlipidemic potential of the isolates/fractions of A. reticulata, which were found enriched in polyphenolics, including Quercetin and Gallic acid; and provided logistic behind the traditional use of the A. reticulata against diabetes and obesity.

Active Ingredients from Euodia ruticarpa Steam Distilled Essential Oil Inhibit PC-3 Prostate Cancer Cell Growth via Direct Action and Indirect Immune Cells Conditioned Media In Vitro.

Active constituents isolated from Euodia ruticarpa (ER) steam distilled essential oil (SDEO) against PC-3 prostate cancer cell growth remain unclear. To clarify the puzzle, ER SDEO was extracted and further resolved into six isolated fractions ERF1–F6 with Sephadex LH-20 gel filtration chromatography to analyze their biological activities. Active ingredients in the isolated fractions were analyzed with GC-MS. Potential isolated fractions were selected to treat PC-3 cells with direct action and indirect treatment by mouse splenocyte- (SCM) and macrophage-conditioned media (MCM). The relationship between PC-3 cell viabilities and corresponding total polyphenols, flavonoid contents as well as Th1/Th2 cytokine profiles in SCM was analyzed using the Pearson product–moment correlation coefficient (r). As a result, ERF1–F3 was abundant in total polyphenols and flavonoids contents with diverse active ingredients. Treatments with ERF1–F3 at appropriate concentrations more or less inhibit PC-3 cell growth in a direct action manner. Only SCM, respectively, cultured with ER SDEO and ERF1–F3 markedly enhanced the effects to inhibit PC-3 cell growth, suggesting that secretions by splenocytes might involve anti-PC-3 effects. There are significantly negative correlations between PC-3 cell viabilities and IL-2, IL-10 as well as IL-10/IL-2 ratios in the corresponding SCM. Total polyphenol and flavonoid contents in the media cultured with ER SDEO isolated fractions positively correlated with IL-10 (Th2) and IL-10/IL-2 (Th2/Th1) cytokine secretion ratios by splenocytes, indicating that polyphenol and flavonoid components in ER SDEO isolated fractions promote Th2-polarized and anti-inflammatory characteristics. These new findings concluded that the inhibitory effects against PC-3 prostate cancer cell growth are attributed to active anti-inflammatory ingredients in ER SDEO and its active ERF1–F3 fractions through direct action and indirect treatment by modulating splenocytes’ cytokine secretion profiles.

Water-holding capacity of enzymatic protein hydrolysates: A study on the synergistic effects of peptide fractions

The synergistic effects of peptides in bovine skin gelatin hydrolysate (BSGH) on the water-holding capacity (WHC), measured as cooking loss, were studied. These peptides were isolated from the hydrolysate in fractions with different hydrophobicities by reversed-phase chromatography on C18 separation columns. Each peptide fraction was included in the meat matrix at various concentrations for the study of water retention. The fraction with the highest WHC (F5) was obtained. The experimental design consisted of a central compound design with axial points; the independent variables were X1 (ingredient concentration in the meat sample) and X2 (concentration of fraction F5 in the ingredient). The dependent variable was cooking loss. The WHC studies showed substantial synergistic effects occurred between the fractions within the BSGH, and the peptide fractions reduced the cooking loss by 62.96% with respect to a sample without the ingredient. However, a decrease of 42.26% was observed for the F5 fraction. This fraction corresponds to the highest WHC applied for the concentration of 3 g/100 g. The results suggest a synergistic WHC effect of the peptides present in the BSGH since this effect was significantly reduced (p ≤ 0.05) when the peptide fractions were separated.

Novel Antihypertension Bioactive Compounds from Rusip

Bioactive compounds from rusip has function as a antihypertension which inhibit Angiotensin Converting Enzyme (ACE). They was obtained by fractionation methode. Fractionation by molecular weight of rusip was to separate its bioactive compounds. Steps taken for this research was extraction and fractionation of rusip, assayed their ACE inhibition and peptide profile analysis. The results showed that there were four fractions from rusip extract, i.e F0 (free supernatant), F1 (Molecular wight fraction > 10 kD), F2 (Molecular wight fraction 1-10 kD), and F3 (Molecular wight fraction > 1 kD) . There are three similar peptides in the Fo and F1 rusip fraction, i.e 44.78 kD, 25.90 kD and 17.18 kD, however there were no peptide in the F2 and F3 fractions. The ACE inhibitor activity of rusip fraction range for 34.40% to 97.52%, where F2 Fraction had highest ACE inhibitor activity. This research revealed those bioactive compounds which have highest activity of ACE inhibition from rusip was low molecular weight (under 10 kD) peptides or other biaoctives compounds.

Tityus serrulatus (Scorpion): From the Crude Venom to the Construction of Synthetic Peptides and Their Possible Therapeutic Application Against Toxoplasma gondii Infection.

Toxoplasmosis, caused by Toxoplasma gondii, is a major public concern owing to its neurotropic nature and high morbidity and mortality rates in immunocompromised patients and newborns. Current treatment for this disease is inefficient and produces side effects. Inflammatory mediators produced during T. gondii infection (e.g., cytokines and nitric oxide) are crucial in controlling parasite replication. In this context, Tityus serrulatus venom (TsV) induces the production of inflammatory mediators by immune cells. Thus, this study aimed to isolate and identify the components of TsV with potential anti-T. gondii activity. TsV was extracted from scorpions and lyophilized or loaded onto a column to obtain its fractions. TsV subfractions were obtained using chromatography, and its amino acid sequence was identified and applied to peptide design using bioinformatics tools. The C57BL/6 mice and their harvested macrophages were used to test the anti-Toxoplasma activity of TsV components and peptides. TsV and its fraction F6 attenuated the replication of tachyzoites in macrophages and induced nitric oxide and cytokine (IL-12, TNF, and IL-6) production by infected cells, without host cell toxicity. Moreover, Su6-B toxin, a subfraction of F6, demonstrated anti-T. gondii activity. The partially elucidated and characterized amino acid sequence of Sub6-B demonstrated 93% similarity with T. serrulatus 2 toxin (Ts2). Ts2 mimetic peptides (“Pep1,” “Pep2a,” and “Pep2b”) were designed and synthesized. Pep1 and Pep2a, but not Pep2b, reduced the replication of tachyzoites in macrophages. In vivo, treatment of T. gondii-infected mice with Pep1, Pep2a, or Pep2b decreased the number of cerebral cysts and did not induce hepatotoxicity in the animals. Taken together, our data show promising immunomodulatory and antiparasitic activity of TsV that could be explored and applied in future therapies for treating infectious parasitic diseases such as toxoplasmosis.

Effect of organosolv extraction on the structure and antioxidant activity of eucalyptus kraft lignin

In this study, three lignin fractions F1, F2, and F3 were obtained from eucalyptus kraft lignin (EKL) by solvent extraction with ethyl acetate, acetone, and acetone/water. The antioxidant activity of these lignin fractions were measured and evaluated by using the DPPH radical scavenging activity, ABTS radical cation scavenging activity and ferric reducing antioxidant power analysis. These lignin fractions were also characterized by applying Gel permeation chromatography, Fourier transform infrared, 31P NMR and 2D HSQC NMR techniques. The three different lignin fractions had rather different average molecular sizes, as well as different phenolic and methoxy functional contents. The ethyl acetate fraction (F1) with the lowest average molecular weight (2342 g/mol) and the highest phenolic hydroxyl content (4.2457 mmol/g) and methoxy groups (6.2714 mmol/g) showed high homogeneity and the highest antioxidant activity. Its DPPH scavenging activity, ABTS+ scavenging activity, and ferric reducing the antioxidant power were 68.67%, 75.57%, and 91.89 μmol/g, respectively. Moreover, the antioxidant activity of F1 and F2 was found to be higher than that of butylated hydroxytoluene. Therefore, solvent extraction was shown to be an effective way to separate lignin fractions with high homogeneity, high antioxidant activity, which could lead to application of lignin with higher value.