Search results for: DPPH assays

Authors: Benhouda Afaf, Yahia Massinissa, Paolo Grieco

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Hyoscyamus albus L. is a small genus from Solanaceae family known by its use in old traditional medicine in the east of Algeria. Aim: This study aimed to characterize bioactive molecules from H. albus, evaluate their anticancer activity in several cancer cells and investigate their possible molecular mechanism. Materials and Methods: Different compounds (Peak h of fraction F), (Peak 3 of Fraction F), (Peak 1 of fraction C) were isolated from H.albus L by using high-performance chromatography (HPLC), mass spectrometry (MS) and proton NMR (NMR H1). All isolated compounds were subjected to cytotoxicity and antiproliferative assays against a panel of the four cell lines: DU-145, U-2 OS, U-87 MG and LN-229 cell lines and were determined using MTT assay, Annexin V and propodium iodide were used to evaluate apoptosis. Results: The phytochemical study of H. albus Fractions led to the isolation of quercetin-3-O-β-dglucopyranosyl-( 1 → 6)-β-d-glucopyranosid, N-trans-feruloyltyramine, Hydrocaffeoyl-N8- caffeoylspermidine.The biological results indicated that all cell lines were consistently sensitive to P1 FC in a dose-dependent manner. This difference in cytotoxic sensitivity was more pronounced in osteosarcoma cell line, U-2 OS, when compared to prostate cancer and U-87 MG. Cell viability data also demonstrated that only U-87 MG cells were responsive to treatment with Ph FF. compounds P1 FC and Ph FF have induced necrosis and apoptosis in a large part of LN-229 cells. Conclusion: The overall results of the present study provided evidence that isolated compounds are potential therapeutic entities against cancer.

Keywords: hyoscyamus albus, cancer cells, coumpounds, HPLC

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Authors: Lila Boulekbache-Makhlouf, Fatiha Brahmi

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This study was carried out with the aim of enriching fresh cheese with the food by-products, which are the leaves of shallots and the peels of potatoes. Firstly, the conditions for extracting the total polyphenols using ultrasound are optimized. Then, the contents of total polyphenols PPT , flavonoids and antioxidant activity were evaluated for the extracts obtained by adopting the optimal parameter. On the other hand, we have carried out some physicochemical, microbiological and sensory analyzes of the cheese produced. The maximum total polyphenols value of 70.44 mg GAE gallic acid equivalent / g of dry matter DM of shallot leaves was reached with 40% (v/v) ethanol, an extraction time of 90 min and a temperature of 10 °C. While, the maximum TPP total polyphenols content of potato peels of 45.03 ± 4.16 mg gallic acid equivalent / g of dry matter DM was obtained using an ethanol /water mixture (40%, v/v), a time of 30 min and a temperature of 60 °C and the flavonoid contents were 13.99 and 7.52 QE quercetin equivalent/g dry matter DM, respectively. From the antioxidant tests, we deduced that the potato peels present a higher antioxidant power with the concentration of extracts causing a 50% inhibition IC50s of 125.42 ± 2.78 μg/mL for 2,2-diphényl 1-picrylhydrazyle DPPH, of 87.21 ± 7.72 μg/mL for phosphomolybdate and 200.77 ± 13.38 μg/mL for iron chelation, compared with the results obtained for shallot leaves which were 204.29 ± 0.09, 45.85 ± 3,46 and 1004.10 ± 145.73 μg/mL, respectively. The results of the physicochemical analyzes have shown that the formulated cheese was compliant with standards. Microbiological analyzes show that the hygienic quality of the cheese produced was satisfactory. According to the sensory analysis, the experts liked the cheese enriched with the powder and pieces of the leaves of the shallots.

Keywords: shallots leaves, potato peels, ultrasound extraction, phenolics, cheese

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Authors: Carolina Pereira Lobato Costa, Cristina Pascual-González, Monica Echeverry, Javier LLorca, Carlos Gonzáléz, Juan Pedro Fernández-Bláquez

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Although the potential of PLA self-reinforced composites for bone applications, not much literature addresses optimal manufacturing conditions. In this regard, this paper describes the woven self-reinforced PLA composites manufacturing processes: the commingling of yarns, weaving, and hot pressing and characterizes the manufactured laminates. Different structures and properties can be achieved by varying the hot compaction process parameters (pressure, holding time, and temperature). The specimens manufactured were characterized in terms of thermal properties (DSC), microstructure (C-scan optical microscope and SEM), strength (tensile test), and biocompatibility (MTT assays). Considering the final device, 155 ℃ for 10 min at 2 MPa act as the more appropriate hot pressing parameters. The laminate produced with these conditions has few voids/porosity, a tensile strength of 30.39 ± 1.21 MPa, and a modulus of 4.09 ± 0.24 GPa. Subsequently to the tensile testing was possible to observe fiber pullout from the fracture surfaces, confirming that this material behaves as a composite. From the results, no single laminate can fulfill all the requirements, being necessary to compromise in function of the priority property. Further investigation is required to improve materials' mechanical performance. Subsequently, process parameters and materials configuration can be adjusted depending on the place and type of implant to suit its function.

Keywords: woven fabric, self-reinforced polymer composite, poly(lactic acid), biodegradable

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Authors: Livy Febria Tedjamulia, Aas Nurasyiah, Ivana Josephin Purnama, Monika Diah Maharani Kusumastuti, Achmad Ridwan Ariyantoro

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Papua's candy is one of Indonesia’s indigenous consumption consisting of areca nut (Areca catechu), forest betel fruit (Piper aduncum), and CaCO3. This research aims to determine the concentration of tannins in areca nut, alkaloids in areca nut, flavonoids in forest betel fruit; detect their interaction and CaCO3; also toform a standardize consumption recommendation. The research methodwas includingDPPH assay for papua’s candy mixture, which resulted in IC50 value. Data analysis used is mathematical linear regression for each experiment. The test result of alkaloid is a Rf value of 0.773, while concentration of tannin and flavonoidare 0.603 mgGAE/g and 125.402 gQE/g, respectively. The IC50 value shows number of 3.0403, showing high antioxidant capacity.Other antioxidant assays were being studied using literature review, namely trolox and oxygen radical absorbance capacity, to figure out interaction among the bioactive compounds. It turned out that the interaction detected is antagonistic, which means the compound that is joined already has a stable molecular structure so that could reduce free radicals by donating hydrogen atoms. The recommendation consumptions given are 4 areca nuts, 5 forest betels, and 1 gram of lime betel. Therefore, papua's candy has its potential to be developed into functional food.

Keywords: antioxidant, bioactive compounds interaction, free radical, papua’s candy

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Authors: Monika Szymańska-Czerwińsk, Kinga Zaręba-Marchewka, Krzysztof Niemczuk

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Chlamydiaceae are Gram-negative bacteria distributed worldwide in animals and humans. One of them is Chlamydia gallinacea recently discovered. Available data show that C. gallinacea is dominant chlamydial agent found in poultry in European and Asian countries. The aim of the studies was screening of poultry flocks in order to evaluate frequency of C. gallinacea shedding and genetic diversity. Sampling was conducted in different regions of Poland in 2019-2020. Overall, 1466 cloacal/oral swabs were collected in duplicate from 146 apparently healthy poultry flocks including chickens, turkeys, ducks, geese and quails. Dry swabs were used for DNA extraction. DNA extracts were screened using a Chlamydiaceae 23S rRNA real-time PCR assay. To identify Chlamydia species, specific real-time PCR assays were performed. Furthermore, selected samples were used for sequencing based on ompA gene fragments and variable domains (VD1-2, VD3-4). In total, 10.3% of the tested flocks were Chlamydiaceae-positive (15/146 farms). The presence of Chlamydiaceae was confirmed mainly in chickens (13/92 farms) but also in turkey (1/19 farms) and goose (1/26 farms) flocks. Eleven flocks were identified as C. gallinacea-positive while four flocks remained unclassified. Phylogenetic analysis revealed at least 16 genetic variants of C. gallinacea. Research showed that Chlamydiaceae occur in a poultry flock in Poland. The strains of C. gallinacea as dominant species show genetic variability.

Keywords: C. gallinacea, emerging agent, poultry, real-time PCR

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Authors: M. Eckert, D. Mendes, J P. Gonçalves, C. Moço, M. Oliveira

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The intensive extraction of natural aggregates leads to both depletion of natural resources and unwanted environmental impacts. On the other hand, uncontrolled disposal of Construction and Demolition Wastes (C&DW) causes the lifetime reduction of landfills. It is known that the European Union produces, each year, about 850 million tons of C&DW. For all the member States of the European Union, one of the milestones to be reached by 2020, according to the Resource Efficiency Roadmap (COM (2011) 571) of the European Commission, is to recycle 70% of the C&DW. In this work, properties of different types of recycled C&DW aggregates and natural aggregates were compared. Assays were performed according to European Standards (EN 13285; EN 13242+A1; EN 12457-4; EN 12620; EN 13139) for the characterization of there: physical, mechanical and chemical properties. Not standardized tests such as water absorption over time, mass stability and post compaction sieve analysis were also carried out. The tested recycled C&DW aggregates were classified according to the requirements of the European Standards regarding there potential use in concrete, mortar, unbound layers of road pavements and embankments. The results of the physical and mechanical properties of recycled C&DW aggregates indicated, in general, lower quality properties when compared to natural aggregates, particularly, for concrete preparation and unbound layers of road pavements. The results of the chemical properties attested that the C&DW aggregates constitute no environmental risk. It was concluded that recycled aggregates produced from C&DW have the potential to be used in many applications.

Keywords: recycled aggregate, sustainability, aggregate properties, European Standard Classification

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Authors: Kina Kim

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Background: There are two types of interferon-gamma release assays (IGRAs) in use for the detection of latent tuberculosis infection (LTBI), QuantiFERON-TB Gold In-tube (QFT-GIT) and T-SPOT.TB. There are some reports that IGRA results are affected by the patient's age. This study aims to compare the results of both IGRA tests according to age groups. Methods: We reviewed 54,882 samples referred to an independent reference laboratory (Seegene Medical Foundation, Seoul, Korea) for the diagnosis of LTBI from January 1, 2021, to December 31, 2021. This retrospective study enrolled 955 patients tested using QFT-GIT and 53,927 patients tested using T-SPOT.TB. The results of both IGRAs were divided in three age groups (0-9, 10-17, and ≥18-year old). The positive rates and the indeterminate rates between QFT-GIT and T-SPOT.TB were compared. We also evaluated the differences in positive and indeterminate rates by age-group. Results: The positive rate of QFT-GIT was 20.1% (192/955) and that of T-SPOT.TB was 8.7% (4704/53927) in overall patients. The positive rates of QFT-GIT in individuals aged 0-9, 10-17, and over 18-year old were 15.4%, 13.3%, and 22.0%, respectively. The positive rates of T-SPOT.TB were 8.9%, 2.0% and 8.8%,in each agegroup, respectively.The overall prevalence of indeterminate results was 2.1% (20/955) of QFT-GIT and 0.5% (270/53927) of T-SPOT.TB. The indeterminate rates of QFT-GIT in individuals aged 0-9, 10-17, and over 18 years were 0.4%, 6.7%, and 2.6%, respectively. The indeterminate rate of T-SPOT.TB were 0.5%, 0.7% and 0.5%,in each age group, respectively. Conclusion: Our findings suggest that T-SPOT.TB has a lower rate of positive results in overall patients and a lower rate of indeterminate results than those of QFT-GIT. The highest positive rate was found in the over 18 years group for QFT-GIT, but the positive rates of T-SPOT.TB was not significantly different among groups by age. QFT-GIT showed variable and higher indeterminate rates according to age group, but T-SPOT.TB showed lower rates in all age groups(<1%).

Keywords: LTBI, IGRA, QFT-GIT, T-SPOT. TB

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Authors: Chiagoziem A. Otuechere, Ebenezer O. Farombi

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The protective effect of dichloromethane: methanol extract of Pterocarpus mildbraedii (PME), a widely consumed Nigerian leafy vegetable, on the toxicity of propanil was investigated in male rats. Animals were distributed into eight groups of five each. Group 1 served as control and received normal saline while rats in groups 2, 3, and 4 received 100 mg/kg, 200 mg/kg, and 400 mg/kg extract doses respectively. Group 5 rats were orally administered 200 mg/kg propanil while groups 6, 7, and 8 rats were given propanil plus extract. Oral administration of propanil elicited a 14.8%, 5%, 122%, and 78% increase in the activity of serum enzymes; alanine aminotransferase (AST), alanine aminotransferase(ALT), Alkaline phoshatase (ALP) and Gamma glutamyl transferase (ﻻGT). There were also increase in Lactate dehydrogenase (LDH) activity, direct bilirubin and lipid peroxidation levels. Furthermore, PME significantly attenuated the marked hepatic oxidative damage that accompanied propanil treatment. The extract significantly decreased LDH activity and bilirubin levels following propanil treatment. Furthermore, propanil-induced alterations in the activities of antioxidant enzymes: Superoxide dismutase (SOD), catalase (CAT) and glutathione s-transferase (GST) in these rats were modulated by the extract. The percentage DPPH Radical Scavenging Activity of the extract was determined as 55% and compared to those of Gallic acid (49%). Hepatic histology examination further confirmed the damage to the liver as it revealed severe periportal cellular infiltration of the hepatocytes. These biochemical and morphological alterations were attenuated in rats pre-treated with 100 mg/kg and 200 mg/kg doses of the extract. These results suggest that PME possesses protective effect against propanil-induced hepatotoxicity.

Keywords: antioxidant, hepatoprotection, Pterocarpus mildbraedii, propanil

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Authors: Vitheejongjaroen P., Jaisin Y., Pachekrepapol U., Taweechotipatr M.

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Lactic acid bacteria (LAB) are the most common type of microorganisms that had been used as probiotics also known for many beneficial health effects. The antioxidant activity of LAB is associated with numerous health-protective effects. This research aimed to investigate the antioxidant activity of lactic acid bacteria isolated from Thai sour pork sausage for their application in fermented milk products. Antioxidant activity determined by DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging assay showed that the isolate FN33-7, as 1 of 8 isolated exhibited scavenging activity in intact cell 5-7%, and supernatant 13-16%, intracellular cell free extract 42-48% respectively. This isolate was identified using 16S ribosomal DNA sequence analysis as Lactobacillus plantarum. The effect of milk fermented with L. plantarum FN33-7 on microbial count, pH and syneresis was assessed during refrigerated storage period of 28 days. The strain showed increased viability, pH level decreased, while syneresis increased. These results are similar to dairy products fermented with commercial starter cultures. Additionally, microstructure analysis of fermented milk by fluorescent microscopy showed that curd structure appeared to be dense and less porous in this fermented milk than commercial yogurt. The results of this study indicated that L. plantarum FN33-7 was a good probiotic candidate to be used in cultured milk products to reduce the risk of diseases caused by oxidative stress.

Keywords: Lactobacillus plantarum, probiotics, free radical, antioxidant, oxidative stress, fermented milk products

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Authors: Fadilah Aleanizy

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Colicins are a family of bacterial toxins that kill Escherichia coli and other closely related species. The mode of action of colicins involves binding to an outer membrane receptor and translocation across the cell envelope, leading to cytotoxicity through specific targets. The mechanism of colicin cytotoxicity includes a non-specific endonuclease activity or depolarization of the cytoplasmic membrane by pore-forming activity. For Group A colicins, translocation requires an interaction between the N-terminal domain of the colicin and a series of membrane- bound and periplasmic proteins known as the Tol system (TolB, TolR, TolA, TolQ, and Pal and the active domain must be translocated through the outer membranes. Protein-protein interactions are intrinsic to virtually every cellular process. The transient protein-protein interactions of the colicin include the interaction with much more complicated assemblies during colicin translocation across the cellular membrane to its target. The potassium release assay detects variation in the K+ content of bacterial cells (K+in). This assays is used to measure the effect of pore-forming colicins such as ColA on an indicator organism by measuring the changes of the K+ concentration in the external medium (K+out ) that are caused by cell killing with a K+ selective electrode. One of the goals of this work is to employ a quantifiable in-vivo method to spot which Tol protein are more implicated in the interaction with colicin A as it is translocated to its target.

Keywords: K+ efflux, Colicin A, Tol-proteins, E. coli

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Authors: Ana P. R. N. Barroso, Maurício N. Kleinberg, Frederico R. Silva, Rodrigo F. Guimarães, Marcelo M. V. Parente, Walney S. Araújo

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The fight against accelerated wear of the equipment used in the oil and gas sector is a challenge for minimizing maintenance costs. Corrosion being one of the main agents of equipment deterioration, we seek alternative materials that exhibit improved corrosion resistance at low cost of production. This study aims to evaluate the corrosion behavior of experimental alloys containing 15% and 17% of chromium (Cr) and 5% of molybdenum (Mo) in comparison with an AISI 444 commercial alloy. Microstructural analyzes were performed on samples of the alloys before and after the electrochemical tests. Two samples of each solubilized alloy were also taken for analysis of the corrosion behavior by testing potentiodynamic polarization (PP) and Electrochemical Impedance Spectroscopy (EIS) with immersion time of 24 hours in electrolytic solution with acidic character. The graphics obtained through electrochemical tests of PP and EIS indicated that among the experimental alloys, the alloy with higher chromium content (17%) had a higher corrosion resistance, confirming the beneficial effect of adding chromium. When comparing the experimental alloys with the AISI 444 commercial alloy, it is observed that the AISI 444 commercial alloy showed superior corrosion resistance to that of the experimental alloys for both assays, PP and EIS. The microstructural analyzes performed after the PP and EIS tests confirmed the results previously described. These results suggest that the addition of these levels of molybdenum did not favor the electrochemical behavior of experimental ferritic alloys for the electrolytic medium studied.

Keywords: corrosion, molybdenum, electrochemical tests, experimental alloys

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Authors: J. Švarc-Gajić, A. Cvetanović

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Apigenin is a flavone synthetized by many plants and quite abundant in chamomile (Matricaria recutita) in its free form and in the form of its glucoside and different acylated forms. Many beneficial health effects have been attributed to apigenin, such as chemo-preventive, anxiolytic, anti-inflammatory, antioxidant and antispasmodic. It is reported that free apigenin is much more bioactive in comparison to its bound forms. Subcritical water offers numerous advantages in comparison to conventional extraction techniques, such as good selectivity, low price and safety. Superheated water exhibits high hydrolytical potential which must be carefully balanced when using this solvent for the extraction of bioactive molecules. Moderate hydrolytical potential can be exploited to liberate apigenin from its bound forms, thus increasing biological potential of obtained extracts. The polarity of pressurized water and its hydrolytical potential are highly dependent on the temperature. In this research chamomile ligulate flowers were extracted by pressurized hot water in home-made subcritical water extractor in conditions of convective mass transfer. The influence of the extraction temperature was investigated at 30 bars. Extraction yields of total phenols, total flavonoids and apigenin depending on the operational temperature were calculated based on spectrometric assays. Optimal extraction temperature for maximum yields of total phenols and flavonoids showed to be 160°C, whereas apigenin yield was the highest at 120°C.

Keywords: superheated water, temperature, chamomile, apigenin

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Authors: Esma Hande Alıcı, Gülnur Arabacı

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Antioxidants are compounds or systems that can safely interact with free radicals and terminate the chain reaction before vital molecules are damaged. The anti-oxidative effectiveness of these compounds depends on their chemical characteristics and physical location within a food (proximity to membrane phospholipids, emulsion interfaces, or in the aqueous phase). Antioxidants (e.g., flavonoids, phenolic acids, tannins, vitamin C, vitamin E) have diverse biological properties, such as antiinflammatory, anti-carcinogenic and anti-atherosclerotic effects, reduce the incidence of coronary diseases and contribute to the maintenance of gut health by the modulation of the gut microbial balance. Plants are excellent sources of antioxidants especially with their high content of phenolic compounds. Raphanus raphanistrum L., the wild radish, is a flowering plant in the family Brassicaceae. It grows in Asia and Mediterranean region. It has been introduced into most parts of the world. It spreads rapidly, and is often found growing on roadsides or in other places where the ground has been disturbed. It is an edible plant, in Turkey its fresh aerial parts are mostly consumed as a salad with olive oil and lemon juice after boiled. The leaves of the plant are also used as anti-rheumatic in traditional medicine. In this study, we determined the antioxidant capacity of two different solvent fractions (methanol and ethyl acetate) obtained from Raphanus raphanistrum L. plant leaves. Antioxidant capacity of the plant was introduced by using three different methods: DPPH radical scavenging activity, CUPRAC (Cupric Ion Reducing Antioxidant Capacity) activity and Reducing power activity.

Keywords: antioxidant activity, antioxidant capacity, Raphanis raphanistrum L., wild radish

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Authors: E. Kayitesi, S. Moyo, V. Mavumengwana

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Cleome gynandra (spider plant) is an African green leafy vegetable categorized as an indigenous, underutilized and has been reported to contain essential phenolic compounds. Phenolic compounds play a significant role in human diets due to their proposed health benefits. These compounds however may be affected by different processing methods such as cooking and drying. Cleome gynandra was subjected to boiling, steam blanching, and drying processes and analysed for Total Phenolic Content (TPC), Total Flavonoid Content (TFC), antioxidant activity and flavonoid composition. Cooking and drying significantly (p < 0.05) increased the levels of phenolic compounds and antioxidant activity of the vegetable. The boiled sample filtrate exhibited the lowest TPC followed by the raw sample while the steamed sample depicted the highest TPC levels. Antioxidant activity results showed that steamed sample showed the highest DPPH, FRAP and ABTS with mean values of 499.38 ± 2.44, 578.68 ± 5.19, and 214.39 ± 12.33 μM Trolox Equivalent/g respectively. An increase in quercetin-3-rutinoside, quercetin-rhamnoside and kaempferol-3-rutinoside occurred after all the cooking and drying methods employed. Cooking and drying exerted positive effects on the vegetable’s phenolic content, antioxidant activity as a whole, but with varied effects on the individual flavonoid molecules. The results obtained help in defining the importance of African green leafy vegetable and resultant processed products as functional foods and their potential to exert health promoting properties.

Keywords: Cleome gynandra, phenolic compounds, cooking, drying, health promoting properties

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Authors: Farideh Tabatabei-Yazdi, Fereshteh Falah, Alireza Vasiee

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Nowadays, production of functional foods has become very essential. Inulin is one of the most functional hydrocolloid compounds used in such products. In the present study, the production of a synbiotic yogurt containing 1, 2.5, and 5% (w/v) inulin has been investigated. The yogurt was fermented with Lactobacillus brevis PML1 derived from Tarkhineh, an Iranian cereal-dairy fermented food. Furthermore, the physicochemical properties, antioxidant activity, sensory attributes, and microbial viability properties were investigated on the 0th, 7th, and 14th days of storage after fermentation. The viable cells of L. brevis PML1 reached 108 CFU/g, and the product resisted to simulated digestive juices. Moreover, the synbiotic yogurt impressively increased the production of antimicrobial compounds and had the most profound antimicrobial effect on S. typhimurium. The physiochemical properties were in the normal range, and the fat content of the synbiotic yogurt was reduced remarkably. The antioxidant capacity of the fermented yogurt was significantly increased (p<0:05), which was equal to those of DPPH (69:18±1:00%) and BHA (89:16±2:00%). The viability of L. brevis PML1 was increased during storage. Sensory analysis showed that there were significant differences in terms of the impressive parameters between the samples and the control (p<0:05). Addition of 2.5% inulin not only improved the physical properties but also retained the viability of the probiotic after 14 days of storage, in addition to the viability of L. brevis with a viability count above 6 log CFU/g in the yogurt. Therefore, a novel synbiotic product containing L. brevis PML1, which can exert the desired properties, can be used as a suitable carrier for the delivery of the probiotic strain, exerting its beneficial health effects.

Keywords: functional food, lactobacillus brevis, symbiotic yogurt, physiochemical properties

Procedia PDF Downloads 92

Authors: Pornrut Rabintossaporn, Suphaket Saenthaweesuk, Amornnat Thuppia, Nuntiya Somparn

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Several dietary and herbal plants have been shown to possess cytoprotective and antioxidant effects with various mechanisms of action. The aim of this study was to determine the antioxidant effects and its mechanism of aqueous leaves extract of Ocimum americanum (OA), commonly known as American basil or 'hoary basil', in rat. The extract was screened for its phytochemical contents and antioxidant activity in vitro. Moreover, the extract was studied in rats to evaluate its effects in vivo. Rats were orally administered with the extract at the dose of 100, 200 and 400 mg/kg for 28 days. Phytochemical screening of plant extracts revealed the presence of alkaloid, cardiac glycosides, tannin and steroid compounds. The extract contained phenolic compounds 36.91 ± 0.66 mg of gallic acid equivalents per gram OA extract. The free radical scavenging activity assessed by DPPH assay gave IC50 of 41.27 ± 1.86 µg/mL, which is relatively lower than that of BHT with IC50 of 12.34 ± 1.14µg/mL. In the animals, the extract was well tolerated by the animals throughout the 28 days of study as shown by normal serum levels AST, ALP, ALT, BUN and Cr as well as normal histology of liver and pancreatic and kidney tissue. The protein expression of antioxidant enzymes, γ-glutamylcysteine ligase (γ-GCL) in liver was significantly increased compared with normal control. Consistent with the induction of γ-GCL protein expression significantly reduction of serum oxidative stress marker malondialdehyde (MDA) was found in rat treated with OA extract compared with control. Taken together, this study provides evidence that Ocimum americanum exhibits direct antioxidant properties and can induce cytoprotective enzyme in vivo.

Keywords: antioxidant, γ-glutamylcysteine ligase, MDA, Ocimum americanum

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Authors: Alok Raghav, Jamal Ahmad

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Background: Serum albumin glycation and advanced glycation end products (AGE) formation correlates in diabetes and its associated complications. Extensive modified human serum albumin is used to study the biochemical, electrochemical and functional properties in hyperglycemic environment with relevance to diabetes. We evaluate Spectroscopic, side chain modifications, amino acid analysis, biochemical and functional group properties in four glucose modified samples. Methods: A series four human serum albumin samples modified with glucose was characterized in terms of amino acid analysis, spectroscopic properties and side chain modifications. The diagnostic technique employed incorporates UV Spectroscopy, Fluorescence Spectroscopy, biochemical assays for side chain modifications, amino acid estimations, electrochemical and optical characterstic of glycated albumin. Conclusion: Glucose modified human serum albumin confers AGEs formation alters biochemical, electrochemical, optical, and functional property that depend on the reactivity of glucose and its concentration used for in-vitro glycation. A biochemical, electrochemical, optical, and functional characterization of modified albumin in-vitro produced AGE product that will be useful to interpret the complications and pathophysiological significance in diabetes.

Keywords: human serum albumin, glycated albumin, adavanced glycation end products, associated pathologies

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Authors: Rafael Estrada, Cristian Ruiz Rueda

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Carbapenems are last-resort antibiotics used in clinical settings to treat antibiotic-resistant bacterial infections. Thus, the emergence and spread of resistance to carbapenems is a major public health concern. Here, we have studied a carbapenem-resistant Aeromonas veronii strain previously isolated from a water sample from Sam Simeon Creek (Hearst San Simeon State Park, CA). Analysis of this isolate using disk-diffusion, CarbaNP, eCIM and mCIM assays revealed that it was resistant to amoxicillin-clavulanic acid and all carbapenems tested and that this isolate produced a potentially novel carbapenemase of the Metallo-β-lactamase family. Whole genome sequencing analysis revealed that this A. veronii isolate carries a novel variant of the blacₚₕₐ class β-carbapenemase gene that was closely related to the blacₚₕₐ₇ gene of Aeromonas jandaei. This isolate also carried a novel variant of the blaₒₓₐ class D carbapenemase gene that was most closely related to the blaₒₓₐ-₉₁₂ gene found in other Aeromonas veronii isolates. Finally, we also identified a novel class C β-lactamase gene moderately related to the blaFₒₓ-₁₇ gene of Providencia stuartii and other blaFₒₓ variants identified in Klebsiella pneumoniae, Escherichia coli and other Enterobacteriaceae. Overall, our findings reveal that environmental isolates are an important reservoir of multiple carbapenemases and other β-lactamases of clinical significance.

Keywords: β-lactamases, carbapenem, antibiotic-resistant, aeromonas veronii

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Authors: Abdelouahab Dehimati, Fatiha Bedjou

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This study focused on the antioxidant and antimicrobial activity of four extracts from the plant Matricaria pubscens (Asteraceae) harvest in the region of Ghardaia, the northern Sahara of Algeria. The different extracts were analyzed for their content of phenolic compounds and their biological activities. The ethanol extract expresses a better extraction yield (44.22%). We have first performed the quantitative colorimetric methods for total polyphenols. Wherein the aqueous extract shows the highest total polyphenol content and total flavonoid (216.66±2.58 mg Eq GA/g and 111.04±0.49 mg Eq Q/g E, respectively) and ethanol extract 50% total tannins content (68.88±2.72 mg Eq AT/g E). The evaluation of the antioxidant activity of extracts of Matricaria pubscens by the arbitrary value IC50. The ethanol 50% extract is expressed strong activity with an IC50 14.19±1.25 mg/m against the DPPH radical and 11.66±0.53 mg/ml against the ABTS radical). In addition, the aqueous extract showed strong reducing power with an IC50 (48.61±1.14 mg/ml). However, the results obtained by the reducing power of phosphomolybdat the test are calculated by the iron maximum absorbance where ethanol extract 50% gives an absorbance of about 1.641 ± 0.01nm. Otherwise, methanol 70% and butanol 80% extracts gave a very large chelating effect of iron with an IC50 (38.38±0.01 μg/ml and 38.58±0.04 μg/ml respectively). By the method of disc Diffuson, the results of the antimicrobial activity are achieved butanolic extract 80% shows high activity towards MRSA (MIC: 3.51mg/ml; BMC>100 mg/ml). Their shares, the extracts were the most active for the antifungal test, the butanol 80% extract was the most active against A. niger (MIC: 12.5 mg/ml; FMC>100 mg/ml). These preliminary results could be used to justify the traditional use of this plant and their phenolic compounds could be exploited for therapeutic purposes, such as antioxidants and antimicrobial effects.

Keywords: Matricaria pubscens, phenolic compounds, antioxidant activity, antimicrobial activity, IC50, MIC

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Authors: Junie B. Billones, Maria Constancia O. Carrillo, Voltaire G. Organo, Stephani Joy Y. Macalino, Inno A. Emnacen, Jamie Bernadette A. Sy

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One of the major interferences in the Philippines’ tuberculosis control program is the widespread prevalence of Mtb strains that are resistant to known drugs, such as the MDR-TB (Multi Drug Resistant Tuberculosis) and XDR-TB (Extensively Drug Resistant Tuberculosis). Therefore, there is a pressing need to search for novel Mtb drug targets in order to be able to combat these drug resistant strains. The enzyme 7,8-diaminopelargonic acid aminotransferase enzyme, or more commonly known as BioA, is one such ideal target, as it is known that humans do not possess this enzyme. BioA primarily plays a key role in Mtb’s lipid biosynthesis pathway; more specifically in the synthesis of the enzyme cofactor biotin. In this study, structure-based pharmacophore screening, docking, and ADMET evaluation of compounds obtained from the DrugBank chemical database were performed against the MtbBioA enzyme. Results of the screening, docking, ADMET, and TOPKAT calculations revealed that out of the 6,516 compounds in the library, only 7 compounds indicated more favorable binding energies as compared to the enzyme’s known inhibitor, amiclenomycin (ACM), as well as good solubility and toxicity properties. Moreover, out of these 7 compounds, Molecule 6 exhibited the best solubility and toxicity properties. In the future, these lead compounds may then be subjected to bioactivity assays in vitro or in vivo for further evaluation of its therapeutic efficacy.

Keywords: 7, 8-diaminopelargonic acid aminotransferase, BioA, pharmacophore, molecular docking, ADMET, TOPKAT

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Authors: Roman Khanferyan

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Endurance exercises and strenuous muscle activity are accompanied by multiple immune dysfunctions due to the activation of cytokines and chemokines synthesis. This study assesses changes in the synthesis of immune regulatory mediators in elite athletes during endurance sports activity. The concentrations of cytokines/chemokines (IL-2, IL-6, IL-8, IL-10, IL-18, MIP-1 beta, GRO-alpha, RANTES, SDF-1a, VEGF) in sera of hockey athletes (n=33) and in supernatants of 24-h cultivated peripheral blood mononuclear cells (PBMC) of boxers (n=6) assayed by ELISA and Luminex xMAP multiplex assays. Estimation of body composition studied by using bioimpedance technology. The dietary energy consumption per person has been estimated using an album of different sizes of portions of the most frequently consumed foods. It has been demonstrated that endurance sports activity enhances the secretions of most pro- and anti-inflammatory cytokines and chemokines in more than 2-6 fold. The study demonstrated that the high increase of more than 3-4 times in the concentration of IL-18 in sera of athletes (327.86 + 45.67 pg/ml) didn’t correlate with BMI (p=0.040) but demonstrated a low correlation with MMI (p=0.234) and BMR (p=0,231). The opposite impact on the concentration of IL-10 has been demonstrated in athletes. It has been shown a negative correlation between its concentration and BMI (p= - 0.251), MMI (p= - 0.327), and BMR (p= - 0.301). In vitro studies in boxers showed greater amounts of chemokines in the PBMC supernatants, including MIP-1β, GRO-α, RANTES, SDF-1α, and IL-8 (P<0.05). At the same time, healthy controls had greater supernatant levels of MCP-1, Eotaxin, and MIP-1α. The study demonstrated a high correlation between physical activity, usual athletes' diet, and consumption of specialized sports nutrition products.

Keywords: sport nutrition, cytokines, chemokines, endurace exercises

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Authors: Nacera Baali, Zahia Belloum, Souad Ameddah, Fadila Benayache, Samir Benayache, Chantal Wrutniak-Cabello

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Allurement of herbs as health beneficial foods and as a source material for the development of new drugs, has led to greater furtherance in the study of herbal medicines during recent years. In the present study, in vitro antioxidant, free radical scavenging capacity, and hepatoprotective activity of butanolic extract from Genista quadriflora Munby (G.quadriflora) were evaluated using established in vitro models such as DPPH radical and hydrogen peroxide radical scavenging activities and antilipidperoxidation ability. Interestingly, the extract showed considerable in vitro antioxidant and free radical scavenging activities in a dose-dependent manner when compared to the standard antioxidant which verified the presence of antioxidant compound in extract tested. The hepatoprotective potential of G.quadriflora extract was also evaluated in male Wistar rats against paracetamol (APAP) induced liver damage. Therapy of G.quadriflora showed the liver protective effect on biochemical and histopathological alterations. Moreover, histological studies also supported the biochemical finding, that is, the maximum improvement in the histoarchitecture of the liver. Results revealed that G.quadriflora extract could protect the liver against APAP-induced oxidative damage by possibly increasing the antioxidant protection mechanism in rats. These findings are of great importance in view of the availability of the plant and its observed possible diverse applications in medicine and nutrition.

Keywords: genista quadriflora munby, antioxidant, liver, paracetamol, oxidative stress

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Authors: Richard I. Licayan, Aisle Janne B. Dagpin, Romeo M. Del Rosario, Nenita D. Palmes

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Hiptage benghalensis, Antigonon leptopus, Macroptillium atropurpureum, and Dioscorea bulbifera L. are herbal weeds that have been used by traditional healers in rural communities in the Philippines as medicine. In this study, the basic pharmacological components of the crude secondary metabolites extracted from the four herbal weeds and their in vitro antioxidant properties was investigated to provide baseline data for the possible development of these metabolites in pharmaceutical products. Qualitative screening of the secondary metabolites showed that alkaloids, tannins, saponins, steroids, and flavonoids were present in their leaf extracts. All of the plant extracts showed varied antioxidant activity. The greatest DPPH radical scavenging activity was observed in H. begnhalensis (84.64%), followed by A. leptopus (68.21%), M. atropurpureum (26.62%), and D. bulbifera L. (19.04%). The FRAP assay revealed that H. benghalensis had the highest antioxidant activity (8.32 mg/g) while ABTS assay showed that M. atropurpureum had the strongest scavenging ability of free radicals (0.0842 mg Trolox/g). The total flavonoid content (TFC) analysis showed that D. bulbifera L. had the highest TFC (420.35 mg quercetin per gram-dried material). The total phenolic content (TPC) of the four herbal weeds showed large variations, between 26.56±0.160 and 55.91±0.087 mg GAE/g dried material. The plant leaf extracts arranged in increasing values of TPC are H. benghalensis (26.565) < A. leptopus (37.29) < D. bulbifera L. (46.81) < M. atropurpureum (55.91). The obtained results may support their use in herbal medicine and as baseline data for the development of new drugs and standardized phytomedicines.

Keywords: antioxidant properties, total flavonoids, total phenolics, creeping herbal weeds

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Authors: İ. Çelik, Goksel Demirer

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Turkey is the third largest producer of pistachio (Pistacia vera L.) after Iran and United States. Harvested pistachio nuts are covered with organic hull which is removed by de-hulling process. Most of the pistachio by-products which are produced during de-hulling process are considered as agricultural waste and often mixed with soil, to a lesser extent are used as feedstuff by local livestock farmers and a small portion is used as herbal medicine. Due to its high organic and phenolic content as well as high solids concentration, pistachio processing wastes create significant waste management problems unless they are properly managed. However, there is not a well-established waste management method compensating the waste generated during the processing of pistachios. This study investigated the anaerobic treatability and biogas generation potential of pistachio hull waste. The effect of pre-treatment on biogas generation potential was investigated. For this purpose, Biochemical Methane Potential (BMP) Assays were conducted for two Chemical Oxygen Demand (COD) concentrations of 22 and 33 g tCOD l-1 at the absence and presence of chemical and thermal pre-treatment methods. The results revealed anaerobic digestion of the pistachio de-hulling wastes and subsequent biogas production as a renewable energy source are possible. The observed percent COD removal and methane yield values of the pre-treated pistachio de-hulling waste samples were significantly higher than the raw pistachio de-hulling waste. The highest methane yield was observed as 213.4 ml CH4/g COD.

Keywords: pistachio de-hulling waste, biogas, renewable energy, pre-treatment

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Authors: Wanwisa Deenin, Abdulhadee Yakoh, Chahya Kreangkaiwal, Orawon Chailapakul, Kanitha Patarakul, Sudkate Chaiyo

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LipL32 is an outer membrane protein present only on pathogenic Leptospira species, which are the causative agent of leptospirosis. Leptospirosis symptoms are often misdiagnosed with other febrile illnesses as the clinical manifestations are non-specific. Therefore, an accurate diagnostic tool for leptospirosis is indeed critical for proper and prompt treatment. Typical diagnosis via serological assays is generally performed to assess the antibodies produced against Leptospira. However, their delayed antibody response and complicated procedure are undoubtedly limited the practical utilization especially in primary care setting. Here, we demonstrate for the first time an early-stage detection of LipL32 by an integrated lateral-flow immunoassay with electrochemical readout (eLFIA). A ferrocene trace tag was monitored via differential pulse voltammetry operated on a smartphone-based device, thus allowing for on-field testing. Superior performance in terms of the lowest detectable limit of detection (LOD) of 8.53 pg/mL and broad linear dynamic range (5 orders of magnitude) among other sensors available thus far was established. Additionally, the developed test strip provided a straightforward yet sensitive approach for diagnosis of leptospirosis using the collected human sera from patients, in which the results were comparable to the real-time polymerase chain reaction technique.

Keywords: leptospirosis, electrochemical detection, lateral flow immunosensor, point-of-care testing, early-stage detection

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Authors: Salma Baig, Bakrudeen Ali Ahmad, Ainnul Hamidah Syahadah Azizan, Hapipah Mohd Ali, Elham Rouhollahi, Mahmood Ameen Abdulla

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Free radical damage induced by reactive oxygen species (ROS) contributes to etiology of many chronic diseases, cancer being one of them. Recent studies have been successful in ROS targeted therapies via antioxidants using mouse models in cancer therapeutics. The present study was designed to scrutinize anticancer activity, antioxidant activity of 5 different extracts of Thymus serpyllum in MDA-MB-231, MCF-7, HepG2, HCT-116, PC3, and A549. Identification of the phytochemicals present in the most active extract of Thymus serpyllum was conducted using gas chromatography coupled with mass spectrophotometry and antioxidant activity was measured by using DPPH radical scavenging and FRAP assay. Anticancer impact of the extract in terms of IC50 was evaluated using MTT cell viability assay. Results revealed that the hexane extract showed the best anticancer activity in HepG2 (Liver Carcinoma Cell Line) with an IC50 value of 23 ± 0.14 µg/ml followed by 25 µg/ml in HCT-116 (Colon Cancer Cell Line), 30 µm/ml in MCF-7 (Breast Cancer Cell Line), 35 µg/ml in MDA-MB-231 (Breast Cancer Cell Line), 57 µg/ml in PC3 (Prostate Cancer Cell Line) and 60 µg/ml in A549 (Lung Carcinoma Cell Line). GC-MS profile of the hexane extract showed the presence of 31 compounds with carvacrol, thymol and thymoquione being the major compounds. Phenolics such as Vitamin E, terpinen-4-ol, borneol and phytol were also identified. Hence, here we present the first report on cytotoxicity of hexane extract of Thymus serpyllum extract in HepG2 cell line with a robust anticancer activity with an IC50 of 23 ± 0.14 µg/ml.

Keywords: Thymus serpyllum L., hexane extract, GC-MS profile, antioxidant activity, anticancer activity, HepG2 cell line

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Authors: Berredjem Hajira, Benlaifa Meriem, Becheker Imene, Bardi Rafika, Djebar Med Reda

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Recent acquired or reactivation T.gondii infection is a serious complication in HIV patients. Classical serological diagnosis relies on the detection of anti-Toxoplasma immunoglobulin ; however, serology may be unreliable in HIV immunodeficient patients who fail to produce significant titers of specific antibodies. PCR assays allow a rapid diagnosis of Toxoplasma infection. In this study, we compared the value of the PCR for diagnosing active toxoplasmosis in cerebrospinal fluid and blood samples from HIV patients. Anti-Toxoplasma antibodies IgG and IgM titers were determined by ELISA. In parallel, nested PCR targeting B1 gene and conventional PCR-ELISA targeting P30 gene were used to detect T. gondii DNA in 25 blood samples and 12 cerebrospinal fluid samples from patients in whom toxoplasmic encephalitis was confirmed by clinical investigations. A total of 15 negative controls were used. Serology did not contribute to confirm toxoplasmic infection, as IgG and IgM titers decreased early. Only 8 out 25 blood samples and 5 out 12 cerebrospinal fluid samples PCRs yielded a positive result. 5 patients with confirmed toxoplasmosis had positive PCR results in either blood or cerebrospinal fluid samples. However, conventional nested B1 PCR gave best results than the P30 gene one for the detection of T.gondii DNA in both samples. All samples from control patients were negative. This study demonstrates the unusefulness of the serological tests and the high sensitivity and specificity of PCR in the diagnosis of toxoplasmic encephalitis in HIV patients.

Keywords: cerebrospinal fluid, HIV, Toxoplasmosis, PCR

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Authors: Saqib Ali, Man-Qun Wang

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The Japanese pine sawyer, Monochamus alternatus Hope (Coleoptera: Cerambycidae), is a major pest of pines and it is also the key vector of the exotic pinewood nematode in China. In the present study, we cloned, expressed, and purified a chemosensory protein (CSP) in M. alternatus. We surveyed its expression in various developmental stages of male and female adult tissues and determined its binding affinities for different pine volatiles using a competitive binding fluorescence assay. A CSP known as CSP5 in M. alternatus was obtained from an antennal cDNA library and expressed in Escherichia coli. Quantitative reverse transcription polymerase chain reaction results indicated that the CSP5 gene was mainly expressed in male and female antennae. Competitive binding assays were performed to test the binding affinity of recombinant CSP5 to 13 odour molecules of pine volatiles. The results showed that CSP5 showed very strong binding abilities to myrcene, (+)-β-pinene, and (−)-isolongifolene, whereas the volatiles 2-methoxy-4-vinylphenol, p-cymene, and (+)-limonene oxide have relatively weak binding affinity at pH 5.0. Three volatiles myrcene, (+)-β-pinene, and (−)-isolongifolene may play crucial roles in CSP5 binding with ligands, but this needs further study for confirmation. The sensitivity of insect to host plant volatiles can effectively be used to control and monitor the population through mass trapping as part of integrated pest management programs.

Keywords: olfactory-specific protein, volatiles, competitive binding assay, expression characteristics, qPCR

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Authors: Elena K. Schneider, Johnny X. Huang, Vincenzo Carbone, Mark Baker, Mohammad A. K. Azad, Matthew A. Cooper, Jian Li, Tony Velkov

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Ivacaftor is a novel CF trans-membrane conductance regulator (CFTR) potentiator that improves the pulmonary function for cystic fibrosis patients bearing a G551D CFTR-protein mutation. Because ivacaftor is highly bound (>97%) to plasma proteins, there is the strong possibility that co-administered CF drugs that compete for the same plasma protein binding sites and impact the free drug concentration. This in turn could lead to drastic changes in the in vivo efficacy of ivacaftor and therapeutic outcomes. This study compares the binding affinity of ivacaftor and co-administered CF drugs for human serum albumin (HSA) and α1-acid glycoprotein (AGP) using surface plasmon resonance and fluorimetric binding assays that measure the displacement of site selective probes. Due to their high plasma protein binding affinities, drug-drug interactions between ivacaftor are to be expected with ducosate, montelukast, ibuprofen, dicloxacillin, omeprazole and loratadine. The significance of these drug-drug interactions is interpreted in terms of the pharmacodynamic/pharmacokinetic parameters and molecular docking simulations. The translational outcomes of the data are presented as recommendations for a staggered treatment regimen for future clinical trials which aims to maximize the effective free drug concentration and clinical efficacy of ivacaftor.

Keywords: human α-1-acid glycoprotein, binding affinity, human serum albumin, ivacaftor, cystic fibrosis

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Authors: Dian Muzdalifah, Anastasia F. Devi, Zatil A. Athaillah, Linar Z. Udin

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Tempe is a functional food prepared from soybeans through Rhizopus spp fermentation. It is well known as functional food, originated from Indonesia. Most studies on tempe functionalities refer to ripe (48 h fermentation) tempe and only limited studies discuss overripe tempe while longer fermentation time possibly increased tempe health benefit. Hence, the present study was performed to investigate the cytotoxic activity againts MCF-7 breast cancer cells and antioxidant property of tempe prepared from 0–156 h of fermentation. Tempe samples were dried and extracted with acetone and ethanol, respectively. Their extracts were used for subsequent analysis. The cytotoxic activity was assessed on MCF 7 breast cancer cells using Alamar Blue method. The antioxidant activity was determined by DPPH free radical scavenging assay. The results indicated that acetone extracts of 108 h tempe had a potent cytotoxic activity against MCF-7 breast cancer cells (IC50 = 2.54 ± 0,30 μg/mL). Ethanol extracts of 108 h tempe also showed the potency, but at slightly higher IC50 (5.20 ± 1.01 μg/mL). Both acetone and ethanol extracts of 108 and 120 h tempe showed high antioxidant activity expressed as percent inhibition with no significant difference. However, acetone extracts of 120 h tempe (81.31 ± 3.70 %) had better ability to inhibit oxidation reaction than that of ethanol extracts (75.77 ± 6.00 %). It can be concluded that the cytotoxic activity of tempe from 0–156 h of fermentation is positively correlated to their corresponding antioxidant property. Longer fermentation time, up to 108 h, increased the ability of tempe to inhibit the growth of MCF-7 breast cancer cells and oxidative reaction. But extended fermentation time, up to 156 h, tends to decrease its ability. Further studies are encouraged to identify the active components contained in each extract.

Keywords: antioxidant property, cytotoxic activity, extracts, overripe tempeh

Procedia PDF Downloads 277