Search results for: headspace gas chromatography

Authors: Akanksha Jain, Santosh J. Passi, William Selvamurthy, Archna Singh

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Heating/frying at elevated temperatures cause numerous physiochemical reactions including oxidative deterioration and trans fatty acid (TFA) formation; however Indian data on these parameters are scanty. The present study was designed to assess the effect of constant heating/frying on formation of TFAs and oxidative stability in groundnut oil. 750 mL of the oil was heated in a large iron karahi (utensil similar to a wok) and freshly cut potato strips were fried constantly at varying temperatures (160ºC, 180ºC, 200ºC, 220ºC, 230ºC). In each case, the oil sample was drawn after one hour and stored at –20ºC until analysed. While TFA was estimated using gas chromatography with flame ionisation detector (AOCS official method Ce 1h–05), other chemical parameters were assessed by AOCS official methods. Oil samples subjected to heating/frying at varying temperatures demonstrated a significant increase in TFAs (p < 0.01) and saturated fatty acids (p < 0.01) while there was a corresponding decrease in cis-unsaturated fatty acids (p < 0.01). Frying process demonstrated greater TFA formation (mean TFA at 160ºC being 0.11±0.01g/100g; at 230ºC it being 2.33±0.05g/100g) as compared to heating alone (mean TFA at 160ºC being 0.07g±0.01/100g; at 230ºC it being 0.47±0.02g/100g), indicating that there was a significant difference in the generation of TFAs during the two thermal treatments (heating vs. frying; p=0.05). With increasing temperatures, acid value, p-anisidine value and total oxidation (TOTOX) value registered a significant increase (p < 0.01); however, peroxide value was found to be inconsistent. Thus, the formation of TFA and various oxidative parameters (except peroxide value) is directly influenced by the temperature of heating/frying. Since TFA formation and poor oxidative stability of oils can pose serious health concerns, food safety agencies/organizations need to devise appropriate policies, stringent food laws/standards and impose necessary safety regulations to curb oil abuse during the process of heating and frying. There is a dire need to raise consumer awareness regarding deleterious health effects of TFA and oxidative deterioration of oils at elevated temperatures employed during heating/frying procedures.

Keywords: cis-unsaturated fatty acid, oxidative stability, saturated fatty acid, trans fatty acid

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Authors: N. Chougui, R. Larbat

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The first economic importance of Opuntia ficus-indica relies on the production of edible fruits. This food transformation generates a large amount of by-products (seeds and peels) in addition to cladodes produced by the plant. Several studies showed the richness of these products with bioactive substances like phenolics that have potential applications. Indeed, phenolics have been associated with protection against oxidation and several biological activities responsible of different pathologies. Consequently, there has been a growing interest in identifying natural antioxidants from plants. This study falls within the framework of the industrial exploitation of by-products of the plant. The study aims to investigate the metabolic profile of three by-products (cladodes, peel seeds) regarding total phenolic content by liquid chromatography coupled to mass spectrometry approach (LC-MSn). The byproducts were first washed, crushed and stored at negative temperature. The total phenolic compounds were then extracted by aqueous-ethanolic solvent in order to be quantified and characterized by LC-MS. According to the results obtained, the peel extract was the richest in phenolic compounds (1512.58 mg GAE/100 g DM) followed by the cladode extract (629.23 GAE/100 g DM) and finally by the seed extract (88.82 GAE/100 g DM) which is mainly used for its oil. The LC-MS analysis revealed diversity in phenolics in the three extracts and allowed the identification of hydroxybenzoic acids, hydroxycinnamic acids and flavonoids. The highest complexity was observed in the seed phenolic composition; more than twenty compounds were detected that belong to acids esters among which three feruloyl sucrose isomers. Sixteen compounds belonging to hydroxybenzoic acids, hydroxycinnamic acids and flavonoids were identified in the peel extract, whereas, only nine compounds were found in the cladode extract. It is interesting to highlight that the phenolic composition of the cladode extract was closer to that of the peel exact. However, from a quantitative viewpoint, the peel extract presented the highest amounts. Piscidic and eucomic acids were the two most concentrated molecules, corresponding to 271.3 and 121.6 mg GAE/ 100g DM respectively. The identified compounds were known to have high antioxidant and antiradical potential with the ability to inhibit lipid peroxidation and to exhibit a wide range of biological and therapeutic properties. The findings highlight the importance of using the Opuntia ficus-indica by-products.

Keywords: characterization, LC-MSn analysis, Opuntia ficus-indica, phenolics

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Authors: Azziz Assoumani, Francois Lestremau, Celine Ferret, Benedicte Lepot, Morgane Salomon, Helene Budzinski, Marie-Helene Devier, Pierre Labadie, Karyn Le Menach, Patrick Pardon, Laure Wiest, Emmanuelle Vulliet, Pierre-Francois Staub

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Seven French wastewater treatment plants (WWTP) were monitored for 53 contaminants of emerging concern within a nation-wide monitoring campaign in surface waters, which took place in 2018. The overall objective of the 2018 campaign was to provide the exercise of prioritization of emerging substances, which is being carried out in 2021, with monitoring data. This exercise should make it possible to update the list of relevant substances to be monitored (SPAS) as part of future water framework directive monitoring programmes, which will be implemented in the next water body management cycle (2022). One sampling campaign was performed in October 2018 in the seven WWTP, where affluent and sludge samples were collected. Surface water samples were collected in September 2018 at three to five sites upstream and downstream the point of effluent discharge of each WWTP. The contaminants (36 biocides and 17 surfactants, selected by the Prioritization Experts Committee) were determined in the seven WWTP effluent and sludge samples and in surface water samples by liquid or gas chromatography coupled with tandem mass spectrometry, depending on the contaminant. Nine surfactants and three biocides were quantified at least in one WWTP effluent sample. Linear alkylbenzene sulfonic acids (LAS) and fipronil were quantified in all samples; the LAS were quantified at the highest median concentrations. Twelve surfactants and 13 biocides were quantified in at least one sludge sample. The LAS and didecyldimethylammonium were quantified in all samples and at the highest median concentrations. Higher concentration levels of the substances quantified in WWTP effluent samples were observed in the surface water samples collected downstream the effluents discharge points, compared with the samples collected upstream, suggesting a contribution of the WWTP effluents in the contamination of surface waters.

Keywords: contaminants of emerging concern, effluent, monitoring, river water, sludge

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Authors: Vadim V. Yanshole, Lyudmila V. Yanshole, Alexey S. Kiryutin, Timofey D. Verkhovod, Yuri P. Tsentalovich

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Cataract, or clouding of the eye lens, is the leading cause of vision impairment in the world. The lens tissue have very specific structure: It does not have vascular system, the lens proteins – crystallins – do not turnover throughout lifespan. The protection of lens proteins is provided by the metabolites which diffuse inside the lens from the aqueous humor or synthesized in the lens epithelial layer. Therefore, the study of changes in the metabolite composition of a cataractous lens as compared to a normal lens may elucidate the possible mechanisms of the cataract formation. Quantitative metabolomic profiles of normal and cataractous human lenses were obtained with the combined use of high-frequency nuclear magnetic resonance (NMR) and ion-pairing high-performance liquid chromatography with high-resolution mass-spectrometric detection (LC-MS) methods. The quantitative content of more than fifty metabolites has been determined in this work for normal aged and cataractous human lenses. The most abundant metabolites in the normal lens are myo-inositol, lactate, creatine, glutathione, glutamate, and glucose. For the majority of metabolites, their levels in the lens cortex and nucleus are similar, with the few exceptions including antioxidants and UV filters: The concentrations of glutathione, ascorbate and NAD in the lens nucleus decrease as compared to the cortex, while the levels of the secondary UV filters formed from primary UV filters in redox processes increase. That confirms that the lens core is metabolically inert, and the metabolic activity in the lens nucleus is mostly restricted by protection from the oxidative stress caused by UV irradiation, UV filter spontaneous decomposition, or other factors. It was found that the metabolomic composition of normal and age-matched cataractous human lenses differ significantly. The content of the most important metabolites – antioxidants, UV filters, and osmolytes – in the cataractous nucleus is at least ten fold lower than in the normal nucleus. One may suppose that the majority of these metabolites are synthesized in the lens epithelial layer, and that age-related cataractogenesis might originate from the dysfunction of the lens epithelial cells. Comprehensive quantitative metabolic profiles of the human eye lens have been acquired for the first time. The obtained data can be used for the analysis of changes in the lens chemical composition occurring with age and with the cataract development.

Keywords: cataract, lens, NMR, LC-MS, metabolome

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Authors: Yu-Mei Hsueh, Cheng-Shiuan Tsai, Chao-Yuan Huang

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Prostate Cancer (PC) is a malignant tumor originated in prostate and is a second common male’s cancer in the world. Incidence of PC in Asia countries, have still been rising over the past few decades. As an antioxidant, selenium can slow down prostate cancer tumor progression, but the association between plasma selenium levels and risk of aggressive prostate cancer may be modified by different genotype of selenoprotein. The aim of this study is to determine the relationship between plasma selenium, polymorphism of selenoprotein, urinaty total arsenic, and prostate cancer. Two hundred ninety five pathologically-confirmed cases of PC and 295 cancer-free controls were individually matched to case subjects by age (± 5 years) were recruited from Department of Urology of National Taiwan University Hospital, Taipei Municipal Wan Fang Hospital and Taipei Medical University Hospital. Personal interview and biospeciment of urine and blood collection from participants were conducted by well-trained interviewers after participants’ informed consent was obtained. Plasma selenium was measured by an inductively coupled plasma mass. Urinary arsenic concentration was detected using high-performance liquid chromatography-linked hydride generator and atomic absorption spectrometry. The polymorphism of SEPP1rs3797310 and SEP15 rs5859 were determined using polymerase chain reaction-restriction fragment length polymorphism method. The higher plasma selenium was the lower OR of PC with a dose-response relationship. Prostate cancer patients with high plasma selenium had low tumor stage and grade. Participants carried SEPP1rs3797310 CT+TT genotype compared to those with CC genotype had a lower OR of PC in crude model; then this relationship was disappeared after confounder was adjusted. Prostate cancer patients with high urinary total arsenic concentration had high tumor stage and grade. Urinary total arsenic concentration was significantly positively related with plasma selenium and prostate specific antigen concentration. Participants with lower plasma selenium concentration and higher urinary total arsenic concentration compared to those with higher plasma selenium concentration and lower urinary total arsenic concentration had a higher OR of PC with a dose-response relationship.

Keywords: prostate cancer, plasma selenium concentration, urinary arsenic concentration, prostate specific antigen

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Authors: Thanaa Abdel Moghny, Mohamed Keshawy, Mahmoud Fathy, Abdul-Raheim M. Abdul-Raheim, Khalid I. Kabel, Ahmed F. El-Kafrawy, Mahmoud Ahmed Mousa, Ahmed E. Awadallah

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Ecological pollution is of great concern for human health and the environment. Numerous organic and inorganic pollutants usually discharged into the water caused carcinogenic or toxic effect for human and different life form. In this respect, this work aims to treat water contaminated by organic and inorganic waste using sorbent based on polystyrene. Therefore, two different series of adsorbent material were prepared; the first one included the preparation of polymeric sorbent from the reaction of styrene acrylate ester and alkyl acrylate. The second series involved syntheses of composite ion exchange resins of waste polystyrene and   amorphous carbon thin film (WPS/ACTF) by solvent evaporation using micro emulsion polymerization. The produced ACTF/WPS nanocomposite was sulfonated to produce cation exchange resins ACTF/WPSS nanocomposite. The sorbents of the first series were characterized using FTIR, ¹H NMR, and gel permeation chromatography. The thermal properties of the cross-linked sorbents were investigated using thermogravimetric analysis, and the morphology was characterized by scanning electron microscope (SEM). The removal of organic pollutant was determined through absorption tests in a various organic solvent. The chemical and crystalline structure of nanocomposite of second series has been proven by studies of FTIR spectrum, X-rays, thermal analysis, SEM and TEM analysis to study morphology of resins and ACTF that assembled with polystyrene chain. It is found that the composite resins ACTF/WPSS are thermally stable and show higher chemical stability than ion exchange WPSS resins. The composite resin was evaluated for calcium hardness removal. The result is evident that the ACTF/WPSS composite has more prominent inorganic pollutant removal than WPSS resin. So, we recommend the using of nanocomposite resin as new potential applications for water treatment process.

Keywords: nanocomposite, sorbent materials, waste water, waste polystyrene

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Authors: Mwafaq Ibdah, Mossab, Yahyaa, Dor Rachmany, Yoram Gerchman, Doron Holland, Liora Shaltiel-Harpaz

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Pear Psylla is the most important pest of pear in all pear-growing regions, in Asian, European, and the USA. Pear psylla damages pears in several ways: high-density populations of these insects can cause premature leaf and fruit drop, diminish plant growth, and reduce fruit size. In addition, their honeydew promotes sooty mold on leaves and russeting on fruit. Pear psyllas are also considered vectors of pear pathogens such as Candidatus Phytoplasma pyri causing pear decline that can lead to loss of crop and tree vigor, and sometimes loss of trees. Psylla control is a major obstacle to efficient integrated pest management. Recently we have identified two naturally resistance pear accessions (Py.760-261 and Py.701-202) in the Newe Ya’ar live collection. GC-MS volatile metabolic profiling identified several volatile compounds common in these accessions but lacking, or much less common, in a sensitive accession, the commercial Spadona variety. Among these volatiles were styrene and its derivatives. When the resistant accessions were used as inter-stock, the volatile compounds appear in commercial Spadona scion leaves, and it showed reduced susceptibility to pear psylla. Laboratory experiments and applications of some of these volatile compounds were very effective against psylla eggs, nymphs, and adults. The genes and enzymes involved in the specific reactions that lead to the biosynthesis of styrene in plant are unknown. We have identified a phenolic acid decarboxylase that catalyzes the formation of p-hydroxystyrene, which occurs as a styrene analog in resistant pear genotypes. The His-tagged and affinity chromatography purified E. coli-expressed pear PyPAD1 protein could decarboxylate p-coumaric acid and ferulic acid to p-hydroxystyrene and 3-methoxy-4-hydroxystyrene. In addition, PyPAD1 had the highest activity toward p-coumaric acid. Expression analysis of the PyPAD gene revealed that its expressed as expected, i.e., high when styrene levels and psylla resistance were high.

Keywords: pear Psylla, volatile, GC-MS, resistance

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Authors: Beyza Sukran Isik, Gokce Altin, Ipek Yalcinkaya, Evren Demircan, Asli Can Karaca, Beraat Ozcelik

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Turmeric (Curcuma longa L.), is used as a food additive (spice), preservative and coloring agent in Asian countries, including China and South East Asia. It is also considered as a medicinal plant. Traditional Indian medicine evaluates turmeric powder for the treatment of biliary disorders, rheumatism, and sinusitis. It has rich polyphenol content. Turmeric has yellow color mainly because of the presence of three major pigments; curcumin 1,7-bis(4-hydroxy-3-methoxyphenyl)-1, 6-heptadiene-3,5-dione), demethoxy-curcumin and bis demothoxy-curcumin. These curcuminoids are recognized to have high antioxidant activities. Curcumin is the major constituent of Curcuma species. Method: To prepare turmeric tea, 0.5 gram of turmeric powder was brewed with 250 ml of water at 90°C, 10 minutes. 500 grams of fresh turmeric washed and shelled prior to squeezing. Both turmeric tea and turmeric juice pass through 45 lm filters and stored at -20°C in the dark for further analyses. Curcumin was extracted from 20 grams of turmeric powder by 70 ml ethanol solution (95:5 ethanol/water v/v) in a water bath at 80°C, 6 hours. Extraction was contributed for 2 hours at the end of 6 hours by addition of 30 ml ethanol. Ethanol was removed by rotary evaporator. Remained extract stored at -20°C in the dark. Total phenolic content and phenolic profile were determined by spectrophotometric analysis and ultra-fast liquid chromatography (UFLC), respectively. Results: The total phenolic content of ethanolic extract of turmeric, turmeric juice, and turmeric tea were determined 50.72, 31.76 and 29.68 ppt, respectively. The ethanolic extract of turmeric, turmeric juice, and turmeric tea have been injected into UFLC and analyzed for curcumin contents. The curcumin content in ethanolic extract of turmeric, turmeric juice, and turmeric tea were 4067.4, 156.7 ppm and 1.1 ppm, respectively. Significance: Turmeric is known as a good source of curcumin. According to the results, it can be stated that its tea is not sufficient way for curcumin consumption. Turmeric juice can be preferred to turmeric tea for higher curcumin content. Ethanolic extract of turmeric showed the highest content of turmeric in both spectrophotometric and chromatographic analyses. Nonpolar solvents and carriers which have polar binding sites have to be considered for curcumin consumption due to its nonpolar nature.

Keywords: phenolic compounds, spectrophotometry, turmeric, UFLC

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Authors: Geetika Vajpayee, Sugandha Asthana, Pratibha Kumari, Shanthy Sundaram

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Pseudomonas species possess a variety of promising properties of antifungal and growth promoting activities in the wheat plant. In the present study, Pseudomonas fluorescens MTCC-9768 is tested against plant pathogenic fungus Tilletia indica, causing Karnal bunt, a quarantine disease of wheat (Triticum aestivum) affecting kernels of wheat. It is one of the 1/A1 harmful diseases of wheat worldwide under EU legislation. This disease develops in the growth phase by the spreading of microscopically small spores of the fungus (teliospores) being dispersed by the wind. The present chemical fungicidal treatments were reported to reduce teliospores germination, but its effect is questionable since T. indica can survive up to four years in the soil. The fungal growth inhibition tests were performed using Dual Culture Technique, and the results showed inhibition by 82.5%. The interaction of antagonist bacteria-fungus causes changes in the morphology of hyphae, which was observed using Lactophenol cotton blue staining and Scanning Electron Microscopy (SEM). The rounded and swollen ends, called ‘theca’ were observed in interacted fungus as compared to control fungus (without bacterial interaction). This bacterium was tested for its antagonistic activity like protease, cellulose, HCN production, Chitinase, etc. The growth promoting activities showed increase production of IAA in bacteria. The bacterial secondary metabolites were extracted in different solvents for testing its growth inhibiting properties. The characterization and purification of the antifungal compound were done by Thin Layer Chromatography, and Rf value was calculated (Rf value = 0.54) and compared to the standard antifungal compound, 2, 4 DAPG (Rf value = 0.54). Further, the in vivo experiments showed a significant decrease in the severity of disease in the wheat plant due to direct injection method and seed treatment. Our results indicate that the extracted and purified compound from the antagonist bacteria, P. fluorescens MTCC-9768 may be used as a potential biocontrol agent against T. indica. This also concludes that the PGPR properties of the bacteria may be utilized by incorporating it into bio-fertilizers.

Keywords: antagonism, Karnal bunt, PGPR, Pseudomonas fluorescens

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Authors: Avijit Dey, Shyam S. Paul, Satbir S. Dahiya, Balbir S. Punia, Luciano A. Gonzalez

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Methane emitted from ruminant livestock not only reduces the efficiency of feed energy utilization but also contributes to global warming. Vegetable oils, a source of poly unsaturated fatty acids, have potential to reduce methane production and increase conjugated linoleic acid in the rumen. However, characteristics of oils, level of inclusion and composition of basal diet influences their efficacy. Therefore, this study was aimed to investigate the effects of sunflower (SFL) and cottonseed (CSL) oils on methanogenesis, volatile fatty acids composition and feed fermentation pattern by in vitro gas production (IVGP) test. Four concentrations (0, 0.1, 0.2 and 0.4ml /30ml buffered rumen fluid) of each oil were used. Fresh rumen fluid was collected before morning feeding from two rumen cannulated buffalo steers fed a mixed ration. In vitro incubation was carried out with sorghum hay (200 ± 5 mg) as substrate in 100 ml calibrated glass syringes following standard IVGP protocol. After 24h incubation, gas production was recorded by displacement of piston. Methane in the gas phase and volatile fatty acids in the fermentation medium were estimated by gas chromatography. Addition of oils resulted in increase (p<0.05) in total gas production and decrease (p<0.05) in methane production, irrespective of type and concentration. Although the increase in gas production was similar, methane production (ml/g DM) and its concentration (%) in head space gas was lower (p< 0.01) in CSL than in SFL at corresponding doses. Linear decrease (p<0.001) in degradability of DM was evident with increasing doses of oils (0.2ml onwards). However, these effects were more pronounced with SFL. Acetate production tended to decrease but propionate and butyrate production increased (p<0.05) with addition of oils, irrespective of type and doses. The ratio of acetate to propionate was reduced (p<0.01) with addition of oils but no difference between the oils was noted. It is concluded that both the oils can reduce methane production. However, feed degradability was also affected with higher doses. Cotton seed oil in small dose (0.1ml/30 ml buffered rumen fluid) exerted greater inhibitory effects on methane production without impeding dry matter degradability. Further in vivo studies need to be carried out for their practical application in animal ration.

Keywords: buffalo, methanogenesis, rumen fermentation, vegetable oils

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Authors: Aruma Baduge Kithma De Silva

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Haskap (Lonicera caerulea L.), also known as blue honeysuckle, is a newly commercialized berry crop in Canada. Haskap berries are rich in polyphenols, including, anthocyanins, which are known for potential health-promoting properties. Cyanidin-3-O-glucoside (C3G) is the most abundant anthocyanin of haskap berries. The compound C3G has the ability to reduce the risk of type 2 diabetes (T2D), which has become an increasingly common health issue around the world. The T2D is characterized as a metabolic disorder of hyperglycemia and insulin resistance. It has been demonstrated that C3G has anti-diabetic effects through several ways, including inhibition of dipeptidyl peptidase-4 (DPP-4), reduction of gluconeogenesis, improvement in insulin sensitivity, and inhibition of activities of carbohydrate hydrolyzing enzymes, including α-amylase and α-glucosidase. The goal of this study was to investigate the influence of variety and harvests maturity of haskap on C3G, other fruit quality characteristics and anti-diabetic activities of haskap berries using in vitro studies. The polyphenols present in four commercially grown haskap cultivars, Aurora, Rebecca, Larissa, and Evie harvested at five harvesting dates (H1-H5) apart from 2-3 days, were extracted separately. High-performance liquid chromatography electrospray ionization mass spectrometry (HPLC-ESI-MS) analyzes of polyphenols revealed that haskap berries contain predominantly anthocyanins, flavonols, flavan-3-ols, and phenolic acids. The compound C3G was the most prominent anthocyanin, which is available in approximately 79% of total anthocyanin in four cultivars. The Larissa at H5 contained the highest C3G content. The antioxidant capacity of Evie at H5 was greater than other cultivars. Furthermore, Larissa H5 showed the greatest inhibition of carbohydrate hydrolyzing enzymes including alpha-glucosidase and alpha-amylase. In conclusion, the haskap variety and harvesting date influenced the polyphenol composition and biological properties. The variety Larissa, at H5 harvesting date, contained the highest polyphenol content and the ability of inhibition of the carbohydrate hydrolyzing enzyme as well as DPP4 enzyme in order to reduce type 2 diabetes.

Keywords: anthocyanin, Haskap, type 2 diabetes, polyphenol

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Authors: Mengfei Peng, Serajus Salaheen, Debabrata Biswas

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Lactobacillus casei found in the human intestine and mouth is commonly applied for dairy production. Recently, it was found that some byproducts produced by Lactobacillus exhibited antimicrobial activities against multiple bacteria. Meanwhile, introduction of prebiotic-like foods (e.g. cocoa) or probiotics or both of them as food supplements in human diets as well as in farm animal feeds is believed to be an effective ways in control/reduce the colonization of foodborne bacterial pathogens infection in the gut environment. We hypothesized that cocoa may stimulate the production antimicrobial components of Lactobacillus casei and may potentially inhibit/reduce the colonization and infection of foodborne bacterial pathogens in the gut. Mixed culture of L. casei (LC) with enterohemorrhagic E. coli EDL933 (EHEC), Salmonella Typhimurium LT2 (ST), or Listeria monocytogenes LM2 (LM) showed that LC could competitively exclude (100%) them within 72 h. Further, investigation of cell-free culture supernatant (CFCS) revealed that the antimicrobial effects of LC came from CFCS. CFCS of LC eliminated (100%) EHEC, ST, and LM within 72 h, and 2 h CFCS treatment increased the hydrophobicity of EHEC (5.10 folds), ST (8.48 folds), and LM (2.03 folds). In addition, LC cells exhibited more inhibitive effects than CFCS on cell adhesive and invasive activities of EHEC (52.14% & 90.45%), ST (66.89% & 93.83%), and LM (61.10% & 83.40%). Two clusters of poly-peptides in CFCS were identified by SDS-PAGE, the molecular weights of which are ≈5 KD and 40-45 KD. LC CFCS with overnight growth in the presence of 3% strengthened all of the antimicrobial activities (growth inhibition, outer membrane disruption, and cell infective ability reduction). Liquid chromatography/Mass spectrometry analysis detected 5 unique components in class of flavonoids in LC CFCS with overnight 3% cocoa supplement. Furthermore, qPCR results showed that CFCSs up-regulated the expression level of genes responsible for flagellin synthesis and motility, but down-regulated genes for specific binding and invasion-associated proteins synthesis. The stimulatory effects of cocoa in producing bioactive components of probiotics may aid prevention of foodborne illness caused by major foodborne enteric bacterial pathogens.

Keywords: foodborne pathogens, probiotics, prebiotics, pathogen exclusion

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Authors: Niki Mougiou, Spyros Didos, Ioanna Bouzouka, Athina Theodorakopoulou, Michael Kornaros, Anagnostis Argiriou

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Grains from traditional old Greek cereal varieties were evaluated and compared to commercial cultivars, like Simeto and Mexicali 81, in an effort to valorize local products and assess the nutritional benefits of ancient grains. The samples studied in this research included common wheat, durum wheat, emmer (Triticum dicoccum) and einkorn (Triticum monococcum), as well as barley, oats and rye grains. The Internal Transcribed Spacer 2 (ITS2) nuclear region was amplified and sequenced as a barcode for species identification, allowing the verification of the label of each product. After that, the total content of bound and free polyphenols and flavonoids, as well as the antioxidant activity of bound and free compounds, was measured by classic colorimetric assays using Folin- Ciocalteu, AlCl₃ and DPPH‧ (2,2-diphenyl-1-picrylhydrazyl) reagents, respectively. Moreover, the level of variation of fatty acids was determined in all samples by gas chromatography. The results showed that local old landraces of emmer and einkorn had the highest polyphenol content, 2.4 and 3.3 times higher than the average value of 5 durum wheat samples, respectively. Regarding the total flavonoid content, einkorn had 2.6-fold and emmer 2-fold higher values than common wheat. The antioxidant activity of free or bound compounds was at the same level, at about 20-30% higher in both einkorn and emmer compared to common wheat. Five main fatty acids were detected in all samples, in order of decreasing amounts: linoleic (C18:2) > palmitic (C16:0) ≈ , oleic (C18:1) > eicosenoic (C20:1, cis-11) > stearic (C18:0). Emmer and einkorn showed a higher diversity of fatty acids and a higher content of mono-unsaturated fatty acids compared to common wheat. The results of this study demonstrate the high nutritional value of old local landraces that have been put aside by more productive, yet with lower qualitative characteristics, commercial cultivars, underlining the importance of maintaining sustainable agricultural practices to ensure their continued cultivation.

Keywords: biochemical analysis, nutritional value, plant barcoding, wheat

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Authors: A.Assariotakis, P. Vahamidis, P. Tarantilis, G. Economou

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Greece, due to its geographical location and the particular climatic conditions, presents high biodiversity of Medicinal and Aromatic Plants. Among them, the genus Origanum not only presents a wide distribution, but it also has great economic importance. After extensive surveys in Ikaria Island (Greece), 3 species of the genus Origanum were identified, namely, Origanum vulgare ssp. hirtum (Greek oregano), Origanum onites (Turkish oregano) and Origanum x intercedens (hybrid), a naturally occurring hybrid between O. hirtum and O. onites. The purpose of this study was to determine their morphological as well as their temporal variability in essential oil yield and composition under field conditions. For this reason, a plantation of each species was created using vegetative propagation and was established at the experimental field of the Agricultural University of Athens (A.U.A.). From the establishment year and for the following two years (3 years of observations), several observations were taken during each growing season with the purpose of identifying the morphological differences among the studied species. Each year collected plant (at bloom stage) material was air-dried at room temperature in the shade. The essential oil content was determined by hydrodistillation using a Clevenger-type apparatus. The chemical composition of essential oils was investigated by Gas Chromatography-Mass Spectrometry (GC – MS). Significant differences were observed among the three oregano species in terms of plant height, leaf size, inflorescence features, as well as concerning their biological cycle. O. intercedens inflorescence presented more similarities with O. hirtum than with O. onites. It was found that calyx morphology could serve as a clear distinction feature between O. intercedens and O. hirtum. The calyx in O. hirtum presents five isometric teeth whereas in O. intercedens two high and three shorter. Essential oil content was significantly affected by genotype and year. O. hirtum presented higher essential oil content than the other two species during the first year of cultivation, however during the second year the hybrid (O. intercedens) recorded the highest values. Carvacrol, p-cymene and γ-terpinene were the main essential oil constituents of the three studied species. In O. hirtum carvacrol content varied from 84,28 - 93,35%, in O. onites from 86,97 - 91,89%, whereas in O. intercedens it was recorded the highest carvacrol content, namely from 89,25 - 97,23%.

Keywords: variability, oregano biotypes, essential oil, carvacrol

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Authors: Carla Idely Palencia-Aguilar

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Artificial wetlands play an important role at Guasca Municipality in Colombia, not only because they are used for the agroindustry, but also because more than 45 species were found, some of which are endemic and migratory birds. Remote sensing was used to determine the changes in the area occupied by water of artificial wetlands by means of Aster and Modis images for different time periods. Evapotranspiration was also determined by three methods: Surface Energy Balance System-Su (SEBS) algorithm, Surface Energy Balance- Bastiaanssen (SEBAL) algorithm, and Potential Evapotranspiration- FAO. Empirical equations were also developed to determine the relationship between Normalized Difference Vegetation Index (NDVI) versus net radiation, ambient temperature and rain with an obtained R2 of 0.83. Groundwater level fluctuations on a daily basis were studied as well. Data from a piezometer placed next to the wetland were fitted with rain changes (with two weather stations located at the proximities of the wetlands) by means of multiple regression and time series analysis, the R2 from the calculated and measured values resulted was higher than 0.98. Information from nearby weather stations provided information for ordinary kriging as well as the results for the Digital Elevation Model (DEM) developed by using PCI software. Standard models (exponential, spherical, circular, gaussian, linear) to describe spatial variation were tested. Ordinary Cokriging between height and rain variables were also tested, to determine if the accuracy of the interpolation would increase. The results showed no significant differences giving the fact that the mean result of the spherical function for the rain samples after ordinary kriging was 58.06 and a standard deviation of 18.06. The cokriging using for the variable rain, a spherical function; for height variable, the power function and for the cross variable (rain and height), the spherical function had a mean of 57.58 and a standard deviation of 18.36. Threatens of eutrophication were also studied, given the unconsciousness of neighbours and government deficiency. Water quality was determined over the years; different parameters were studied to determine the chemical characteristics of water. In addition, 600 pesticides were studied by gas and liquid chromatography. Results showed that coliforms, nitrogen, phosphorous and prochloraz were the most significant contaminants.

Keywords: DEM, evapotranspiration, geostatistics, NDVI

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Authors: Xiaojie Cheng, Ulrike Frank, Feng Zhao, Karin Pritsch

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Ragweed (Ambrosia artemisiifolia) is an invasive weed that has become an increasing global problem. In addition to affecting land use and crop yields, ragweed has a strong impact on human health as it produces highly allergenic pollen. Global warming will result in an earlier and longer pollen season enhanced pollen production and an increase in pollen allergenicity with a negative effect on atopic patients. The aims of this study were to investigate the effects of increasing temperature, the future climate scenario in the Munich area, southern Germany, predicted on the basis of RCP8.5 until the end of 2050s, or/and NO₂, a major air pollutant, 1) on the vegetative and reproductive characteristics of ragweed plants, 2) on the total allergenicity of ragweed pollen, 3) on the total pollen proteomic patterns. Ragweed plants were cultivated for the whole plant vegetation period under controlled conditions either under ambient climate conditions or 4°C higher temperatures with or without additional NO₂. Higher temperature resulted in bigger plant sizes, longer male inflorescences, and longer pollen seasons. The total allergenic potential of the pollen was accessed by dot blot using serum from ragweed pollen sensitized patients. The comparative immunoblot analysis revealed that the in vivo fumigation of ragweed plants with elevated NO₂-concentrations significantly increased the allergenic potential of the pollen, and in combination with increased temperature, the allergenic potential was even higher. On the other hand, label-free protein quantification by liquid chromatography-tandem mass spectrometry (LC-MS/MS) was performed. The results showed that more proteins were significantly up- and down-regulated under higher temperatures with/without elevated NO₂ conditions. Most of the highly expressed proteins were participating intensively in the metabolic process, the cellular process, and the stress defense process. These findings suggest that rising temperature and elevated NO₂ are important environmental factors for higher abiotic stress activities, catalytic activities, and thus higher allergenic potential observed in pollen proteins.

Keywords: climate change, NO₂, pollen proteome, ragweed, temperature

Procedia PDF Downloads 164

Authors: Ashkan Zargar, Ali Taheri Mirghaed, Zein Talal Barakat, Alireza Khosravi, Hamed Paknejad

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With the increase of bacterial resistance to the chemical antibiotics, replacing it with ecofriendly herbal materials and with no adverse effects in the host body is very important. Therefore, in this study, the effect of combined essential oil (Thymus vulgaris-Origanum magorana and Ziziphora clinopodioides) on the growth behavior of Yersinia ruckeri, Aeromonas hydrophila and Lactococcus garvieae was evaluated. The compositions of the herbal essential oils used in this study were determined by gas chromatography-mass spectrometry (GC-MS) while, the investigating of antimicrobial effects was conducted by the agar-disc diffusion method, determination of minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC), and bacterial growth curves determination relied on optical density (OD) at 630 nm. The main compounds were thymol (40.60 %) and limonene (15.98 %) for Thymus vulgaris while carvacrol (57.86 %) and thymol (13.54 %) were the major compounds in Origanum magorana. As regards Ziziphora clinopodiodes, α-pinene (22.6 %) and carvacrol (21.1 %) represented the major constituents. Concerning Yersinia ruckeri, disc-diffusion results showed that t.O.z (50 % Origanum majorana) combined essential oil was presented the best inhibition zone (30.66 mm) but it was exhibited no significant differences with other tested commercial antibiotics except oxytetracycline (P <0/05). The inhibitory activity and the bactericidal effect of the t.O.z, unveiled by the MIC= 0.2 μL /mL and MBC= 1.6 μL /mL values, were clearly the best between all combined oils. The growth behaviour of Yersinia ruckeri was affected by this combined essential oil and changes in temperature and pH conditions affected herbal oil performance. As regard Aeromonas hydrophila, its results were so similar to Yersinia ruckeri results and t.O.z (50 % Origanum majorana) was the best between all combined oils (inhibition zone= 26 mm, MIC= 0.4 μL /mL and MBC= 3.2 μL /mL, combined essential oil was affected bacterial growth behavior). Also for Lactococcus garvieae, t.O.z (50 % Origanum majorana) was the best between all combined oils having the best inhibition zone= 20.66 mm, MIC= 0.8 μL /mL and MBC= 1.6 μL /mL and best effect on inhibiting bacterial growth. Combined herbal essential oils have a good and noticeable effect on the growth behavior of pathogenic bacteria in the laboratory, and by continuing research in the host, they may be a suitable alternative to control, prevent and treat diseases caused by these bacteria.

Keywords: bacterial pathogen, herbal medicine, growth behavior, fish

Procedia PDF Downloads 49

Authors: Thomas Chinedu Aniokete, Gordian Onyebuchukwu Mbah, Michael Daramola

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A sustainable NaOH integrated hydrothermal protocol was developed for the synthesis of waste-derived hydroxy sodalite catalysts for transesterification of waste animal fat (WAF) with a high per cent free fatty acid (FFA) to biodiesel. In this work, hydroxy sodalite catalyst was synthesized from two complex waste materials namely coal fly ash (CFA) and waste industrial brine (WIB). Measured amounts of South African CFA and WIB obtained from a coal mine field were mixed with NaOH solution at different concentrations contained in secured glass vessels equipped with magnetic stirrers and formed consistent slurries after aging condition at 47 oC for 48 h. The slurries were then subjected to hydrothermal treatments at 140 oC for 48 h, washed thoroughly and separated by the action of a centrifuge on the mixture. The resulting catalysts were calcined in a muffle furnace for 2 h at 200 oC and subsequently characterized for different effects using X-ray diffraction (XRD), scanning electron microscopy (SEM), Fourier transform infrared (FT-IR), and Bennett Emmet Teller (BET) adsorption-desorption techniques. The produced animal fat methyl ester (AFME) was analyzed using the gas chromatography-mass spectrometry (GC-MS) method. Results of the investigation indicate profoundly an enhanced catalyst purity, textural property and desired morphology due to the action of NaOH. Similarly, the performance evaluation with respect to catalyst activity reveals a high catalytic conversion efficiency of 98 % of the high FFA WAF to biodiesel under the following reaction conditions; a methanol-to-WAF ratio of 15:1, amount of SOD catalyst of 3 wt % with a stirring speed of 300-500 rpm, a reaction temperature of 60 oC and a reaction time of 8 h. There was a recovered 96 % stable catalyst after reactions and potentially recyclable, thus contributing to the economic savings to the process that had been a major bottleneck to the production of biodiesel. This NaOH route for synthesizing waste-derived hydroxy sodalite (SOD) catalyst is a sustainable and eco-friendly technology that speaks directly to the global quest for renewable-fossil fuel controversy enforcing sustainable development goal 7.

Keywords: coal fly ash, waste industrial brine, waste-derived hydroxy sodalite catalyst, sodium hydroxide, biodiesel, transesterification, biomass conversion

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Authors: Josline Salib, Sayed El-Toumy, Abeer Salama, Enayat Omara, Emad Hassan

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Diabetic nephropathy is an important cause of morbidity and mortality and is now among the most common causes of end-stage renal failure (ESRF) in developed countries. Thus, the aim of the present study was to investigate the phenolic compounds content of Cichorium intybus aerial parts extracts as well as the therapeutic effects on diabetic nephropathy, oxidative stress, and anti-inflammatory by characterizing biochemical, histopathological changes and immunohistochemistry in an experimental diabetic rat model as compared with Amaryl. Ten known compounds of flavonoids, coumarins and phenolic acid derivatives were isolated from the C. intybus aqueous methanolic extract. Structures of the isolated compounds were established by chromatography, UV and 1D⁄2D 1H⁄ 13C spectroscopy. The aqueous methanol extract of C. intybus aerial parts was administered to Streptozotocin diabetes rats at doses (100 and 200 mg/kg) for 21 days. After treatment, blood glucose, serum insulin, urea, creatinine, and TNF-α were evaluated. Enzymatic scavengers including catalase (CAT), glutathione (GSH), malondialdehyde (MDA) and nitric oxide (NO) were determined to evaluate the oxidative status in the renal tissue. Diabetic rats treated with C. intybus extract showed a dose-dependent reduction of fasting blood glucose and kidney antioxidant status in comparison to the diabetic control group. The extract was able to enhance the antioxidant defenses of the kidney by increasing the reduced GSH and CAT content and decreasing MDA content in addition to significantly decreasing kidney nitric oxide content compared to diabetic control rats. Furthermore, the histopathological findings in C. intybus extract administered rats were observed at markedly lesser extent than the diabetic control group. Also, inducible nitric oxide synthase (iNOS) levels were decreased significantly after the administration of high-dose C. intybus extract in diabetic rats. Showing significant antihyperglycemic and antioxidant properties of C. intybus aerial parts extract, which is attributed to its polyphenolic content, may offer a potential source for the treatment of diabetes.

Keywords: antioxidant activity, anti-diabetic nephropathy, cichorium intybus aerial parts, phenolic compounds

Procedia PDF Downloads 99

Authors: Bharti Jain, Rajeev Jain, Abuzar Kabir, Torki Zughaibi, Shweta Sharma

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Drug-facilitated crimes (DFCs) entail the use of a single drug or a mixture of drugs to render a victim unable. Traditionally, biological samples have been gathered from victims and conducted analysis to establish evidence of drug administration. Nevertheless, the rapid metabolism of various drugs and delays in analysis can impede the identification of such substances. For this, the present article describes a rapid, sustainable, highly efficient and miniaturized protocol for the identification and quantification of three sedative-hypnotic drugs, namely diazepam, chlordiazepoxide and ketamine in alcoholic beverages and complex food samples (cream of biscuit, flavored milk, juice, cake, tea, sweets and chocolate). The methodology involves utilizing fabric phase sorptive extraction (FPSE) to extract diazepam (DZ), chlordiazepoxide (CDP), and ketamine (KET). Subsequently, the extracted samples are subjected to analysis using gas chromatography-mass spectrometry (GC-MS). Several parameters, including the type of membrane, pH, agitation time and speed, ionic strength, sample volume, elution volume and time, and type of elution solvent, were screened and thoroughly optimized. Sol-gel Carbowax 20M (CW-20M) has demonstrated the most effective extraction efficiency for the target analytes among all evaluated membranes. Under optimal conditions, the method displayed linearity within the range of 0.3–10 µg mL–¹ (or µg g–¹), exhibiting a coefficient of determination (R2) ranging from 0.996–0.999. The limits of detection (LODs) and limits of quantification (LOQs) for liquid samples range between 0.020-0.069 µg mL-¹ and 0.066-0.22 µg mL-¹, respectively. Correspondingly, the LODs for solid samples ranged from 0.056-0.090 µg g-¹, while the LOQs ranged from 0.18-0.29 µg g-¹. Notably, the method showcased better precision, with repeatability and reproducibility both below 5% and 10%, respectively. Furthermore, the FPSE-GC-MS method proved effective in determining diazepam (DZ) in forensic food samples connected to drug-facilitated crimes (DFCs). Additionally, the proposed method underwent evaluation for its whiteness using the RGB12 algorithm.

Keywords: drug facilitated crime, fabric phase sorptive extraction, food forensics, white analytical chemistry

Procedia PDF Downloads 47

Authors: Brahim Sow

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Background: Throughout West Africa, Balanites aegyptiaca (BA), or Zygophyllaceae, is widely used in traditional medicine to treat diabetes, hypertension, inflammation, malaria and liver disorders. In our recent research, we found that BA has nephroprotective potential against diabetes mellitus, hypertension and kidney disorders. However, to our knowledge, no systematic studies have been carried out on its derivative (toxicity) profile. Aim of the study: The study was conducted to assess the potential potency of the hydroalcoholic extract of BA bark in rats by the acute and sub-acute oral route. Materials and methods: Male and female rats in the acute depression study received BA extract orally at single doses of 500 mg/kg, 2000 mg/kg, 3000 mg/kg and 5000 mg/kg (n = 6 per group/sex). To assess acute depression, abnormal behaviour, toxic symptoms, weight and death were observed for 14 consecutive days. For the subacute impairment study, Wistar rats received the extract orally at doses of 125, 250 and 500 mg/kg (n=6 per group/sex) per day for 28 days. Behaviour and body weight were monitored daily. At the end of the treatment period, biochemical, haematological and histopathological examinations were performed, and gross and histopathological examinations of several organs were carried out. To determine the presence or absence of phytochemicals, the BA extract was subjected to gage phage chromatographic examination. Results: The absence of absorption chromatography of BA indicates the absence of cyanide groups. This suggests that the BA extract does not contain toxic substances. No mortality or adverse effects were observed at 5000 mg/kg in the acute depression test. With regard to body weight, general behaviour, relative organ weights, haematological and biochemical parameters, BA extract did not induce any mortality or potentially treatment-related effects in the sub-acute study. The normal architecture of the vital organs was revealed by histopathological examination, indicating the absence of morphological alterations. Conclusion: BA extract administered orally for 28 days at doses up to 500 mg/kg did not cause toxicological damage in rats in the present study. The median lethal dose (LD50) of the extract was estimated to be over 5000 mg/kg in an acute hyperglycaemia study.

Keywords: Balanites aegyptiaca L Delile, haematology, biochemistry, rat

Procedia PDF Downloads 51

Authors: L. V. Lokesha, Jagadeesh S. Sanganal, Yogesh S. Gowda, Shekhar, N. B. Shridhar, N. Prakash, Prashantkumar Waghe, H. D. Narayanaswamy, Girish V. Kumar

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Organochlorine (OC) insecticides are among the most important organotoxins and make a large group of pesticides. Physicochemical properties of these toxins, especially their lipophilicity, facilitate the absorption and storage of these toxins in the meat thus possess public health threat to humans. The presence of these toxins in broiler meat can be a quantitative and qualitative index for the presence of these toxins in animal bodies, which is attributed to Waste water of irrigation after spraying the crops, contaminated animal feeds with pesticides, polluted air are the potential sources of residues in animal products. Fifty broiler meat samples were collected from different retail outlets of Bengaluru city, Karnataka state, in ice cold conditions and later stored under -20°C until analysis. All the samples were subjected to Gas Chromatograph attached to Electron Capture Detector(GC-ECD, VARIAN make) screening and quantification of OC pesticides viz; Alachlor, Aldrin, Alpha-BHC, Beta-BHC, Dieldrin, Delta-BHC, o,p-DDE, p,p-DDE, o,p-DDD, p,p-DDD, o,p-DDT, p,p-DDT, Endosulfan-I, Endosulfan-II, Endosulfan Sulphate and Lindane(all the standards were procured from Merck). Extraction was undertaken by blending fifty grams (g) of meat sample with 50g Sodium Sulphate anahydrous, 120 ml of n-hexane, 120 ml acetone for 15 mins, extract is washed with distilled water and sample moisture is dried by sodium sulphate anahydrous, partitioning is done with 25 ml petroleum ether, 10 ml acetonitrile and 15 ml n-hexane shake vigorously for two minutes, sample clean up was done with florosil column. The reconstituted samples (using n-hexane) (Merck chem) were injected to Gas Chromatograph–Electron Capture Detector(GC-ECD). The present study reveals that, among the fifty chicken samples subjected for analysis, 60% (15/50), 32% (8/50), 28% (7/50), 20% (5/50) and 16% (4/50) of samples contaminated with DDTs, Delta-BHC, Dieldrin, Aldrin and Alachlor respectively. DDT metabolites, Delta-BHC were the most frequently detected OC pesticides. The detected levels of the pesticides were below the levels of MRL(according to Export Council of India notification for fresh poultry meat).

Keywords: accuracy, gas chromatography, meat, pesticide, petroleum ether

Procedia PDF Downloads 298

Authors: J. Vitku, L. Kolatorova, T. Chlupacova, J. Heracek, M. Hill, M. Duskova, L. Starka

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Background: 7β-hydroxy-epiandrosterone (7β–OH-EpiA) is an endogenous steroid, that has been shown to exert neuroprotective and anti-inflammatory effects in vitro as well as in animal models. However, to the best of our knowledge no information is available about concentration of this androgen metabolite in human population. The aim of the study was to measure and compare levels of 7β–OH-EpiA in men and pregnant women in different biological fluids and evaluate the relationship between 7β–OH-EpiA in men and their sperm quality. Methods: First, a sensitive isotope dilution high performance liquid chromatography-mass spectrometry method for measurement of 7β–OH-EpiA in different biological fluids was developed. Validation of the method met the requirements of FDA guidelines. Afterwards 7β–OH-EpiA in plasma and seminal plasma of 191 men with different degree of infertility (healthy men, lightly infertile men, moderately infertile men, severely infertile men) was analysed. Furthermore, the levels of 7β–OH-EpiA in plasma of 34 pregnant women in 37th week of gestation and corresponding cord plasma that reflects steroid levels in the fetus were measured. Results: Concentrations of 7β–OH-EpiA in seminal plasma were significantly higher in severely infertile men in comparison with healthy men and lightly infertile men. The same trend was observed when blood plasma was evaluated. Furthermore, plasmatic 7β –OH-EpiA negatively correlated with concentration (-0.215; p < 0.01) and total count (-0.15; p < 0.05). Seminal 7β–OH-EpiA was negatively associated with motility (-0.26; p < 0.01), progressively motile sperms (-0.233; p < 0.01) and nonprogressively motile sperms (-0.188; p < 0.05). Plasmatic 7β –OH-EpiA levels in men were generally higher in comparison with pregnant women. Levels 7β–OH-EpiA were under the lower limit of quantification (LLOQ) in majority of samples of pregnant women and cord plasma. Only 4 plasma samples of pregnant women and 7 cord blood plasma samples were above LLOQ and where in range of units of pg/ml. Conclusion: Based on available information, this is the first study measuring 7β–OH-EpiA in human samples. 7β–OH-EpiA is associated with lower sperm quality and certainly it is worth to explore its role in this field thoroughly. Interestingly, levels of 7β–OH-EpiA in pregnant women were extremely low despite the fact that steroid levels including androgens are generally higher during pregnancy. Acknowledgements: This work was supported by the project MH CR 17-30528 A from the Czech Health Research Council, MH CZ - DRO (Institute of Endocrinology - EU, 00023761) and by the MEYS CR (OP RDE, Excellent research - ENDO.CZ).

Keywords: 7β-hydroxy-epiandrosterone, steroid, sperm quality, pregnancy

Procedia PDF Downloads 231

Authors: Ane Escobar, Paula Angelomé, Mihaela Delcea, Marek Grzelczak, Sergio Enrique Moya

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The antibacterial capacity of bone-anchoring implants can be improved by the use of antibiotics that can be delivered to the media after the surgery. Mesoporous films have shown great potential in drug delivery for orthopedic applications, since pore size and thickness can be tuned to produce different surface area and free volume inside the material. This work shows the synthesis of mesoporous titania films (MTF) by sol-gel chemistry and evaporation-induced self-assembly (EISA) on top of glass substrates. Pores with a diameter of 12nm were observed by Transmission Electron Microscopy (TEM). A film thickness of 100 nm was measured by Scanning Electron Microscopy (SEM). Gentamicin was used to study the antibiotic delivery from the film by means of High-performance liquid chromatography (HPLC). The Staphilococcus aureus strand was used to evaluate the effectiveness of the penicillin loaded films toward inhibiting bacterial colonization. MC3T3-E1 pre-osteoblast cell proliferation experiments proved that MTFs have a good biocompatibility and are a suitable surface for MC3T3-E1 cell proliferation. Moreover, images taken by Confocal Fluorescence Microscopy using labeled vinculin, showed good adhesion of the MC3T3-E1 cells to the MTFs, as well as complex actin filaments arrangement. In order to improve cell proliferation Bone Morphogenetic Protein-2 (BMP-2) was adsorbed on top of the mesoporous film. The deposition of the protein was proved by measurements in the contact angle, showing an increment in the hydrophobicity while the protein concentration is higher. By measuring the dehydrogenase activity in MC3T3-E1 cells cultured in dually functionalized mesoporous titatina films with gentamicin and BMP-2 is possible to find an improvement in cell proliferation. For this purpose, the absorption of a yellow-color formazan dye, product of a water-soluble salt (WST-8) reduction by the dehydrogenases, is measured. In summary, this study proves that by means of the surface modification of MTFs with proteins and loading of gentamicin is possible to achieve an antibacterial effect and a cell growth improvement.

Keywords: antibacterial, biocompatibility, bone morphogenetic protein-2, cell proliferation, gentamicin, implants, mesoporous titania films, osteoblasts

Procedia PDF Downloads 144

Authors: Ayman El Sabagh, AbdElhamid Omar, Dekoum Assaha, Khair Mohammad Youldash, Akihiro Ueda, Celaleddin Barutçular, Hirofumi Saneoka

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Drought and salinity stress are a worldwide problem, constraining global crop production seriously. Hence, soybean is susceptible to yield loss from water deficit and salinity stress. Therefore, different approaches have been suggested to solve these issues. Osmoprotectants play an important role in protection the plants from various environmental stresses. Moreover, organic fertilization has several beneficial effects on agricultural fields. Presently, efforts to maximize nitrogen fixation in soybean are critical because of widespread increase in soil degradation in Egypt. Therefore, a greenhouse research was conducted at plant nutritional physiology laboratory, Hiroshima University, Japan for assessing the impact of exogenous osmoregulators and compost application in alleviating the adverse effects of salinity and water stress on soybean. Treatments was included (i) water stress treatments (different soil moisture levels consisting of (100%, 75%, and 50% of field water holding capacity), (ii) salinity concentrations (0 and 15 mM) were applied in fully developed trifoliolate leaf node (V1), (iii) compost treatments (0 and 24 t ha-1) and (iv) the exogenous, proline and glycine betaine concentrations (0 mM and 25 mM) for each, was applied at two growth stages (V1 and R1). The seeds of soybean cultivar Giza 111, was sown into basin from wood (length10 meter, width 50cm, height 50cm and depth 350cm) containing a soil mixture of granite regosol soil and perlite (2:1 v/v). The nitrogen-fixing activity was estimated by using gas chromatography and all measurements were made in three replicates. The results showed that water deficit and salinity stress reduced biological nitrogen fixation and specific nodule activity than normal irrigation conditions. Exogenous osmoprotectants were improved biological nitrogen fixation and specific nodule activity as well as, applying of compost led to improving many of biological nitrogen fixation and specific nodule activity with superiority than stress conditions. The combined application compost fertilizer and exogenous osmoprotectants were more effective in alleviating the adverse effect of stress to improve biological nitrogen fixation and specific nodule activity of Soybean.

Keywords: a biotic stress, biological nitrogen fixation, compost, osmoprotectants, specific nodule activity, soybean

Procedia PDF Downloads 290

Authors: Temesgen Sidamo, Prakruti S. Rao, Eleni Akllilu, Workineh Shibeshi, Yumi Park, Yong-Soon Cho, Jae-Gook Shin, Scott K. Heysell, Stellah G. Mpagama, Ephrem Engidawork

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The fluoroquinolones (FQs) are used off-label for the treatment of multidrug-resistant tuberculosis (MDR-TB), and for evaluation in shortening the duration of drug-susceptible TB in recently prioritized regimens. Within the class, levofloxacin (LFX) and moxifloxacin (MXF) play a substantial role in ensuring success in treatment outcomes. However, sub-therapeutic plasma concentrations of either LFX or MXF may drive unfavorable treatment outcomes. To the best of our knowledge, the pharmacokinetics of LFX and MXF in Ethiopian patients with MDR-TB have not yet been investigated. Therefore, the aim of this study was to develop a population pharmacokinetic (PopPK) model of levofloxacin (LFX) and moxifloxacin (MXF) and assess the percent probability of target attainment (PTA) as defined by the ratio of the area under the plasma concentration-time curve over 24-h (AUC0-24) and the in vitro minimum inhibitory concentration (MIC) (AUC0-24/MIC) in Ethiopian MDR-TB patients. Steady-state plasma was collected from 39 MDR-TB patients enrolled in the programmatic treatment course and the drug concentrations were determined using optimized liquid chromatography-tandem mass spectrometry. In addition, the in vitro MIC of the patients' pretreatment clinical isolates was determined. PopPK and simulations were run at various doses, and PK parameters were estimated. The effect of covariates on the PK parameters and the PTA for maximum mycobacterial kill and resistance prevention was also investigated. LFX and MXF both fit in a one-compartment model with adjustments. The apparent volume of distribution (V) and clearance (CL) of LFX were influenced by serum creatinine (Scr), whereas the absorption constant (Ka) and V of MXF were influenced by Scr and BMI, respectively. The PTA for LFX maximal mycobacterial kill at the critical MIC of 0.5 mg/L was 29%, 62%, and 95% with the simulated 750 mg, 1000 mg, and 1500 mg doses, respectively, whereas the PTA for resistance prevention at 1500 mg was only 4.8%, with none of the lower doses achieving this target. At the critical MIC of 0.25 mg/L, there was no difference in the PTA (94.4%) for maximum bacterial kill among the simulated doses of MXF (600 mg, 800 mg, and 1000 mg), but the PTA for resistance prevention improved proportionately with dose. Standard LFX and MXF doses may not provide adequate drug exposure. LFX PopPK is more predictable for maximum mycobacterial kill, whereas MXF's resistance prevention target increases with dose. Scr and BMI are likely to be important covariates in dose optimization or therapeutic drug monitoring (TDM) studies in Ethiopian patients.

Keywords: population PK, PTA, moxifloxacin, levofloxacin, MDR-TB patients, ethiopia

Procedia PDF Downloads 94

Authors: Bruno Baptista, Andreia S. R. Oliveira, Patricia V. Mendonca, Jorge F. J. Coelho, Fani Sousa

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Drug delivery systems are designed to allow adequate protection and controlled delivery of drugs to specific locations. These systems aim to reduce side effects and control the biodistribution profile of drugs, thus improving therapeutic efficacy. This study involved the synthesis of polymeric nanoparticles, based on amphiphilic diblock copolymers, comprising a biocompatible, poly (oligo (ethylene oxide) methyl ether methacrylate (POEOMA) as hydrophilic segment and a pH-sensitive block, the poly (2-diisopropylamino)ethyl methacrylate) (PDPA). The objective of this work was the development of polymeric pH-responsive nanoparticles to encapsulate and carry small RNAs as a model to further develop non-coding RNAs delivery systems with therapeutic value. The responsiveness of PDPA to pH allows the electrostatic interaction of these copolymers with nucleic acids at acidic pH, as a result of the protonation of the tertiary amine groups of this polymer at pH values below its pKa (around 6.2). Initially, the molecular weight parameters and chemical structure of the block copolymers were determined by size exclusion chromatography (SEC) and nuclear magnetic resonance (1H-NMR) spectroscopy, respectively. Then, the complexation with small RNAs was verified, generating polyplexes with sizes ranging from 300 to 600 nm and with encapsulation efficiencies around 80%, depending on the molecular weight of the polymers, their composition, and concentration used. The effect of pH on the morphology of nanoparticles was evaluated by scanning electron microscopy (SEM) being verified that at higher pH values, particles tend to lose their spherical shape. Since this work aims to develop systems for the delivery of non-coding RNAs, studies on RNA protection (contact with RNase, FBS, and Trypsin) and cell viability were also carried out. It was found that they induce some protection against constituents of the cellular environment and have no cellular toxicity. In summary, this research work contributes to the development of pH-sensitive polymers, capable of protecting and encapsulating RNA, in a relatively simple and efficient manner, to further be applied on drug delivery to specific sites where pH may have a critical role, as it can occur in several cancer environments.

Keywords: drug delivery systems, pH-responsive polymers, POEOMA-b-PDPA, small RNAs

Procedia PDF Downloads 240

Authors: Ahmed Auda, Manjree Agarwala, Giles Hardya, Yonglin Rena

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Botrytis cinerea is a major pest for many plants. It can attack a wide range of plant parts. It can attack buds, flowers, and leaves, stems, and fruit. However, B. cinerea can be mixed with other diseases that cause the same damage. There are many species of botrytis and more than one different strains of each. Botrytis might infect the foliage of nursery stock stored through winter in damp conditions. There are no known resistant plants. Botrytis must have nutrients or food source before it infests the plant. Nutrients leaking from wounded plant parts or dying tissue like old flower petals give the required nutrients. From this food, the fungus becomes more attackers and invades healthy tissue. Dark to light brown rot forms in the ill tissue. High humidity conditions support the growth of this fungus. However, we suppose that selection pressure can act on the morphological and neurophysiologic filter properties of the receiver and on both the biochemical and the physiological regulation of the signal. Communication is implied when signal and receiver evolves toward more and more specific matching, culminating. In other hand, receivers respond to portions of a body odor bouquet which is released to the environment not as an (intentional) signal but as an unavoidable consequence of metabolic activity or tissue damage. Each year Botrytis species can cause considerable economic losses to plant crops. Even with the application of strict quarantine and control measures, these fungi can still find their way into crops and cause the imposition of onerous restrictions on exports. Blueberry fruit mould caused by a fungal infection usually results in major losses during post-harvest storage. Therefore, the management of infection in early stages of disease development is necessary to minimize losses. The overall purpose of this study will develop sensitive, cheap, quick and robust diagnostic techniques for the detection of B. cinerea in blueberry. The specific aim was designed to investigate the performance of volatile organic compounds (VOCs) in the detection and discrimination of blueberry fruits infected by fungal pathogens with an emphasis on Botrytis in the early storage stage of post-harvest.

Keywords: botrytis cinerea, blueberry, GC/MS, VOCs

Procedia PDF Downloads 221

Authors: Vuiswa Lucia Sethunya, Tonderayi Matambo, Diane Hildebrandt

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South African’s informal settlements produce tonnes of kitchen waste (KW) per year which is dumped into the landfill. These landfill sites are normally located in close proximity to the household of the poor communities; this is a problem in which the young children from those communities end up playing in these landfill sites which may result in some health hazards because of methane, carbon dioxide and sulphur gases which are produced. To reduce this large amount of organic materials being deposited into landfills and to provide a cleaner place for those within the community especially the children, an energy conversion process such as anaerobic digestion of the organic waste to produce biogas was implemented. In this study, the digestion of various kitchen waste was investigated in order to understand and develop a system that is suitable for household use to produce biogas for cooking. Three sets of waste of different nutritional compositions were digested as per acquired in the waste streams of a household at mesophilic temperature (35ᵒC). These sets of KW were co-digested with cow dung (CW) at different ratios to observe the microbial behaviour and the system’s stability in a laboratory scale system. The gas chromatography-flame ionization detector analyses have been performed to identify and quantify the presence of organic compounds in the liquid samples from co-digested and mono-digested food waste. Acetic acid, propionic acid, butyric acid and valeric acid are the fatty acids which were studied. Acetic acid (1.98 g/L), propionic acid (0.75 g/L) and butyric acid (2.16g/L) were the most prevailing fatty acids. The results obtained from organic acids analysis suggest that the KW can be an innovative substituent to animal manure for biogas production. The faster degradation period in which the microbes break down the organic compound to produce the fatty acids during the anaerobic process of KW also makes it a better feedstock during high energy demand periods. The C/N ratio analysis showed that from the three waste streams the first stream containing vegetables (55%), fruits (16%), meat (25%) and pap (4%) yielded more methane-based biogas of 317mL/g of volatile solids (VS) at C/N of 21.06. Generally, this shows that a household will require a heterogeneous composition of nutrient-based waste to be fed into the digester to acquire the best biogas yield to sustain a households cooking needs.

Keywords: anaerobic digestion, biogas, kitchen waste, household

Procedia PDF Downloads 171

Authors: Anshuman Srivastava, Shiv Singh, Sheelendra Pratap Singh

Abstract:

A simple, sensitive and effective gas chromatography tandem mass spectrometry (GC-MS/MS) method was developed for simultaneous analysis of multi pesticide residues (organophosphate, organochlorines, synthetic pyrethroids and herbicides) in food commodities using phenolic resin based activated carbon fibers (ACFs) as reversed-dispersive solid phase extraction (r-DSPE) sorbent in modified QuEChERS (Quick Easy Cheap Effective Rugged Safe) method. The acetonitrile-based QuEChERS technique was used for the extraction of the analytes from food matrices followed by sample cleanup with ACFs instead of traditionally used primary secondary amine (PSA). Different physico-chemical characterization techniques such as Fourier transform infrared spectroscopy, scanning electron microscopy, X-ray diffraction and Brunauer-Emmet-Teller surface area analysis were employed to investigate the engineering and structural properties of ACFs. The recovery of pesticides and herbicides was tested at concentration levels of 0.02 and 0.2 mg/kg in different commodities such as cauliflower, cucumber, banana, apple, wheat and black gram. The recoveries of all twenty-six pesticides and herbicides were found in acceptable limit (70-120%) according to SANCO guideline with relative standard deviation value < 15%. The limit of detection and limit of quantification of the method was in the range of 0.38-3.69 ng/mL and 1.26 -12.19 ng/mL, respectively. In traditional QuEChERS method, PSA used as r-DSPE sorbent plays a vital role in sample clean-up process and demonstrates good recoveries for multiclass pesticides. This study reports that ACFs are better in terms of removal of co-extractives in comparison of PSA without compromising the recoveries of multi pesticides from food matrices. Further, ACF replaces the need of charcoal in addition to the PSA from traditional QuEChERS method which is used to remove pigments. The developed method will be cost effective because the ACFs are significantly cheaper than the PSA. So the proposed modified QuEChERS method is more robust, effective and has better sample cleanup efficiency for multiclass multi pesticide residues analysis in different food matrices such as vegetables, grains and fruits.

Keywords: QuEChERS, activated carbon fibers, primary secondary amine, pesticides, sample preparation, carbon nanomaterials

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