Search results for: amino acid reagents

Authors: Iddrisu Ibrahim, Joseph Atia Ayariga, Junhuan Xu, Daniel A. Abugri, Robertson K. Boakai, Olufemi S. Ajayi

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The recalcitrance of pathogenic bacteria indicates that millions of people who are at risk of infection arising from chronic diseases, surgery, organ transplant, diabetes, and several other debilitating diseases present an aura of potentially untreatable illness due to resistance development. Antimicrobial resistance has successfully become a global health menace, and resistances are often acquired by bacteria through health-care-related incidence (HRI) orchestrated by multi-drug resistant (MDR) and extended drug-resistant pathogens (EDRP). To understand the mechanisms S. Typhimurium uses to resist CDB, we study the abundance of LPS modification, Ergosterols, Mysristic palmitic resistance, Oleic acid resistance of susceptible and resistant S. Typhimurium. Using qPCR, we also analyzed the expression of selected genes known for enabling resistance in S. Typhimurium. We found high abundance of LPS, Ergosterols, Mysristic palmitic resistance, Oleic acid resistance of and high expression of resistant genes in S. Typhimurium compared to the susceptible strain. LPS modification, Ergosterols, Mysristic palmitic resistance, Oleic acid and genes such as Fims, integrons, blaTEM are important indicators of resistance development of S. typhimurium.

Keywords: antimicrobials, resistance, Cannabidiol, Salmonella, blaTEM, fimA, Lipopolysaccharide, Ergosterols

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Authors: Lucas Kurth de Azambuja, Loise Silveira da Silva, Gean Vitor Salmoria, Darlan Dallacosta, Carlos Rodrigo de Mello Roesler

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This work provides a physicochemical and thermal characterization assessment of three different hyaluronic acid (HA)-based injections used for rejuvenation skin treatments. The three products analyzed are manufactured by the same manufacturer and commercialized for application on different skin levels. According to the manufacturer, all three HA-based injections are crosslinked and have a concentration of 23 mg/mL of HA, and 0.3% of lidocaine. Samples were characterized by Fourier-transformed infrared (FTIR), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), and scanning electron microscope (SEM) techniques. FTIR analysis resulted in a similar spectrum when comparing the different products. DSC analysis demonstrated that the fusion points differ in each product, with a higher fusion temperature observed in specimen A, which is used for subcutaneous applications, when compared with B and C, which are used for the middle dermis and deep dermis, respectively. TGA data demonstrated a considerable mass loss at 100°C, which means that the product has more than 50% of water in its composition. TGA analysis also showed that Specimen A had a lower mass loss at 100°C when compared to Specimen C. A mass loss of around 220°C was observed on all samples, characterizing the presence of hyaluronic acid. SEM images displayed a similar structure on all samples analyzed, with a thicker layer for Specimen A when compared with B and C. This series of analyses demonstrated that, as expected, the physicochemical and thermal properties of the products differ according to their application. Furthermore, to better characterize the crosslinking degree of each product and their mechanical properties, a set of different techniques should be applied in parallel to correlate the results and, thereby, relate injection application with material properties.

Keywords: hyaluronic acid, characterization, soft-tissue fillers, injectable gels

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Authors: M. Zasada, A. Markiewicz, A. Erkiert-Polguj, E. Budzisz

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The epidermis is a keratinized stratified squamous epithelium covered by the hydro-lipid barrier. Therefore, active substances should be able to penetrate through this hydro-lipid coating. L-ascorbic acid is one of the vitamins which plays an important role in stimulation fibroblast to produce collagen type I and in hyperpigmentation lightening. Vitamin C is a water-soluble antioxidant, which protects skin from oxidation damage and rejuvenates photoaged skin. No-needle mesotherapy is a non-invasive rejuvenation technique depending on electric pulses, electroporation, and ultrasounds. These physicals factors result in deeper penetration of cosmetics. It is important to increase the penetration of L-ascorbic acid, thereby increasing the spectrum of its activity. The aim of the work was to assess the effectiveness of pure L-ascorbic acid activity in anti-aging therapy using a needle-free and micro-needling mesotherapy. The study was performed on a group of 35 healthy volunteers in accordance with the Declaration of Helsinki of 1964 and agreement of the Ethics Commissions no RNN/281/16/KE 2017. Women were randomized to mesotherapy or control group. Control group applied topically 2,5 ml serum containing 20% L-ascorbic acid with hydrate from strawberries, every 10 days for a period of 9 weeks. No-needle mesotherapy, on the left half of the face and micro-needling on the right with the same serum, was done in mesotherapy group. The pH of serum was 3.5-4, and the serum was prepared directly prior to the facial treatment. The skin parameters were measured at the beginning and before each treatment. The measurement of the forehead skin was done using Cutometer® (measurement of skin elasticity and firmness), Corneometer® (skin hydration measurement), Mexameter® (skin tone measurement). Also, the photographs were taken by Fotomedicus system. Additionally, the volunteers fulfilled the questionnaire. Serum was tested for microbiological purity and stability after the opening of the cosmetic. During the study, all of the volunteers were taken care of a dermatologist. The regular application of the serum has caused improvement of the skin parameters. Respectively, after 4 and 8 weeks improvement in hydration and elasticity has been seen (Corneometer®, Cutometer® results). Moreover, the number of hyper-pigmentated spots has decreased (Mexameter®). After 8 weeks the volunteers has claimed that the tested product has smoothing and moisturizing features. Subjective opinions indicted significant improvement of skin color and elasticity. The product containing the L-ascorbic acid used with intercellular penetration promoters demonstrates higher anti-aging efficiency than control. In vivo studies confirmed the effectiveness of serum and the impact of the active substance on skin firmness and elasticity, the degree of hydration and skin tone. Mesotherapy with pure L-ascorbic acid provides better diffusion of active substances through the skin.

Keywords: anti-aging, l-ascorbic acid, mesotherapy, promoters

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Authors: Ming-Kwei Lee, Chih-Feng Yen

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The electrical characteristics of liquid phase deposited silicon oxynitride film on ammonium sulfide treated p-type (100) gallium arsenide substrate were investigated. Hydrofluosilicic acid, ammonia and boric acid aqueous solutions were used as precursors. The electrical characteristics of silicon oxynitride film are much improved on gallium arsenide substrate with ammonium sulfide treatment. With post-metallization annealing, hydrogen ions can further passivate defects in SiON/GaAs film and interface. The leakage currents can reach 7.1 × 10-8 and 1.8 × 10-7 at ± 2 V. The dielectric constant and effective oxide charges are 5.6 and -5.3 × 1010 C/cm2, respectively. The hysteresis offset of hysteresis loop is merely 0.09 V.

Keywords: liquid phase deposition, SiON, GaAs, PMA, (NH4)2S

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Authors: Kyung Hwa Hong, Eunmi Koh

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Wool and cotton fabrics are pretreated by a tannic acid aqueous solution to increase their dyeability and then dyed by Purple-Fleshed Sweet Potato (PSP) extract. The dyed fabrics are then investigated by various analysis techniques. The results revealed that wool and cotton fabrics can be dyed bluish red through the pretreatment and dyeing process. Both wool and cotton fabrics only pretreated with tannic acid display decreased L* value but no significant changes in a* and b* values as the concentration of tannic acid increases. And, as expected, the pretreated fabrics are even darker and show a richer purple color after the dyeing process with the PSP extract. With regard to the colorfastness of wool and cotton fabrics dyed by PSP extract in cleaning circumstances, such as dry-cleaning (for wool) and washing (for cotton), the wool and cotton fabrics had a 4.0 and 4.0 grade of colorfastness to dry-cleaning and washing, respectively. However, they both exhibited significantly inferior colorfastness to light (grade of 1.5). Thus, it was found that there is still a need for improvement with regard to color fastness, particularly against light. On the other hand, the wool and cotton fabrics also showed antibacterial and antioxidant characteristics. In addition, both the wool and cotton fabrics showed potential antibacterial ability (>99%) against Staphylococcus aureus; however, they showed somewhat insufficient antibacterial ability (60.8% for wool and 94.8% for cotton) against Klebsiella pneumoniae. Also, their antioxidant abilities increased up to ca. 90% with an increase in the tannic acid concentration (up to 0.5%). However, after the dyeing process, the antibacterial and antioxidant ability tended to decrease. This is assumed to have occurred because functional moieties such as phenolic acids were detached from the pretreated fabrics into the hot water (the dyeing solution) during the dyeing process. Therefore, further study would be necessary to derive the optimum treatment and dyeing conditions so as to maximize the coloring effect and functionalities of the fabrics.

Keywords: antibacterial activity, antioxidant activity, purple-fleshed sweet potato, fabrics

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Authors: H. Boucheloukh, S. Rouissa, N. Aoun, M. Beloucifa, T. Sehili, F. Parrino, V. Loddo

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To optimize water purification and wastewater treatment by heterogeneous photocatalysis, we used NiFe₂O₄ as a catalyst and solar irradiation as a source of energy. In this concept, an organic substance present in many industrial effluents was chosen: naproxen ((S)-6-methoxy-α-methyl-2-naphthaleneacetic acid or 2-(6-methoxynaphthalenyl) propanoic), a non-steroidal anti-inflammatory drug. The main objective of this study is to degrade naproxen by an iron and nickel catalyst, the degradation of this organic pollutant by nickel ferrite has been studied in a heterogeneous aqueous medium, with the study of the various factors influencing photocatalysis such as the concentration of matter and the acidity of the medium. The photocatalytic activity was followed by HPLC-UV andUV-Vis spectroscopy. A first-order kinetic model appropriately fitted the experimental data. The degradation of naproxen was also studied in the presence of H₂O₂ as well as in an aqueous solution. The new hetero-system NiFe₂O₄/oxalic acid is also discussed. The fastest naproxen degradation was obtained with NiFe₂O₄/H₂O₂. In a first-place, we detailed the characteristics of the material NiFe₂O₄, which was synthesized by the sol-gel methods, using various analytical techniques: visible UV spectrophotometry, X-ray diffraction, FTIR, cyclic voltammetry, luminescent discharge optical emission spectroscopy.

Keywords: naproxen, nickelate, photocatalysis, oxalic acid

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Authors: Fedili Benamar, Belloulou Mohamed Lamine, Ouahes Hassane, Ghattas Samir

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Introduction: Perioperative blood loss is a frequent cause of complications in total hip replacement (THR). The present prospective study assessed the efficacy of tranexamic acid (Exacyl(®)) in reducing blood loss in primary THR. Hypothesis: Tranexamic acid reduces blood loss in THR. Material and method: -This is a prospective randomized study on the effectiveness of Exacyl (tranexamic acid) in total hip replacement surgery performed on a standardized technique between 2019 and September 2022. -It involved 154 patients, of which 84 received a single injection of Exacyl (group 1) at a dosage of 10 mg/kg over 20 minutes during the perioperative period. -All patients received postoperative thromboprophylaxis with enoxaparin 0.4 ml subcutaneously. -All patients were admitted to the post-interventional intensive care unit for a duration of 24 hours for monitoring and pain management as per the service protocol. Results: 154 patients, of which 84 received a single injection of Exacyl (group 1) and 70 patients patients who did not receive Exacyl perioperatively : (Group 2 ) The average age is 57 +/- 15 years The distribution by gender was nearly equal with 56% male and 44% female; "The distribution according to the ASA score was as follows: 20.2% ASA1, 82.3% ASA2, and 17.5% ASA3. "There was a significant difference in the average volume of intraoperative and postoperative bleeding during the 48 hours." The average bleeding volume for group 1 (received Exacyl) was 614 ml +/- 228, while the average bleeding volume for group 2 was 729 +/- 300, with a chi-square test of 6.35 and a p-value < 0.01, which is highly significant. The ANOVA test showed an F-statistic of 7.11 and a p-value of 0.008. A Bartlett test revealed a chi-square of 6.35 and a p-value < 0.01." "In Group 1 (patients who received Exacyl), 73% had bleeding less than 750 ml (Group A), and 26% had bleeding exceeding 750 ml (Group B). In Group 2 (patients who did not receive Exacyl perioperatively), 52% had bleeding less than 750 ml (Group A), and 47% had bleeding exceeding 750 ml (Group B). "Thus, the use of Exacyl reduced perioperative bleeding and specifically decreased the risk of severe bleeding exceeding 750 ml by 43% with a relative risk (RR) of 1.37 and a p-value < 0.01. The transfusion rate was 1.19% in the population of Group 1 (Exacyl), whereas it was 10% in the population of Group 2 (no Exacyl). It can be stated that the use of Exacyl resulted in a reduction in perioperative blood transfusion with an RR of 0.1 and a p-value of 0.02. Conclusions: The use of Exacyl significantly reduced perioperative bleeding in this type of surgery.

Keywords: acid tranexamic, blood loss, anesthesia, total hip replacement, surgery

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Authors: Dmitry Nosik, Nickolay Nosik, Elli Kaplina, Olga Lobach, Marina Chataeva, Lev Rasnetsov

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Background and aims: Human Immunodeficiency Virus (HIV) infection is very often associated with Herpes Simplex Virus (HSV) infection but HIV patients are treated with a cocktail of antiretroviral drugs which are toxic. The use of an antiviral drug which will be active against both viruses like ferrovir found in our previous studies is rather actual. Earlier we had shown that Fullerene poly-amino capronic acid (FPACA) was active in case of monoinfection of HIV-1 or HSV-1. The aim of the study was to analyze the efficiency of FPACA against mixed infection of HIV and HSV. Methods: The peripheral blood lymphocytes, CEM, MT-4 cells were simultaneously infected with HIV-1 and HSV-1. FPACA was added 1 hour before infection. Cells viability was detected by MTT assay, virus antigens detected by ELISA, syncytium formation detected by microscopy. The different multiplicity of HIV-1/HSV-1 ratio was used. Results: The double viral HIV-1/HSV-1 infection was more cytopathic comparing with monoinfections. In mixed infection by the HIV-1/HSV-1 concentration of HIV-1 antigens and syncytium formations increased by 1,7 to 2,3 times in different cells in comparison with the culture infected with HIV-1 alone. The concentration of HSV-1 increased by 1,5-1,7 times, respectively. Administration of FPACA (1 microg/ml) protected cells: HIV-1/HSV-1 (1:1) – 80,1%; HIV-1/HSV-1 (1:4) – 57,2%; HIV-1/HSV-1 (1:8) – 46,3 %; HIV-1/HSV-1 (1:16) – 17,0%. Virus’s antigen levels were also reduced. Syncytium formation was totally inhibited in all cases of mixed infection. Conclusion: FPACA showed antiviral activity in case of mixed viral infection induced by Human Immunodeficiency Virus and Herpes Simplex Virus. The effect of viral inhibition increased with the multiplicity of HIV-1 in the inoculum. The mechanism of FPACA action is connected with the blocking of the virus particles adsorption to the cells and it could be suggested that it can have an antiviral activity against some other viruses too. Now FPACA could be considered as a potential drug for treatment of HIV disease complicated with opportunistic herpes viral infection.

Keywords: antiviral drug, human immunodeficiency virus (hiv), herpes simplex virus (hsv), mixed viral infection

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Authors: Alanna Goodman, Kayla Murray, Keith Warriner

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The following reports on a comparative study on the efficacy of different decontamination technologies to decrease Listeria monocytogenes inoculated onto white sliced mushrooms and assesses the fate of residual levels during posttreatment storage under aerobic conditions at 8uC. The treatments were chemical (hydrogen peroxide, peroxyacetic acid, ozonated water, electrolyzed water, chitosan, lactic acid), biological (Listeria bacteriophages), and physical (UV-C, UV:hydrogen peroxide). None of the treatments achieved .1.2 log CFU reduction in L. monocytogenes levels; bacteriophages at a multiplicity of infection of 100 and 3% (vol/vol) hydrogen peroxide were the most effective of the treatments tested. However, growth of residual L. monocytogenes during posttreatment storage attained levels equal to or greater than levels in the nontreated controls. The growth of L. monocytogenes was inhibited on mushrooms treated with chitosan, electrolyzed water, peroxyacetic acid, or UV. Yet, L. monocytogenes inoculated onto mushrooms and treated with UV:hydrogen peroxide decreased during posttreatment storage, through a combination of sublethal injury and dehydration of the mushroom surface. Although mushrooms treated with UV:hydrogen peroxide became darker during storage, the samples were visually acceptable relative to controls. In conclusion, of the treatments evaluated, UV:hydrogen peroxide holds promise to control L. monocytogenes on mushroom surfaces.

Keywords: listeria monocytogenes, sliced mushrooms, bacteriophages, UV, sanitizers

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Authors: Nayeon, Lee, Won-Young, Cho, Hyun Joo, Jang, Chi-Ho, Lee

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This study investigated the effects of the dietary supplementation of the Allium hookeri root, and processed sulfur, on the growth performance of guinea pigs. The guinea pigs were fed a control diet (CON), as well as the control diet including 1% freeze-dried Allium hookeri root (AH), or 0.1% processed sulfur (S), or including both the freeze-dried Allium hookeri root and the processed sulfur (AHS). The weight of perirenal adipose tissue (PAT) and the epididymal adipose tissue (EAT) in the AH were significantly lower than CON (p < 0.05). The serum cholesterols levels of the AH and the AHS were significantly lower than the S (p < 0.05). While the total saturated fatty acid content in the serum of the AH and AHS groups showed a tendency to decrease, the total monounsaturated fatty acid increased. The results of this study suggested that dietary consumption of Allium hookeri root may help to decrease fat accumulation, lower serum cholesterol levels, and control serum free fatty acid contents in the guinea pigs.

Keywords: Allium hookeri, dietary supplementation, growth performance, processed sulfur, Guinea pig

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Authors: Shivam Yadav, Neelam Atri

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Cyanobacteria are photoautotrophic prokaryotes able to grow in diverse ecological habitats, originated 2.5 - 3.5 billion years ago and brought oxygenic photosynthesis. Since then superoxide dismutases (SODs) acquired great significance due to their ability to catalyze detoxification of byproducts of oxygenic photosynthesis, i.e. superoxide radicals. Sequence information from several cyanobacterial genomes offers a unique opportunity to conduct a comprehensive comparative analysis of the superoxide dismutases family. In the present study, we extracted information regarding SODs from species of sequenced cyanobacteria and investigated their diversity, conservation, domain structure, and evolution. 144 putative SOD homologues were identified. SODs are present in all cyanobacterial species reflecting their significant role in survival. However, their distribution varies, fewer in unicellular marine strains whereas abundant in filamentous nitrogen-fixing cyanobacteria. Motifs and invariant amino acids typical in eukaryotic SODs were conserved well in these proteins. These SODs were classified into three major families according to their domain structures. Interestingly, they lack additional domains as found in proteins of other family. Phylogenetic relationships correspond well with phylogenies based on 16S rRNA and clustering occurs on the basis of structural characteristics such as domain organization. Similar conserved motifs and amino acids indicate that cyanobacterial SODs make use of a similar catalytic mechanism as eukaryotic SODs. Gene gain-and-loss is insignificant during SOD evolution as evidenced by absence of additional domain. This study has not only examined an overall background of sequence-structure-function interactions for the SOD gene family but also revealed variation among SOD distribution based on ecophysiological and morphological characters.

Keywords: comparative genomics, cyanobacteria, phylogeny, superoxide dismutases

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Authors: M. El-haj, Z. Olama, H. Holail

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Single cell oils (SCOs) accumulated by oleaginous fungi have emerged as a potential alternative feedstock for biodiesel production. Five fungal strains were isolated from the Lebanese environment namely Fusarium oxysporum, Mucor hiemalis, Penicillium citrinum, Aspergillus tamari, and Aspergillus niger that have been selected among 39 oleaginous strains for their potential ability to accumulate lipids (lipid content was more than 40% on dry weight basis). Wide variations were recorded in the environmental factors that lead to maximum lipid production by fungi under test and were cultivated under submerged fermentation on medium containing glucose as a carbon source. The maximum lipid production was attained within 6-8 days, at pH range 6-7, 24 to 48 hours age of seed culture, 4 to 6.107 spores/ml inoculum level and 100 ml culture volume. Eleven culture conditions were examined for their significance on lipid production using Plackett-Burman factorial design. Reducing sugars and nitrogen source were the most significant factors affecting lipid production process. Maximum lipid yield was noticed with 15.62, 14.48, 12.75, 13.68 and 20.41g/l for Fusarium oxysporum, Mucor hiemalis, Penicillium citrinum, Aspergillus tamari, and Aspergillus niger respectively. A verification experiment was carried out to examine model validation and revealed more than 94% validity. The profile of extracted lipids from each fungal isolate was studied using thin layer chromatography (TLC) indicating the presence of monoacylglycerols, diaacylglycerols, free fatty acids, triacylglycerols and sterol esters. The fatty acids profiles were also determined by gas-chromatography coupled with flame ionization detector (GC-FID). Data revealed the presence of significant amount of oleic acid (29-36%), palmitic acid (18-24%), linoleic acid (26.8-35%), and low amount of other fatty acids in the extracted fungal oils which indicate that the fatty acid profiles were quite similar to that of conventional vegetable oil. The cost of lipid production could be further reduced with acid-pretreated lignocellulotic corncob waste, whey and date molasses to be utilized as the raw material for the oleaginous fungi. The results showed that the microbial lipid from the studied fungi was a potential alternative resource for biodiesel production.

Keywords: agro-industrial waste products, biodiesel, fatty acid, single cell oil, Lebanese environment, oleaginous fungi

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Authors: Azmawati Mohammed Nawi, Siok-Fong Chin, Shamsul Azhar Shah, Rahman Jamal

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In recent years, trace elements have gained importance as biomarkers in many chronic diseases. Unfortunately, the requirement for sample volume increases according to the extent of investigation for diagnosis or elucidating the mechanism of the disease. Here, we describe the method development and validation for simultaneous determination of 25 trace elements (lithium (Li), beryllium (Be), magnesium (Mg), aluminium (Al), vanadium (V), chromium (Cr), manganese (Mn), iron (Fe), cobalt (Co), nickel (Ni), copper (Cu), zinc (Zn), gallium (Ga), arsenic (As), selenium (Se), rubidium (Rb), strontium (Sr), silver (Ag), cadmium (Cd), caesium (Cs), barium (Ba), mercury (Hg), thallium (Tl), lead (Pb), uranium (U)) using just 20 µL of human serum. Serum samples were digested with nitric acid and hydrochloric acid (ratio 1:1, v/v) and analysed using inductively coupled plasma–mass spectrometry (ICP-MS). Seronorm®, a human-derived serum control material was used as quality control samples. The intra-day and inter-day precisions were consistently < 15% for all elements. The validated method was later applied to 30 human serum samples to evaluate its suitability. In conclusion, we have successfully developed and validated a precise and accurate analytical method for determining 25 trace elements requiring very low volume of human serum.

Keywords: acid digestion, ICP-MS, trace element, serum

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Authors: Feng-Tsai Weng, Rick Wang, Yan-Cong Liao

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Passivation is a kind of surface treatment for material to reinforce the corrosion resistance specially the stainless alloy. Passive film, is to getting more potential compared to their status before passivation. An oxidation film can be formed on the surface of stainless steel, which has a strong corrosion resistance ability after passivation treatment. In this research, a new passivation technology is proposed for a special stainless alloy which contains a 12-14% Chromium. This method includes the A-A-A (alkaline-acid-alkaline) process basically, which was developed by Carpenter that can neutralize trapped acid. Besides, a corrosion resistant coating layer was obtained by immersing the parts in a water bath of mineral oil at high temperature. Salt spray test ASTM B368 was conducted to investigated performance of corrosion resistant of the passivated stainless steel alloy parts. Results show much better corrosion resistant that followed a coating process after A-A-A Passivation process, than only using A-A-A process. The passivation time is with more than 380 hours of salt spray test ASTM B368, which is equal to 3000 hours of Salt spray test ASTM B117. Proposed passivation method of stainless steel can be completed in about 3 hours.

Keywords: passivation, alkaline-acid-alkaline, stainless steel, salt spray test

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Authors: Mohit Batra, Noel Sarkar, Jayeeta Mitra

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In this study silica nanoparticles were synthesized using different methods and different silica sources namely Tetraethyl ortho silicate (TEOS), Sodium Silicate, Rice husk while chitosan nanoparticles were prepared with ionic gelation method using Sodium tripolyphosphate (TPP). Size and texture of silica nanoparticles were studied using field emission scanning electron microscopy (FESEM) and transmission electron microscopy (TEM) along with the effect of change in concentration of various reagents in different synthesis processes. Size and dispersion of Silica nanoparticles prepared from TEOS using stobber’s method were found better than other methods while nanoparticles prepared using rice husk were cheaper than other ones. Catalyst found to play a very significant role in controlling the size of nanoparticles in all methods.

Keywords: silica nanoparticles, gelatin, bio-nanocomposites, SEM, TEM, chitosan

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Authors: Sudhanshu Sharma

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Lisinopril, 1-[N-{(s)-I-carboxy-3 phenyl propyl}-L-proline dehydrate is a lysine analog of enalaprilat, the active metabolite of enalapril. It is long-acting, non-sulhydryl angiotensin-converting enzyme (ACE) inhibitor that is used for the treatment of hypertension and congestive heart failure in daily dosage 10-80 mg. Pharmacological activity of lisinopril has been proved in various experimental and clinical studies. Owing to its importance and widespread use, efforts have been made towards the development of simple and reliable analytical methods. As per our literature survey, lisinopril in pharmaceutical formulations has been determined by various analytical methodologies like polaragraphy, potentiometry, and spectrophotometry, but most of these analytical methods are not too suitable for the Identification of lisinopril from clinical samples because of the interferences caused by the amino acids and amino groups containing metabolites present in biological samples. This report is an attempt in the direction of developing a simple and reliable method for on plate identification and quantification of lisinopril in pharmaceutical formulations as well as from human urine samples using silica gel H layers developed with a new mobile phase comprising of micellar solutions of N-cetyl-N, N, N-trimethylammonium bromide (CTAB). Micellar solutions have found numerous practical applications in many areas of separation science. Micellar liquid chromatography (MLC) has gained immense popularity and wider applicability due to operational simplicity, cost effectiveness, relatively non-toxicity and enhanced separation efficiency, low aggressiveness. Incorporation of aqueous micellar solutions as mobile phase was pioneered by Armstrong and Terrill as they accentuated the importance of TLC where simultaneous separation of ionic or non-ionic species in a variety of matrices is required. A peculiarity of the micellar mobile phases (MMPs) is that they have no macroscopic analogues, as a result the typical separations can be easily achieved by using MMPs than aqueous organic mobile phases. Previously MMPs were successfully employed in TLC based critical separations of aromatic hydrocarbons, nucleotides, vitamin K1 and K5, o-, m- and p- aminophenol, amino acids, separation of penicillins. The human urine analysis for identification of selected drugs and their metabolites has emerged as an important investigation tool in forensic drug analysis. Among all chromatographic methods available only thin layer chromatography (TLC) enables a simple fast and effective separation of the complex mixtures present in various biological samples and is recommended as an approved testing for forensic drug analysis by federal Law. TLC proved its applicability during successful separation of bio-active amines, carbohydrates, enzymes, porphyrins, and their precursors, alkaloid and drugs from urine samples.

Keywords: lisnopril, surfactant, chromatography, micellar solutions

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Authors: M. M. Hefnawy, A. M. A. Homoda, M. A. Abounassif, A. M. Alanazia, A. Al-Majed, Gamal A. E. Mostafa

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PVC membrane sensors using different approach e.g. ion-pair, ionophore, and Schiff-base has been used as testing membrane sensor. Analytical applications of membrane sensors for direct measurement of variety of different ions in complex biological and environmental sample are reported. The most important step of such PVC membrane sensor is the sensing active material. The potentiometric sensors have some outstanding advantages including simple design, operation, wide linear dynamic range, relative fast response time, and rotational selectivity. The analytical applications of these techniques to pharmaceutical compounds in dosage forms are also discussed. The construction and electrochemical response characteristics of Poly (vinyl chloride) membrane sensors for moxifloxacin HCl (MOX) are described. The sensing membranes incorporate ion association complexes of moxifloxacin cation and sodium tetraphenyl borate (NaTPB) (sensor 1), phosphomolybdic acid (PMA) (sensor 2) or phosphotungstic acid (PTA) (sensor 3) as electroactive materials. The sensors display a fast, stable and near-Nernstian response over a relative wide moxifloxacin concentration range (1 ×10-2-4.0×10-6, 1 × 10-2-5.0×10-6, 1 × 10-2-5.0×10-6 M), with detection limits of 3×10-6, 4×10-6 and 4.0×10-6 M for sensor 1, 2 and 3, respectively over a pH range of 6.0-9.0. The sensors show good discrimination of moxifloxacin from several inorganic and organic compounds. The direct determination of 400 µg/ml of moxifloxacin show an average recovery of 98.5, 99.1 and 98.6 % and a mean relative standard deviation of 1.8, 1.6 and 1.8% for sensors 1, 2, and 3 respectively. The proposed sensors have been applied for direct determination of moxifloxacin in some pharmaceutical preparations. The results obtained by determination of moxifloxacin in tablets using the proposed sensors are comparable favorably with those obtained using the US Pharmacopeia method. The sensors have been used as indicator electrodes for potentiometric titration of moxifloxacin.

Keywords: potentiometry, PVC, membrane sensors, ion-pair, ionophore, schiff-base, moxifloxacin HCl, sodium tetraphenyl borate, phosphomolybdic acid, phosphotungstic acid

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Authors: Ravi R. Patel, Vasudev R. Thakkar

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Use of plant growth-promoting rhizobacteria (PGPR) to increase the production and decrees disease occurrence is a recent method in agriculture. An RA2 rhizospheric culture was isolated from peanut rhizosphere from Junagadh region of Gujarat, India and showed different direct and indirect plant growth promoting activity like indole acetic acid, gibberellic acid, siderophore, hydrogen cyanide, Ammonia and (1-Aminocyclopropane-1-Carboxylate) deaminase production, N2 fixation, phosphate and potassium solubilization in vitro. RA2 was able to protect peanut germinating seedling from A. niger infection and reduce collar rot disease incidence 60-35% to 72-41% and increase germination percentage from 70-82% to 75-97% in two varieties GG20 and GG2 of peanut. RA2 was found to induce resistance in A. hypogaea L. seedlings via induction of different defense-related enzymes like phenylalanine ammonia lyase, peroxidase, polyphenol oxidase, lipoxygenase and pathogenesis related protein like chitinase, ß – 1,3- glucanase. Jasmonic acid one of the major signaling molecules of inducing systemic resistance was also found to induced due to RA2 treatments. RA2 bacterium was also promoting peanut growth and reduce A. niger infection in pot studies. 16S rDNA sequence of RA2 showed 99 % homology to Azotobacter species.

Keywords: plant growth promoting rhizobacteria, peanut, aspergillus niger, induce systemic resistance

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Authors: Tímea Gerzsenyi, Ágnes M. Ilosvai, László Vanyorek, Emma Szőri-Dorogházi

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DNA isolation is a crucial step in many molecular biological applications for diagnostic and research purposes. However, traditional extraction requires toxic reagents, and commercially available kits are expensive, this leading to the recently wide-spread method, the magnetic nanoparticle (MNP)-based DNA isolation. Different ferrite containing MNPs were examined and compared in their plasmid DNA isolation efficiency. Among the tested MNPs, one has never been used for the extraction of plasmid molecules, marking a distinct application. pDNA isolation process was optimized for each type of nanoparticle and the best protocol was selected based on different criteria: DNA quantity, quality and integrity. With the best-performing magnetic nanoparticle, which excelled in all aspects, further tests were performed to recover genomic DNA from bacterial cells and a protocol was developed.

Keywords: DNA isolation, nanobiotechnology, magnetic nanoparticles, protocol optimization, pDNA, gDNA

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Authors: Monika Patel, Kazuaki Matsumura

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Synthetic hydrogels, with their unique properties such as porosity, strength, and swelling in aqueous environment, are being used in many fields from food additives to regenerative medicines, from diagnostic and pharmaceuticals to drug delivery systems (DDS). But, hydrogels also have some limitations in terms of homogeneity of drug distribution and quantity of loaded drugs. As an alternate, polymeric micelles are extensively used as DDS. With the ease of self-assembly, and distinct stability they remarkably improve the solubility of hydrophobic drugs. However, presently, combinational therapy is the need of time and so are systems which are capable of releasing more than one drug. And it is one of the major challenges towards DDS to control the release of each drug independently, which simple DDS cannot meet. In this work, we present an amphiphilic polypeptide based micelle hydrogel composite to study the dual drug release for wound healing purposes using Amphotericin B (AmpB) and Curcumin as model drugs. Firstly, two differently charged amphiphilic polypeptide chains were prepared namely, poly L-Lysine-b-poly phenyl alanine (PLL-PPA) and poly Glutamic acid-b-poly phenyl alanine (PGA-PPA) through ring opening polymerization of amino acid N-carboxyanhydride. These polymers readily self-assemble to form micelles with hydrophobic PPA block as core and hydrophilic PLL/PGA as shell with an average diameter of about 280nm. The thus formed micelles were loaded with the model drugs. The PLL-PPA micelle was loaded with curcumin and PGA-PPA was loaded with AmpB by dialysis method. Drug loaded micelles showed a slight increase in the mean diameter and were fairly stable in solution and lyophilized forms. For forming the micelles hydrogel composite, the drug loaded micelles were dissolved and were cross linked using genipin. Genipin uses the free –NH2 groups in the PLL-PPA micelles to form a hydrogel network with free PGA-PPA micelles trapped in between the 3D scaffold formed. Different composites were tested by changing the weight ratios of the both micelles and were seen to alter its resulting surface charge from positive to negative with increase in PGA-PPA ratio. The composites with high surface charge showed a burst release of drug in initial phase, were as the composites with relatively low net charge showed a sustained release. Thus the resultant surface charge of the composite can be tuned to tune its drug release profile. Also, while studying the degree of cross linking among the PLL-PPA particles for effect on dual drug release, it was seen that as the degree of crosslinking increases, an increase in the tendency to burst release the drug (AmpB) is seen in PGA-PPA particle, were as on the contrary the PLL-PPA particles showed a slower release of Curcumin with increasing the cross linking density. Thus, two different pharmacokinetic profile of drugs were seen by changing the cross linking degree. In conclusion, a unique charged amphiphilic polypeptide based micelle hydrogel composite for dual drug delivery. This composite can be finely tuned on the basis of need of drug release profiles by changing simple parameters such as composition, cross linking and pH.

Keywords: amphiphilic polypeptide, dual drug release, micelle hydrogel composite, tunable DDS

Procedia PDF Downloads 207

Authors: Saeed Ahmed, Muhammad Imran, Yasir Allah Ditta, Shahid Mehmood, Zahid Rasool

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A study was conducted to investigate the effect of different levels of microencapsulated butyric (MEB) on growth performance, gut health and immunity in commercial broiler chickens. In total, 336 day-old Hubbard classic broilers chicks were randomly assigned to 4 dietary treatments (Control, 0.25, 0.35 and 0.45g/kg of butyric acid) under completely randomized design. Each treatment was replicated 3 times with 28 birds in each replicate. Feed intake, body weight gain, feed conversion ratio, intestinal morphology, apparent ileal digestibility of protein and immunity parameters were evaluated. At the end of the experiment (35-d) 3 birds/replicate in each group were randomly selected and slaughtered to collect blood, duodenal samples and ileal digesta. The data were analyzed by using ANOVA technique. The results indicated improved body weight gain (P = 0.0222), feed conversion ratio (P = 0.0056), duodenal villus height (P = 0.0512), AID (P = 0.0098) antibody titer against Newcastle disease improved (P = 0.0326). Treatments remained unresponsive with respect to feed intake (P = 0.9685).

Keywords: butyric acid, broilers, gut health, ileal digestibility

Procedia PDF Downloads 324

Authors: Muhammad Iqbal, Hafiz Muhammad Tauqeer, Hamza Rafaqat, Muhammad Naveed, Muhammad Awais Irshad

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Soil pollution with chromium (Cr) has become one of the most important concerns due to its toxicity for humans. To date, various remediation approaches have been employed for the remediation and management of Cr contaminated soils. Phytoextraction is an eco-friendly and emerging remediation approach which has gained attention due to several advantages over conventional remediation approach. The use of energy crops for phytoremediation is an emerging trend worldwide. These energy crops have high tolerance against various environmental stresses, the potential to grow in diverse ecosystems and high biomass production make them a suitable candidate for phytoremediation of contaminated soils. The removal efficiency of plants in phytoextraction depends upon several soil and plant factors including solubility, bioavailability and metal speciation in soils. A pot scale experiment was conducted to evaluate the phytoextraction potential of Arundo donax L. with the application of acetic acid (A.A) in Cr contaminated soils. Plants were grown in pots filled with 5 kg soils for 90 days. After 30 days plants acclimatization in pot conditions, plants were treated with various levels of Cr (2.5 mM, 5 mM, 7.5 mM, 10 mM) and A.A (Cr 2.5 mM + A.A 2.5 mM, Cr 5 mM + A.A 2.5 mM, Cr 7.5 mM + A.A 2.5 mM, Cr 10 mM + A.A 2.5 mM). The application of A.A significantly increased metal uptake and in roots and shoots of A. donax. This increase was observed at Cr 7.5 mM + A.A 2.5 mM but at high concentrations, visual symptoms of Cr toxicity were observed on leaves. Similarly, A.A applications also affect the activities of key enzymes including catalase (CAT), superoxidase dismutase (SOD), and ascorbate peroxidase (APX) in leaves of A. donax. Based on results it is concluded that the applications of A.A acid for phytoextraction is an alternative approach for the management of Cr affected soils and synthetic chelators should be replaced with organic acids.

Keywords: acetic acid, A. donax, chromium, energy crop, phytoextraction

Procedia PDF Downloads 388

Authors: Beata Pospiech

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Separation of metal ions from aqueous solutions is important as well as difficult process in hydrometallurgical technology. This process is necessary for obtaining of clean metals. Solvent extraction and membrane processes are well known as separation methods. Recently, ionic liquids (ILs) are very often applied and studied as extractants and carriers of metal ions from aqueous solutions due to their good extractability properties for various metals. This work discusses a method to separate copper(II) and iron(III) from hydrochloric acid solutions by solvent extraction and transport across polymer inclusion membranes (PIM) with the selected ionic liquids as extractants/ion carriers. Cyphos IL 101 (trihexyl(tetradecyl)phosphonium chloride), Cyphos IL 104 (trihexyl(tetradecyl)phosphonium bis(2,4,4 trimethylpentyl)phosphi-nate), trioctylmethylammonium thiosalicylate [A336][TS] and trihexyl(tetradecyl)phosphonium thiosalicylate [PR4][TS] were used for the investigations. Effect of different parameters such as hydrochloric acid concentration in aqueous phase on iron(III) and copper(II) extraction has been investigated. Cellulose triacetate membranes with the selected ionic liquids as carriers have been prepared and applied for transport of iron(IIII) and copper(II) from hydrochloric acid solutions.

Keywords: copper, iron, ionic liquids, solvent extraction

Procedia PDF Downloads 279

Authors: Payam Rasoulnia, Seyyed Mohammad Mousavi

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Heavy fuel oil firing power plants produce huge amounts of ashes as solid wastes, which seriously need to be managed and processed. Recycling precious metals of V and Ni from these oil-fired ashes which are considered as secondary sources of metals recovery, not only has a great economic importance for use in industry, but also it is noteworthy from the environmental point of view. Vanadium is an important metal that is mainly used in the steel industry because of its physical properties of hardness, tensile strength, and fatigue resistance. It is also utilized in oxidation catalysts, titanium–aluminum alloys and vanadium redox batteries. In the present study bioleaching of vanadium and nickel from an oil-fired ash sample was conducted using Aspergillus niger fungus. The experiments were carried out using spent-medium bioleaching method in both Erlenmeyer flasks and also bubble column bioreactor, in order to compare them together. In spent-medium bioleaching the solid waste is not in direct contact with the fungus and consequently the fungal growth is not retarded and maximum organic acids are produced. In this method the metals are leached through biogenic produced organic acids present in the medium. In shake flask experiments the fungus was cultured for 15 days, where the maximum production of organic acids was observed, while in bubble column bioreactor experiments a 7 days fermentation period was applied. The amount of produced organic acids were measured using high performance liquid chromatography (HPLC) and the results showed that depending on the fermentation period and the scale of experiments, the fungus has different major lixiviants. In flask tests, citric acid was the main produced organic acid by the fungus and the other organic acids including gluconic, oxalic, and malic were excreted in much lower concentrations, while in the bioreactor oxalic acid was the main lixiviant and it was produced considerably. In Erlenmeyer flasks during 15 days fermentation of Aspergillus niger, 8080 ppm citric acid and 1170 ppm oxalic acid was produced, while in bubble column bioreactor over 7 days of fungal growth, 17185 ppm oxalic acid and 1040 ppm citric acid was secreted. The leaching tests using the spent-media obtained from both of fermentation experiments, were performed at the same conditions of leaching duration of 7 days, leaching temperature of 60 °C and pulp density up to 3% (w/v). The results revealed that in Erlenmeyer flask experiments 97% of V and 50% of Ni were extracted while using spent medium produced in bubble column bioreactor, V and Ni recoveries were achieved to 100% and 33%, respectively. These recovery yields indicate that in both scales almost total vanadium can be recovered, while nickel recovery was lower. With help of the bioreactor spent-medium nickel recovery yield was lower than that of obtained from the flask experiments, which it could be due to precipitation of some values of Ni in presence of high levels of oxalic acid existing in its spent medium.

Keywords: Aspergillus niger, bubble column bioreactor, oil-fired ash, spent-medium bioleaching

Procedia PDF Downloads 229

Authors: Taiwo O. Margaret, Olaoluwa O. Olaoluwa

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Moraceae family has immense phytochemical constituents and significant pharmacological properties, hence have great medicinal values. The aim of this study was to screen and quantify phytochemicals as well as the antioxidant activities of the leaf and stem bark extracts and fractions (crude ethanol extracts, n-hexane, ethyl acetate and aqueous ethanol fractions) of Ficus sagittifolia. Leaf and stem bark of F. sagittifolia were extracted by maceration method using ethanol to give ethanol crude extract. The ethanol crude extract was partitioned by n-hexane and ethyl-acetate to give their respective fractions. All the extracts were screened for their phytochemicals using standard methods. The total phenolic, flavonoid, tannin, saponin contents and antioxidant activity were determined by spectrophotometric method while the alkaloid content was evaluated by titrimetric method. The amount of total phenolic in extracts and fractions were estimated in comparison to gallic acid, whereas total flavonoids, tannins and saponins were estimated corresponding to quercetin, tannic acid and saponin respectively. 2, 2-diphenylpicryl hydrazyl radical (DPPH)* and phosphomolybdate methods were used to evaluate the antioxidant activities of leaf and stem bark of F. sagittifolia. Phytochemical screening revealed the presence of flavonoids, saponins, terpenoids/steroids, alkaloids for both extracts of leaf and stem bark of F. sagittifolia. The phenolic content of F. sagittifolia was most abundant in leaf ethanol crude extract as 3.53 ± 0.03 mg/g equivalent of gallic acid. Total flavonoids and tannins content were highest in stem bark aqueous ethanol fraction of F. sagittifolia estimated as 3.41 ± 0.08 mg/g equivalent of quercetin and 1.52 ± 0.05 mg/g equivalent of tannic acid respectively. The hexane leaf fraction of F. sagittifolia had the utmost saponin and alkaloid content as 5.10 ± 0.48 mg/g equivalent of saponins and 0.171 ± 0.39 g of alkaloids. Leaf aqueous ethanol fraction of F. sagittifolia showed high antioxidant activity (IC₅₀ value of 63.092 µg/mL) and stem ethanol crude extract (227.43 ± 0.78 mg/g equivalent of ascorbic acid) for DPPH and phosphomolybdate method respectively and the least active was found to be the stem hexane fraction using both methods (313.32 µg/mL; 16.21 ± 1.30 mg/g equivalent of ascorbic acid). The presence of these phytochemicals in the leaf and stem bark of F. sagittifolia are responsible for their therapeutic importance as well as the ability to scavenge free radicals in living systems.

Keywords: Moraceae, Ficus sagittifolia, phytochemicals, antioxidant

Procedia PDF Downloads 230

Authors: Ezaldin A. M. Mohammed, Youssef K. H. Abdalhafid, Masoud. M. Zatout

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The study aims to clarify the toxic effect of plant hormones, which are widely used in agriculture. One of these is the plant hormones (indole acetic acid); has been ٳata hormone to rats at 100 ppm salt solution of 0.2 per day after day for a period of forty days before conception until the fourteenth day or sixteenth or childbirth. Treatment brought about a marked shortage in the rate of increase in the weight of mice., And a percentage of the weight of the liver there was a distinct increase in the relative weight of the liver. As well as the increase in pathological changes and increase the size of the nuclei and Kupffer cell, as noted widespread and dense clusters of inflammatory cells accompanied by about the erosion of liver tissue and blood ٳrchah. Biochemical analyzes showed a marked decrease of the liver in antioxidant enzymes and an increase in the rate of free radicals. It was also noted an increase in cases of abortion. The owner of so many birth defects. It was also noted the lack of body weight in fetuses and increase the absorption rate of embryos in fetuses of mothers treatment compared to the control group. Showed microscopic examinations of the liver of mice born in the transaction and the decay in the presence of hepatic cells and edema, blood vessels and increase the rate of cell death.

Keywords: indol acetic acid, liver, pathological changes, albino rats

Procedia PDF Downloads 400

Authors: M. Koksal, T. Ozyazici, E. Gurdal, M. Yarım, E. Demirpolat, M. B. Y. Aycan

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The discovery of new drugs in cancer chemotherapy is still a major topic because of severe side effects, selectivity problems and resistance development potential of existing drugs. In recent years, combined anticancer therapies or multi-acting drugs are clinically preferred over traditional cytotoxic treatment, with the aim of avoiding resistance and toxic side effects. Arrangement of multi-acting targets can be carried out either by combination of several drugs with different mechanisms or by usage of a single chemical compound capable of regulating several targets of a disease with multiple factors. In literature, several pyrimidine and piperazine derivatives have been involved in the structure of many compounds which have been used as chemotherapeutic agents along with wide clinical applications. The aim of this study is to combine pyrimidine and piperazine core structures to research and develop novel piperazinylpyrimidine derivatives with selective cytotoxicity over cancer cells. In this study, a group of novel 6-fluorophenyl-3-[2-(substitutedpiperazinyl)ethyl] hexahydropyrimidine-2,4-dione derivatives designed to observe the desired anticancer activity due to pyrimidine and piperazine based scaffolds. Target compounds were obtained by the reaction of appropriate piperazine derivatives and 6-(2/4-fluorophenyl)-3-(2-chloroethyl)hexahydropyrimidine-2,4-dione. The synthetic pathway of 6-(2/4-fluorophenyl)-3-(2-chloroethyl)hexahydropyrimidine-2,4-dione was started with Rodionov reaction using aldehyde, malonic acid and ammonium acetate in ethanol. Isolated β-fluorophenyl-β-amino acids were treated with 2-chloroethylisocyanate in the presence of an aqueous sodium hydroxide solution at room temperature to yield the sodium salts of the corresponding ureido acids. By addition of a mineral acid, ureido acids were precipitated. Later, these ureido acids were refluxed in thionyl chloride to give the 6-(2/4-fluorophenyl)-3-(2-chloroethyl)hexahydropyrimidine-2,4-di-one which were furthermore treated with secondary amines. Structures of purified compounds were characterized with IR, 1H-NMR, 13C-NMR, mass spectroscopies and elemental analysis. All of the compounds gave satisfactory analytical and spectroscopic data, which were in full accordance with their depicted structures. In IR spectra of the compounds, N-H group was seen at 3230-3213 cm⁻¹. C-H was seen at 3100-2820 cm⁻¹ and C=O vibrational peaks were observed approximately at 1725 and 1665 cm⁻¹ in accordance with literature. In the NMR spectra of target compounds, the methylene protons of piperazine give two separate multiplet peaks around 3.5 and 4.5 ppm representing the successful N-alkylation of the structure. The cytotoxic activity of the synthesized compounds was investigated on human bronchial epithelial (BEAS 2B), lung (A549), colon adenocarcinoma (COLO205) and breast (MCF7) cell lines, by means of sulphorhodamine B (SRB) assays in triplicate. IC₅₀ values of the screened derivatives were found in range of 11.8-78 µM. This project was supported by The Scientific and Technological Research Council of Turkey (TUBITAK, Project no: 215S157).

Keywords: cytotoxicity, hexahydropyrimidine, piperazine, sulphorhodamine B assay

Procedia PDF Downloads 152

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

Procedia PDF Downloads 184

Authors: M. L. Mangena, N. Mokoena, K. Rashamuse, M. G. Tlou

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Tertiary alcohol ester (TAE) hydrolases are a group of esterases (EC 3.1.1.-) that catalyze the kinetic resolution of TAEs and as a result, they are sought-after for the production of optically pure tertiary alcohols (TAs) which are useful as building blocks for number biologically active compounds. What sets these enzymes apart is, the presence of a GGG(A)X-motif in the active site which appears to be the main reason behind their activity towards the sterically demanding TAEs. The genome of Pseudomonas syringae pv. maculicola (Psm) comprises a multitude of genes that encode esterases. We therefore, hypothesize that some of these genes encode TAE hydrolases. In this study, Psm was screened for TAE hydrolase activity using the linalyl acetate (LA) plate assay and a positive reaction was observed. As a result, the genome of Psm was screened for esterases with a GGG(A)X-motif using the motif search tool and two potential TAE hydrolase genes (PsmEST1 and 2, 1100 and 1000bp, respectively) were identified, PsmEST1 was amplified by PCR and the gene sequenced for confirmation. Analysis of the sequence data with the SingnalP 4.1 server revealed that the protein comprises a signal peptide (22 amino acid residues) on the N-terminus. Primers specific for the gene encoding the mature protein (without the signal peptide) were designed such that they contain NdeI and XhoI restriction sites for directional cloning of the PCR products into pET28a. The gene was expressed in E. coli JM109 (DE3) and the clones screened for TAE hydrolase activity using the LA plate assay. A positive clone was selected, overexpressed and the protein purified using nickel affinity chromatography. The activity of the esterase towards LA was confirmed using thin layer chromatography.

Keywords: hydrolases, tertiary alcohol esters, tertiary alcohols, screening, Pseudomonas syringae pv., maculicola genome, esterase activity, linalyl acetate

Procedia PDF Downloads 355

Authors: Eleftheria Barouni, Antonia Terpou, Maria Kanellaki, Argyro Bekatorou, Athanasios A.Koutinas

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The aim of the present work was to evaluate the production of cheese using a composite filter of tubular cellulose (TC) with [a] entrapped rennin enzyme and [b] immobilized L.casei and entrapped enzyme. Tubular cellulose from sawdust was prepared after lignin removal with 1% NaOH. The biocatalysts were thermally dried at 38oC and used for milk coagulation. The effect of temperature (5,20,37 oC) of the first dried biocatalyst on the pH kinetics of milk coagulation was examined. The optimum temperature (37oC) of the first biocatalyst was used for milk coagulation with the second biocatalyst prepared by entrapment of both rennin enzyme and probiotic lactic acid bacteria in order to introduce a sour taste in cheeses. This co-biocatalyst was used for milk coagulation. Samples were studied as regards its effect on lactic acid formation and its correlation with taste test results in cheeses. For both biocatalysts samples were analyzed for total acidity and lactic acid formation by HPLC. The quality of the produced cheeses was examined through the determination of volatile compounds by SPME GC/MS analysis. Preliminary taste tests and microbiological analysis were performed and encourage us for further research regarding scale up.

Keywords: tubular cellulose, Lactobacillus casei, rennin enzyme, cheese production

Procedia PDF Downloads 358