Search results for: antioxidant enzyme

Authors: Michela Terlizzi, Chiara Colarusso, Aldo Pinto, Rosalinda Sorrentino

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Sphingosine-1-phosphate (S1P) is a membrane-derived bioactive phospholipid exerting a multitude of effects on respiratory cell physiology and pathology through five S1P receptors (S1PR1-5). Higher levels of S1P have been registered in a broad range of respiratory diseases, including inflammatory disorders and cancer, although its exact role is still elusive. Based on our previous study in which we found that S1P/S1PR3 is involved in an inflammatory pattern via the activation of Toll-like Receptor 9 (TLR9), highly expressed on lung cancer cells, the main goal of the current study was to better understand the involvement of S1P/S1PR3 pathway/signaling during lung carcinogenesis, taking advantage of a mouse model of first-hand smoke exposure and of carcinogen-induced lung cancer. We used human samples of Non-Small Cell Lung Cancer (NSCLC), a mouse model of first-hand smoking, and of Benzo(a)pyrene (BaP)-induced tumor-bearing mice and A549 lung adenocarcinoma cells. We found that the intranuclear, but not the membrane, localization of S1PR3 was associated to the proliferation of lung adenocarcinoma cells, the mechanism that was correlated to human and mouse samples of smoke-exposure and carcinogen-induced lung cancer, which were characterized by higher utilization of S1P. Indeed, the inhibition of the membrane S1PR3 did not alter tumor cell proliferation after TLR9 activation. Instead, according to the nuclear localization of sphingosine kinase (SPHK) II, the enzyme responsible for the catalysis of the S1P last step synthesis, the inhibition of the kinase completely blocked the endogenous S1P-induced tumor cell proliferation. These results prove that the endogenous TLR9-induced S1P can on one side favor pro-inflammatory mechanisms in the tumor microenvironment via the activation of cell surface receptors, but on the other tumor progression via the nuclear S1PR3/SPHK II axis, highlighting a novel molecular mechanism that identifies S1P as one of the crucial mediators for lung carcinogenesis-associated inflammatory processes and that could provide differential therapeutic approaches especially in non-responsive lung cancer patients.

Keywords: sphingosine-1-phosphate (S1P), S1P Receptor 3 (S1PR3), smoking-mice, lung inflammation, lung cancer

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Authors: Vandana Deopanée Tomar, Gurpreet Kaur Sidhu, Panchsheela Nogia, Rajesh Mehrotra, Sandhya Mehrotra

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The cyanobacterial CO₂ concentrating mechanism (CCM) operates to raise the levels of CO₂ in the vicinity of the main carboxylation enzyme Rubisco which is encapsulated in protein micro compartments called carboxysomes. Thus, due to the presence of CCM, cyanobacterial cells are able to work with high photosynthetic efficiency even at low Ci conditions and can accumulate 1000 folds high internal concentrations of Ci than external environment. Engineering of some useful CCM components into higher plants is one of the plausible approaches to improve their photosynthetic performance. The first step and the simplest approach for attaining this objective would be the transfer of cyanobacterial bicarbonate transporter such as SbtA to inner chloroplast envelope of C₃ plants. For this, SbtA transporter gene from Synechococcus elongatus PCC 7942 was fused to a transit peptide element to generate chimeric constructs in order to direct it to chloroplast inner envelope. Two transit peptides namely, TnaXTP (transit peptide from AT3G56160) and TMDTP (transit peptide from AT2G02590) were shortlisted from Arabidopsis thaliana genome and cloned in plant expression vector pCAMBIA1302 having mgfp5 as a reporter gene. Plant transformation was done by agro infiltration and Agrobacterium mediated co-culture. DNA, RNA, and protein were isolated from the leaves four days post infiltration, and the presence of transgene was confirmed by gene specific PCR (Polymerase Chain Reaction) analysis and by RT-PCR (Reverse Transcription Polymerase Chain Reaction). The expression was confirmed at the protein level by western blotting using anti-GFP primary antibody and horseradish peroxidase (HRP) conjugated secondary antibody. The localization of the protein was detected by confocal microscopy of isolated protoplasts. We observed chloroplastic expression for both the fusion constructs which suggest that the transit peptide sequences are capable of taking the cargo protein to the chloroplasts. These constructs are now being used to generate stable transgenic plants by Agrobacterium mediated transformation. The stability of transgene expression will be analyzed from T₀ to T₂ generation.

Keywords: agro infiltration, bicarbonate transporter, carbon concentrating mechanisms, cyanobacteria, SbtA

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Authors: Manal M. Shehata, Kawther M. Abdel-Hamid, Nashwa A. Mohamed, Marwa H. Bakr, Maged S. Mahmoud, Hala M. Elbadre

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Introduction: The worldwide rapid increase in diabetes poses a significant challenge to current therapeutic approaches. Therapeutic utility of bone marrow transplantation in diabetes is an attractive approach. However, the oxidative stress generated by hyperglycemia may hinder β-cell regeneration. Curcumin, is a dietary spice with antioxidant activity. Aim of work: The present study was undertaken to investigate the therapeutic potential of curcumin, bone marrow transplantation, and their combined effects in the reversal of experimental diabetes. Material and Methods: Fifty adult male healthy albino rats were included in the present study.They were divided into two groups: Group І: (control group) included 10 rats. Group П: (diabetic group): included 40 rats. Diabetes was induced by single intraperitoneal injection of streptozotocin (STZ). Group II will be further subdivided into four groups (10 rats for each): Group II-a (diabetic control). Group II-b: rats were received single intraperitoneal injection of bone marrow suspension (un-fractionated bone marrow cells) prepared from rats of the same family. Group II-c: rats were treated with curcumin orally by gastric intubation for 6 weeks. Group II-d: rats were received a combination of single bone marrow transplantation and curcumin for 6 weeks. After 6 weeks, blood glucose, insulin levels were measured and the pancreas from all rats were processed for Histological, Immunohistochemical and morphometric examination. Results: Diabetic group, showed progressive histological changes in the pancreatic islets. Treatment with either curcumin or bone marrow transplantation improved the structure of the islets and reversed streptozotocin-induced hyperglycemia and hypoinsulinemia. Combination of curcumin and bone marrow transplantation elicited more profound alleviation of streptozotocin-induced changes including islet regeneration and insulin secretion. Conclusion: The use of natural antioxidants combined with bone marrow transplantation to induce pancreatic regeneration is a promising strategy in the management of diabetes.

Keywords: diabtes, panceatic islets, bone marrow transplantation, curcumin

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Authors: Daniela Nedeltcheva-Antonova, Kamelia Getchovska, Vera Deneva, Stanislav Bozhanov, Liudmil Antonov

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Rosa damascena Mill. (Damask rose) is one of the most important plants belonging to the Rosaceae family, with a long historical use in traditional medicine and as a valuable oil-bearing plant. Many pharmacological effects have been reported from this plant, including anti-inflammatory, hypnotic, analgesic, anticonvulsant, anti-depressant, antianxiety, antitussive, antidiabetic, relaxant effects on tracheal chains, laxative, prokinetic and hepatoprotective activities. Pharmacological studies have shown that the various health effects of R. damascena flowers can mainly be attributed to its large amount of polyphenolic components. Phenolics possess a wide range of pharmacological activities, such as antioxidants, free-radical scavengers, anticancer, anti-inflammatory, antimutagenic, and antidepressant, with flavonoids being the most numerous group of natural polyphenolic compounds. According to the technological process in the production of rose concrete (solvent extraction with non-polar solvents of fresh rose flowers), it can be assumed that the resulting plant residue would be as rich of polyphenolics, as the plant itself, and could be used for the development of novel products with promising health-promoting effect. Therefore, an optimisation of the extraction procedure of the by-product from the rose concrete production was carried out. An assay of the extracts in respect of their total polyphenols and total flavonoids content was performed. HPLC analysis of quercetin and kaempferol, the two main flavonoids found in R. damascena, was also carried out. The preliminary results have shown that the flavonoid content in the rose extracts is comparable to that of the green tea or Gingko biloba, and they could be used for the development of various products (food supplements, natural cosmetics and phyto-pharmaceutical formulation, etc.). The fact that they are derived from the by-product of industrial plant processing could add the marketing value of the final products in addition to the well-known reputation of the products obtained from Bulgarian roses (R. damascena Mill.).

Keywords: gas chromatography-mass-spectromrtry, dry extract, flavonoids, Rosa damascena Mill

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Authors: Amina Omer, Ikram Elsiddig

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Arctostaphylos uvaursi is a plant of the family Ericaceae, it’s used in skin care products mostly for its depigmenting action, due to the presence of hydroquinones that are well known inhibitors of tyrosinase, an enzyme involved in melanin biosynthesis in humans. The main hydroquinone found within the A. uvaursi is arbutin, which is found with varying percentage within the plant depending on the season, and area from which the plant is harvested. An in vitro experiment has shown that the arbutin found within the bearberry leaf extract inhibited the biosynthesis of melanin in human melanoma cells and in three-dimensional human skin model. Madurella mycetomatis is filamentous fungus that causes the fungal form of mycetoma known as eumycetoma, with existing anti-fungals and surgery, only 35% of people living eumycetoma are treated, M. mycetomatis has been found to shield itself against the antifungal therapy through the production of melanin decreasing the effectiveness of the therapy, therefore there is a need for a new and more effective therapy. The aim of the study was to investigate and compare the effect of arbutin powder and aqueous extract of A. uvaursi containing arbutin on the biosynthesis of melanin by M. mycetomatis. The experiment was carried out by culturing M. mycetomatis on minimal media composed of 2% agar, 15 mM glucose, 10 mM MgSO4, 29.4 mM KH2PO4, 13 mM glycin and 80mg/l gentamicin, the media was supplied with different concentration of arbutin solution (5, 25 50,and 75mg) and aqueous extract of A. uvaursi to contain arbutin with concentrations (5, 25 50,and 75mg), the plates were incubated for two month and the result was observed by the naked eye. The results revealed that the arbutin powder had an inhibitory effect on melanin synthesis by M. mycetomatis that correlated with its established inhibitory effect on melanin synthesis in humans. The inhibitory effect of arbutin on melanin synthesis by M. mycetomatis was found to be dose dependent. A. uvaursi aqueous leaf extract containing arbutin was also found to decrease melanin production by M. mycetomatis, however plates containing high concentrations of aqueous extract couldn’t be assessed for its melanin inhibitory effect due to the high content of carbohydrates in the extract that promoted the growth of fungi Asperigullus niger rendering the plates unsuitable for visual inspection. In conclusion inhibition of melanin synthesis was observed on the arbutin powder as well as the aqueous extract containing arbutin. A. uvaursi is known to exhibit anti-inflammatory activity, which can aid in wound healing that is beneficial in the chronic inflammation caused by M. mycetomatis.

Keywords: arbutin, arctostaphylos, Madurella, melanin

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Authors: Hoda M. Eid, Abir Nachar, Farah Thong, Gary Sweeney, Pierre S. Haddad

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Caffeic acid methyl ester (CAME) and caffeic ethyl esters (CAEE) were previously reported to potently stimulate glucose uptake in cultured C2C12 skeletal muscle cells via insulin-independent mechanisms involving the activation of adenosine monophosphate-activated protein kinase (AMPK). In the present study, we investigated the effect of the two compounds on the translocation of glucose transporter GLUT4 in L6 skeletal muscle cells. The cells were treated with the optimum non-toxic concentration (50 µM) of either CAME or CAEE for 18 h. Levels of GLUT4myc at the cell surface were measured by O-phenylenediamine dihydrochloride (OPD) assay. The effects of CAME and CAEE on GLUT1 and GLUT4 protein content were also measured by western immunoblot. Our results show that CAME and CAEE significantly increased glucose uptake, GLUT4 translocation and GLUT4 protein content. Furthermore, the effect of the two CA esters on two insulin-sensitive cell lines: H4IIE rat hepatoma and 3T3-L1 adipocytes were investigated. CAME and CAEE reduced the enzymatic activity of the key hepatic gluconeogenic enzyme glucose-6-phosphatase in a concentration-dependent manner. In addition, they exerted a concentration-dependent antiadipogenic effect on 3T3-L1 cells. Mitotic clonal expansion (MCE), a prerequisite for adipocytes differentiation was also concentration-dependently inhibited. The two compounds abrogated lipid droplet accumulation, blocked MCE and maintained cells in fibroblast-like state when applied at the maximum non-toxic concentration (100 µM). In addition, the expression of the early key adipogenic transcription factors CCAAT enhancer-binding protein beta (C/EBP-β) and the master regulator of adipogenesis peroxisome-proliferator-activated receptor gamma (PPAR-γ) were inhibited. We, therefore, conclude that CAME and CAEE exert pleiotropic benefits in several insulin-sensitive cell lines through insulin-independent mechanisms involving AMPK, hence they may treat obesity, diabetes and other metabolic diseases.

Keywords: type 2 diabetes mellitus, insulin resistance, GLUT4, Akt, AMPK.

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Authors: Zainab S. Ahmed, Mohammed S. Ali, Nadia A. Elshiekh, Sami Adam Ibrahim, Ghada M. El-Tayeb, Ahmed H. Elsadig, Rihab A. Omer, Sofia B. Mohamed

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Maple syrup urine (MSUD) is an autosomal recessive disease that causes a deficiency in the enzyme branched-chain alpha-keto acid (BCKA) dehydrogenase. The development of disease has been associated with SNPs in the DBT gene. Despite that, the computational analysis of SNPs in coding and noncoding and their functional impacts on protein level still remains unknown. Hence, in this study, we carried out a comprehensive in silico analysis of missense that was predicted to have a harmful influence on DBT structure and function. In this study, eight different in silico prediction algorithms; SIFT, PROVEAN, MutPred, SNP&GO, PhD-SNP, PANTHER, I-Mutant 2.0 and MUpo were used for screening nsSNPs in DBT including. Additionally, to understand the effect of mutations in the strength of the interactions that bind protein together the ELASPIC servers were used. Finally, the 3D structure of DBT was formed using Mutation3D and Chimera servers respectively. Our result showed that a total of 15 nsSNPs confirmed by 4 software (R301C, R376H, W84R, S268F, W84C, F276C, H452R, R178H, I355T, V191G, M444T, T174A, I200T, R113H, and R178C) were found damaging and can lead to a shift in DBT gene structure. Moreover, we found 7 nsSNPs located on the 2-oxoacid_dh catalytic domain, 5 nsSNPs on the E_3 binding domain and 3 nsSNPs on the Biotin Domain. So these nsSNPs may alter the putative structure of DBT’s domain. Furthermore, we detected all these nsSNPs are on the core residues of the protein and have the ability to change the stability of the protein. Additionally, we found W84R, S268F, and M444T have high significance, and they affected Leucine, Isoleucine, and Valine, which reduces or disrupt the function of BCKD complex, E2-subunit which the DBT gene encodes. In conclusion, based on our extensive in-silico analysis, we report 15 nsSNPs that have possible association with protein deteriorating and disease-causing abilities. These candidate SNPs can aid in future studies on Maple Syrup Urine Disease type II base in the genetic level.

Keywords: DBT gene, ELASPIC, in silico analysis, UCSF chimer

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Authors: A. O. Omonona, T. A. Jarikre

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Increase specialization in agriculture and use of pesticides may inadvertently cause ecosystem degradation and eventually loss of biodiversity. The populations of numerous wildlife species have undergone a precipitous decline. Many of these problems have been attributed directly to habitat loss and over exploitation resulting from unregulated pesticide uses. Carbendazim a broad spectrum benzimidazole fungicide and a metabolite of benomyl, is used to control plant disease in cereals and fruit. The effect of carbendazim exposure and the ameliorative effect of tocopherol (vitamin E) were assessed on African giant rat AGR. Hematological, biochemical and histological changes were used to determine the health condition of the animals exposed to pesticide. Sixteen AGR were stabilized, weighed and then divided into four experimental groups (A to D). Two groups were pretreated with vitamin. Group A was exposed to carbendazim only, B- carbendazim + vitamin, C- vitamin only, and D- blank (control). Packed cell volume PCV was estimated by the microhematocrit method, Leucocyte and Platelet counts were determined using the hemocytometric method. Cholinesterase (AchE) and markers of oxidative stress were quantified, and tissue changes examined microscopically. There were no behavioral changes observed in the animals, but there was a decrease in body weight and abortion after 23 days of exposure to carbendazim. There was significant differences in the packed cell volume, the hemoglobin concentration and the red blood cell counts (p < 0.05). The increases in malonyl aldehyde MDA was significant (p < 0.05) in the pesticide intoxicated rats compared to control. Vitamin E supplementation reduced MDA level significantly (p < 0.05). There was a sharp remarkable decrease in acetylcholinesterase levels in the pesticide intoxicated rats (p < 0.05). Vitamin E supplementation normalise the AchE levels comparable to that in control. Grossly, the vital organs appeared normal in the pesticide exposed and control groups except moderate pulmonary congestion. Microscopically, there was severe diffuse hepatocellular swelling in carbendazim exposed group. The severity of hepatocellular injury was reduced in the rats with vitamin E. This study ascertained the toxic effect of carbendazim and antioxidative properties of vitamins in the Africa giant rat.

Keywords: African giant rat, antioxidant, carbendazim, pesticides, toxicity

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Authors: Mesih Kocamuftuoglu, Gonca Ozan, Enver Ozan, Nalan Kaya, Sema Temizer Ozan

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Metabolic Syndrome, one of the common metabolic disorder, occurs with co-development of insulin resistance, obesity, dislipidemia and hypertension problems. Insulin resistance appears to play a pathogenic role in the metabolic syndrome. Also, there is a relationship between insulin resistance and infertility as known. Turmeric (Curcuma longa L.) a polyphenolic chemical is widely used for its coloring, flavoring, and medicinal properties, and exhibits a strong antioxidant activity. In this study, we assess the effects of turmeric on rat uterine tissue in metabolic syndrome model induced by high fructose diet. Thirty-two adult female Wistar rats weighing 220±20 g were randomly divided into four groups (n=8) as follows; control, fructose, turmeric, and fructose plus turmeric. Metabolic syndrome was induced by fructose solution 20% (w/v) in tap water, and turmeric (C.Longa) administered at the dose of 80 mg/kg body weight every other day by oral gavage. After the experimental period of 8 weeks, rats were decapitated, serum and uterine tissues were removed. Serum lipid profile, glucose, insülin levels were measured. Uterine tissues were fixed for histological analyzes. The uterine tissue sections were stained with hematoxylin-eosin (H & E) stain, then examined and photographed on a light microscope (Novel N-800Mx20). As a result, fructose consumption effected serum lipids, insulin levels, and insulin resistance significantly. Endometrium and myometrium layers were observed in normal structure in control group of uterine tissues. Perivascular edema, peri glandular fibrosis, and inflammatory cell increase were detected in fructose group. Sections of the fructose plus turmeric group showed a significant improvement in findings when compared to the fructose group. Turmeric group cell structures were observed similar with the control group. These results demonstrated that high-fructose consumption could change the structure of the uterine tissue. On the other hand, turmeric administration has beneficial effects on uterine tissue at that dose and duration when administered with fructose.

Keywords: metabolic syndrome, rat, turmeric, uterus

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Authors: Eman M. Sarhan, Doaa A. Ghareeb, Gabriella Ortore, Amr A. Amara, Mohamed M. El-Sayed

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Drug salting and nanoparticle-based drug delivery formulations are considered to be an effective means for rendering the hydrophobic drugs’ nano-scale dispersion in aqueous media, and thus circumventing the pitfalls of their poor solubility as well as enhancing their membrane permeability. The current study aims to increase the bioavailability of quaternary ammonium berberine through acid salting and biodegradable bovine serum albumin (BSA)-based nanoparticulate drug formulation. Berberine hydroxide (BBR-OH) that was chemically synthesized by alkalization of the commercially available berberine hydrochloride (BBR-HCl) was then acidified to get Di-berberine sulfate (BBR)₂SO₄. The purified crystals were spectrally characterized. The desolvation technique was optimized for the preparation of size-controlled BSA-BBR-HCl, BSA-BBR-OH, and BSA-(BBR)₂SO₄ nanoparticles. Particle size, zeta potential, drug release, encapsulation efficiency, Fourier transform infrared spectroscopy (FTIR), tandem MS-MS spectroscopy, energy-dispersive X-ray spectroscopy (EDX), scanning and transmitting electron microscopic examination (SEM, TEM), in vitro bioactivity, and in silico drug-polymer interaction were determined. BSA (PDB ID; 4OR0) protonation state at different pH values was predicted using Amber12 molecular dynamic simulation. Then blind docking was performed using Lamarkian genetic algorithm (LGA) through AutoDock4.2 software. Results proved the purity and the size-controlled synthesis of berberine-BSA-nanoparticles. The possible binding poses, hydrophobic and hydrophilic interactions of berberine on BSA at different pH values were predicted. Antioxidant, anti-hemolytic, and cell differentiated ability of tested drugs and their nano-formulations were evaluated. Thus, drug salting and the potentially effective albumin berberine nanoparticle formulations can be successfully developed using a well-optimized desolvation technique and exhibiting better in vitro cellular bioavailability.

Keywords: berberine, BSA, BBR-OH, BBR-HCl, BSA-BBR-HCl, BSA-BBR-OH, (BBR)₂SO₄, BSA-(BBR)₂SO₄, FTIR, AutoDock4.2 Software, Lamarkian genetic algorithm, SEM, TEM, EDX

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Authors: Olarinde Olaniran, Oluyomi A. Sowemimo

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Toxoplasmosis is frequently misdiagnosed or underdiagnosed, and it is the third most common cause of hospitalization due to food-borne infection. Intra-uterine infection with Toxoplasma gondii due to active parasitaemia during pregnancy can cause severe and often fatal cerebral damage, abortion, and stillbirth of a fetus. The aim of the study was to investigate the prevalence of T. gondii infection in women of childbearing age in selected communities of Osun State with a view to determining the risk factors which predispose to the T. gondii infection. Five (5) ml of blood was collected by venopuncture into a plain blood collection tube by a medical laboratory scientist. Serum samples were separated by centrifuging the blood samples at 3000 rpm for 5 mins. The sera were collected with Eppendorf tubes and stored at -20°C analysis for the presence of IgG and IgM antibodies against T. gondii by commercially available enzyme-linked immunosorbent assay (ELISA) kit (Demeditec Diagnostics GmbH, Germany) conducted according to the manufacturer’s instructions. The optical densities of wells were measured by a photometer at a wavelength of 450 nm. Data collected were analysed using appropriate computer software. The overall seroprevalence of T. gondii among the women of child-bearing age in selected seven communities in Osun state was 76.3%. Out of 76.3% positive for Toxoplasma gondii infection, 70.0% were positive for anti- T. gondii IgG, and 32.3% were positive for IgM, and 26.7% for both IgG and IgM. The prevalence of T. gondii was lowest (58.9%) among women from Ile Ife, a peri-urban community, and highest (100%) in women residing in Alajue, a rural community. The prevalence of infection was significantly higher (P= 0.000) among Islamic women (87.5%) than in Christian women (70.8%). The highest prevalence (86.3%) was recorded in women with primary education, while the lowest (61.2%) was recorded in women with tertiary education (p =0.016). The highest prevalence (79.7%) was recorded in women that reside in rural areas, and the lowest (70.1%) was recorded in women that reside in peri-urban area (p=0.025). The prevalence of T. gondii infection was highest (81.4%) in women with one miscarriage, while the prevalence was lowest in women with no miscarriages (75.9%). The age of the women (p=0.042), Islamic religion (p=0.001), the residence of the women (p=0.001), and water source were all positively associated with T. gondii infection. The study concluded that there was a high seroprevalence of T. gondii recorded among women of child-bearing age in the study area. Hence, there is a need for health education and create awareness of the disease and its transmission to women of reproductive age group in general and pregnant women in particular to reduce the risk of T. gondii in pregnant women.

Keywords: seroepidemiology, Toxoplasma gondii, women, child-bearing, age, communities, Ile -Ife, Nigeria

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Authors: Tadele Araya, Tsehaye Asmelash, Girmatsion Fiseha

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Background: Hepatitis B virus (HBV), Hepatitis C virus (HCV) and Human Immunodeficiency Virus (HIV) constitute serious healthcare problems worldwide. Blood-borne pathogens HBV, HCV and HIV are commonly associated with infections among substance or Injection Drug Users (IDUs). The objective of this study was to determine the prevalence of HBV, HCV, and HIV infections among substance users in Mekelle Substance users Treatment and Rehabilitation Centers. Methods: A cross-sectional study design was used from Dec 2020 to Sep / 2021 to conduct the study. A total of 600 substance users were included. Data regarding the socio-demographic, clinical and sexual behaviors of the substance users were collected using a structured questionnaire. For laboratory analysis, 5-10 ml of venous blood was taken from the substance users. The laboratory analysis was performed by Enzyme-Linked Immunosorbent Assay (ELISA) at Mekelle University, Department of Medical Microbiology and Immunology Research Laboratory. The Data was analyzed using SPSS and Epi-data. The association of variables with HBV, HCV and HIV infections was determined using multivariate analysis and a P value < 0.05 was considered statistically significant. Result: The overall prevalence rate of HBV, HCV and HIV infections were 10%, 6.6%, and 7.5%, respectively. The mean age of the study participants was 28.12 ± 6.9. A higher prevalence of HBV infection was seen in participants who were users of drug injections and in those who were infected with HIV. HCV was comparatively higher in those who had a previous history of unsafe surgical procedures than their counterparts. Homeless participants were highly exposed to HCV and HIV infections than their counterparts. The HBV/HIV Co-infection prevalence was 3.5%. Those doing unprotected sexual practices [P= 0.03], Injection Drug users [P= 0.03], those who had an HBV-infected person in their family [P=0.02], infected with HIV [P= 0.025] were statistically associated with HBV infection. HCV was significantly associated with Substance users and previous history of unsafe surgical procedures [p=0.03, p=0.04), respectively. HIV was significantly associated with unprotected sexual practices and being homeless [p=0.045, p=0.05) respectively. Conclusion-The highly prevalent viral infection was HBV compared to others. There was a High prevalence of HBV/HIV co-infection. The presence of HBV-infected persons in a family, unprotected sexual practices and sharing of needles for drug injection were the risk factors associated with HBV, HIV, and HCV. Continuous health education and screening of the viral infection coupled with medical and psychological treatment is mandatory for the prevention and control of the infections.

Keywords: hepatitis b virus, hepatitis c virus, HIV, substance users

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Authors: M. Mirmohammadi, S. Taghdisi, F. Anali

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Gossypol is a yellow anti-nutritional compound found in the cotton plant. This substance exists in the cottonseed and other parts of the cotton plant, such as bark, leaves, and stems. Chemically, gossypol is a very active polyphenolic aldehyde compound, and due to this polyphenolic structure, it has antioxidant and therapeutic properties. On the other hand, this compound, especially in free form, has many toxic effects, that its excessive consumption can be very dangerous for humans and animals. In this study, gossypol was extracted as a derivative compound of gossypol acetic acid from cottonseed using the n-hexane solvent with an efficiency of 0.84 ± 0.04, which compared to the Gossypol extracted from cottonseed oil with the same method (cold press) showed a significant difference with its efficiency of 1.14 ± 0.06. Therefore, it can be suggested to use cottonseed oil to extract this valuable compound. In the other part of this research, cottonseed extracts and cotton bolls extracts were obtained by two methods of soaking and Soxhlet with hydroalcoholic solvent taken with a ratio of (25:75), then by using extracts and corn starch powder, four herbal medicine code was created and after receiving the code of ethics (IR.SSU.REC.1398.136) the therapeutic effect of each one on the Cutaneous leishmaniasis resistant to drugs (caused by the leishmaniasis parasite) was investigated in real patients and its results was compared with the common drug glucantime (local ampoule) (n = 36). Statistical studies showed that the use of herbal medicines prepared with cottonseed extract and cotton bolls extract has a significant positive effect on the treatment of the disease’s wounds (p-value > 0.05) compared to the control group (only ethanol). Also, by comparing the average diameter of the wounds after a two-month treatment period, no significant difference was found between the use of ointment containing extracts and local glucantime ampoules (p-value < 0.05). Bolls extract extracted with the Soxhlet method showed the best therapeutic effects, although there was no significant difference between them (p-value < 0.05). Therefore, there is acceptable reliability to recommend this medicine for the treatment of Cutaneous leishmaniasis resistant to drugs without the side effects of the chemical drug glucantime and the pain of injecting the ampoule.

Keywords: cottonseed oil, gossypol, cotton boll, cutaneous leishmaniasis

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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: Ariel Vilchez, Francisca Acevedo, Rodrigo Navia

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The wound healing process can be accelerated and improved by the action of antioxidants such as curcumin (Cur) over the tissues; however, the efficacy of curcumin used through the digestive system is not enough to exploit its benefits. Electrospinning presents an alternative to carry curcumin directly to the wounds, and polyhydroxybutyrate (PHB) is proposed as the matrix to load curcumin owing to its biodegradable and biocompatible properties. PHB is among 150 types of Polyhydroxyalkanoates (PHAs) identified, it is a natural thermoplastic polyester produced by microbial fermentation obtained from microorganisms. The proposed objective is to develop electrospun bacterial PHB-based microfibers containing curcumin for possible biomedical applications. Commercial PHB was solved in Chloroform: Dimethylformamide (4:1) to a final concentration of 7% m/V. Curcumin was added to the polymeric solution at 1%, and 7% m/m regarding PHB. The electrospinning equipment (NEU-BM, China) with a rotary collector was used to obtain Cur-PHB fibers at different voltages and flow rate of the polymeric solution considering a distance of 20 cm from the needle to the collector. Scanning electron microscopy (SEM) was used to determine the diameter and morphology of the obtained fibers. Thermal stability was obtained from Thermogravimetric (TGA) analysis, and Fourier Transform Infrared Spectroscopy (FT-IR) was carried out in order to study the chemical bonds and interactions. A preliminary curcumin release to Phosphate Buffer Saline (PBS) pH = 7.4 was obtained in vitro and measured by spectrophotometry. PHB fibers presented an intact chemical composition regarding the original condition (dust) according to FTIR spectra, the diameter fluctuates between 0.761 ± 0.123 and 2.157 ± 0.882 μm, with different qualities according to their morphology. The best fibers in terms of quality and diameter resulted in sample 2 and sample 6, obtained at 0-10kV and 0.5 mL/hr, and 0-10kV and 1.5 mL/hr, respectively. The melting temperature resulted near 178 °C, according to the bibliography. The crystallinity of fibers decreases while curcumin concentration increases for the studied interval. The curcumin release reaches near 14% at 37 °C at 54h in PBS adjusted to a quasi-Fickian Diffusion. We conclude that it is possible to load curcumin in PHB to obtain continuous, homogeneous, and solvent-free microfibers by electrospinning. Between 0% and 7% of curcumin, the crystallinity of fibers decreases as the concentration of curcumin increases. Thus, curcumin enhances the flexibility of the obtained material. HPLC should be used in further analysis of curcumin release.

Keywords: antioxidant, curcumin, polyhydroxybutyrate, wound healing

Procedia PDF Downloads 131

Authors: Maria Miranda-Medina, Sara Salopek, Andras Vernes, Martin Jech

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Sliding lubricated surfaces induce the formation of tribofilms that reduce friction, wear and prevent large-scale damage of contact parts. Engine oils and lubricants use antiwear and antioxidant additives such as zinc dialkyldithiophosphate (ZDDP) from where protective tribofilms are formed by degradation. The ZDDP tribofilms are described as a two-layer structure composed of inorganic polymer material. On the top surface, the long chain polyphosphate is a zinc phosphate and in the bulk, the short chain polyphosphate is a mixed Fe/Zn phosphate with a gradient concentration. The polyphosphate chains are partially adherent to steel surface through a sulfide and work as anti-wear pads. In this contribution, ZDDP tribofilms formed on gray cast iron surfaces are studied. The tribofilms were generated in a reciprocating sliding tribometer with a piston ring-cylinder liner configuration. Fully formulated oil of SAE grade 5W-30 was used as lubricant during two tests at 40Hz and 50Hz. For the estimation of the tribofilm thicknesses, spectroscopic ellipsometry was used due to its high accuracy and non-destructive nature. Ellipsometry works under an optical principle where the change in polarisation of light reflected by the surface, is associated with the refractive index of the surface material or to the thickness of the layer deposited on top. Ellipsometrical responses derived from tribofilms are modelled by effective medium approximation (EMA), which includes the refractive index of involved materials, homogeneity of the film and thickness. The materials composition was obtained from x-ray photoelectron spectroscopic studies, where the presence of ZDDP, O and C was confirmed. From EMA models it was concluded that tribofilms formed at 40 Hz are thicker and more homogeneous than the ones formed at 50 Hz. In addition, the refractive index of each material is mixed to derive an effective refractive index that describes the optical composition of the tribofilm and exhibits a maximum response in the UV range, being a characteristic of glassy semitransparent films.

Keywords: effective medium approximation, reciprocating sliding tribometer, spectroscopic ellipsometry, zinc dialkyldithiophosphate

Procedia PDF Downloads 251

Authors: Sabrina Sebbane, Alina Elena Parvu

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Introduction: Santolina rosmarinifolia is a plant of the Santolina genus, a family made of medicinal plants widely used. Some of the Santolina species have been proven to have potent anti-inflammatory and anti-oxidant effects. However, no in vivo study has been made to demonstrate this in Santolina rosmarinifolia. The aim of our study is to experimentally evaluate the potential anti-inflammatory and anti-oxidant effects of Santolina rosmarinifolia plant extracts on acute inflammation in rats. These effects are defined by measuring the modifications on nitric oxide, reactive oxygen species and anti-oxidant response in serum. Materials and Methods: Rats were divided into 5 groups (n=6). Three groups were given Santolina rosmarinifolia extract by gavage in different concentrations(100%, 50%, 25%) for a week. Inflammation was induced by i.m injection of turpentine oil on the 8th day. One group was only given turpentine oil and the fifth group acted as control and was given only saline solution. Blood was collected and serum separated. Global tests were used to measure the oxidative stress, total oxidative status (TOS), total antioxidant reactivity (TAR) and the modified method of Griess assay to measure NO synthesis. Malondilaldehyde (MDA) and thiols levels were also assessed. Results: Santolina rosmarinifolia did not significantly change the TOS levels (p > 0.05). Santolina rosmarinifolia 25% and 50% decreased significantly the TAR levels (p < 0.001). Santolina 100% didn't have a significant effect on TAR (p > 0.05). All concentrations of Santolina rosmarinifolia increased the oxidative stress index (OSI) significantly(p < 0.05). Santolina rosmarinifolia 100% significantly decreased NO synthesis (p value < 0.05). In the diluted Santolina groups, no significant effect on NO synthesis was observed. In the groups treated with Santolina 100% and Santolina rosmarinifolia 50%, thiols concentration were significantly higher compared to the inflammation group (p < 0.02). A higher stimulatory effect was found in the Santolina 25% group (p value < 0.05). MDA levels were not significantly modified by the administration of Santolina rosmarinifolia (p > 0.05). Conclusion: All three solutions of Santolina rosmarinifolia had no important effect on oxidant production. However, Santolina rosmarinifolia solutions had a positive effect by increasing the thiols concentration in the serum of the models. The sum of all the effects produced by the administration of Santolina did not show a significant decrease of nitro-oxidative stress. Further experiments including smaller concentrations of Santolina rosmarinifolia will be made. Santolina rosmarinifolia should also be tested as a curative treatment.

Keywords: inflammation, MDA, nitric oxide, santolina rosmarinifolia, thiols, TAR, TOS

Procedia PDF Downloads 260

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

Procedia PDF Downloads 472

Authors: Rahil Malekipoor, Zora Singh, Alan Payne

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Ethylene management during storage life of organically grown apples is a challenging issue due to limited available options. The objective of this investigation was to examine the effects of lemon and cinnamon oils fumigation on storage life, the incidence of superficial scald and quality of Granny Smith apple which were kept in cold storage with and without ozone. The fruit was fumigated with 3µl L⁻¹ lemon or cinnamon oil for 24 h and untreated fruit was kept as a control. Following the treatments, the fruit was stored at (0.5 to -1°C) with and without ozone for 100 and 150 days. After each storage period, ethylene production and respiration rate, superficial scald and various fruit quality parameters were estimated. Lemon oil fumigated fruit showed significantly reduced the mean climacteric peak ethylene production rate in both 100 and 150 days stored fruit. Mean climacteric peak ethylene production rate was significantly reduced in the apples which were kept in an ozonated as compared to cold stored without ozone for 100 days only. The climacteric ethylene peak was delayed only in 100 days cold stored fruit with ozone (8.78 d) as compared to without ozone (3.89 d). Firmness was significantly higher in the fruit fumigated with lemon or cinnamon oil compared to control for both storage time. The fruit stored for 150 days in cold storage without ozone exhibited higher mean firmness than those stored in ozonated. Lemon or cinnamon oil fumigation significantly reduced superficial scald in both cold stored fruit with or without ozone. Levels of total phenols were significantly higher in cinnamon oil treated fruit and stored for 100 days as compared to all other treatments. In 150 days stored fruit fumigated with lemon oil showed the significantly higher level of total phenols compared to cinnamon oil fumigation and control. The fruit fumigated with lemon oil or cinnamon oil following 150 days cold storage resulted in significantly higher levels of ascorbic acid and antioxidant capacity as compared to the control fruit. In conclusion, lemon oil fumigation was more effective in suppressing ethylene production in 100-150 days cold stored fruit than cinnamon oil. Whilst, fumigation of both lemon or cinnamon oil were effective in reducing superficial scald and maintaining quality in 100-150 days cold stored fruit.

Keywords: apple, cold storage, organic oil, ozone

Procedia PDF Downloads 147

Authors: Mohammadjavad Sotoudeheian

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COVID-19, which began in December 2019, uses the angiotensin-converting enzyme 2 (ACE2) receptor to enter and spread through the cells. ACE2 mRNA is present in almost every organ, including nasopharynx, lung, as well as the brain. Ports of entry of SARS-CoV-2 into the central nervous system (CNS) may include arterial circulation, while viremia is remarkable. However, it is imperious to develop neurological symptoms evaluation CSF analysis in patients with COVID-19, but theoretically, ACE2 receptors are expressed in cerebellar cells and may be a target for SARS-CoV-2 infection in the brain. Recent evidence agrees that SARS-CoV-2 can impact the brain through direct and indirect injury. Two biomarkers for CNS injury, glial fibrillary acidic protein (GFAP) and neurofilament light chain (NFL) detected in the plasma of patients with COVID-19. NFL, an axonal protein expressed in neurons, is related to axonal neurodegeneration, and GFAP is over-expressed in CNS inflammation. GFAP cytoplasmic accumulation causes Schwan cells to misfunction, so affects myelin generation, reduces neuroskeletal support over NfLs during CNS inflammation, and leads to axonal degeneration. Interleukin-6 (IL-6), which extensively over-express due to interleukin storm during COVID-19 inflammation, regulates gene expression, as well as GFAP through STAT molecular pathway. IL-6 also impresses the phosphoinositide 3-kinase (PI3K)/STAT/smads pathway. The PI3K/ protein kinase B (Akt) pathway is the main modulator upstream of the mammalian target of rapamycin (mTOR), and alterations in this pathway are common in neurodegenerative diseases. Most neurodegenerative diseases show a disruption of autophagic function and display an abnormal increase in protein aggregation that promotes cellular death. Therefore, induction of autophagy has been recommended as a rational approach to help neurons clear abnormal protein aggregates and survive. The mTOR is a major regulator of the autophagic process and is regulated by cellular stressors. The mTORC1 pathway and mTORC2, as complementary and important elements in mTORC1 signaling, have become relevant in the regulation of the autophagic process and cellular survival through the extracellular signal-regulated kinase (ERK) pathway.

Keywords: mTORC1, COVID-19, PI3K, autophagy, neurodegeneration

Procedia PDF Downloads 86

Authors: Nima Samie, Batoul Sadat Haerian, Sekaran Muniandy, M. S. Kanthimathi

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Colon and breast cancers are categorized as the most prevalent types of cancer worldwide. Recently, the broad clinical application of metal complex compounds has led to the discovery of potential therapeutic drugs. The aim of this study was to evaluate the cytotoxic action of a selected nickel complex compound (NCC) against human colon and breast cancer cells. In this context, we determined the potency of the compound in the induction of apoptosis, cell cycle arrest, and cytoskeleton rearrangement. HT-29, WiDr, CCD-18Co, MCF-7 and Hs 190.T cell lines were used to determine the IC50 of the compound using the MTT assay. Analysis of apoptosis was carried out using immunofluorescence, acridine orange/ propidium iodide double staining, Annexin-V-FITC assay, evaluation of the translocation of NF-kB, oxygen radical antioxidant capacity, quenching of reactive oxygen species content , measurement of LDH release, caspase-3/-7, -8 and -9 assays and western blotting. The cell cycle arrest was examined using flowcytometry and gene expression was assessed using qPCR array. Results showed that our nickel complex compound displayed a potent suppressive effect on HT-29, WiDr, MCF-7 and Hs 190.T after 24 h of treatment with IC50 value of 2.02±0.54, 2.13±0.65, 3.76±015 and 3.14±0.45 µM respectively. This cytotoxic effect on normal cells was insignificant. Dipping in the mitochondrial membrane potential and increased release of cytochrome c from the mitochondria indicated induction of the intrinsic apoptosis pathway by the nickel complex compound. Activation of this pathway was further evidenced by significant activation of caspase 9 and 3/7.The nickel complex compound (NCC) was also shown activate the extrinsic pathways of apoptosis by activation of caspase-8 which is linked to the suppression of NF-kB translocation to the nucleus. Cell cycle arrest in the G1 phase and up-regulation of glutathione reductase, based on excessive ROS production were also observed. The results of this study suggest that the nickel complex compound is a potent anti-cancer agent inducing both intrinsic and extrinsic pathways as well as cell cycle arrest in colon and breast cancer cells.

Keywords: nickel complex, apoptosis, cytoskeletal rearrangement, colon cancer, breast cancer

Procedia PDF Downloads 313

Authors: Youcef Amir

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The consumption of beverages continues to grow worldwide due to increasing demography, but pure fruit juices and high-quality nectars can induce protective effects on human health because of their natural bioactive components. In contrast, sodas and gaseous drinks containing synthetic food additives are considered as responsible for consumers of several pathologies such as obesity, diabetes, and non-alcoholic fatty liver disease. The nutritional and therapeutic virtues of fruit juices are generally a remarkable antioxidant power, anti-cancer activity linked to their richness of indigestible and indigestible sugars, vitamins, mineral salts, carotenoids and phenolic compounds. The main reasons, which led us to produce these fruit derivatives, are the non-availability of the fresh fruits mentioned above all along the year and also the existence of variations in the chemical composition of these different fruits as well as for the major or minor components. We tested, therefore, the physicochemical characteristics of each fruit juice and pulp apart and afterward those of the cocktails formulated. The fresh juices used during our experiments were obtained from the following fruits from north-central Algeria: prickly pear, pomegranate, melon, red oranges. The formulations of these fruit juices were tested after several trials comprising sensorial analysis, physicochemical factors (pH, titratable acidity, Brix degree, formal index, water content, total ash, total and reducing sugars, vitamin C, carotenoids, phenolic compounds) and microbial analysis after a storage period. To the pure juices proportions, citric acid E330, sucrose, and water were added followed by pasteurisation. These products were analysed from the physicochemical, microbial and sensorial viewpoints after a storage period of one month according to national legislation to evaluate their stability. The results of the physicochemical parameters of the prepared beverages had shown good physicochemical results, acceptable sensorial characteristics and microbial stability and safety before and after a storage period. We measured appreciable amounts of minor compounds with health properties.

Keywords: fruit juices, microbial analyses, nectars, physico chemical characteristics, sensorial analysis, storage period

Procedia PDF Downloads 229

Authors: Natan C. G. Silva, Carlos A. C. Girao Neto, Marcele M. S. Vasconcelos, Luciana R. B. Goncalves, Maria Valderez P. Rocha

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Galactooligosaccharides (GOS) are sugars with prebiotic function that can be synthesized chemically or enzymatically, and this last one can be promoted by the action of β-galactosidases. In addition to favoring the transgalactosylation reaction to form GOS, these enzymes can also catalyze the hydrolysis of lactose. A highly studied type of GOS is lactulose because it presents therapeutic properties and is a health promoter. Among the different raw materials that can be used to produce lactulose, whey stands out as the main by-product of cheese manufacturing, and its discarded is harmful to the environment due to the residual lactose present. Therefore, its use is a promising alternative to solve this environmental problem. Thus, lactose from whey is hydrolyzed into glucose and galactose by β-galactosidases. However, in order to favor the transgalactosylation reaction, the medium must contain fructose, due this sugar reacts with galactose to produce lactulose. Then, the glucose-isomerase enzyme can be used for this purpose, since it promotes the isomerization of glucose into fructose. In this scenario, the aim of the present work was first to develop β-galactosidase biocatalysts of Kluyveromyces lactis and to apply it in the integrated reactions of hydrolysis, isomerization (with the glucose-isomerase from Streptomyces murinus) and transgalactosylation reaction, using whey as a substrate. The immobilization of β-galactosidase in chitosan previously functionalized with 0.8% glutaraldehyde was evaluated using different enzymatic loads (2, 5, 7, 10, and 12 mg/g). Subsequently, the hydrolysis and transgalactosylation reactions were studied and conducted at 50°C, 120 RPM for 20 minutes. In parallel, the isomerization of glucose into fructose was evaluated under conditions of 70°C, 750 RPM for 90 min. After, the integration of the three processes for the production of lactulose was investigated. Among the evaluated loads, 7 mg/g was chosen because the best activity of the derivative (44.3 U/g) was obtained, being this parameter determinant for the reaction stages. The other parameters of immobilization yield (87.58%) and recovered activity (46.47%) were also satisfactory compared to the other conditions. Regarding the integrated process, 94.96% of lactose was converted, achieving 37.56 g/L and 37.97 g/L of glucose and galactose, respectively. In the isomerization step, conversion of 38.40% of glucose was observed, obtaining a concentration of 12.47 g/L fructose. In the transgalactosylation reaction was produced 13.15 g/L lactulose after 5 min. However, in the integrated process, there was no formation of lactulose, but it was produced other GOS at the same time. The high galactose concentration in the medium probably favored the reaction of synthesis of these other GOS. Therefore, the integrated process proved feasible for possible production of prebiotics. In addition, this process can be economically viable due to the use of an industrial residue as a substrate, but it is necessary a more detailed investigation of the transgalactosilation reaction.

Keywords: beta-galactosidase, glucose-isomerase, galactooligosaccharides, lactulose, whey

Procedia PDF Downloads 141

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: Recep Karadag

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Natural dyeing had started since pre-historical times for dyeing of textile materials. The natural dyeing had continued to beginning of 20th century. At the end of 19th century some synthetic dyes were synthesized. Although development of dyeing technologies and methods, natural dyeing was not developed in recent years. Despite rapid advances of synthetic dyestuff industries, natural dye processes have not developed. Therefore natural dyeing was not competed against synthetic dyes. At the same time, it was very difficult that large quantities of coloured textile was dyed with natural dyes And it was very difficult to get reproducible results in the natural dyeing using classical and traditional processes. However, natural dyeing has used slightly in the textile handicraft up to now. It is very important view that re-using of natural dyes to create awareness in textiles in recent years. Natural dyes have got many awareness and sustainability properties. Natural dyes are more eco-friendly than synthetic dyes. A lot of natural dyes have got antioxidant, antibacterial, antimicrobial, antifungal and anti –UV properties. It had been known that were obtained limited numbers colours with natural dyes in the past. On the contrary, colour scale is too wide with natural dyes. Except fluorescent colours, numerous colours can be obtained with natural dyes. Fastnesses of dyed textiles with natural dyes are good that there are light, washing, rubbing, etc. The fastness values can be improved depend on dyeing processes. Thanks to these properties mass production can be made with natural dyes in textiles. Therefore fabric dyeing machine was designed. This machine is too suitable for natural dyeing and mass production. Also any dyeing machine can be modified for natural dyeing. Although dye extraction and dyeing are made separately in the traditional natural dyeing processes and these procedures are become by designed this machine. Firstly, colouring compounds are extracted from natural dye resources, then dyeing is made with extracted colouring compounds. The colouring compounds are moderately dissolved in water. Less water is used in the extraction of colouring compounds from dye resources and dyeing with this new technique on the contrary much quantity water needs to use for dissolve of the colouring compounds in the traditional dyeing. This dyeing technique is very useful method for mass productions with natural dyes in traditional natural dyeing that use less energy, less dye materials, less water, etc. than traditional natural dyeing techniques. In this work, cotton, silk, linen and wool fabrics were dyed with some natural dye plants by the technique. According to the analysis very good results were obtained by this new technique. These results are shown sustainability and awareness of natural dyes for textiles.

Keywords: antibacterial, antimicrobial, natural dyes, sustainability

Procedia PDF Downloads 523

Authors: R. H. Begaydarova, B. Zh. Kultanov, B. T. Esilbaeva, G. E. Nasakaeva, Y. Yukhnevich, G. K. Alshynbekova, A. E. Dyusembaeva

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Background: The level of middle molecules of peptides (MMP) allows to evaluate the severity and prognosis of the disease and is a criterion for the effectiveness of the treatment. The detection the products of lipidperoxidation cascade, such as conjugated dienes, malondialdehyde in biological material, has an important role in the development of pathogenesis, the diagnosis and prognosis in different parasitic diseases. Purpose of the study was to evaluate the state of endogenous intoxication and indicators of lipid peroxidation in patients with giardiasis before and after treatment. Materials and methods: Endogenous intoxication was evaluated in patients with giardiasis in the level of middle molecules of peptides (MMP) in the blood. The amount of MMP and products of lipid peroxidation were determined in the blood of 198 patients with giardiasis, 129 of them were women (65%), 69 were men (35%). The MMP level was detected for comparison in the blood of 84 healthy volunteers. The lipid peroxidation were determined in 40 healthy men and women without giardiasis and history of chronic diseases. Data were processed by conventional methods of variation statistics, we calculated the arithmetic mean (M) and standard dispersion (m). t-test (t) was used to assess differences. Results: The level of MMP in the blood was significantly higher in patients with giardiasis in comparison with group of healthy men and women. MMP concentration in the blood of women with Giardia was 2.5 times greater than that of the comparison groups of women. The level of MMP exceeds more than 6 times in men with giardiasis. The decrease in the intensity of endogenous intoxication was two weeks after antigiardia therapy, both men and women. According to the study, a statistically significant increase in the level of all the studied parameters lipid peroxidation cascade was observed in the blood of men with giardiasis, with the exception of the total primary production (NGN). The treatment of giardiasis helped to stabilize the level of almost all metabolites of lipid peroxidation cascade. The exception was level of malondialdehyde, it was significantly elevated to compare with the control group and after treatment. Conclusion: Thus, the MMP level was significantly higher in blood of patients with giardiasis than in comparison group. This is evidence of severe endogenous intoxication caused by giardia infection. The accumulation of primary and secondary products of lipid peroxidation was observed in the blood of men and women. These processes tend to be more active in men than in women. Antigiardiasis therapy contributed to the normalization of almost all the studied indicators of lipid peroxidation in the blood of participants, except the level malondialdehyde in the blood of men.

Keywords: enzymes of antioxidant protection, giardiasis, blood, treatment

Procedia PDF Downloads 238

Authors: Mohamad Ammar Ayass, Natalya Griko, Victor Pashkov, Wanying Cao, Kevin Zhu, Jin Zhang, Lina Abi Mosleh

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Respiratory syndrome coronavirus 2 (SARS-CoV-2) is the causative agent for coronavirus disease 2019 (COVID-19). Early evidence pointed at the angiotensin-converting enzyme 2 (ACE-2) expressed on the epithelial cells of the lung as the main entry point of SARS-CoV-2 into the cells. The viral entry is mediated by the binding of the Receptor Binding Domain (RBD) of the spike protein that is expressed on the surface of the virus to the ACE-2 receptor. As the number of SARS-CoV-2 variants continues to increase, mutations arising in the RBD of SARS-CoV-2 may lead to the ineffectiveness of RBD targeted neutralizing antibodies. To address this limitation, the objective of this study is to develop a combination of aptamers that target different regions of the RBD, preventing the binding of the spike protein to ACE-2 receptor and subsequent viral entry and replication. A safe and innovative biomedical tool was developed to inhibit viral infection and reduce the harms of COVID-19. In the present study, DNA aptamers were developed against a recombinant trimer S protein using the Systematic Evolution of Ligands by Exponential enrichment (SELEX). Negative selection was introduced at round number 7 to select for aptamers that bind specifically to the RBD domain. A series of 9 aptamers (ADI2010, ADI2011, ADI201L, ADI203L, ADI205L, ADIR68, ADIR74, ADIR80, ADIR83) were selected and characterized with high binding affinity and specificity to the RBD of the spike protein. Aptamers (ADI25, ADI2009, ADI203L) were able to bind and pull down endogenous spike protein expressed on the surface of SARS-CoV-2 virus in COVID-19 positive patient samples and determined by liquid chromatography- tandem mass spectrometry analysis (LC-MS/MS). LC-MS/MS data confirmed that aptamers can bind to the RBD of the spike protein. Furthermore, results indicated that the combination of the 9 best aptamers inhibited the binding of the purified trimer spike protein to the ACE-2 receptor found on the surface of Vero E6 cells. In the same experiment, the combined aptamers displayed a better neutralizing effect than antibodies. The data suggests that the selected aptamers could be used in therapy to neutralize the effect of the SARS-CoV-2 virus by inhibiting the interaction between the RBD and ACE-2 receptor, preventing viral entry into target cells and therefore blocking viral replication.

Keywords: aptamer, ACE-2 receptor, binding inhibitor, COVID-19, spike protein, SARS-CoV-2, treatment

Procedia PDF Downloads 185

Authors: Adnan Bekhit, Eskedar Lodebo, Ariaya Hymete, Hanan Ragab, Alaa El-Din Bekhit

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Malaria and leishmaniasis have emerged as serious universal health problems throughout history of mankind. According to the WHO 2008 malarial report, half of the world population is at risk of malarial infection with an estimate of 1 million deaths occurring annually mainly in the African region. Furthermore, 12-15 million people are infected with Leishmaniasis worldwide. Despite the continuous introduction of a large number of agents for the treatment of malaria, there is still unmet medical needs due to the emergence of resistance. Resistance has occurred for almost all therapeutic agents approved for the treatment of malaria. Accordingly, it was the aim of this work to design and synthesis a group of antimalarial-antileshmanial agents that would show inhibitory activity against chloroquine-resistant strain of Plasmodium falciparum. The synthesized compounds were designed to contain a pyrazolylpyrazoline moiety having an aromatic group (p-tolyl or p-chlorophenyl) at N1-position of one pyrazoline ring due to the reports of promising activities of such compounds. A formyl or acyl substituent was introduced at the N1-position of the other pyrazoline ring, to investigate the effect of bulkiness of acyl substituents at this position. The synthesized compounds were evaluated for their in-vivo antimalarial activity against Plasmodium berghei infected mice at dose levels of 20 and 30 mg/Kg. the two most active compounds were evaluated for their antimalarial activity against chloroquin-resistant strain (RKL9) of Plasmodium falciparum. In addition, the synthesized compounds were tested for their in-vitro antileshmanial activity against Leishmania aethiopica promastigotes and amastigotes. For both antimalarial and antileishmanial activities, compounds having an N1-p-tolyl group at the first pyrazoline ring did not require bulkiness at the second pyrazoline ring nitrogen where the compound bearing an acetyl group proved to be the most active of the whole series. On the other hand, bulkiness at the N1-position of the second pyazoline ring was necessary in case of compounds carrying the p-chlorophenyl group, where the two derivatives having an N1-butanoyl and an N1-benzoyl moieties at the second pyrazoline showed the best activity. Furthermore, the toxicity of the active compounds were tested and were proved to be non-toxic at 125, 250 and 500 mg/Kg. In addition, docking of the most active compound (having a p-tolyl group at the first pyrazoline-N and an acetyl moiety on the other pyrazoline-N) was performed against dihydrofolate reductase enzyme.

Keywords: pyrazoline derivatives, in-vivo antimalarial activity, docking, dihydrofolate reductase

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Authors: Mansi Patel, Priti Mehta

Abstract:

Background: Ionizing radiations have detrimental effects on humans, and the growing technological encroachment has increased human exposure to it enormously. So, the safety issues have emphasized researchers to develop radioprotector from natural resources having minimal toxicity. A substance having anti-inflammatory, antioxidant, and immunomodulatory activity can be a potential candidate for radioprotection. One such plant with immense potential i.e. Bamboo was selected for the present study. Purpose: The study aims to evaluate the potential of Indian bamboo leaves for protection against the clastogenic effect of gamma radiation. Methods: The protective effect of bamboo leaf extract against gamma radiation-induced genetic damage in human peripheral blood lymphocytes (HPBLs) was evaluated in vitro using Cytokinesis blocked micronuclei assay (CBMN). The blood samples were pretreated with varying concentration of extract 30 min before the radiation exposure (4Gy & 6Gy). The reduction in the frequency of micronuclei was observed for the irradiated and control groups. The effect of various concentration of bamboo leaf extract (400,600,800 mg/kg) on the development of radiation induced sickness and altered mortality in mice exposed to 8 Gy of whole-body gamma radiation was studied. The developed symptoms were clinically scored by multiple endpoints for 30 days. Results: Treatment of HPBLs with varying concentration of extract before exposure to a different dose of γ- radiation resulted in significant (P < 0.0001) decline of radiation induced micronuclei. It showed dose dependent and concentration driven activity. The maximum protection ~ 70% was achieved at nine µg/ml concentration. Extract treated whole body irradiated mice showed 50%, 83.3% and 100% survival for 400, 600, and 800mg/kg with 1.05, 0.43 and 0 clinical score respectively when compared to Irradiated mice having 6.03 clinical score and 0% survival. Conclusion: Our findings indicate bamboo leaf extract reduced the radiation induced cytogenetic damage. It has also increased the survival ratio and reduced the radiation induced sickness and mortality when exposed to a lethal dose of gamma radiation.

Keywords: bamboo leaf extract, Cytokinesis blocked micronuclei (CBMN) assay, ionizing radiation, radio protector

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Authors: Sinethemba Yakobi, Lindiwe Zuma, Ofentse Pooe

Abstract:

Gonococcal infections present a notable public health issue, and the major approach for treatment involves using β-lactam antibiotics that specifically target penicillin-binding protein 2 (PBP2) in Neisseria gonorrhoeae. This study examines the influence of flavonoids, namely rutin, on the structural changes of PBP2 in both penicillin-resistant (FA6140) and penicillin-susceptible (FA19) strains. The research clarifies the structural effects of particular mutations, such as inserting an aspartate residue at position 345 (Asp-345a) in the PBP2 protein. The strain FA6140, which is resistant to penicillin, shows specific changes that lead to a decrease in penicillin binding. These mutations, namely P551S and F504L, significantly impact the pace at which acylation occurs and the stability of the strain under high temperatures. Molecular docking analyses investigate the antibacterial activities of rutin and other phytocompounds, emphasizing its exceptional binding affinity and potential as an inhibitor of PBP2. Quercetin and protocatechuic acid have encouraging antibacterial effectiveness, with quercetin displaying characteristics similar to those of drugs. Molecular dynamics simulations offer a detailed comprehension of the interactions between flavonoids and PBP2, highlighting rutin's exceptional antioxidant effects and strong affinity for the substrate binding site. The study's wider ramifications pertain to the pressing requirement for antiviral treatments in the context of the ongoing COVID-19 epidemic. Flavonoids have a strong affinity for binding to PBP2, indicating their potential as inhibitors to impair cell wall formation in N. gonorrhoeae. Ultimately, this study provides extensive knowledge on the interactions between proteins and ligands, the dynamics of the structure, and the ability of flavonoids to combat penicillin-resistant N. gonorrhoeae bacteria. The verified simulation outcomes establish a basis for creating potent inhibitors and medicinal therapies to combat infectious illnesses.

Keywords: phytochemicals, penicillin-binding protein 2, gonococcal infection, ligand-protein interaction, binding energy, neisseria gonorrhoeae FA19, neisseria gonorrhoeae FA6140, flavonoids

Procedia PDF Downloads 69