Search results for: enzyme-linked immunosorbent assay

Authors: Mustafa M. Donma, Orkide Donma

Abstract:

Zinc (Zn) is a vital element required for growth and development particularly in children. It exhibits some protective effects against cardiovascular diseases (CVDs). Zn may be a potential biomarker of cardiovascular health. High sensitive cardiac troponin T (hs-cTnT) and cardiac myosin binding protein C (cMyBP-C) are new generation markers used for prediagnosis, diagnosis and prognosis of CVDs. The aim of this study is to determine Zn as well as new generation cardiac markers’ profiles in children with normal body mass index (N-BMI), obese (OB), morbid obese (MO) children and children with metabolic syndrome (MetS) findings. The association among them will also be investigated. Four study groups were constituted. The study protocol was approved by the institutional Ethics Committee of Tekirdag Namik Kemal University. Parents of the participants filled informed consent forms to participate in the study. Group 1 is composed of 44 children with N-BMI. Group 2 and Group 3 comprised 43 OB and 45 MO children, respectively. 45 MO children with MetS findings were included in Group 4. World Health Organization age- and sex-adjusted BMI percentile tables were used to constitute groups. These values were 15-85, 95-99 and above 99 for N-BMI, OB and MO, respectively. Criteria for MetS findings were determined. Routine biochemical analyses including Zn were performed. hs-cTnT and cMyBP-C concentrations were measured by enzyme-linked immunosorbent assay. Data were analyzed by using SPSS software. p < 0.05 was accepted as significant. Four groups were matched for age and gender. Decreased Zn concentrations were measured in Groups 2, 3 and 4 compared to Group 1. Groups did not differ from one another in terms of hs-cTnT. There were statistically significant differences between cMyBP-C levels of MetS group and N-BMI as well as OB groups. There was an increasing trend going from N-BMI group to MetS group. There were statistically significant negative correlations between Zn and hs-cTnT as well as cMyBP-C concentrations in MetS group. In conclusion, inverse correlations detected between Zn and new generation cardiac markers (hs-TnT and cMyBP-C) have pointed out that decreased levels of Zn accompany increased levels of hs-cTnT as well as cMyBP-C in children with MetS. This finding emphasizes that both Zn and these new generation cardiac markers may be evaluated as biomarkers of cardiovascular health during severe childhood obesity precipitated with MetS findings and also suggested as the messengers of the future risk in the adulthood periods of children with MetS.

Keywords: Cardiac myosin binding protein-C, cardiovascular diseases, children, high sensitive cardiac troponin T, obesity.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 442

Authors: M. Shahar Yar, M. Mustaqeem Abdullah, Jaseela Majeed

Abstract:

A series of 1-(1H-benzimidazol-2-yl)-3-(substituted phenyl)-2-propen-1-one were allowed to react with hydrazine hydrate and phenyl hydrazine in submitted reactions to get pyrazoline and phenyl pyrazoline derivatives. All the compounds entered for screening at the Tuberculosis Antimicrobial Acquisition and Coordinating Facility (TAACF) for their in vitro antibacterial activity against Mycobacterium tuberculosis H37Rv strain (ATCC 27294) using Microplate Alamar Blue Assay (MABA) susceptibility test. The results expressed as MIC (minimum inhibitory concentration) in μg/mL. Among the fifteen compounds, eight compounds were found to have MIC values less than 10 μg/mL. These were subjected for cytotoxicity assay in VERO cells to determine CC50 (cytotoxic concentration 50%) values and finally SI (Selectivity Index) were calculated. Compound (XV) 2-[5-(4- fluorophenyl)-1-phenyl-4,5-dihydro-1H-3-pyrazolyl]-1Hbenzimidazole was considered the best candidate of the series that could be a good starting point to develop new lead compounds in the fight against tuberculosis.

Keywords: anti-tubercular activity, benzimidazole, pyrazoline.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 1855

Authors: Mustafa M. Donma, Orkide Donma

Abstract:

Spexin, expressed in the central nervous system, has attracted much interest in feeding behavior, obesity, diabetes, energy metabolism and cardiovascular functions. Fetuin A is known as the negative acute phase reactant synthesized in the liver. Eosinophils are early indicators of cardiometabolic complications. Patients with elevated platelet count, associated with hypercoagulable state in the body, are also more liable to cardiovascular diseases (CVDs). In this study, the aim is to examine the profiles of spexin and fetuin A concomitant with the course of variations detected in eosinophil as well as platelet counts in morbid obese children. 34 children with normal-body mass index (N-BMI) and 51 morbid obese (MO) children participated in the study. Written-informed consent forms were obtained prior to the study. Institutional ethics committee approved the study protocol. Age- and sex-adjusted BMI percentile tables prepared by World Health Organization were used to classify healthy and obese children. Mean age ± SEM of the children were 9.3 ± 0.6 years and 10.7 ± 0.5 years in N-BMI and MO groups, respectively. Anthropometric measurements of the children were taken. BMI values were calculated from weight and height values. Blood samples were obtained after an overnight fasting. Routine hematologic and biochemical tests were performed. Within this context, fasting blood glucose (FBG), insulin (INS), triglycerides (TRG), high density lipoprotein-cholesterol (HDL-C) concentrations were measured. Homeostatic model assessment for insulin resistance (HOMA-IR) values were calculated. Spexin and fetuin A levels were determined by enzyme-linked immunosorbent assay. Data were evaluated from the statistical point of view. Statistically significant differences were found between groups in terms of BMI, fat mass index, INS, HOMA-IR and HDL-C. In MO group, all parameters increased as HDL-C decreased. Elevated concentrations in MO group were detected in eosinophils (p < 0.05) and platelets (p > 0.05). Fetuin A levels decreased in MO group (p > 0.05). However, decrease was statistically significant in spexin levels for this group (p < 0.05). In conclusion, these results have suggested that increases in eosinophils and platelets exhibit behavior as cardiovascular risk factors. Decreased fetuin A behaved as a risk factor suitable to increased risk for cardiovascular problems associated with the severity of obesity. Along with increased eosinophils, increased platelets and decreased fetuin A, decreased spexin was the parameter, which reflects best its possible participation in the early development of CVD risk in MO children.

Keywords: Cardiovascular diseases, eosinophils, fetuin A, pediatric morbid obesity, platelets, spexin.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 600

Authors: N. Hoosain, S. Nene, B. Pearce, C. Jacobs, M. Du Plessis, M. Benjeddou

Abstract:

Organic cation transporter (OCT) 1could influence an individual’s response to various treatments and increase their susceptibility to diseases.Genotypic and allelic frequencies of nineteen non-synonymous and one intronic Single Nucleotide Polymorphism (SNP) from the OCT1 gene were determined in 101 unrelated healthy Zulu participants, using a SNaPshot^® multiplex assay. Minor allele frequencies (MAF)were compared to representative populations of Africa, Asia and Europe, from Ensembl. MAFs for S14F, V519F, rs622342 and P341L were 2.0%, 6.0%, 6.0% and 1.0%, respectively. Sixteen of nineteen investigated non-synonymous SNPs were monomorphic. No study participant harbored variant alleles for S189L, G220V, P283L, G401S, M420V, M440I, G465R, I542V, R61C, R287G, C88S, A306T, A413V, I421F, C436F and V501E. Haplotype, CGTCGCCGCGCAAGAGGTGA, was most frequently observed (81.23%).Further investigations are encouraged to evaluate potential roles these SNPs could play in the therapeutic efficacy of clinically important drugs and in the development of various diseases in the Zulu population.

Keywords: OCT1, PCR, SNaPshot assay, Zulu population.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 2230

Authors: S. Ibrahim, U. B. Abubakar, S. Danbirni, A. Usman, F. M. Ballah, C. A. Kudi, L. Lawson, G. H. Abdulrazak, I. A. Abdulkadir

Abstract:

Performing culture and characterization of mycobacteria in low resource settings like Nigeria is a very difficult task to undertake because of the very few and limited laboratories carrying out such an experiment; this is a largely due to stringent and laborious nature of the tests. Hence, a rapid, simple and accurate test for characterization is needed. The “SD BIOLINE TB Ag MPT 64 Rapid ®” is a simple and rapid immunochromatographic test used in differentiating Mycobacteria into Mycobacterium tuberculosis (NTM). The 100 sputa were obtained from patients suspected to be infected with tuberculosis and presented themselves to hospitals for check-up and treatment were involved in the study. The samples were cultured in a class III Biosafety cabinet and level III biosafety practices were followed. Forty isolates were obtained from the cultured sputa, and there were identified as Acid-fast bacilli (AFB) using Zeihl-Neelsen acid-fast stain. All the isolates (AFB positive) were then subjected to the SD BIOLINE Analyses. A total of 31 (77.5%) were characterized as MTBC, while nine (22.5%) were NTM. The total turnaround time for the rapid assay was just 30 minutes as compared to a few days of phenotypic and genotypic method. It was simple, rapid and reliable test to differentiate MTBC from NTM.

Keywords: Culture, mycobacteria, non-tuberculous mycobacteria, SD bioline.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 1148

Authors: Mustafa M. Donma, Orkide Donma

Abstract:

The impact of metabolic syndrome (MetS) on cognition and functions of the brain is being investigated. Iron deficiency and deficiencies of B9 (folate) as well as B12 (cobalamin) vitamins are best-known nutritional anemias. They are associated with cognitive disorders and learning difficulties. The antidepressant effects of vitamin D are known and the deficiency state affects mental functions negatively. The aim of this study is to investigate possible correlations of MetS with serum brain-derived neurotrophic factor (BDNF), iron, folate, cobalamin and vitamin D in pediatric patients. 30 children, whose age- and sex-dependent body mass index (BMI) percentiles vary between 85 and 15, 60 morbid obese children with above 99^th percentiles constituted the study population. Anthropometric measurements were taken. BMI values were calculated. Age- and sex-dependent BMI percentile values were obtained using the appropriate tables prepared by the World Health Organization (WHO). Obesity classification was performed according to WHO criteria. Those with MetS were evaluated according to MetS criteria. Serum BDNF was determined by enzyme-linked immunosorbent assay. Serum folate was analyzed by an immunoassay analyzer. Serum cobalamin concentrations were measured using electrochemiluminescence immunoassay. Vitamin D status was determined by the measurement of 25-hydroxycholecalciferol [25-hydroxy vitamin D3, 25(OH)D] using high performance liquid chromatography. Statistical evaluations were performed using SPSS for Windows, version 16. The p values less than 0.05 were accepted as statistically significant. Although statistically insignificant, lower folate and cobalamin values were found in MO children compared to those observed for children with normal BMI. For iron and BDNF values, no alterations were detected among the groups. Significantly decreased vitamin D concentrations were noted in MO children with MetS in comparison with those in children with normal BMI (p ≤ 0.05). The positive correlation observed between iron and BDNF in normal-BMI group was not found in two MO groups. In THE MetS group, the partial correlation among iron, BDNF, folate, cobalamin, vitamin D controlling for waist circumference and BMI was r = -0.501; p ≤ 0.05. None was calculated in MO and normal BMI groups. In conclusion, vitamin D should also be considered during the assessment of pediatric MetS. Waist circumference and BMI should collectively be evaluated during the evaluation of MetS in children. Within this context, BDNF appears to be a key biochemical parameter during the examination of obesity degree in terms of mental functions, cognition and learning capacity. The association observed between iron and BDNF in children with normal BMI was not detected in MO groups possibly due to development of inflammation and other obesity-related pathologies. It was suggested that this finding may contribute to mental function impairments commonly observed among obese children.

Keywords: Brain-derived neurotrophic factor, iron, Vitamin B9, Vitamin B12, Vitamin D.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 685

Authors: Eszter Tóth, Ervin Welker

Abstract:

Shadoo protein (Sho) was described in 2003 as the newest member of Prion protein superfamily [1]. Sho has similar structural motifs like prion protein (PrP) that is known for its central role in transmissible spongiform enchephalopathies. Although a great number of functions have been proposed, the exact physiological function of PrP is not known yet. Investigation of the function and localization of Sho may help us to understand the function of the Prion protein superfamily. Analyzing the subcellular localization of YFP-tagged forms of Sho, we detected the protein in the plasma membrane and in the nucleus of various cell lines. To reveal the localization of the endogenous protein we generated antibodies against Shadoo as well as employed commercially available anti-Shadoo antibodies: i) EG62 anti-mouse Shadoo antibody generated by Eurogentec Ltd.; ii) S-12 anti-human Shadoo antibody by Santa Cruz Biotechnology Inc.; iii) R-12 anti-mouse Shadoo antibody by Santa Cruz Biotechnology Inc.; iv) SPRN antibody against human Shadoo by Abgent Inc. We carried out immunocytochemistry on non-transfected HeLa, Zpl 2-1, Zw 3-5, GT1-1, GT1-7 and SHSY5Y cells as well as on YFP-Sho, Sho-YFP, and YFP-GPI transfected HeLa cells. Their specificity (in antibody-peptide competition assay) and co-localization (with the YFP signal) were assessed.

Keywords: Shadoo, prion protein, immunocytochemistry, antibody-peptide competition assay, antibody.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 1667

Authors: Mustafa M. Donma, Orkide Donma, Savas Guzel

Abstract:

Iron is physiologically essential. However, it also participates in the catalysis of free radical formation reactions. Its deficiency is associated with amplified health risks. This trace element establishes some links with another physiological process related to cell death, apoptosis. Both iron deficiency and iron overload are closely associated with apoptosis. Soluble tumor necrosis factor-like weak inducer of apoptosis (sTWEAK) has the ability to trigger apoptosis and plays a dual role in the physiological versus pathological inflammatory responses of tissues. The aim of this study was to investigate the status of these parameters as well as the associations among them in children with obesity, a low-grade inflammatory state. The study was performed on groups of children with normal body mass index (N-BMI) and obesity. 43 children were included in each group. Based upon age- and sex-adjusted BMI percentile tables prepared by the World Health Organization, children whose values varied between 85 and 15 were included in N-BMI group. Children, whose BMI percentile values were between 99 and 95, comprised obese (OB) group. Institutional ethical committee approval and informed consent forms were taken prior to the study. Anthropometric measurements (weight, height, waist circumference, hip circumference, head circumference, neck circumference) and blood pressure values (systolic blood pressure and diastolic blood pressure) were recorded. Routine biochemical analyses, including serum iron, total iron binding capacity (TIBC), transferrin saturation percent (Tf Sat %) and ferritin, were performed. sTWEAK levels were determined by enzyme-linked immunosorbent assay. study data were evaluated using appropriate statistical tests performed by the statistical program SPSS. Serum iron levels were 91 ± 34 mcrg/dl and 75 ± 31 mcrg/dl in N-BMI and OB children, respectively. The corresponding values for TIBC, Tf Sat %, ferritin were 265 mcrg/dl vs. 299 mcrg/dl, 37.2 ± 19.1% vs. 26.7 ± 14.6%, and 41 ± 25 ng/ml vs 44 ± 26 ng/ml. In N-BMI and OB groups, sTWEAK concentrations were measured as 351 ng/L and 325 ng/L, respectively (p > 0.05). Correlation analysis revealed significant associations between sTWEAK levels and iron related parameters (p < 0.05) except ferritin. In conclusion, iron contributes to apoptosis. Children with iron deficiency have decreased apoptosis rate in comparison with that of healthy children. sTWEAK is an inducer of apoptosis. OB children had lower levels of both iron and sTWEAK. Low levels of sTWEAK are associated with several types of cancers and poor survival. Although iron deficiency state was not observed in this study, the correlations detected between decreased sTWEAK and decreased iron as well as Tf Sat % values were valuable findings, which point out decreased apoptosis. This may induce a proinflammatory state, potentially leading to malignancies in the future lives of OB children.

Keywords: Apoptosis, children, iron-related parameters, obesity, soluble tumor necrosis factor-like weak inducer of apoptosis.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 444

Authors: Eva Tvrdá, Hana Greifová, Peter Ivanič, Norbert Lukáč

Abstract:

The aim of this study was to evaluate the dose- and time-dependent in vitro effects of berberine (BER), a natural alkaloid with numerous biological properties on bovine spermatozoa during three time periods (0 h, 2 h, 24 h). Bovine semen samples were diluted and cultivated in physiological saline solution containing 0.5% DMSO together with 200, 100, 50, 10, 5, and 1 μmol/L BER. Spermatozoa motility was assessed using the computer assisted semen analyzer. The viability of spermatozoa was assessed by the metabolic (MTT) assay, production of superoxide radicals was quantified using the nitroblue tetrazolium (NBT) test, and chemiluminescence was used to evaluate the generation of reactive oxygen species (ROS). Cell lysates were prepared and the extent of lipid peroxidation (LPO) was evaluated using the TBARS assay. The results of the movement activity showed a significant increase in the motility during long term cultivation in case of concentrations ranging between 1 and 10 μmol/L BER (P < 0.01; P < 0.001; 24 h). At the same time, supplementation of 1, 5 and 10 μmol/L BER led to a significant preservation of the cell viability (P < 0.001; 24 h). BER addition at a range of 1-50 μmol/L also provided a significantly higher protection against superoxide (P < 0.05) and ROS (P < 0.001; P < 0.01) overgeneration as well as LPO (P < 0.01; P<0.05) after a 24 h cultivation. We may suggest that supplementation of BER to bovine spermatozoa, particularly at concentrations ranging between 1 and 50 μmol/L, may offer protection to the motility, viability and oxidative status of the spermatozoa, particularly notable at 24 h.

Keywords: Berberine, bulls, motility, oxidative profile, spermatozoa, viability.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 647

Authors: Mustafa M. Donma, Orkide Donma

Abstract:

Childhood obesity is an important clinical problem, because it may lead to chronic diseases during the adulthood period of the individual. Obesity is a metabolic disease associated with low-grade inflammation. The liver occurs at the center of metabolic pathways. Adropin, fibroblast growth factor-21 (FGF-21) and fetuin A are hepatokines. Due to the immense participation of the liver in glucose metabolism, these liver derived factors may be associated with insulin resistance (IR), which is a phenomenon discussed within the scope of obesity problems. The aim of this study is to determine the concentrations of adropin, FGF-21 and fetuin A in childhood obesity, to point out possible differences between the obesity groups and to investigate possible associations among these three hepatokines in obese and morbid obese children. A total of 132 children were included in the study. Two obese groups were constituted. The groups were matched in terms of mean±SD values of ages. Body mass index values of the obese and morbid obese groups were 25.0±3.5 kg/m2 and 29.8±5.7 kg/m2, respectively. Anthropometric measurements including waist circumference, hip circumference, head circumference, and neck circumference were recorded. Informed consent forms were taken from the parents of the participants and the Ethics Committee of the institution approved the study protocol. Blood samples were obtained after an overnight fasting. Routine biochemical tests including glucose- and lipid-related parameters were performed. Concentrations of the hepatokines (adropin, FGF-21, fetuin A) were determined by enzyme-linked immunosorbent assay. Insulin resistance indices such as homeostasis model assessment for IR (HOMA-IR), alanine transaminase-to aspartate transaminase ratio (ALT/AST), diagnostic obesity notation model assessment laboratory index, diagnostic obesity notation model assessment metabolic syndrome index as well as obesity indices such as diagnostic obesity notation model assessment-II index, and fat mass index were calculated using the previously derived formulas. Statistical evaluation of the study data as well as findings of the study were performed by SPSS for Windows. Statistical difference was accepted significant when p < 0.05. Statistically significant differences were found for insulin, triglyceride, high density lipoprotein cholesterol levels of the groups. A significant increase was observed for FGF-21 concentrations in the morbid obese group. Higher adropin and fetuin A concentrations were observed in the same group in comparison with the values detected in the obese group (p > 0.05). There was no statistically significant difference between the ALT/AST values of the groups. In all of the remaining IR and obesity indices, significantly increased values were calculated for morbid obese children. Significant correlations were detected between HOMA-IR and each of the hepatokines. The highest one was the association with fetuin A (r = 0.373, p = 0.001). In conclusion, increased levels observed in adropin, FGF-21 and fetuin A have shown that these hepatokines possess increasing potential going from the obese to morbid obese state. Out of the correlations found with IR index, the most affected hepatokine was fetuin A, the parameter possibly used as the indicator of the advanced obesity stage.

Keywords: adropin, fetuin A, fibroblast growth factor-21, insulin resistance, pediatric obesity

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 450

Authors: T. C. Machaba, S. M. Mahlo

Abstract:

The current study investigates the antifungal properties of crude plant extracts from selected medicinal plant species. Eight plant species used by the traditional healers and local people to treat fungal infections were selected for further phytochemical analysis and biological assay. The selected plant species were extracted with solvent of various polarities such as acetone, methanol, ethanol, hexane, dichloromethane, ethyl acetate and water. Leaf, roots and bark extracts of Maerua juncea Pax, Albuca seineri (Engl & K. Krause) J.C Manning & Goldblatt, Senna italica Mill., Elephantorrhiza elephantina (Burch.) Skeels, Indigofera circinata Benth., Schinus molle L., Asparagus buchananii Bak., were screened for antifungal activity against three animal fungal pathogens (Candida albicans, Aspergillus fumigatus and Cryptococcus neoformans). All plant extracts were active against the tested microorganisms. Acetone, dichloromethane, hexane and ethanol extracts of Senna italica and Elephantorrhiza elephantine had excellent activity against Candida albicans and A. fumigatus with the lowest MIC value of 0.02 mg/ml. Bioautography assay was used to determine the number of antifungal compounds presence in the plant extracts. No active compounds were observed in plant extracts of Indigofera circinnata, Schinus molle and Pentarrhinum insipidum with good antifungal activity against C. albicans and A. fumigatus indicating possible synergism between separated metabolites.

Keywords: Antifungal activity, minimum inhibitory concentration, bioautography.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 1353

Authors: Perumal Siddhuraju, Arumugam Abirami, Gunasekaran Nagarani, Marimuthu Sangeethapriya

Abstract:

Aqueous ethanol and aqueous acetone extracts of Moringa oleifera (outer pericarp of immature fruit and flower) and Sesbania grandiflora white variety (flower and leaf) were examined for radical scavenging capacities and antioxidant activities. Ethanol extract of S. grandiflora (flower and leaf) and acetone extract of M. oleifera (outer pericarp of immature fruit and flower) contained relatively higher levels of total dietary phenolics than the other extracts. The antioxidant potential of the extracts were assessed by employing different in vitro assays such as reducing power assay, DPPH˙, ABTS˙+ and ˙OH radical scavenging capacities, antihemolytic assay by hydrogen peroxide induced method and metal chelating ability. Though all the extracts exhibited dose dependent reducing power activity, acetone extract of all the samples were found to have more hydrogen donating ability in DPPH˙ (2.3% - 65.03%) and hydroxyl radical scavenging systems (21.6% - 77.4%) than the ethanol extracts. The potential of multiple antioxidant activity was evident as it possessed antihemolytic activity (43.2 % to 68.0 %) and metal ion chelating potency (45.16 - 104.26 mg EDTA/g sample). The result indicate that acetone extract of M. oleifera (OPIF and flower) and S. grandiflora (flower and leaf) endowed with polyphenols, could be utilized as natural antioxidants/nutraceuticals.

Keywords: Antioxidant activity, Moringa oleifera, Polyphenolics, Sesbania grandiflora, Underutilized vegetables.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 2362

Authors: Ch. Sarojnalini, Wahengbam Sarjubala Devi

Abstract:

Antioxidant properties and nutritive values of raw and cooked Pool barb, Puntius sophore (Hamilton-Buchanan) of Eastern Himalayas, India were determined. Antioxidant activity of the methanol extract of the raw, steamed, fried and curried Pool barb was evaluated by using 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging assay. In DPPH scavenging assay the IC₅₀ value of the raw, steamed, fried and curried Pool barb was 1.66 micro-gram/ml, 16.09 micro-gram/ml, 8.99 micro-gram/ml, 0.59 micro-gram/ml whereas the IC₅₀ of the reference ascorbic acid was 46.66miro-gram/ml. These results showed that the fish have high antioxidant activity. Protein content was found highest in raw (20.50±0.08%) and lowest in curried (18.66±0.13%). Moisture content in raw, fried and curried was 76.35±0.09, 46.27±0.14 and 57.46±0.24 respectively. Lipid content was recorded 2.46±0.14% in raw and 21.76±0.10% in curried. Ash content varied from 12.57±0.11 to 22.53±0.07%. The total amino acids varied from 36.79±0.02 and 288.43±0.12 mg/100g. Eleven essential mineral elements were found abundant in all the samples. The samples had considerable amount of Fe ranging from 152.17 to 320.39 milli-gram/100gram, Ca 902.06 to 1356.02 milli-gram/100gram, Zn 91.07 to 138.14 milli-gram/100gram, K 193.25 to 261.56 milli-gram/100gram, Mg 225.06 to 229.10 milli-gram/100gram. Ni was not detected in the curried fish. The Mg and K contents were significantly decreased in frying method; however the Fe, Cu, Ca, Co and Mn contents were increased significantly in all the cooked samples. The Mg and Na contents were significantly increased in curried sample and the Cr content was decreased significantly (p<0.05) in all the cooked samples.

Keywords: Antioxidant property, Pool barb, minerals, amino acids, proximate composition, cooking methods.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 2675

Authors: Morteza Elsa, Amirhossein Moghanian

Abstract:

The major aim of this study was to evaluate the effect of CaO content on in vitro hydroxyapatite formation, MC3T3 cells cytotoxicity and proliferation as well as antibacterial efficiency of sol-gel derived SiO₂–CaO–P₂O₅ ternary system. For this purpose, first two grades of bioactive glass (BG); BG-58s (mol%: 60%SiO₂–36%CaO–4%P₂O₅) and BG-68s (mol%: 70%SiO₂–26%CaO–4%P₂O₅)) were synthesized by sol-gel method. Second, the effect of CaO content in their composition on in vitro bioactivity was investigated by soaking the BG-58s and BG-68s powders in simulated body fluid (SBF) for time periods up to 14 days and followed by characterization inductively coupled plasma atomic emission spectrometry (ICP-AES), Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), and scanning electron microscopy (SEM) techniques. Additionally, live/dead staining, 3-(4,5dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), and alkaline phosphatase (ALP) activity assays were conducted respectively, as qualitatively and quantitatively assess for cell viability, proliferation and differentiations of MC3T3 cells in presence of 58s and 68s BGs. Results showed that BG-58s with higher CaO content showed higher in vitro bioactivity with respect to BG-68s. Moreover, the dissolution rate was inversely proportional to oxygen density of the BG. Live/dead assay revealed that both 58s and 68s increased the mean number live cells which were in good accordance with MTT assay. Furthermore, BG-58s showed more potential antibacterial activity against methicillin-resistant Staphylococcus aureus (MRSA) bacteria. Taken together, BG-58s with enhanced MC3T3 cells proliferation and ALP activity, acceptable bioactivity and significant high antibacterial effect against MRSA bacteria is suggested as a suitable candidate in order to further functionalizing for delivery of therapeutic ions and growth factors in bone tissue engineering.

Keywords: Antibacterial, bioactive glass, hydroxyapatite, proliferation, sol-gel processes.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 776

Authors: Suttijit Sriwatcharakul

Abstract:

The bioactivity studies from the weed ethanolic crude extracts from leaf, stem, pod and root of wild spider flower; Cleoma viscosa Linn. were analyzed for the growth inhibition of 6 bacterial species; Salmonella typhimurium TISTR 5562, Pseudomonas aeruginosa ATCC 27853, Staphylococcus aureus TISTR 1466, Streptococcus epidermidis ATCC 1228, Escherichia coli DMST 4212 and Bacillus subtilis ATCC 6633 with initial concentration crude extract of 50 mg/ml. The agar well diffusion results found that the extracts inhibit only gram positive bacteria species; S. aureus, S. epidermidis and B. subtilis. The minimum inhibition concentration study with gram positive strains revealed that leaf crude extract give the best result of the lowest concentration compared with other plant parts to inhibit the growth of S. aureus, S. epidermidis and B. subtilis at 0.78, 0.39 and lower than 0.39 mg/ml, respectively. The determination of total phenolic compounds in the crude extracts exhibited the highest phenolic content was 10.41 mg GAE/g dry weight in leaf crude extract. Analyzed the efficacy of free radical scavenging by using DPPH radical scavenging assay with all crude extracts showed value of IC50 of leaf, stem, pod and root crude extracts were 8.32, 12.26, 21.62 and 35.99 mg/ml, respectively. Studied cytotoxicity of crude extracts on human breast adenocarcinoma cell line by MTT assay found that pod extract had the most cytotoxicity CC50 value, 32.41 µg/ml. Antioxidant activity and cytotoxicity of crude extracts exhibited that the more increase of extract concentration, the more activities indicated. According to the bioactivities results, the leaf crude extract of Cleoma viscosa Linn. is the most interesting plant part for further work to search the beneficial of this weed.

Keywords: Antimicrobial, antioxidant activity, Cleoma viscosa Linn., cytotoxicity test, total phenolic compound.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 1726

Authors: G. Spada, E. Gavini, P. Giunchedi

Abstract:

Aim of this work was to compare the efficacy of two loading methods of proteins onto polymeric nanocarriers: adsorption and encapsulation methods. Preliminary studies of protein loading were done using Bovine Serum Albumin (BSA) as model protein. Nanocarriers were prepared starting from polylactic co-glycolic acid (PLGA) polymer; production methods used are two different variants of emulsion evaporation method. Nanoparticles obtained were analyzed in terms of dimensions by Dynamic Light Scattering and Loading Efficiency of BSA by Bradford Assay. Loaded nanoparticles were then submitted to in-vitro protein dissolution test in order to study the effect of the delivery system on the release rate of the protein.

Keywords: Drug delivery, nanoparticles, PLGA, proteinadsorption, protein encapsulation.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 2464

Authors: Zhazgul Kelgenbaeva, Emil Omurzak, Saadat Sulaimankulova, Tsutomu Mashimo

Abstract:

Spherical shaped magnetite (Fe3O4) and Au@Fe3O4 nanoparticles were successfully synthesized from Fe electrodes immersed in water with CTAB surfactant and HAuCl4 solution using simple method-pulsed plasma in liquid, without the use of dopants or special conditions for stabilization. Vibrating sample magnetometer indicated ferromagnetic behavior of particles at room temperature with coercivity and saturation magnetization of (Hc=105 Oe, Ms=6.83 emu/g) for Fe3O4 and (Hc=175, Ms=3.56emu/g) for Au@Fe3O4 nanoparticles. Structure and morphology of nanoparticles were characterized by X-ray Diffraction analysis and HR-TEM measurements. The cytotoxicity of nanoparticles was indicated using a XTT assay to be very low (cell viability: 98-89% with Fe3O4 and 99-91% for Au@Fe3O4 NPs).

Keywords: Magnetite, Gold coated magnetite, Nanoparticles, Pulsed Plasma in Liquid, Cytotoxicity.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 2366

Authors: Samar Saadeldin Abdelmotalab Omer, Ikram Mohamed Eltayeb Elsiddig, Amna Beshir Medani Ahmed, Saad Mohamed Hussein Ayoub

Abstract:

Many herbal medicinal products are considered potential anti-hypercholesterolemic agents with encouraging safety profiles, however only a limited amount of clinical research exists to support their efficacy. The present study was designed to compare the antihypercholesterolemic and antioxidant activities of the crude ethanolic extracts of Citrus reticulata fruit peel, Zingiber officinale rhizome and Sesamum indicum seeds. Forty-five rats were used throughout the experiment which are extended for four weeks. These were divided into nine groups, five rats per each group as follows; group 1 was the normal control group (rats only fed standard normal rat diet), group 2 was the hypercholesterolemic control group (rats fed only hypercholesterolemic diet which contained 1% cholesterol plus 10% saturated animal fat added to the normal rat diet), groups 3 and 4 were fed hypercholesterolemic diet in addition to Citrus reticulata ethanolic extract at doses of (250mg/kg (group 3) and 500mg/kg (group 4)) administered daily via oral route, groups 5 and 6 were given hypercholesterolemic diet in addition to Zingiber officinale ethanolic extract at doses of (250mg/kg (group 5) and 500mg/kg (group 6)) daily through oral route, groups 7 and 8 fed on hypercholesterolemic diet in addition to Sesamum indicum ethanolic extract at doses of (250mg/kg (group 7) and 500mg/kg (group 8)) daily orally; and group 9 rats were given hypercholesterolemic diet in addition to atorvastatin (0.18mg/kg) daily via oral route as a standard reference antihypercholesterolemic drug. Blood samples from all groups were drawn from the retro-orbital venous plexus four weeks following treatment after overnight fasting and the lipid profile (total cholesterol (TC), high density lipoprotein-cholesterol (HDL-C), low density lipoprotein-cholesterol (LDL-C) and triglyceride levels) were measured and the risk ratio (TC/HDL-C) was assessed. The antioxidant activity of the three plants extracts was determined using DPPH free-radical antioxidant assay. Results of in vivo and in vitro antihypercholesterolemic and antioxidant assay respectively, revealed that the three extracts possess comparable antioxidant and antihypercholesterolemic activities.

Keywords: Antihypercholesterolemic effects, Antioxidant activity, HDL, LDL, TC, TGs, Citrus reticulata, Sesamum indicum, Zingiber officinale.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 2634

Authors: M. Ghanadi, M. Rezazadeh*, M. Ardeshiri, R. Gholipour Peyvandi, M. Jafarzadeh, M. Shahriari, M.Rezaei Rad, Z. Gholamzadeh

Abstract:

A 3D industrial computed tomography (CT) manufactured based on a first generation CT systems, single-source – single-detector, was evaluated. Operation accuracy assessment of the manufactured system was achieved using simulation in comparison with experimental tests. 137Cs and 60Co were used as a gamma source. Simulations were achieved using MCNP4C code. Experimental tests of 137Cs were in good agreement with the simulations

Keywords: Gamma source, Industrial CT, MCNP4C, Operation assessment

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 1617

Authors: Mashayekh Amir Shahriar, Akhlaghi Morteza, Rajaee Hajar, Khani Mohammad Reza, Shokri Babak

Abstract:

A new and novel approach in medicine is the use of cold plasma for various applications such as sterilization blood coagulation and cancer cell treatment. In this paper a pin-to-hole plasma jet suitable for biological applications is investigated and characterized and the possibility and feasibility of cancer cell treatment is evaluated. The characterization includes power consumption via Lissajous method, thermal behavior of plasma using Infra-red camera as a novel method, Optical Emission Spectroscopy (OES) to determine the species that are generated. Treatment of leukemia cancer cells is also implemented and MTT assay is used to evaluate viability.

Keywords: Atmospheric Pressure Plasma Jet (APPJ), Plasma Medicine, Cancer cell treatment, leukemia.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 2195

Authors: Balgabay Sadepovich Maikanov, Laura Tyulegenovna Auteleyeva, Seidenova Simbat Polatbekovna

Abstract:

In this article reduced nutritional value of the rabbits’ meat at 1, 1 dimethylhydrazine experimental toxicosis is shown. The assay was performed on liquid chromatograph SHIMADZU LC-20 Prominence (Japan) with fluorometric and spectrophotometric detector. This research has revealed that samples of rabbit meat of the experimental group had significant differences from the control group:in amino acids concentration from 1.2% to 9.1%; vitamin concentration from 11.2% to 60.5%, macro – minerals concentration from 17.4% to 78.1% and saturated fatty acids concentration from 17,1% to 34.5%, respectively. The decrease in the chemical composition of rabbits’ meat at 1,1 dimethylhydrazine toxicosis may be due to changes in the internal processes associated with impaired metabolic homeostasis of animals.

Keywords: 1, 1-dimethylhydrazine, metabolic homeostasis, nutritional value, rabbit meat.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 724

Authors: Michael Russelle S. Alvarez, Noel S. Quiming, Francisco M. Heralde

Abstract:

This study aims to: a) obtain ethanolic (95% EtOH) and aqueous extracts of Selaginella elmeri, Christella dentata, Elatostema sinnatum, Curculigo capitulata, Euphorbia hirta, Murraya koenigii, Alpinia speciosa, Cymbopogon citratus, Eucalyptus globulus, Jatropha curcas, Psidium guajava, Gliricidia sepium, Ixora coccinea and Capsicum frutescens and screen them for larvicidal activities against Aedes aegypti (Linn.) and Aedes albopictus (Skuse) larvae; b) to fractionate the most active extract and determine the most active fraction; c) to determine the larvicidal properties of the most active extract and fraction against by computing their percentage mortality, LC50, and LC90 after 24 and 48 hours of exposure; and d) to determine the nature of the components of the active extracts and fractions using phytochemical screening. Ethanolic (95% EtOH) and aqueous extracts of the selected plants will be screened for potential larvicidal activity against Ae. aegypti and Ae. albopictus using standard procedures and 1% malathion and a Piper nigrum based ovicide-larvicide by the Department of Science and Technology as positive controls. The results were analyzed using One-Way ANOVA with Tukey’s and Dunnett’s test. The most active extract will be subjected to partial fractionation using normal-phase column chromatography, and the fractions subsequently screened to determine the most active fraction. The most active extract and fraction were subjected to dose-response assay and probit analysis to determine the LC50 and LC90 after 24 and 48 hours of exposure. The active extracts and fractions will be screened for phytochemical content. The ethanolic extracts of C. citratus, E. hirta, I. coccinea, G. sepium, M. koenigii, E globulus, J. curcas and C. frutescens exhibited significant larvicidal activity, with C. frutescens being the most active. After fractionation, the ethyl acetate fraction was found to be the most active. Phytochemical screening of the extracts revealed the presence of alkaloids, tannins, indoles and steroids. A formulation using talcum powder–300 mg fraction per 1 g talcum powder–was made and again tested for larvicidal activity. At 2 g/L, the formulation proved effective in killing all of the test larvae after 24 hours.

Keywords: Larvicidal activity screening, partial purification, dose-response assay, Capsicum frutescens.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 1674

Authors: Kakali Bhadra

Abstract:

Harmalol administration caused remarkable reduction in proliferation of HepG₂ cells with GI₅₀ of 14.2 mM, without showing much cytotoxicity in embryonic liver cell line, WRL-68. Data from circular dichroism and differential scanning calorimetric analysis of harmalol-CT DNA complex shows conformational changes with prominent CD perturbation and stabilization of CT DNA by 8 ^oC. Binding constant and stoichiometry was also calculated using the above biophysical techniques. Further, dose dependent apoptotic induction ability of harmalol was studied in HepG₂ cells using different biochemical assays. Generation of ROS, DNA damage, changes in cellular external and ultramorphology, alteration of membrane, formation of comet tail, decreased mitochondrial membrane potential and a significant increase in Sub G_o/G₁ population made the cancer cell, HepG₂, prone to apoptosis. Up regulation of p53 and caspase 3 further indicated the apoptotic role of harmalol.

Keywords: Apoptosis, beta carboline alkaloid, comet assay, cytotoxicity, ROS.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 1129

Authors: Rizna Triana Dewi, Sanro Tachibana, Ahmad Darmawan

Abstract:

The bioassay-guided isolation and purification of an ethyl acetate extract of Aspergillus terreus MC751 led to the characterization of butyrolactone I as an antidiabetic and antioxidant. The antidiabetic activity of butyrolactone I was evaluated by α- glucosidase and α-amylase inhibition assays. Butyrolactone I demonstrated significant concentration-dependent, mixed-type inhibitory activity against yeast α-glucosidase with an IC50 of 54μM. However, the compound exhibited less activity against rat intestinal α-glucosidase and α-amylase. This is the first report on α-glucosidase inhibitory activity of butyrolactone I. The antioxidative activity of butyrolactone I was evaluated based on scavenging effects on 1,1- diphenyl-2-picrylhydrazyl (DPPH) (IC50 =51 μM) and hydrogen peroxide (IC50= 141 μM) radicals as well as a reducing power assay. The results suggest that butyrolactone I is a promising antidiabetic as well as antioxidant and should be considered for clinical trials.

Keywords: Aspergillus terreus MC751, antidiabetic, antioxidant, Butyrolactone I.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 2774

Authors: Y. Baba hamed, A. Medjdoub, H. Merzouk, M. Narce

Abstract:

The aim of this work was to study the in vitro effects of δ-lactam 1 and its 4-chlorophenyl derivative 2, on the proliferative responses of human lymphocytes and Th1 and Th2 cytokine secretion. The possible protective role of vitamin E on intracellular stress oxidative induced by these compounds was also investigated. Peripheral blood lymphocytes were isolated using differential centrifugation on a density gradient of Histopaque. They were cultured with mitogen concanavalin A, vitamin E (10 μM) and with different concentrations of the compounds 1 and 2 (0.1 to 10 μM). Proliferation (MTT assay), IL-2, INFγ and IL-4 (Elisa kits), intracellular superoxide anion were determined. 1 and 2 were immunostimulant and increased cytokine secretion with a shift away from Th1 response to Th2. These properties were however accompanied by an increase in intracellular oxidative stress. The presence of vitamin E exhibited protective effects by reducing δ- lactam-induced superoxide anion generation in lymphocytes.

Keywords: Cytokines, δ-Lactams, In vitro Lymphocyte Proliferation, Superoxide Anion

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 3835

Authors: Majid Forghani, Michael Khachay

Abstract:

In genetics, the impact of neighboring amino acids on a target site is referred as the nearest-neighbor effect or simply neighbor effect. In this paper, a new method called wavelet particle decomposition representing the one-dimensional neighbor effect using wavelet packet decomposition is proposed. The main idea lies in known dependence of wavelet packet sub-bands on location and order of neighboring samples. The method decomposes the value of a signal sample into small values called particles that represent a part of the neighbor effect information. The results have shown that the information obtained from the particle decomposition can be used to create better model variables or features. As an example, the approach has been applied to improve the correlation of test and reference sequence distance with titer in the hemagglutination inhibition assay.

Keywords: Antigenic variants, neighbor effect, wavelet packet, wavelet particle decomposition.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 728

Authors: Achara Dholvitayakhun, Nathanon Trachoo

Abstract:

In this study, the forty Thai medicinal plants were used to screen the antibacterial activity against Campylobacter jejuni. Crude 95% ethanolic extracts of each plant were prepared. Antibacterial activity was investigated by the disc diffusion assay, and MICs and MBCs were determined by broth microdilution. The results of antibacterial screening showed that five plants have activity against C.jejuni including Adenanthera pavonina L., Moringa oleifera Lam., Annona squamosa L., Hibiscus sabdariffa L. and Eupotorium odortum L. The extraction of A. pavonina L. and A. squamosa L. produced an outstanding against C. jejuni, inhibiting growth at 62.5-125 and 250-500 μg/mL, respectively. The MBCs of two extracts were just 4-fold higher than MICs against C. jejuni, suggesting the extracts are bactericidal against this species. These results indicate that A. pavonina and A. squamosa could potentially be used in modern applications aimed at treatment or prevention of foodborne disease from C. jejuni.

Keywords: Antibacterial activity, Thai medicinal plants, Campylobacter jejuni

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 2629

Authors: A. Chennaf, M. Yahia, W. Bouafia, S. Benbia, D. Khellaf

Abstract:

Our study was designed to highlight changes in  certain biochemical parameters (CH, TG, HDL, GOT, GPT, LDL and  CRP), obese women infertile fertile witnesses and research potential  pathophysiological link between obesity and infertility in this  population of women. This practical work was focused on a  population of 24 obese women infertile, compared to controls,  subjects without any pathology causing disruption of parameters to  be studied to determine the contribution of obesity in the etiology of  infertility. The assay results revealed a highly significant difference  between the two groups in serum CH, TG, HDL, TGO and TGP (P  <0.0001) and in the rate of LDL (p = 0.0017) and CRP (p = 0.02).  The present study indicates that obesity is associated with infertility,  but no direct pathophysiological link between obesity and infertility  has been determined. Further in-depth studies are needed to  determine the exact mechanism by which overweight leads to female  infertility.

 

Keywords: Obesity, fertility, infertility, biochemical, women.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 2341

Authors: Nneka N. Uchegbu, Ndidi F. Amulu

Abstract:

The work studied the effect of germination on proximate, phenol and flavonoid content and antioxidant activities (AOA) of African Yam been (AYB). Germination was done in controlled dark chamber (100% RH, 28oC). The proximate, phenol and flavonoid content and antioxidant activities before and after germination were investigated. The crude protein, moisture, and crude fiber content of germinated AYB were significantly higher (P<0.05) than that of ungermianated seed, while the fat, Ash and carbohydrate content of ungerminated were higher than the germinated seed. Germination increased the phenol and flavoniod content by 19.14% and 14.53% respectively. The results of AOA assay showed that the DPPH, reducing power and FRAP of germinated AYB seed gave high values: 48.92 ±1.22 μg/ml, 0.75± 0.15μg/ml and 98.60±0.04 μmol/g while that of ungerminated seed were: 31.33μ/ml, 0.56±1.52μg/ml and 96.11±1.13μmol/g respectively. Germinated AYB has phytochemicals with potential AOA for disease prevention.

Keywords: Antioxidant, flovonoid, germination, phenol.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 2312

Authors: Amr A. El Hanafy, Yasir Anwar, Saleh A. Mohamed, Saleh Mohamed Saleh Al-Garni, Jamal S. M. Sabir, Osama A. H. Abu Zinadah, Mohamed Morsi Ahmed

Abstract:

In the vicinity of red sea about 15 fungi species were isolated from oil contaminated sites. On the basis of aptitude to degrade the crude oil and DCPIP assay, two fungal isolates were selected amongst 15 oil degrading strains. Analysis of ITS-1, ITS-2 and amplicon pyrosequencing studies of fungal diversity revealed that these strains belong to Penicillium and Aspergillus species. Two strains that proved to be the most efficient in degrading crude oil was Aspergillus niger (54%) and Penicillium commune (48%) Subsequent to two weeks of cultivation in BHS medium the degradation rate were recorded by using spectrophotometer and GC-MS. Hence, it is cleared that these fungal strains has capability of degradation and can be utilize for cleaning the Saudi Arabian environment.

Keywords: Fungal strains, hydrocarbon contaminants, molecular identification, biodegradation, GC-MS.

Procedia APA BibTeX Chicago EndNote Harvard JSON MLA RIS XML ISO 690 PDF Downloads 2869