Search results for: spectrophotometer assay

Authors: Siobhan O’Shea, Sangeetha Vijaysri Nair, Hee Cheol Kim, Charles Thomas Nugent, Cheuk Yan William Tong, Sam Douthwaite, Andrew Worlock

Abstract:

The Aptima® HIV-1 Quant Dx Assay is a fully automated assay on the Panther system. It is based on Transcription-Mediated Amplification and real time detection technologies. This assay is intended for monitoring HIV-1 viral load in plasma specimens and for the detection of HIV-1 in plasma and serum specimens. Nine-hundred and seventy nine specimens selected at random from routine testing at St Thomas’ Hospital, London were anonymised and used to compare the performance of the Aptima HIV-1 Quant Dx assay and Roche COBAS® AmpliPrep/COBAS® TaqMan® HIV-1 Test, v2.0. Two-hundred and thirty four specimens gave quantitative HIV-1 viral load results in both assays. The quantitative results reported by the Aptima Assay were comparable those reported by the Roche COBAS AmpliPrep/COBAS TaqMan HIV-1 Test, v2.0 with a linear regression slope of 1.04 and an intercept on -0.097. The Aptima assay detected HIV-1 in more samples than the Roche assay. This was not due to lack of specificity of the Aptima assay because this assay gave 99.83% specificity on testing plasma specimens from 600 HIV-1 negative individuals. To understand the reason for this higher detection rate a side-by-side comparison of low level panels made from the HIV-1 3rd international standard (NIBSC10/152) and clinical samples of various subtypes were tested in both assays. The Aptima assay was more sensitive than the Roche assay. The good sensitivity, specificity and agreement with other commercial assays make the HIV-1 Quant Dx Assay appropriate for both viral load monitoring and detection of HIV-1 infections.

Keywords: HIV viral load, Aptima, Roche, Panther system

Procedia PDF Downloads 348

Authors: Meltem Betül Sağlam, Halil İbrahim Güler, Aykut Sağlam

Abstract:

In this work, phthalocyanine compounds containing α-naphtholbenzeinunits have been synthesized. Mutagenicity and DNA binding properties of the compounds were investigated by Salmonella/Microsome Assay and spectrophotometer. According to the results of the preliminary range finding tests, the compounds gave no toxic effect to all tester strain S. typhimurium TA98 and TA100 at doses of 500, 1100, 350, 500 and 750 µg/plate in the presence and absence of S9, respectively. This study showed that all compounds exhibited efficient DNA-binding activity. In conclusion, these non-toxic compounds may be used as effective DNA dyes for molecular biology studies.

Keywords: dye, mutagenicity, phthalocyanine, toxicity

Procedia PDF Downloads 207

Authors: K. C. Gayithri, S. K. Naveen Kumar

Abstract:

ZnO is one of the most important semiconductor materials, non toxic, biocompatible, antibacterial properties for research and it is used in many biomedical applications. MnxZn1-xO nano thin films were prepared by a spin coating sol-gel method on silicon substrate. The structural, optical, electrical properties of Mn Doped ZnO are studied by using X-rd, FESEM, UV-Visible spectrophotometer. The X-rd reveals that the sample shows hexagonal wurtzits structure. Surface morphology and thickness of the sample are characterized by field emission scanning electron microscopy. Absorption and transmission spectra are studied by UV-Visible spectrophotometer. The electrical properties are measured by TCR meter.

Keywords: transition metals, Mn doped ZnO, Sol-gel, x-ray diffraction

Procedia PDF Downloads 372

Authors: Ali Bahri Lubis

Abstract:

Tryptophan oxidation by Heme-dioxygenase enzyme is initial important stepTryptophan oxidation by Heme-dioxygenase enzyme is initial important step in kynurenine pathway implicating to several severe diseases such as Parkinson’s Disease, Huntington Disease, poliomyelitis and cataract. It is crucial to comprehend the oxidation mechanism with the hope to find decent treatment upon abovementioned diseases. The mechanism has been debatable since no one has been yet proved the mechanism obviously. In this research we have attempted to prove mechanistic steps of tryptophan oxidation via human indoleamine dioxygenase (h-IDO) using various substrates: L-tryptophan, L-tryptophan (indole-ring-2-13C), L-fully-labelled13C-tryptophan, L-N-methyl-tryptophan, L-tryptophan and 2-amino-3-(benzo(b)thiophene-3-yl) propanoic acid. All enzyme assay experiments were measured using a UV-Vis spectrophotometer, LC-MS, 1H-NMR, and HSQC. We also successfully synthesized enzyme products as our control in NMR measurements. The result exhibited that the distinct substrates produced N-formyl kynurenine (NFK) and hydroxypyrrolloindoleamine carboxylate acid (HPIC) in different concentrations and isomers, correlated to the proposal of considered mechanism reaction in kynurenine pathway implicating to several severe diseases such as Parkinson’s Disease, Huntington Disease, poliomyelitis and cataract. It is crucial to comprehend the oxidation mechanism with the hope to find decent treatment for the abovementioned diseases. The mechanism has been debatable since no one has yet proven the mechanism obviously. In this research we have attempted to prove mechanistic steps of tryptophan oxidation via human indoleamine dioxygenase (h-IDO) using various substrates: L-tryptophan, L-tryptophan (indole-ring-2-13C), L-fully-labelled13C-tryptophan, L-N-methyl-tryptophan, L-tryptophan and 2-amino-3-(benzo(b)thiophene-3-yl) propanoic acid. All enzyme assay experiments were measured using a UV-Vis spectrophotometer, LC-MS, 1H-NMR and HSQC. We also successfully synthesized enzyme products as our control in NMR measurements. The result exhibited that the distinct substrates produced N-formyl kynurenine (NFK) and hydroxypyrrolloindoleamine carboxylate acid (HPIC) in different concentrations and isomers, correlated to the proposal of considered mechanism reaction.

Keywords: heme-dioxygenase enzyme, tryptophan oxidation, kynurenine pathway, n-formyl kynurenine

Procedia PDF Downloads 59

Authors: P. Singh, R. M. Banik

Abstract:

Artificial neural network (ANN) was employed to optimize assay parameters viz., time, temperature, pH of reaction mixture, enzyme volume and substrate concentration of L-glutaminase from Bacillus cereus MTCC 1305. ANN model showed high value of coefficient of determination (0.9999), low value of root mean square error (0.6697) and low value of absolute average deviation. A multilayer perceptron neural network trained with an error back-propagation algorithm was incorporated for developing a predictive model and its topology was obtained as 5-3-1 after applying Levenberg Marquardt (LM) training algorithm. The predicted activity of L-glutaminase was obtained as 633.7349 U/l by considering optimum assay parameters, viz., pH of reaction mixture (7.5), reaction time (20 minutes), incubation temperature (35˚C), substrate concentration (40mM), and enzyme volume (0.5ml). The predicted data was verified by running experiment at simulated optimum assay condition and activity was obtained as 634.00 U/l. The application of ANN model for optimization of assay conditions improved the activity of L-glutaminase by 1.499 fold.

Keywords: Bacillus cereus, L-glutaminase, assay parameters, artificial neural network

Procedia PDF Downloads 410

Authors: K. Khaerunnisa, D. Ryangga, S. A. Pawiro

Abstract:

Over the past few decades, film dosimetry has become a tool which is used in various radiotherapy modalities, either for clinical quality assurance (QA) or dose verification. The response of the film to irradiation is usually expressed in optical density (OD) or net optical density (netOD). While the film's response to radiation is not linear, then the use of film as a dosimeter must go through a calibration process. This study aimed to compare the function of the calibration curve of various measurement methods with various densitometer, using a flat bad scanner, point densitometer and spectrophotometer. For every response function, a radichromic film calibration curve is generated from each method by performing accuracy, precision and sensitivity analysis. netOD is obtained by measuring changes in the optical density (OD) of the film before irradiation and after irradiation when using a film scanner if it uses ImageJ to extract the pixel value of the film on the red channel of three channels (RGB), calculate the change in OD before and after irradiation when using a point densitometer, and calculate changes in absorbance before and after irradiation when using a spectrophotometer. the results showed that the three calibration methods gave readings with a netOD precision of doses below 3% for the uncertainty value of 1σ (one sigma). while the sensitivity of all three methods has the same trend in responding to film readings against radiation, it has a different magnitude of sensitivity. while the accuracy of the three methods provides readings below 3% for doses above 100 cGy and 200 cGy, but for doses below 100 cGy found above 3% when using point densitometers and spectrophotometers. when all three methods are used for clinical implementation, the results of the study show accuracy and precision below 2% for the use of scanners and spectrophotometers and above 3% for precision and accuracy when using point densitometers.

Keywords: Callibration Methods, Film Dosimetry EBT3, Flat Bad Scanner, Densitomete, Spectrophotometer

Procedia PDF Downloads 110

Authors: Tahisa M. Pedroso, Hérida R. N. Salgado

Abstract:

The microbiological turbidimetric assay allows the determination of potency of the drug, by measuring the turbidity (absorbance), caused by inhibition of microorganisms by ertapenem sodium. Ertapenem sodium (ERTM), a synthetic antimicrobial agent of the class of carbapenems, shows action against Gram-negative, Gram-positive, aerobic and anaerobic microorganisms. Turbidimetric assays are described in the literature for some antibiotics, but this method is not described for ertapenem. The objective of the present study was to develop and validate a simple, sensitive, precise and accurate microbiological assay by turbidimetry to quantify ertapenem sodium injectable as an alternative to the physicochemical methods described in the literature. Several preliminary tests were performed to choose the following parameters: Staphylococcus aureus ATCC 25923, IAL 1851, 8 % of inoculum, BHI culture medium, and aqueous solution of ertapenem sodium. 10.0 mL of sterile BHI culture medium were distributed in 20 tubes. 0.2 mL of solutions (standard and test), were added in tube, respectively S1, S2 and S3, and T1, T2 and T3, 0.8 mL of culture medium inoculated were transferred to each tube, according parallel lines 3 x 3 test. The tubes were incubated in shaker Marconi MA 420 at a temperature of 35.0 °C ± 2.0 °C for 4 hours. After this period, the growth of microorganisms was inhibited by addition of 0.5 mL of 12% formaldehyde solution in each tube. The absorbance was determined in Quimis Q-798DRM spectrophotometer at a wavelength of 530 nm. An analytical curve was constructed to obtain the equation of the line by the least-squares method and the linearity and parallelism was detected by ANOVA. The specificity of the method was proven by comparing the response obtained for the standard and the finished product. The precision was checked by testing the determination of ertapenem sodium in three days. The accuracy was determined by recovery test. The robustness was determined by comparing the results obtained by varying wavelength, brand of culture medium and volume of culture medium in the tubes. Statistical analysis showed that there is no deviation from linearity in the analytical curves of standard and test samples. The correlation coefficients were 0.9996 and 0.9998 for the standard and test samples, respectively. The specificity was confirmed by comparing the absorbance of the reference substance and test samples. The values obtained for intraday, interday and between analyst precision were 1.25%; 0.26%, 0.15% respectively. The amount of ertapenem sodium present in the samples analyzed, 99.87%, is consistent. The accuracy was proven by the recovery test, with value of 98.20%. The parameters varied did not affect the analysis of ertapenem sodium, confirming the robustness of this method. The turbidimetric assay is more versatile, faster and easier to apply than agar diffusion assay. The method is simple, rapid and accurate and can be used in routine analysis of quality control of formulations containing ertapenem sodium.

Keywords: ertapenem sodium, turbidimetric assay, quality control, validation

Procedia PDF Downloads 375

Authors: Anurag Gaur, Neha Sharma

Abstract:

In this work, we have synthesized BaTiO3 via sol gel method by sintering at different temperatures (600-1000 0C) and studied their structural, optical and ferroelectric properties through X-Ray diffraction (XRD), UV-Vis spectrophotometer and PE Loop Tracer. X-Ray diffraction patterns of barium titanate samples show that the peaks of the diffractogram are successfully indexed with the tetragonal structure of BaTiO3 along with some minor impurities of BaCO3. The optical band gap calculated through UV Visible spectrophotometer varies from 4.37 to 3.80 eV for the samples sintered at 600 to 1000 0 C, respectively. The particle size calculated through transmission electron microscopy varies from 20 to 60 nm for the samples sintered at 600 to 1000 0C, respectively. Moreover, it has been observed that the ferroelectricity reduces as we increase the sintering temperature.

Keywords: nanostructures, ferroelectricity, sol-gel method, diffractogram

Procedia PDF Downloads 395

Authors: Rajesh S. Mony, Vaidyaratnam Oushadhasala

Abstract:

An attempt is made in this study to evolve a drug development modality based on classical Ayurvedic knowledge base as well as on modern scientific methodology. The present study involves (a) identification of a specific ailment condition, (b) the selection of a polyherbal formulation, (c) deciding suitable extraction procedure, (d) confirming the efficacy of the combination by in-vitro trials and (e) fixing up the recommended dose. The ailment segment selected is arthritic condition. The selected herbal combination is Kunturushka, Vibhitaki, Guggulu, Haridra, Maricha and Nirgundi. They were selected as per Classical Ayurvedic references, Authentified as per API (Ayurvedic Pharmacopeia of India), Extraction of each drug was done by different ratios of Hydroalcoholic menstrums, Invitro assessment of each extract after removing residual solvent for anti-Inflammatory, anti-arthritic activities (by UV-Vis. Spectrophotometer with positive control), Invitro assessment of each extract for COX enzyme inhibition (by UV-Vis. Spectrophotometer with positive control), Selection of the extracts was made having good in-vitro activity, Performed the QC testing of each selected extract including HPTLC, that is the in process QC specifications, h. Decision of the single dose with mixtures of selected extracts was made as per the level of in-vitro activity and available toxicology data, Quantification of major groups like Phenolics, Flavonoids, Alkaloids and Bitters was done with both standard Spectrophotometric and Gravimetric methods, Method for Marker assay was developed and validated by HPTLC and a good resolved HPTLC finger print was developed for the single dosage API (Active Pharmaceutical Ingredient mixture of extracts), Three batches was prepared to fix the in process and API (Active Pharmaceutical Ingredient) QC specifications.

Keywords: drug development, antiinflammatory, quality stardardisation, planar chromatography

Procedia PDF Downloads 75

Authors: Puchita Chokcharoenying

Abstract:

Polyphenols are the most abundant antioxidants found in plants, and they are highly effective at scavenging oxidative free radicals. Antioxidants are substances found in medicinal plants to help prevent heart disease, stroke, and some cancers. This study focused on evaluating the flavonoids content of Chrysanthemum Indicum and determine their antioxidant capacity by using DPPH and ABTS radical scavenging capacity assay. The total flavonoid content of C. indicumextract was determined and expressed as quercetin equivalents (QE)/g measured by an aluminiumchloride colorimetric method. The results showed that the IC50 of C. indicum extract were 83.57μg/mL ± 0.875 and52.57μg/mL ± 0.632for DPPH and ABTS, respectively. C. indicumextract exhibited antioxidant activities as a concentration dependent manner. In the DPPH assay, vitamin C was used as a positive control, whereas Trolox was used as a positive control in the ABTS assay. In summary, C. indicum extract is rich in flavonoids, which have potent antioxidant properties. Thus, C. indicum extract is a good source of antioxidants and can be developed for medicinal purposes. Nevertheless, more research on the antioxidant activity of C. indicum extract and in vivo antioxidant studies are still needed.

Keywords: ABTS assay, antioxidant, chrysanthemum indicum, DPPH assay, total flavonoid content

Procedia PDF Downloads 235

Authors: Ajay Pratap Singh Lodhi, Deepak Kumar

Abstract:

Vegetable oil-based environmentally friendly metalworking fluids (MWFs) are formulated. The tribological performance, cytotoxicity, and corrosion resistance of the formulated fluids (FFs) are evaluated and benchmarked with commercial mineral oil-based MWFs (CF). Results show that FFs exhibited better machining characteristics (roughness, cutting forces, and surface morphology) during machining than CF. MTT assay and Live dead cell assay confirm the cytocompatibility nature of the FFs relative to the toxic CF. Electrochemical analysis shows that FFs and CF exhibited comparable corrosion current density.

Keywords: corrosion inhibitors, cytotoxicity, machining, MTT assay, Taguchi method, vegetable oil

Procedia PDF Downloads 167

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 the 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 the capability of degradation and can be utilized for cleaning the Saudi Arabian environment.

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

Procedia PDF Downloads 498

Authors: Dawit Gebreegzabher Hagos, Yazezew Kebede Kiro, Mahmud Abdulkader, Henk H. D. F. Schallig, Dawit Wolday

Abstract:

Rapid and accurate visceral leishmaniasis (VL) diagnosis is needed to initiate prompt treatment to reduce morbidity and mortality. Here, we evaluated the performance of loop-mediated isothermal amplification (LAMP) assay for the diagnosis of VL from blood in an endemic area in Ethiopia. LAMP was positive in 117/122 confirmed VL cases and negative in 149/152 controls, resulting in a sensitivity of 95.9% (95% CI: 90.69–98.66) and a specificity of 98.0% (95% CI: 94.34–99.59), respectively. The sensitivity of the LAMP assay was 95.0% (95% CI: 88.61–98.34) in HIV-negatives and 100% (95% CI: 85.18–100.0) in HIV-positives. Compared with microscopy, LAMP detected 82/87 (94.3%, 95% CI: 87.10–98.11) of the microscopy1 cases and was negative in 11/27 (40.7%, 95% CI: 22.39–61.20) of the microscopy2 cases. Compared with the rK39 serology, LAMP detected 113/120 (94.2%, 95% CI: 88.35–97.62) of the rK391 cases and was negative in 149/154 (96.8%, 95% CI: 92.59–98.94) of the rK392 cases. However, when compared with microscopy only, rK39 detected 83/87 (95.4%, 95% CI: 88.64–98.73) of the microscopy1 cases and negative in only 12/27 (44.4%, 95% CI: 25.48–64.67) of the microscopy– cases. There was an excellent agreement between rK39 and LAMP (Kappa 5 0.91, 95% CI: 0.86–0.96). Furthermore, an algorithm using rK39 followed by LAMP would yield a sensitivity of 99.2% (95%CI: 95.52–99.89) and a specificity of 98.0% (95% CI: 94.34–99.59). The findings demonstrate that the LAMP assay is an accurate and rapid molecular assay for VL diagnosis, including in HIV-1 co-infected patients, in an endemic setting.

Keywords: visceral leishmaniasis, HIV, diagnosis, LAMP, Ethiopia

Procedia PDF Downloads 69

Authors: Jirathan Pongchababnapa

Abstract:

Polyphenols are the most common antioxidant found in plants and are efficient in capturing oxidative free radicals. Antioxidants are substances found in medicinal plants which may have a protective role to play in certain conditions such as heart disease, stroke and some cancers. By relying on these benefits, we have traced out the presence of antioxidant in Camellia sinensis leaves extract. This study aims to evaluate flavonoids content in C. sinensisextract and investigate antioxidant activities by using DPPH and ABTS radical scavenging capacity assay. The total flavonoid content of C. Sinensis extract was determined and expressed as quercetin equivalents (QE)/g measured by the aluminum chloride colorimetric method. The results showed that the IC₅₀ of C. Sinensis leaves extract were 40.90 μg/mL ± 0.755 and32.96 μg/mL ± 0.679 for DPPH and ABTS, respectively. C. Sinensis extract at increasing concentration showed antioxidant activities as a concentration dependent manner. In the DPPH assay, vitamin C was used as a positive control, whereas Trolox was used as a positive control in the ABTS assay. In conclusion, C. Sinensis extract consisted of a high amount of flavonoids content which possesses potent antioxidant activity. However, further investigation on the identification of pure compound of this plant and molecular antioxidant assays are still required.

Keywords: ABTS assay, antioxidant, camellia sinensis, DPPH assay, total flavonoid content

Procedia PDF Downloads 177

Authors: Tawqeer Ali Syed, Prakash Chandra

Abstract:

This study aims to evaluate the antileishmanial activity of Nigella sativa black seed extract. This well-known plant extract was taken from the botanical garden of Kashmir. Materials and Methods: The methanolic extracts of these plants were screened for their antileishmanial activity against Leishmania major using 3‑(4.5‑dimethylthiazol‑2yl)‑2.5‑diphenyltetrazolium bromide assay or MTT assay. Results: The methanolic extract of Nigella sativa showed potential antileishmanial activity at an inhibition% value of 80.29% ± 0.65%. IC 50 was calculated after 48 hours to be 964.3 µg/ml. Conclusion: Considering these results, these medicinal plants from Kashmir could serve as potential drug sources for antileishmanial compounds.

Keywords: MTT assay, antileishmanial, cell viability, Nigella sativa

Procedia PDF Downloads 185

Authors: Huma Naeem, Saif-Ur-Rehman Kashif, Muhammad Nawaz Chaudhry

Abstract:

Milk plays a significant role in the dietary requirements of human beings as it is a single source that provides various essential nutrients. A study was conducted to evaluate the heavy metal concentration in the raw milk marketed in Data Gunj Baksh Town of Lahore. A total of 180 samples of raw milk were collected in pre-monsoon, monsoon and post-monsoon season from five colonies of Data Gunj Baksh Town, Lahore. The milk samples were subjected to heavy metal analysis (Cr, Cu) by atomic absorption spectrophotometer. Results indicated high levels of Cr and Cu in post-monsoon seasons. Heavy metals were detected in milk in all samples under study and exceeded the standards given by FAO.

Keywords: atomic absorption spectrophotometer, chromium, copper, heavy metal

Procedia PDF Downloads 410

Authors: Ayse Ozge Demir, Nihat Mert

Abstract:

In this research, Kilis and Honamli goats are used, which are specific local genetic resources of Turkey. The herds were independent, but they had similar care and nutrition circumstances. From each breed 30 samples were taken, in all 120 samples were collected. Erytrocyte, all blood and serum samples were used for hemoglobine (Hb), glutathione (GSH) and Tf with Cp analysis, respectively. In the analysis of this samples, Hb and Tf bands were determined by electrophoresis. However, Cp and GSH levels were analyzed by the spectrophotometer. Three Hb phenotypes (AA, BB, AB) and Six Tf phenotypes (AA, AB, AC, BB, BC, CC) were determined in this study. In addition, both the observed and the expected values of polymorphic characteristic for 2 characters were presented according to the Hardy-Weinberg Equilibrium (HWE). Cp levels were detected as 0.822 ± 0.055 mg/dl and 1.793 ± 0.109 mg/dl in Kilis and Honamli herds, respectively. GSH levels were detected as, 42,486 ± 1,034 mg/dl and 33.515 ± 0.345 mg/dl in these breeds, respectively,. On the other hand, the high and low GSH levels (GSHH and GSHh) of herds were presented.

Keywords: electrophoresis, gene resource, goat, spectrophotometer

Procedia PDF Downloads 324

Authors: Hadiatullah, T. S. D. Desak Ketut, A. A. Ayu, A. N. Isna, D. P. Ririn

Abstract:

Spirogyra sp. is genus of microalgae which has a high carbohydrate content that used as a best medium for bacterial fermentation to produce cellulose. This study objective to determine the effect of pulp banana in the fermented paper production process using Spirogyra sp. and characterizing of the paper product. The method includes the production of bacterial cellulose, assay of the effect fermented paper interpolating with banana pulp using Spirogyra sp., and the assay of paper characteristics include gram-mage paper, water assay absorption, thickness, power assay of tensile resistance, assay of tear resistance, density, and organoleptic assay. Experiments were carried out with completely randomized design with a variation of the concentration of sewage treatment in the fermented paper production interpolating banana pulp using Spirogyra sp. Each parameter data to be analyzed by Anova variance that continued by real difference test with an error rate of 5% using the SPSS. Nata production results indicate that different carbon sources (glucose and sugar) did not show any significant differences from cellulose parameters assay. Significantly different results only indicated for the control treatment. Although not significantly different from the addition of a carbon source, sugar showed higher potency to produce high cellulose. Based on characteristic assay of the fermented paper showed that the paper gram-mage indicated that the control treatment without interpolation of a carbon source and a banana pulp have better result than banana pulp interpolation. Results of control gram-mage is 260 gsm that show optimized by cardboard. While on paper gram-mage produced with the banana pulp interpolation is about 120-200 gsm that show optimized by magazine paper and art paper. Based on the density, weight, water absorption assays, and organoleptic assay of paper showing the highest results in the treatment of pulp banana interpolation with sugar source as carbon is 14.28 g/m2, 0.02 g and 0.041 g/cm2.minutes. The conclusion found that paper with nata material interpolating with sugar and banana pulp has the potential formulation to produce super-quality paper.

Keywords: cellulose, fermentation, grammage, paper, Spirogyra sp.

Procedia PDF Downloads 314

Authors: Monthon Tangjitmungman

Abstract:

Numerous diseases have been linked to oxidative stress, in which a disproportion of free radicals in the body leads to tissue or cell damage. Polyphenols are the most abundant antioxidants found in plants, and they are highly effective at scavenging oxidative free radicals. Due to the presence of phenolic compounds in Caesalpinia sappan has been discovered to have antioxidant activity. It has several health benefits, the most important of which is preventing cardiovascular and cancer diseases. This study aimed to determine the phenolic content and antioxidant activity of C. sappan extract using a variety of antioxidant assays. The extract of C. sappan was made using a mixture of solvents (ethyl alcohol: water in ratio 8:2). The total phenolic content of C. sappan extract was determined and expressed as gallic acid equivalents using the Folin-Cioucalteu method (GAE). The antioxidant activity of C. sappan extract was assessed using the 2, 2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging assay and the ABTS radical scavenging capacity assay. An association was found between antioxidant activity and total phenol content. The antioxidant activity of C. sappan extract was also determined by DPPH and ABTS assays. The IC50 values for C. sappan extract from DPPH and ABTS assays were 54.48 μg/mL ± 0.545 and 25.46 μg/mL ± 0.790, respectively, in the DPPH assay. In the DPPH assay, vitamin C was used as a positive control, whereas Trolox was used as a positive control in the ABTS assay. In conclusion, C. sappan extract contains a high level of total phenolics and exhibits significant antioxidant activity. Nevertheless, more research should be done on the antioxidant activity, such as SOD and ROS scavenging assays and in vivo experiments, to determine whether the compound has antioxidant activity.

Keywords: ABTS assay, antioxidant activity, Caesalpinia sappan, DPPH assays, total phenolic content

Procedia PDF Downloads 356

Authors: Ibrahim Aly, Rabab Zalat, Bahaa EL Deen W. El Aswad, Ismail M. Moharm, Basam M. Masoud, Tarek Diab

Abstract:

The objective of the present study was to evaluate the novel nanomagnetic beads based–latex agglutination assay (NMB-LAT) as a simple test for diagnosis of S. haematobium as well as standardize the novel nanomagnetic beads based –ELISA (NMB-ELISA). According to urine examination this study included 85 S. haematobium infected patients, 30 other parasites infected patients and 25 negative control samples. The sensitivity of novel NMB-LAT was 82.4% versus 96.5% and 88.2% for NMB-ELISA and currently used sandwich ELISA respectively. The specificity of NMB-LAT was 83.6% versus 96.3% and 87.3% for NMB-ELISA and currently used sandwich ELISA respectively. In conclusion, the novel NMB-ELISA is a valuable applicable diagnostic technique for diagnosis of human schistosomiasis haematobium. The novel NMB-ELISA assay is a suitable applicable diagnostic method in field survey especially when followed by ELISA as a confirmatory test in query false negative results. Trials are required to increase the sensitivity and specificity of NMB-ELISA assay.

Keywords: diagnosis, iatex agglutination, nanomagnetic beads, sandwich ELISA

Procedia PDF Downloads 354

Authors: Shayma Munqith Al-Baker, Fadhl Ahmed Saeed Al-Gasha’a, Samira Hamid Hanash, Ahmed Ali Al-Hazmi

Abstract:

A total of 200 samples (180 fecal materials and 20 organ samples) were collected from (5 different poultry farms, 10 local poultry shops, 5 houses poultry, 5 Eggs stores shops and 5 hand slaughters centers) in Ibb city, Yemen, 2014. According to morphological, cultural, as well as biochemical characterization and serological tests, 59 29.5% isolates were identified as Salmonella spp. and all Salmonella isolates were categorized by serotype, which comprised of, 37 62.71% Salmonella Typhimurium serovar, 21 35.59%. Salmonella Enteritidis serovar and 11.69% Salmonella Heidelberg serovar. Antibiotic sensitivity test was done for bacterial isolates and the results showed there were clear differences in antibiotic resistant. Antimicrobial susceptibility of the isolates varies as follows: Ofloxacin 79.66%, Ciprofloxacin 67.80%, Colistin 59.32% and Gentamycin 52.54%. All of isolates were resistant to Erythromycin, Penicillin and Lincomycin. Antibacterial activity was done for both aqueous and ethanol extracts of Dodonaea viscosa plant by using well and disc diffusion assay. The results indicated that well diffusion assay had best results than disc diffusion assay, the highest inhibition zone was 22 mm for well diffusion and 15 mm for disc diffusion assay, the results observed that ethanol extract had best antibacterial effect than aqueous extract which the percentage of bacterial isolates affected with ethanol extract was 71.19% comparing with aqueous extract 28.81% by using disc diffusion assay, while the percentage of bacterial isolates affected with ethanol extract was 88.13% comparing with aqueous extract 52.54% by using will diffusion assay.

Keywords: Salmonella spp, Dodonaea viscosa, antimicrobial and salmonellosis

Procedia PDF Downloads 451

Authors: Ayman K. El Essawy, Amal M. Hosny, Hala M. Abu Shady

Abstract:

The rapid detection of TB and drug resistance, both optimizes treatment and improves outcomes. In the current study, respiratory specimens were collected from 155 patients. Conventional susceptibility testing and MIC determination were performed for rifampicin (RIF) and isoniazid (INH). Genotype MTBDRplus assay, which is a molecular genetic assay based on the DNA-STRIP technology and specific gene sequencing with primers for rpoB, KatG, and mab-inhA genes were used to detect mutations associated with resistance to rifampicin and isoniazid. In comparison to other categories, most of rifampicin resistant (61.5%) and isoniazid resistant isolates (47.1%) were from patients relapsed in treatment. The genotypic profile (using Genotype MTBDRplus assay) of multi-drug resistant (MDR) isolates showed missing of katG wild type 1 (WT1) band and appearance of mutation band katG MUT2. For isoniazid mono-resistant isolates, 80% showed katG MUT1, 20% showed katG MUT1, and inhA MUT1, 20% showed only inhA MUT1. Accordingly, 100% of isoniazid resistant strains were detected by this assay. Out of 17 resistant strains, 16 had mutation bands for katG distinguished high resistance to isoniazid. The assay could clearly detect rifampicin resistance among 66.7% of MDR isolates that showed mutation band rpoB MUT3 while 33.3% of them were considered as unknown. One mono-resistant rifampicin isolate did not show rifampicin mutation bands by Genotype MTBDRplus assay, but it showed an unexpected mutation in Codon 531 of rpoB by DNA sequence analysis. Rifampicin resistance in this strain could be associated with a mutation in codon 531 of rpoB (based on molecular sequencing), and Genotype MTBDRplus assay could not detect the associated mutation. If the results of Genotype MTBDRplus assay and sequencing were combined, this strain shows hetero-resistance pattern. Gene sequencing of eight selected isolates, previously tested by Genotype MTBDRplus assay, could detect resistance mutations mainly in codon 315 (katG gene), position -15 in inhA promotes gene for isoniazid resistance and codon 531 (rpoB gene) for rifampicin resistance. Genotyping techniques allow distinguishing between recurrent cases of reinfection or reactivation and supports epidemiological studies.

Keywords: M. tuberculosis, rpoB, KatG, inhA, genotype MTBDRplus

Procedia PDF Downloads 137

Authors: Aqeela Ashraf, Muhammad Imran, Tahir Yaqub, Muhammad Tayyab, Yung Fu Chang

Abstract:

For the identification of bovine mastitic pathogen, an economical, rapid and sensitive molecular diagnostic assay is developed by PCR multiplexing of gene and pathogenic species specific DNA sequences. The multiplex PCR assay is developed for detecting nine important bacterial pathogens causing mastitis Worldwide. The bacterial species selected for this study are Streptococcus agalactiae, Streptococcus dysagalactiae, Streptococcus uberis, Staphylococcus aureus, Escherichia coli, Staphylococcus haemolyticus, Staphylococcus chromogenes Mycoplasma bovis and Staphylococcus epidermidis. A single reaction assay was developed and validated by 27 reference strains and further tested on 276 bacterial strains obtained from culturing mastitic milk. The multiplex PCR assay developed here is further evaluated by applying directly on genomic DNA isolated from 200 mastitic milk samples. It is compared with bacterial culturing method and proved to be more sensitive, rapid, economical and can specifically identify 9 bacterial pathogens in a single reaction. It has detected the pathogens in few culture negative mastitic samples. Recognition of disease is the foundation of disease control and prevention. This assay can be very helpful for maintaining the udder health and milk monitoring.

Keywords: multiplex PCR, bacteria, mastitis, milk

Procedia PDF Downloads 305

Authors: J. Hidalgo de Quintana, I. Stoner, M. Tackett, G. Doran, C. Rafferty, A. Windemuth, J. Tytell, D. Pregibon

Abstract:

We have developed the Multiplexed Circulating microRNA assay that allows the detection of up to 68 microRNA targets per sample. The assay combines particle­based multiplexing, using patented Firefly hydrogel particles, with single­ step RT-PCR signal. Thus, the Circulating microRNA assay leverages PCR sensitivity while eliminating the need for separate reverse transcription reactions and mitigating amplification biases introduced by target­-specific qPCR. Furthermore, the ability to multiplex targets in each well eliminates the need to split valuable samples into multiple reactions. Results from the Circulating microRNA assay are interpreted using Firefly Analysis Workbench, which allows visualization, normalization, and export of experimental data. To aid discovery and validation of biomarkers, we have generated fixed panels for Oncology, Cardiology, Neurology, Immunology, and Liver Toxicology. Here we present the data from several studies investigating circulating and tumor microRNA, showcasing the ability of the technology to sensitively and specifically detect microRNA biomarker signatures from fluid specimens.

Keywords: biomarkers, biofluids, miRNA, photolithography, flowcytometry

Procedia PDF Downloads 340

Authors: Yeimy L. Quintana, Juan G. Zuluaga, Sandra S. Arango

Abstract:

The comet assay is a method based on electrophoresis that is used to measure DNA damage in cells and has shown important results in the identification of substances with a potential risk to the human population as innumerable physical, chemical and biological agents. With this technique is possible to obtain images like a comet, in which the tail of these refers to damaged fragments of the DNA. One of the main problems is that the image has unequal luminosity caused by the fluorescence microscope and requires different processing to condition it as well as to know how many optimal comets there are per sample and finally to perform the measurements and determine the percentage of DNA damage. In this paper, we propose the design and implementation of software using Image Processing Toolbox-MATLAB that allows the automation of image processing. The software chooses the optimum comets and measuring the necessary parameters to detect the damage.

Keywords: artificial vision, comet assay, DNA damage, image processing

Procedia PDF Downloads 279

Authors: Md Fazle Alam, Amaj Ahmed Laskar, Hina Younus

Abstract:

Melamine toxicity which causes renal failure and death of humans and animals have recently attracted worldwide attention. Developing an easy, fast and sensitive method for the routine melamine detection is the need of the hour. Herein, we have developed a rapid, sensitive, one step and selective colorimetric method for the detection of melamine in milk samples based upon in-situ formation of silver nanoparticles (AgNPs) via tannic acid at room temperature. These AgNPs thus formed were characterized by UV-VIS spectrophotometer, transmission electron microscope (TEM), zetasizer and dynamic light scattering (DLS). Under optimal conditions, melamine could be selectively detected within the concentration range of 0.05-1.4 µM with a limit of detection (LOD) of 10.1 nM, which is lower than the strictest melamine safety requirement of 1 ppm. This assay does not utilize organic cosolvents, enzymatic reactions, light sensitive dye molecules and sophisticated instrumentation, thereby overcoming some of the limitations of conventional methods.

Keywords: milk adulteration, melamine, silver nanoparticles, tannic acid

Procedia PDF Downloads 232

Authors: M. Shemis, O. Maher, G. Casterou, F. Gauffre

Abstract:

Hepatitis C virus (HCV) is a major cause of chronic liver disease with a global 170 million chronic carriers at risk of developing liver cirrhosis and/or liver cancer. Egypt reports the highest prevalence of HCV worldwide. Currently, two classes of assays are used in the diagnosis and management of HCV infection. Despite the high sensitivity and specificity of the available diagnostic assays, they are time-consuming, labor-intensive, expensive, and require specialized equipment and highly qualified personal. It is therefore important for clinical and economic terms to develop a low-tech assay for the direct detection of HCV RNA with acceptable sensitivity and specificity, short turnaround time, and cost-effectiveness. Such an assay would be critical to control HCV in developing countries with limited resources and high infection rates, such as Egypt. The unique optical and physical properties of gold nanoparticles (AuNPs) have allowed the use of these nanoparticles in developing simple and rapid colorimetric assays for clinical diagnosis offering higher sensitivity and specificity than current detection techniques. The current research aims to develop a detection assay for HCV RNA using gold nanoparticles (AuNPs). Methods: 200 anti-HCV positive samples and 50 anti-HCV negative plasma samples were collected from Egyptian patients. HCV viral load was quantified using m2000rt (Abbott Molecular Inc., Des Plaines, IL). HCV genotypes were determined using multiplex nested RT- PCR. The assay is based on the aggregation of AuNPs in presence of the target RNA. Aggregation of AuNPs causes a color shift from red to blue. AuNPs were synthesized using citrate reduction method. Different sets of probes within the 5’ UTR conserved region of the HCV genome were designed, grafted on AuNPs and optimized for the efficient detection of HCV RNA. Results: The nano-gold assay could colorimetrically detect HCV RNA down to 125 IU/ml with sensitivity and specificity of 91.1% and 93.8% respectively. The turnaround time of the assay is < 30 min. Conclusions: The assay allows sensitive and rapid detection of HCV RNA and represents an inexpensive and simple point-of-care assay for resource-limited settings.

Keywords: HCV, gold nanoparticles, point of care, viral load

Procedia PDF Downloads 185

Authors: Paiwan Buachan, Maneekarn Namsa-Aid, Suchada Jongrungruangchok, Foengchat Jarintanan, Wanlaya Uthaisang-Tanechpongtamb

Abstract:

Lung cancer or pulmonary carcinoma is the uncontrolled growth of abnormal cells in one or both of the lungs. These abnormal cells can spread to other organs of the body through lymphatic system or bloodstream which is called metastatic stage that leading cause of cancer death. Terrein (C₈H₁₀O₃; MW= 154.06 kDa) is a secondary bioactive fungal metabolite, which was isolated from the Aspergillus terreus. In this study, we investigated the effects of terrein on the inhibition of human lung cancer cell proliferation and metastasis. The A549 human non-small cell lung cancer cell line was used as a model. Terrein significantly inhibited lung cancer cell proliferation measuring by a colorimetric MTT assay (IC₅₀ 0.32 mM) and significantly inhibited metastatic processes including migration, invasion, and adhesion that determined by wound healing assay, transwell assay, and adhesion assay, respectively. These findings indicate that terrein could be a potential therapeutic agent for lung cancer.

Keywords: terrein, lung cancer, anticancer, antimetastatic

Procedia PDF Downloads 147

Authors: Paam Bidaya

Abstract:

The red seaweed Gracilaria fisheri, widely distributed along Thailand's southern coastlines, has been discovered to be edible. Sulfated polysaccharides from G. fisheri were extracted in low-temperature (25 °C) water. Seaweed polysaccharides (SPs) have been shown to have various advantageous biological effects. This study aims to investigate total phenolic content and antioxidant capacity of G. fisheri extract. The total phenolic content of G. fisheri extract was determined using Folin-Cioucalteu method and calculated as gallic acid equivalents (GAE). The antioxidant activity of G. fisheri extract was performed via 2, 2-diphenyl-1- picrylhydrazyl (DPPH) free radical scavenging assay and 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging capacity assays. The findings exhibited a strong correlation between antioxidant activity and the total phenol contents. In addition, DPPH and ABTS assays showed that G. fisheri extract showed antioxidant activities as a concentration-dependent manner. The IC50 values of G. fisheri extract were 902.19 μg/mL ± 0.785 and 727.98 μg/mL ± 0.822 for DPPH and ABTS, respectively. Vitamin C was used as a positive control in DPPH assay, while Trolox was used as a positive control in ABTS assay. To conclude, G. fisheri extract consists of a high amount of total phenolic content, which exhibit a significant antioxidant activity. However, further investigation regarding antioxidant activity should be performed in order to identify the mechanism of Gracilaria fisheri action.

Keywords: ABTS assay, DPPH assay, sulfated polysaccharides, total phenolic content

Procedia PDF Downloads 175

Authors: Iwan Cony Setiadi, Aulia M. T. Nasution

Abstract:

The multispectral imaging (MSI) technique has been used for skin analysis, especially for distant mapping of in-vivo skin chromophores by analyzing spectral data at each reflected image pixel. For ergonomic purpose, our multispectral imaging system is decomposed in two parts: a light source compartment based on LED with 11 different wavelenghts and a monochromatic 8-Bit CCD camera with C-Mount Objective Lens. The software based on GUI MATLAB to control the system was also developed. Our system provides 11 monoband images and is coupled with a software reconstructing hyperspectral cubes from these multispectral images. In this paper, we proposed a new method to build a hyperspectral reflectance cube based on artificial neural network algorithm. After preliminary corrections, a neural network is trained using the 32 natural color from X-Rite Color Checker Passport. The learning procedure involves acquisition, by a spectrophotometer. This neural network is then used to retrieve a megapixel multispectral cube between 380 and 880 nm with a 5 nm resolution from a low-spectral-resolution multispectral acquisition. As hyperspectral cubes contain spectra for each pixel; comparison should be done between the theoretical values from the spectrophotometer and the reconstructed spectrum. To evaluate the performance of reconstruction, we used the Goodness of Fit Coefficient (GFC) and Root Mean Squared Error (RMSE). To validate reconstruction, the set of 8 colour patches reconstructed by our MSI system and the one recorded by the spectrophotometer were compared. The average GFC was 0.9990 (standard deviation = 0.0010) and the average RMSE is 0.2167 (standard deviation = 0.064).

Keywords: multispectral imaging, reflectance cube, spectral reconstruction, artificial neural network

Procedia PDF Downloads 300