scholarly journals SINTESIS DAN KARAKTERISASI NUKLEOTIDA BERTANDA [α-32P]ATP

2019 ◽  
Vol 10 (1) ◽  
pp. 30
Author(s):  
Wira Yusril Rahman ◽  
Endang Sarmini ◽  
H. Herlina ◽  
A. Abidin ◽  
T. Triyanto ◽  
...  
Keyword(s):  
Mobile Phase ◽  
Deoxyribonucleic Acid ◽  
Enzymatic Reaction ◽  
Health Sector ◽  
Reaction Vessel ◽  
Molecular Techniques ◽  
Nuclear Technology ◽  
Test Results ◽  
Nuclease P1 ◽  
Continuous Reaction

The utilization of nuclear technology in health sector with molecular techniques is increasingly developed today, especially in Indonesia.  One of which is nucleotide compound marked with [α-32P]ATP, this compound has been used as tracer for deoxyribonucleic acid (DNA)/ ribonucleic acid (RNA) in the study of various physiological and pathological processes. [α-32P]ATP is synthesized through several stages of continuous reaction in one reaction vessel. It begins with synthesis of [γ-32P]ATP through an enzymatic reaction, using H332PO4 and ADP, and enzymes  of lactate dehydrogenase, 3-phosphoglycerate phosphokinase and glyceraldehide 3-phospho  dehydrogenase; followed by phosphorilation of 3’AMP with T4 polinucleotide kinase enzyme to produce 3’-[5’-32P]ATP. The result is hydrolyzed with nuclease P1 enzyme to produce [5’-32P]AMP. The unreacted [γ-32P] is degraded by the addition of hexokinase enzyme and glucose. At the final stage of the reaction, the [5’-32P]AMP is  phosphorilated using phosphoenol-piruvat, piruvat kinase, and myokinase to produce [α-32P]ATP. The test results show that the every stage of reaction is characterized using TLC method, PEI cellulose paper as stationary phase and KH3PO4 0,5 M pH 3,5 as mobile phase. At the end of reaction, the yield of [α-32P]ATP reaches 71,7%, at Rf = 0,2.

scholarly journals Determination of Tiamulin in Type C Medicated Swine Feeds Using High Throughput Extraction with Liquid Chromatography

2002 ◽  
Vol 85 (3) ◽  
pp. 533-540 ◽  
Author(s):  
Douglas B Moore ◽  
Nancy L Britton ◽  
Robert L Smallidge ◽  
Ken L Riter
Keyword(s):  
Liquid Chromatography ◽  
Mobile Phase ◽  
Test Results ◽  
Improved Method ◽  
Mobile Phase Composition ◽  
Significance Level ◽  
Paired Sample ◽  
Recovery Study ◽  
Type C

Abstract An improved method for extraction and analysis of tiamulin is presented to address issues that arose during routine analysis of Type C medicated swine feeds under the current U.S. Food and Drug Administration-Center for Veterinary Medicine (FDA-CVM) approved method. The issues included the need for higher sample throughput and the ability to accommodate a wider variety of feed matrixes. Changes to the FDA-CVM approved method include reduced sample size and solvent volumes, phosphate buffering of tartaric acid, centrifugation, and use of a new liquid chromatography column and adjusted mobile phase composition. A paired sample study was performed to compare performance of the new and existing methods. The paired sample study showed no statistical difference between sample means of paired sets of 17 samples analyzed by both methods (t = 1.95 at 0.05 significance level, p = 0.068). A recovery study showed the method precision to be 2.06% (coefficient of variation) with an average standard recovery of 95.8%. Ruggedness test results indicated good overall ruggedness of the method.

scholarly journals Anxiety amongst physicians during COVID-19: Cross-sectional study in Pakistan

2020 ◽  
Author(s):  
Qaisar Khalid Mahmood ◽  
Sara Rizvi Jafree ◽  
Aisha Jalil ◽  
Syed Mujtaba Hasnain Nadir ◽  
Florian Fischer
Keyword(s):  
Structural Equation ◽  
Public Support ◽  
Healthcare Providers ◽  
Health Sector ◽  
Cross Sectional Study ◽  
Equation Modeling ◽  
Test Results ◽  
Cross Sectional ◽  
Family Strain ◽  
High Workload

Abstract Background: Ensuring safety and wellbeing of healthcare providers is crucial, particularly during times of a pandemic. In this study, we aim to identify the determinants of anxiety in physicians on duty in coronavirus wards or quarantine centers. Methods: We conducted a cross-sectional quantitative survey with an additional qualitative item. Five constructs of workload, exhaustion, family strain, feeling of protection, and anxiety were measured using items from two validated tools. Modifications were made for regional relevance. Factor analysis was performed showing satisfactory Cronbach alpha results. Overall, 103 physicians completed the questionnaire. Results: T-test results revealed significant associations between gender and anxiety. Structural equation modeling identified that high workload contributed to greater exhaustion (β=0.41, R2=0.17, p<0.001) and greater family strain (β=0.47, R2=0.22, p<0.001). Exhaustion (β=0.17, p<0.005), family strain (β=0.34, p<0.001), and feelings of protection (β=-0.30, p<0.001) significantly explained anxiety (R2=0.28). Qualitative findings further identified specific needs of physicians with regard to protective equipment, compensation, quarantine management, resource allocation, security and public support, governance improvement, and health sector development. Conclusions: It is imperative to improve governmental and social support for physicians and other healthcare providers during the corona pandemic. Immediate attention is needed to reduce anxiety, workload, and family strain in frontline practitioners treating coronavirus patients, and to improve their (perceptions of) protection. This is a precondition for patient safety.

scholarly journals Role of Anti-SARS-CoV-2 antibodies in different cohorts: Can they provide clues for appropriate patient triaging?

2020 ◽  
Author(s):  
Manohar B. Mutnal ◽  
Amin A. Mohammad ◽  
Alejandro C. Arroliga ◽  
Yinan Hua ◽  
Liping Wang ◽  
...  
Keyword(s):  
Molecular Techniques ◽  
Molecular Testing ◽  
Moderate Increase ◽  
Test Results ◽  
Serum Samples ◽  
Serological Tests ◽  
Health Crisis ◽  
Serum Igg ◽  
Inpatient Population

AbstractThe emergence of coronavirus disease 2019 (COVID-19) has become a major global health crisis. Currently, diagnosis is based on molecular techniques, which detect the viral nucleic acids when present at detectable levels. The serum IgG response against SARS-CoV-2 was examined by using an ELISA-based assay. Serum samples, along with nasopharyngeal specimens were collected from various cohorts and analyzed by ELISA and rRT-PCR, respectively. A total of 167 serum samples were tested for serum IgG antibodies against SARS-CoV-2 in outpatient cohorts, 15 (8.9%) were positive by rRT-PCR and the remaining 152 (91%) were negative. We used these data to generate two different assay cutoffs for serum IgG assay and investigated percent concordance with rRT-PCR test results. The emergency department data revealed, out of 151 nasopharyngeal swabs, 4 (2.6%) were positive by rRT-PCR and 18 (11.9%) were positive for serum IgG assay. Among the 18 patients that were positive for serum IgG, 13 (72.2%) exhibited 1-3 symptoms of COVID-19 and 5 (27.7%) patients did not present with any COVID-19 related symptoms, per CDC criteria. All 4 (100%) patients that were positive by rRT-PCR had symptoms of COVID-19 disease. A longitudinal study from the inpatient population suggested there was a sharp increase in the serum IgG titers in 5 patients, a moderate increase in 1 patient and a plateau in 3 patients. Sero-prevalence of COVID-19 disease in pre-procedure patients was 5.5%. Our findings suggest serological tests can be used for appropriate patient triaging when performed as an adjunct to existing molecular testing.

The Utilization of MS-MLPA as the First-Line Test for the Diagnosis of Prader–Willi Syndrome in Thai Patients

2022 ◽  
Author(s):  
Chanita Prapasrat ◽  
Preyaporn Onsod ◽  
Veerawat Korkiatsakul ◽  
Budsaba Rerkamnuaychoke ◽  
Duangrurdee Wattanasirichaigoon ◽  
...  
Keyword(s):  
Genetic Testing ◽  
Genetic Disorder ◽  
Cost Effective ◽  
Molecular Techniques ◽  
Prader Willi Syndrome ◽  
Test Results ◽  
First Line ◽  
Line Test ◽  
Clinical Diagnostic Criteria ◽  
Dependent Probe

AbstractPrader–Willi syndrome (PWS) is a genetic disorder caused by the expression disruption of genes on the paternally inherited allele of chromosome 15q11.2-q13. Apart from clinical diagnostic criteria, PWS is confirmed by genetic testing. Methylation-specific multiplex ligation-dependent probe amplification (MS-MLPA) is one of the molecular techniques used to analyze this syndrome. This study aimed to evaluate the concordance of the test results of MS-MLPA with conventional techniques in the diagnosis of PWS in Thai patients. Forty leftover specimens from routine genetic testing (MS-PCR and FISH) were tested to obtain MS-MLPA results. By comparison, perfect concordance was shown between the result of MS-MLPA and those of conventional techniques. In conclusion, MS-MLPA is an accurate and cost-effective assay that can be used to confirm PWS diagnosis with explicit deletion of affected genes.

scholarly journals Putting Thought to Paper: A μARCS Protease Screen

2003 ◽  
Vol 8 (6) ◽  
pp. 668-675 ◽  
Author(s):  
Duncan R. Groebe ◽  
Mary L. Maus ◽  
Terry Pederson ◽  
Jill Clampit ◽  
Stevan Djuric ◽  
...  
Keyword(s):  
Enzymatic Reaction ◽  
Reaction Vessel ◽  
Throughput Capacity ◽  
Agarose Gel ◽  
Handling System ◽  
Agarose Gels ◽  
The Matrix ◽  
Compound Screening ◽  
Blotter Paper ◽  
Protease Assay

In micro-arrayed compound screening (μARCS), an agarose gel is used as a reaction vessel that maintains humidity and compound location as well as being a handling system for reagent addition. Two or more agarose gels may be used to bring test compounds, targets, and reagents together, relying on the pore size of the gel matrix to regulate diffusion of reactants. It is in the microenvironment of the agarose matrix that all the components of an enzymatic reaction interact and result in inhibitable catalytic activity. In an effort to increase the throughput of μARCS-based screens, reduce the effort involved in manipulating agarose gels, and reduce costs, blotter paper was used rather than a second agarose gel to introduce a substrate to a gel containing a target enzyme. In this assay, the matrix of the blotter paper did not prevent the substrate from diffusing into the enzyme gel. The compound density of the μARCS format, the ease of manipulating sheets of paper for reagent addition, and a scheduled protocol for running multiple gels allowed for a throughput capacity of more than 200,000 tests per hour. A protease assay was developed and run in the μARCS format at a rate of 200,000 tests per hour using blotter paper to introduce the substrate. Picks in the primary screen were retested in the μARCS format at a density of 384 compounds per sheet. IC50 values were confirmed in a 96-well plate format. The screen identified several small molecule inhibitors of the enzyme. The details of the screening format and the analysis of the hits from the screen are presented.

scholarly journals EFFECT OF TRANSFERSOME FORMULATION ON THE STABILITY AND ANTIOXIDANT ACTIVITY OF N-ACETYLCYSTEINE IN ANTI-AGING CREAM

2020 ◽  
pp. 156-162
Author(s):  
HARMITA HARMITA ◽  
ISKANDARSYAH ISKANDARSYAH ◽  
SHOFIYAH FATIN AFIFAH
Keyword(s):  
Antioxidant Activity ◽  
Mobile Phase ◽  
Test Results ◽  
Penetration Test ◽  
Injection Volume ◽  
Accelerated Stability ◽  
Dpph Method ◽  
The Stability ◽  
Franz Diffusion Cells

Objective: N-acetylcysteine is an antioxidant with thiol/sulfhydryl groups and is currently being developed as an active ingredient in anti-agingcreams. The study’s aim was to compare the stability and antioxidant activity of N-acetylcysteine in anti-aging creams formulated with and withouta transfersome carrier system.Methods: Stability was assessed by performing cycling, centrifugal, and accelerated stability tests. In addition, antioxidant activity was measuredby the DPPH method, and in vitro penetration was measured using Franz diffusion cells. The analysis of N-acetylcysteine was performed using highperformanceliquid chromatography with ultraviolet–visible detection at a wavelength of 214 nm and a flow rate of 1.0 mL/min, injection volume of5 μL, and a mobile phase of phosphate buffer pH 3.0.Results: The N-acetylcysteine transfersome and non-transfersome cream preparations did not change color or show phase separation during thecycling and centrifugal tests. The N-acetylcysteine in the transfersome and non-transfersome cream preparations had strong antioxidant activity,with half-maximal inhibitory concentrations of 26.90 μg/mL and 38.63 μg/mL, respectively. The in vitro penetration test using Franz diffusion cellsshowed that the cumulative amount of penetrated N-acetylcysteine was 7355.13 μg/cm2 (flux of 845.67 μg/cm2∙h) in the transfersome cream and4677.61 μg/cm2 (flux of 533.33 μg/cm2∙h) in the non-transfersome cream.Conclusion: The in vitro penetration test results showed that the transfersome formulations in creams were able to increase the cumulative amountand flux of penetrated N-acetylcysteine in anti-aging cream preparations relative to those not formulated with transfersome.

scholarly journals Comparison of Antigen Tests and qPCR in Rapid Diagnostics of Infections Caused by SARS-CoV-2 Virus

Viruses ◽  
2021 ◽  
Vol 14 (1) ◽  
pp. 17
Author(s):  
Adrianna Klajmon ◽  
Aldona Olechowska-Jarząb ◽  
Dominika Salamon ◽  
Agnieszka Sroka-Oleksiak ◽  
Monika Brzychczy-Włoch ◽  
...  
Keyword(s):  
Quantitative Polymerase Chain Reaction ◽  
Molecular Techniques ◽  
Nucleic Acid Amplification ◽  
Laboratory Diagnostics ◽  
Test Results ◽  
Rapid Diagnostics ◽  
Antigen Test ◽  
Qualified Personnel ◽  
Replication Phase ◽  
Antigen Tests

Diagnostics of the coronavirus disease 2019 (COVID-19) using molecular techniques from the collected respiratory swab specimens requires well-equipped laboratory and qualified personnel, also it needs several hours of waiting for results and is expensive. Antigen tests appear to be faster and cheaper but their sensitivity and specificity are debatable. The aim of this study was to compare a selected antigen test with quantitative polymerase chain reaction (qPCR) tests results. Nasopharyngeal swabs were collected from 192 patients with COVID-19 symptoms. All samples were tested using Vitassay qPCR SARS-CoV-2 kit and the Humasis COVID-19 Ag Test (MedSun) antigen immunochromatographic test simultaneously. Ultimately, 189 samples were tested; 3 samples were excluded due to errors in taking swabs. The qPCR and antigen test results were as follows: 47 positive and 142 negative, and 45 positive and 144 negative, respectively. Calculated sensitivity of 91.5% and specificity of 98.6% for the antigen test shows differences which are not statistically significant in comparison to qPCR. Our study showed that effectiveness of the antigen tests in rapid laboratory diagnostics is high enough to be an alternative and support for nucleic acid amplification tests (NAAT) in the virus replication phase in the course of COVID-19.

scholarly journals Penentuan Kadar Kalium Sorbat dan Persen Recovery Pada Selai dengan Metoda High Performance Liquid Chromatography

2020 ◽  
Vol 1 (1) ◽  
pp. 25
Author(s):  
Pevi Riani ◽  
Ulvie Ameinda Fannin
Keyword(s):  
High Performance Liquid Chromatography ◽  
Liquid Chromatography ◽  
Mobile Phase ◽  
High Performance ◽  
Food Additives ◽  
Quality Standard ◽  
Potassium Sorbate ◽  
Test Results ◽  
Standard Set ◽  
The Right

Fruit jam is one of the preserved products that have a soft texture that contains crushed fruit and is mixed with sugar with the addition or without the addition of water. Preservatives are added to prevent damage to jams. A preservative that is usually added to jam is potassium sorbate. Potassium sorbate is commonly used as a preservative because it dissolves more easily when added to food. This study aims to determine the levels of potassium sorbate in jam samples. Potassium sorbate was determined using High Performance Liquid Chromatography (HPLC). From the test results, it was found that potassium sorbate levels ranged from719, 65 mg/kg to 757,24 mg/kg. The results of this test will be compared with the standards set by the Food and Drug Supervisory Agency (BPOM) No. 36 of 2013 concerning the maximum limit for the use of preservative food additives, namely 500 mg/kg. From the test results, it can be seen that the potassium sorbate in all samples exceeds the quality standard set by the BPOM. Therefore, people must be more careful in choosing jam to consume daily. The test results have used the right method, namely by adjusting the stationary phase, mobile phase and the right wavelength so that the% recovery results are 103%.

scholarly journals Analytical Method Validation and Determination of Pyridoxine, Nicotinamide, and Caffeine in Energy Drinks Using Thin Layer Chromatography-Densitometry

2015 ◽  
Vol 15 (1) ◽  
pp. 9-15 ◽  
Author(s):  
Florentinus Dika Octa Riswanto ◽  
RR. Endang Lukitaningsih ◽  
Sudibyo Martono
Keyword(s):  
Mobile Phase ◽  
Energy Drink ◽  
Food Supplement ◽  
Test Results ◽  
Optimal Method ◽  
Analytical Method Validation ◽  
Limit Value ◽  
Tlc Plates ◽  
Layer Chromatography

Food supplement which contains vitamins and stimulants such as caffeine were classified as energy drink. TLC-densitometry method was chosen to determine the pyridoxine, nicotinamide, and caffeine in the energy drink sample. TLC plates of silica gel 60 F254 was used as the stationary phase and methanol : ethyl acetate : ammonia 25% (134:77:10) was used as the mobile phase. The correlation coefficient for each pyridoxine, nicotinamide, and caffeine were 0.9982, 0.9997, and 0.9966, respectively. Detection and quantitation limits of from the three analytes were 4.05 and 13.51 µg/mL; 13.15 and 43.83 µg/mL; 5.43 and 18.11 µg/mL, respectively. The recovery of pyridoxine, nicotinamide, and caffeine were within the required limit range of 95-105%. The percent of RSD were below the limit value of 5.7% for caffeine and nicotinamide and 8% for pyridoxine. The content amount of pyridoxine in the sample 1 and 2 were 33.59 ± 0.981 and 30.29 ± 2.061 µg/mL, respectively. The content amount of nicotinamide in the sample 1 and 2 were 106.53 ± 3.521 and 98.20 ± 3.648 µg/mL, respectively. The content amount of caffeine in the sample 1 and 2 were 249.50 ± 5.080 and 252.80 ± 2.640 µg/mL, respectively. Robustness test results showed that the most optimal method conditions should be applied for the analysis.

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