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41.
The method for virus titer determination of avian infectious bursal disease (IBD) live vaccine, developed long before regulatory validation guidelines is a cell culture based biological assay intended for use in vaccine release testing.The aim of our study was to perform a validation, based on fit-for-purpose principle, of an old 50% tissue culture infectious dose (TCID50) method according to Guidelines of the International Cooperation on Harmonization of Technical Requirements for Registration of Veterinary Medicinal Products (VICH).This paper addresses challenges and discusses some key aspects that should be considered when validating biological methods. A different statistical approach and non-parametric statistics was introduced in validation protocol in order to derive useful information from experimental data. This approach is applicable for a wide range of methods.In conclusion, the previous virus titration method had showed to be precise, accurate, linear, robust and in accordance with current regulatory standards, which indicates that there is no need for additional re-development or upgrades of the method for its suitability for intended use. 相似文献
42.
Jian-Hong Chen Lei-Li Wang Lin Tao Bin Qi Yong Wang Yu-Jie Guo Liu Miao 《Journal of cellular and molecular medicine》2021,25(22):10736-10746
The present study aimed to explore the potential hub genes and pathways of ischaemic cardiomyopathy (ICM) and to investigate the possible associated mechanisms. Two microarray data sets ( GSE5406 and GSE57338 ) were downloaded from the Gene Expression Omnibus (GEO) database. The limma package was used to analyse the differentially expressed genes (DEGs). Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment, Disease Ontology (DO) and Gene Ontology (GO) annotation analyses were performed. A protein-protein interaction (PPI) network was set up using Cytoscape software. Significant modules and hub genes were identified by the Molecular Complex Detection (MCODE) app. Then, further functional validation of hub genes in other microarrays and survival analysis were performed to judge the prognosis. A total of 1065 genes were matched, with an adjusted p < 0.05, and 17 were upregulated and 25 were downregulated with|log2 (fold change)|≥1.2. After removing the lengthy entries, GO identified 12 items, and 8 pathways were enriched at adjusted p < 0.05 (false discovery rate, FDR set at <0.05). Three modules with a score >8 after MCODE analysis and MYH6 were ultimately identified. When validated in GSE23561 , MYH6 expression was lower in patients with CAD than in healthy controls (p < 0.05). GSE60993 data suggested that MYH6 expression was also lower in AMI patients (p < 0.05). In the GSE59867 data set, MYH6 expression was lower in CAD patients than in AMI patients and lower in heart failure (HF) patients than in non-HF patients. However, there was no difference at different periods within half a year, and HF was increased when MYH6 expression was low (p < 0.05–0.01). We performed an integrated analysis and validation and found that MYH6 expression was closely related to ICM and HF. However, whether this marker can be used as a predictor in blood samples needs further experimental verification. 相似文献
43.
Steady‐state and synchronous spectrofluorimetric methods for simultaneous determination of aliskiren hemifumarate and amlodipine besylate in dosage forms 下载免费PDF全文
Walid M. Ebeid Ehab F. Elkady Asmaa A. El‐Zaher Ramzia I. El‐Bagary Gabor Patonay 《Luminescence》2014,29(7):878-883
Aliskiren hemifumarate (ALS) and amlodipine besylate (AML) were simultaneously determined by two different spectrofluorimetric techniques. The first technique depends on direct measurement of the steady‐state fluorescence intensities of ALS and AML at 313 nm and 452 nm upon excitation at 290 and 375 nm, respectively, in a solvent composed of methanol and water (10: 90, v/v) . The second technique utilizes synchronous fluorimetric quantitative screening of the emission spectra of ALS and AML at 272 and 366 nm, respectively using Δλ of 97 nm. Effects of different solvents and surfactants on relative fluorescence intensity were studied. The method was validated according to ICH guidelines. Linearity, accuracy and precision were found to be satisfactory in both techniques over the concentration ranges of 1–15 and 0.4–4 µg/mL for ALS and AML, respectively. In the first technique, limit of detection and limit of quantification were estimated and found to be 0.256 and 0.776 µg/mL for ALS as well as 0.067 and 0.204 µg/mL for AML, respectively. Also, limit of detection and limit of quantification were calculated in the synchronous method and found to be 0.293 and 0.887 µg/mL for ALS as well as 0.034 and 0.103 µg/mL for AML, respectively. The methods were successfully applied for the determination of the two drugs in their co‐formulated tablets. The results were compared statistically with reference methods and no significant difference was found. The developed methods are rapid, sensitive, inexpensive and accurate for the quality control and routine analysis of the cited drugs in bulk and in pharmaceutical preparations without pre‐separation. Copyright © 2014 John Wiley & Sons, Ltd. 相似文献
44.
45.
Six pesticides and two spray oils were tested against Polyphagotarsonemus latus. The chemicals were evaluated under laboratory conditions, requiring the development of a novel bioassay method, which is reported here. The pesticide toxicities fell into three distinct groups, namely abamectin, conventional pesticides and oils. The relative pesticide toxicities at the LC50 level were abamectin 4.9×10-8 g ai l-1, endosulfan 1.1×10-3 g ai l-1, fenpyroximate 2.3×10-3 g ai l-1, pyridaben 4.1×10-3 g ai l-1, tebufenpyrad 4.4×10-3 g ai l-1, dicofol 4.5×10-3 g ai l-1, petroleum spray oil 3.4×10-1 g ai l-1 and canola oil 4.1×10-1 g ai l-1. The calculation of the LC99.9 values allows for resistance monitoring in P. latus and the suggested discriminating concentrations are abamectin 1.0×10-4 g ai l-1; endosulfan, pyridaben and dicofol 1.0×10-1 g ai l-1 fenpyroximate and tebufenpyrad 5.0×10-1 g ai l-1. 相似文献
46.
Jeffrey E. Banikowski Swiatoslav W. Kaczmar John F. Hunt 《Soil & Sediment Contamination》2009,18(3):243-263
This paper reports the results of a field study to evaluate the use of helium as a tracer gas during soil vapor sampling. The authors found that the helium tracer method recommended in regulatory guidance can detect atmospheric leakage. However, the degree of leakage can be underestimated due to: 1) losses of helium from the tracer reservoir during sampling; 2) the entry of atmospheric air from an area outside the tracer reservoir; 3) temporal variability of leakage; and 4) flow-related reduction in helium detector response. Frequent leakage was observed in soil vapor probes installed using conventional direct push techniques. Probes installed at a depth of 8 ft exhibited less leakage than those installed at 4 ft. Probes installed in glacial till exhibited greater and more frequent leakage than those installed in sandy loam. Based on the frequency of observed leakage, refinement and standardization of tracer methods, with routine incorporation of tracers and in-line pressure and flow monitoring, is recommended. 相似文献
47.
Objective: To assess the feasibility of using a questionnaire‐based needs assessment tool (D‐E‐N‐T‐A‐L) to screen for dental treatment need as part of the single assessment process (SAP) for older people in Sheffield. Materials and methods: Test validation study comparing questionnaire‐assessed and normative need in two consecutive samples of older adults: 48 living at home in the transition phase of older age and 29 frail older adults living in care homes. Each answered the six D‐E‐N‐T‐A‐L questions as part of SAP and a dental examination was carried out within 2 weeks in participants’ homes. Question‐defined need was then compared to the normative need. Results: Questionnaire‐defined and normative need were high in both the transitional group (83% and 90% respectively) and the frail group (83% and 62%). These high levels of need meant that the sensitivity and positive predictive values of D‐E‐N‐T‐A‐L were high, but the specificity and negative predictive values were low. Conclusion: The high levels of need in these patient groups suggests that preliminary questionnaire‐based screening is an unnecessary step. A clinical examination of all older people undergoing SAP may be necessary. Further research may be warranted on the use of questionnaires to assess dental treatment needs among people with different attendance patterns. 相似文献
48.
Alan M. Goldberg John M. Frazier David Brusick Michael S. Dickens Oliver Flint Stephen D. Gettings Richard N. Hill Robert L. Lipnick Kevin J. Renskers June A. Bradlaw Robert A. Scala Bellina Veronesi Sidney Green Neil L. Wilcox Rodger D. Curren 《In vitro cellular & developmental biology. Animal》1993,29(9):688-692
Summary The development and application of in vitro alternatives designed to reduce or replace the use of animals, or to lessen the
distress and discomfort of laboratory animals, is a rapidly developing trend in toxicology. However, at present there is no
formal administrative process to organize, coordinate, or evaluate validation activities. A framework capable of fostering
the validation of new methods is essential for the effective transfer of new technologic developments from the research laboratory
into practical use. This committee has identified four essential validation resources: chemical bank(s), cell and tissue banks,
a data bank, and reference laboratories. The creation of a Scientific Advisory Board composed of experts in the various aspects
and endpoints of toxicity testing, and representing the academic, industrial, and regulatory communities, is recommended.
Test validation acceptance is contingent on broad buy-in by disparate groups in the scientific community—academics, industry,
and government. This is best achieved by early and frequent communication among parties and agreement on common goals. It
is hoped that the creation of a validation infrastructure composed of the elements described in this report will facilitate
scientific acceptance and utilization of alternative methodologies and speed implementation of replacement, reduction, and
refinement alternatives in toxicity testing. 相似文献
49.
The June beetle, Hoplia philanthus Füessly (Coleoptera: Scarabaeidae), has become a widespread and destructive insect pest of lawns, sport turf, pastures, and horticultural crops in Belgium. The virulence of 34 entomopathogenic fungal isolates from the genera Metarhizium, Beauveria, and Paecilomyces to third-instar H. philanthus was tested in bioassays by dipping larvae in 10(7)conidia/ml suspensions. Two isolates of Metarhizium anisopliae (CLO 53 and CLO 54) caused maximally 90% mortality 10 weeks post-inoculation while other isolates only caused mortalities between 10 and 62%. The virulence of M. anisopliae CLO 53 was further tested by exposing H. philanthus larvae to conidial serial concentrations of 10(4)-10(9)conidia/g sandy soil for up to 11 weeks at 15, 20 or 25 degrees C. Mortality was dependant on the fungal concentration, exposure time, and temperature. Eleven weeks after inoculation, the LC50 values for this isolate ranged from 1.3 to 4.0 x 10(6), 1.0 to 3.2 x 10(5), and 2.5 x 10(4) to 10(5)conidia/g soil at 15, 20, and 25 degrees C, respectively. The LT50 values for this isolate ranged from 3.5 to 21.7, 2.4 to 18.7, and 2.9 to 16.1 weeks at concentrations of 10(9) and 10(4)conidia/g soil at 15, 20, and 25 degrees C, respectively. In glasshouse pot experiment with perennial ryegrass (Lolium perenne L.), the isolate CLO 53 caused mortalities of 50 and 88% of H. philanthus larvae 10 weeks after application of 10(4) and 10(6)conidia/cm(2) soil surface, respectively. The present results suggest that the Belgian isolate CLO 53 has excellent potential for biological control of H. philanthus. 相似文献
50.
Virus inactivation validation studies have been widely applied in the risk assessment of biogenic material-based medical products, such as biological products, animal tissue-derived biomaterials, and allogeneic biomaterials, to decrease the risk of virus transmission. Traditional virus detection methods in an inactivation validation study utilize cell culture as a tool to quantify the infectious virus by observing cytopathic effects (CPEs) after virus inactivation. However, this is susceptible to subjective factors because CPEs must be observed by experts under a microscope during virus titration. In addition, this method is costly and time- and labor-consuming. Molecular biological technologies such as quantitative polymerase chain reaction (qPCR) have been widely used for virus detection but cannot distinguish infectious and noninfectious viruses. Therefore, qPCR cannot be directly applied to virus inactivation validation studies. In this paper, methods to detect viruses and progress in the challenge of differentiating infectious and noninfectious viruses with the combination of pretreatment and qPCR techniques such as the integrated cell culture-qPCR (ICC-qPCR) method are reviewed. In addition, the advantages and disadvantages of each new method, as well as its prospect in virus inactivation validation studies, are discussed. 相似文献