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1.
Sci Total Environ ; 953: 176073, 2024 Sep 07.
Artículo en Inglés | MEDLINE | ID: mdl-39250965

RESUMEN

Wastewater surveillance for COVID-19 and other pathogens has expanded globally. Rapid development and availability of various assays has facilitated swift adoption of wastewater surveillance in localities with diverse requirements. However, it presents challenges in comparing data due to methodological variations. Using surrogates for recovery control to address quantification biases has limitations as the recovery of surrogates and target pathogens often diverges significantly. Using non-spiked field-obtained wastewater samples as reference samples in an inter-lab study, this article proposes a straightforward, inexpensive, and most representative way of measuring relative quantification biases that occurs in analyzing field wastewater samples. Five labs participated in the study, testing five types of assays, resulting in a total of seven methods of lab-assay combinations. Each method quantified the concentration of SARS-CoV-2 and pepper mild mottle virus (PMMoV) RNAs in two types of reference samples. The results showed significant variations in quantification among methods, but the relative quantification biases were consistent across reference samples. This suggests that relative quantification biases measured with the reference samples are contingent on methods rather than wastewater samples, and that the once-determined method-specific factors can be used to correct for quantification biases in routine wastewater surveillance results. Subsequent data standardization was performed on year-long observational data from seven cities, serving as a preliminary validation of the proposed approach. This process demonstrated the potential for quantitative data comparison through the bias correction factors obtained in this inter-lab study.

2.
Cureus ; 16(5): e59645, 2024 May.
Artículo en Inglés | MEDLINE | ID: mdl-38832145

RESUMEN

Inter-lab quality control (ILQC) is vital for ensuring reliable test results, especially when laboratories are using assays authorized for newly emerging pathogens. The Indian Council of Medical Research (ICMR), New Delhi, had developed a network of laboratories to assess the quality of real-time reverse transcription (RT) polymerase chain reaction (PCR) assays used in India to detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). In a three-tier ILQC lab structure, All India Institute of Medical Sciences (AIIMS) Nagpur, an institute of national importance & a tertiary care hospital, was designated as a state quality control (QC) lab for the region of Maharashtra. ILQC activities were planned biannually. The ICMR had assigned 22 government and 19 private SARS-CoV-2 RT-PCR testing laboratories, under the Department of Microbiology, AIIMS Nagpur. AIIMS Nagpur had conducted four ILQC activities during 2020-2021. The finding of the ILQC assessment (cumulative includes all four ILQC) conducted by AIIMS Nagpur revealed that the results of 77% of laboratories were 100% concordant, the results of 14% of laboratories were 90%, and very few laboratories (i.e. 9%) showed <90% concordant.

3.
ACS Synth Biol ; 12(6): 1823-1835, 2023 06 16.
Artículo en Inglés | MEDLINE | ID: mdl-37246820

RESUMEN

In recent years, a plethora of new synthetic biology tools for use in cyanobacteria have been published; however, their reported characterizations often cannot be reproduced, greatly limiting the comparability of results and hindering their applicability. In this interlaboratory study, the reproducibility of a standard microbiological experiment for the cyanobacterial model organism Synechocystis sp. PCC 6803 was assessed. Participants from eight different laboratories quantified the fluorescence intensity of mVENUS as a proxy for the transcription activity of the three promoters PJ23100, PrhaBAD, and PpetE over time. In addition, growth rates were measured to compare growth conditions between laboratories. By establishing strict and standardized laboratory protocols, reflecting frequently reported methods, we aimed to identify issues with state-of-the-art procedures and assess their effect on reproducibility. Significant differences in spectrophotometer measurements across laboratories from identical samples were found, suggesting that commonly used reporting practices of optical density values need to be supplemented by cell count or biomass measurements. Further, despite standardized light intensity in the incubators, significantly different growth rates between incubators used in this study were observed, highlighting the need for additional reporting requirements of growth conditions for phototrophic organisms beyond the light intensity and CO2 supply. Despite the use of a regulatory system orthogonal to Synechocystis sp. PCC 6803, PrhaBAD, and a high level of protocol standardization, ∼32% variation in promoter activity under induced conditions was found across laboratories, suggesting that the reproducibility of other data in the field of cyanobacteria might be affected similarly.


Asunto(s)
Synechocystis , Reproducibilidad de los Resultados , Biomasa , Synechocystis/genética , Genes Reporteros , Regiones Promotoras Genéticas
4.
Toxicology ; 448: 152637, 2021 01 30.
Artículo en Inglés | MEDLINE | ID: mdl-33220337

RESUMEN

Microphysiological systems (MPS) aim to mimic the dynamic microenvironment and the interaction between tissues. While MPS exist for investigating pharmaceuticals, the applicability of MPS for cosmetics ingredients is yet to be evaluated. The HUMIMIC Chip2 ("Chip2″), is the first multi-organ chip technology to incorporate skin models, allowing for the topical route to be tested. Therefore, we have used this model to analyze the impact of different exposure scenarios on the pharmacokinetics and pharmacodynamics of two topically exposed chemicals, hyperforin and permethrin. The Chip2 incorporated reconstructed human epidermis models (EpiDerm™) and HepaRG-stellate spheroids. Initial experiments using static incubations of single organoids helped determine the optimal dose. In the Chip2 studies, parent and metabolites were analyzed in the circuit over 5 days after application of single and repeated topical or systemic doses. The gene expression of relevant xenobiotic metabolizing enzymes in liver spheroids was measured to reflect toxicodynamics effects of the compounds in liver. The results show that 1) metabolic capacities of EpiDerm™ and liver spheroids were maintained over five days; 2) EpiDerm™ model barrier function remained intact; 3) repeated application of compounds resulted in higher concentrations of parent chemicals and most metabolites compared to single application; 4) compound-specific gene induction e.g. induction of CYP3A4 by hyperforin depended on the application route and frequency; 5) different routes of application influenced the systemic concentrations of both parents and metabolites in the chip over the course of the experiment; 6) there was excellent intra- and inter-lab reproducibility. For permethrin, a process similar to the excretion in a human in vivo study could be simulated which was remarkably comparable to the in vivo situation. These results support the use of the Chip2 model to provide information on parent and metabolite disposition that may be relevant to risk assessment of topically applied cosmetics ingredients.


Asunto(s)
Hígado/efectos de los fármacos , Permetrina/farmacocinética , Floroglucinol/análogos & derivados , Piel/efectos de los fármacos , Terpenos/farmacocinética , Técnicas de Cultivo de Tejidos/métodos , Humanos , Insecticidas/toxicidad , Hígado/citología , Hígado/metabolismo , Técnicas de Cultivo de Órganos/métodos , Permetrina/toxicidad , Floroglucinol/farmacocinética , Floroglucinol/toxicidad , Piel/citología , Piel/metabolismo , Terpenos/toxicidad
5.
Nanomaterials (Basel) ; 10(11)2020 Oct 30.
Artículo en Inglés | MEDLINE | ID: mdl-33143118

RESUMEN

In this manuscript, we explore the sensor properties of epitaxially grown graphene on silicon carbide decorated with nanolayers of CuO, Fe3O4, V2O5, or ZrO2. The sensor devices were investigated in regard to their response towards NH3 as a typical reducing gas and CO, C6H6, CH2O, and NO2 as gases of interest for air quality monitoring. Moreover, the impact of operating temperature, relative humidity, and additional UV irradiation as changes in the sensing environment have been explored towards their impact on sensing properties. Finally, a cross-laboratory study is presented, supporting stable sensor responses, and the final data is merged into a simplified sensor array. This study shows that sensors can be tailored not only by using different materials but also by applying different working conditions, according to the requirements of certain applications. Lastly, a combination of several different sensors into a sensor array leads to a well-performing sensor system that, with further development, could be suitable for several applications where there is no solution on the market today.

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