Good Laboratory Practice Research Articles

Reappearance of effector T cells is associated with recovery from COVID-19.

BackgroundElucidating the role of T cell responses in COVID-19 is of utmost importance to understand the clearance of SARS-CoV-2 infection.Methods30 hospitalized COVID-19 patients and 60 age- and gender-matched healthy controls (HC) participated in this study. We used two comprehensive 11-colour flow cytometric panels conforming to Good Laboratory Practice and approved for clinical diagnostics.FindingsAbsolute numbers of lymphocyte subsets were differentially decreased in COVID-19 patients according to clinical severity. In severe disease (SD) patients, all lymphocyte subsets were reduced, whilst in mild disease (MD) NK, NKT and γδ T cells were at the level of HC. Additionally, we provide evidence of T cell activation in MD but not SD, when compared to HC. Follow up samples revealed a marked increase in effector T cells and memory subsets in convalescing but not in non-convalescing patients.InterpretationOur data suggest that activation and expansion of innate and adaptive lymphocytes play a major role in COVID-19. Additionally, recovery is associated with formation of T cell memory as suggested by the missing formation of effector and central memory T cells in SD but not in MD. Understanding T cell-responses in the context of clinical severity might serve as foundation to overcome the lack of effective anti-viral immune response in severely affected COVID-19 patients and can offer prognostic value as biomarker for disease outcome and control.FundingFunded by State of Lower Saxony grant 14–76,103–184CORONA-11/20 and German Research Foundation, Excellence Strategy – EXC2155“RESIST”–Project ID39087428, and DFG-SFB900/3–Project ID158989968, grants SFB900-B3, SFB900-B8.

Diagnostic tests for COVID-19: the importance of the before and the after

In 2019, in the city of Wuhan, China, patients were diagnosed with a novel coronavirus originally named 2019-nCoV, currently known as SARS-CoV-2. The alarming expansion of this pandemic makes it necessary to find out and implement reliable diagnostic methods in order to properly detect and treat patients, thus contributing to slowing down the spread of the disease. It is extremely important to have trained professionals in sample collection, good clinical laboratory practices, and molecular and hematological techniques to adequately detect any cases of infection with this virus. Moreover, standardized protocols are essential for obtaining, transferring and storing samples to stop the spread of the COVID-19 pandemic caused by SARS-CoV-2.

Developing laboratory capacity for Good Laboratory Practice certification: lessons from a Tanzanian insecticide testing facility

Background: With increasing insecticide resistance in malaria-endemic countries there is an urgent need for safe and effective novel vector control products. To improve the capacity of facilities that test insecticides in sub-Saharan Africa, a programme is supporting seven facilities towards Good Laboratory Practice (GLP) certification, the globally recognized standard for quality management system (QMS) for the conduct of non-clinical and environmental studies. The World Health Organization (WHO) GLP Handbook provides guidance on a stepwise approach to implement a GLP compliant QMS. This study assesses auditor GLP checklists and timings outlined in the WHO GLP Handbook in the real-life context of a Tanzanian insecticide-testing facility, evaluating their implementation in this context. Methods and Principle Findings: We conducted document review and semi-structured interviews with staff at all levels of the test facility to explore factors that influenced progress towards GLP certification. We found that while auditor GLP checklists underemphasised computer systems, they were otherwise broadly applicable. Factors that delayed time to completion of GLP certification included the need for extensive infrastructure improvements, the availability of regional expertise related to GLP, the capacity of national and regional external systems and services to meet GLP compliance requirements, and training development required for Standard Operating Procedure implementation. Conclusion: The standards required for full GLP compliance are rigorous, with an expected completion timeline to implementation of 24 months. This study shows that in low and middle-income countries this timeline may be unrealistic due to challenges related to infrastructure development and lack of regional capacity and expertise. We recommend a comprehensive gap analysis when starting a project, including these areas which are beyond those recommended by the WHO GLP Handbook. These challenges can be successfully overcome and the experience in Tanzania provides key lessons for other facilities seeking GLP certification or the development of similar QMS.

Open Digital Educational Resources for Self-Training Chemistry Lab Safety Rules

Educational resources that cover essential knowledge related to chemistry safety rules are openly accessible on the CHIMACTIV website (http://chimactiv.agroparistech.fr/). Organized into two online tracks, the content covers personal and collective protections, first aid, as well as the handling of hazardous chemicals. Being interactive and abundantly illustrated by photos and videos, these resources may be used by teachers to encourage (or even oblige) their students to self-train before lab sessions. Our user experience shows that students improve their chemical safety knowledge and culture upon consulting these resources: they arrive in the lab more confident and are more attentive to safety rules (especially concerning safety glasses and gloves), with a deeper knowledge of chemical hazards and of waste management options. Thanks to their digital responsiveness, these open resources may also be accessible during the lab sessions, enabling the students to get the information “just-in-time”. In addition, decision trees for liquid and solid waste management are available, helping the students to gain reflexivity as well as to acquire good laboratory practice during lab sessions. In this way, the supervisors of the practical work sessions waste much less time taking back students who do not follow the safety rules properly.

Bioseguridad en laboratorios de diagnóstico molecular de SARS-CoV-2 (COVID-19) mediante RT-qPCR

La pandemia del síndrome respiratorio agudo conocido como COVID-19 y causada por el virus SARS-CoV-2, ha obligado a diversos laboratorios alrededor del mundo a rediseñar y reforzar sus programas de bioseguridad con la finalidad de facilitar el diagnóstico de la enfermedad y colaborar con datos epidemiológicos para la toma de decisiones asociadas a mitigación y control. Una combinación de buenas prácticas de laboratorio y procedimientos claramente definidos, apoyados en la adecuación de la infraestructura, son necesarios para proteger al personal de laboratorio y asegurar la reproducibilidad de los resultados generados. En esta revisión se muestran los lineamientos de bioseguridad fundamentales a implementar en los laboratorios de diagnóstico de la COVID-19, basados en RT-qPCR. El establecimiento del nivel de seguridad biológica a adoptar, así como de los procedimientos operativos estándar, dependerán de la evaluación de riesgo derivada de las actividades intrínsecas del laboratorio. Hasta la fecha, la OMS ha recomendado un nivel 2 de seguridad biológica NBS-2 (BSL-2), con medidas intensificadas de nivel 3 NBS-3 (BSL-3), para actividades de diagnóstico, con prácticas y equipos de protección personal que minimicen la generación de aerosoles y reduzcan la probabilidad de infecciones adquiridas en laboratorio. De igual forma, se ha hecho énfasis en un flujo de trabajo que tome en cuenta el transporte de la muestra bajo reglamentos internacionales y procedimientos para la inactivación del virus, compatibles con la prueba diagnóstica o con los protocolos para la descontaminación de superficies. A pesar del reto que conlleva la adecuación de programas de mitigación ante un patógeno relativamente desconocido como el SARS-CoV-2, es importante destacar que la cultura de bioseguridad permite reducir los riesgos del personal y del medio ambiente y asegurar la calidad de los resultados generados, que permitan contribuir significativamente a lograr el control de la enfermedad.

Assessment of the Biorisk Status of Veterinary Laboratories in Southwest Nigeria: Application of the Food and Agriculture Organization Laboratory Mapping Tool–Safety Module:

Introduction:Because of the nature of work conducted in veterinary laboratories and potential exposures to pathogenic microorganisms, good laboratory practices, risk assessments, biosafety, and bio...

Developing laboratory capacity for Good Laboratory Practice certification: lessons from a Tanzanian insecticide testing facility.

Background: With increasing insecticide resistance in malaria-endemic countries there is an urgent need for safe and effective novel vector control products. To improve the capacity of facilities that test insecticides in sub-Saharan Africa, a programme is supporting seven facilities towards Good Laboratory Practice (GLP) certification, the globally recognized standard for quality management system (QMS) for the conduct of non-clinical and environmental studies. The World Health Organization (WHO) GLP Handbook provides guidance on a stepwise approach to implement a GLP compliant QMS. This study assesses auditor GLP checklists and timings outlined in the WHO GLP Handbook in the real-life context of a Tanzanian insecticide-testing facility, evaluating their implementation in this context. Methods and Principle Findings: We conducted document review and semi-structured interviews with staff at all levels of the test facility to explore factors that influenced progress towards GLP certification. We found that while auditor GLP checklists underemphasised computer systems, they were otherwise broadly applicable. Factors that delayed time to completion of GLP certification included the need for extensive infrastructure improvements, the availability of regional expertise related to GLP, the capacity of national and regional external systems and services to meet GLP compliance requirements, and training development required for Standard Operating Procedure implementation. Conclusion: The standards required for full GLP compliance are rigorous, with an expected completion timeline to implementation of 24 months. This study shows that in low and middle-income countries this timeline may be unrealistic due to challenges related to infrastructure development and lack of regional capacity and expertise. We recommend a comprehensive gap analysis when starting a project, including these areas which are beyond those recommended by the WHO GLP Handbook. These challenges can be successfully overcome and the experience in Tanzania provides key lessons for other facilities seeking GLP certification or the development of similar QMS.

Exploring pre-analytical factors for the optimisation of serum diagnostics: Progressing the clinical utility of ATR-FTIR spectroscopy

Data quality and reproducibility are vital for robust, reliable and consistent diagnostic techniques within clinical environments. Most variance within clinical laboratories occur within the pre-analytical phase, prior to obtaining and analysing data. Herein, we investigate pre-analytical considerations for the spectroscopic analysis of blood serum for the purpose of clinical diagnostics.Variables within sample collection, storage and preparation are explored in order to evaluate their effects on the spectral outcome, including; differences between sample collection tubes, centrifugal speed and time, short and long-term storage conditions, individual analyst technique, sample volumes, environment of sample batches, as well as various drying techniques. Exploratory data analysis using principal component analysis was implemented to unearth spectral variance not immediately observable.Minor spectral variations were observed within each experiment; however, these were not considered significant differences as a result of these varying experimental factors. Variation between different operator techniques when preparing samples was observed, yet can be resolved with appropriate standard operating procedures, regardless of other factors such as sample volume and storage conditions. The findings within this report suggest that experimental variations within a laboratory, or between different laboratories, does not significantly affect the spectral outcome, and with good laboratory practice and careful sample handling results should be consistent and reliable.

Croatian Society of Clinical Embryologists – guidelines on the epidemiological framework for the implementation of medically assisted reproduction (MAR) procedures during the COVID-19 pandemic regarding the safety of patients and medical health workers

Due to the high virulence of the SARS-CoV-2 virus, the infection rate in the community has led to a state of pandemic, leading to the introduction of new emergency measures all over the world. With the aim of controlling and preventing the SARS-CoV-2 viral epidemic, the health institutions performing medically assisted reproduction (MAR) suspended any new MAR treatments in order to reduce the burden on the health care system and implement current social distancing recommendations. Considering the favorable epidemiological situation in Croatia, our perspective is that it is time to conceive, plan and bring forth guidelines for restarting work in MAR centres, taking into account the selection of patients and organization of good laboratory and clinical practices with emphasis on the safety of patients and health workers. In regard to epidemiological knowledge, it is important to establish the reorganization of work in MAR centres including epidemiological measures of reducing unnecessary stays in closed spaces, the usage of protective gear by patients and health workers and disinfection of the working spaces and equipment.

Building a Quality Management System in a Core Facility: A Genomics Core Case Study

Core facilities are key resources supporting the academic research enterprise, providing access to innovative and essential technologies and expertise. Given the constraints placed on core facilities as recharge centers and the ever-changing research environment, an important competitive differentiator that can support rigorous and reproducible approaches in core labs is the implementation of a quality management system (QMS). This paper describes a systematic approach to building a QMS in a genomics core facility at the University of North Carolina School of Medicine. This model is based on principles of the International Organization for Standardization 9001 system with initiatives focused on process mapping, training (communication, customer service, performance management, development of standard operating procedures, and quality audits), root cause analysis, visual control boards, mock quality audits, and continuous improvement through metrics tracking and "voice of the customer" exercises. The goal of this paper is to share practical step-by-step recommendations and outcomes of this core facility QMS that are generally applicable to academic core facilities, regardless of technical focus. Application of these good laboratory practice principles will foster "competitiveness through compliance" and promote outstanding interdisciplinary research between academic cores and their nonacademic pharmaceutical and federal research partners. Additionally, implementation of the QMS qualified this core to apply for federally funded contracts, thereby diversifying its types of projects and sources of revenue.

Weevil Zabrotes subfasciatus (Boheman, 1883) (Chrysomelidae: Bruchinae) Rearing in Dry Bean (Phaseolus vulgaris L.)

Zabrotes subfasciatus (Boheman, 1833) is a cosmopolitan pest, occurring in all countries that grow dry bean, Phaseolus vulgaris L. To assure the development of quality bioassays with this insect, a rearing procedure was implemented according to Standards of Good Laboratory Practices. An average of 2,751.94 adults was produced from 300 couples, in a kilogram of dry bean, in approximately one month. The rearing methodology described herein has been conducted successfully for several generations, providing insects of quality, which have been used routinely for academic and research purposes.

Toxicological evaluation of exosomes derived from human adipose tissue-derived mesenchymal stem/stromal cells

Several studies report that the therapeutic mechanism of action of mesenchymal stem/stromal cells (MSCs) is mainly mediated by paracrine factors that are released from MSCs such as exosomes. Exosomes are nano-sized extracellular vesicles that are transferred to target cells for cell-to-cell communication. Although MSC-derived exosomes (MSC-exosomes) are suggested as novel cell-free therapeutics for various human diseases, evaluation studies for the safety and toxicity of MSC-exosomes are limited. The purpose of our study was to evaluate the toxicological profile, including skin sensitization, photosensitization, eye and skin irritation, and acute oral toxicity using exosomes derived from human adipose tissue-derived MSCs (ASC-exosomes) in accordance with the OECD guidelines and the principles of Good Laboratory Practice. The ASC-exosomes were classified as a potential non-sensitizer in the skin sensitization test, UN GHS no category in the eye irritation test, and as a skin non-irritant in the skin irritation test, and did not induce any toxicity in the phototoxicity test or in acute oral toxicity testing. Our findings are the first to suggest that ASC-exosomes are safe for use as a topical treatment, with no adverse effects in toxicological testing, and have potential application as a therapeutic agent, cosmetic ingredient, or for other biological uses.

Managing the IVF laboratory during a pandemic: international perspectives from laboratory managers

Fertility societies worldwide responded to the COVID-19 pandemic by recommending that fertility clinics close, or sharply reduce, the clinical operation, leading to a shift in the management of IVF laboratories in three phases: shutdown preparation; maintenance during shutdown; and restart. Each of these phases carries distinct risks that need identification and mitigation, forcing laboratory managers to rethink and adapt their procedures in response to the pandemic. The sudden and unprecedented nature of the pandemic forced laboratory managers from around the world to base decisions on opinion and experience when evidence-based response options were unavailable. These perspectives on pandemic response were presented during a virtual international symposium on COVID-19, held on 3 April 2020, and organized by the London Laboratory Managers’ Group. Laboratory managers from seven different countries at different stages of the pandemic (China, Italy, Spain, France, UK, Brazil and Australia) presented their personal experiences to a select audience of experienced laboratory managers from 19 different countries. The intention of this paper is to collect the learnings and considerations from this group of laboratory managers who collaborated to share personal experiences to contribute to the debate surrounding what constitutes good IVF laboratory practice in extraordinary circumstances, such as the COVID-19 pandemic.

The Clinical Pharmacology Sections in Drug Package Inserts: Do We Need to Reexamine the Basis?

The Clinical Pharmacology Sections in Drug Package Inserts: Do We Need to Reexamine the Basis?

Light‐Activated Protein Conjugation and 89Zr‐Radiolabelling with Water‐Soluble Desferrioxamine Derivatives

Protein-conjugates are vital tools in biomedical research, drug discovery and imaging science. For example, functionalised monoclonal antibodies (mAbs) coupled to the desferrioxamine B (DFO) chelate and radiolabelled with 89 Zr4+ ions are used as radiopharmaceuticals for diagnostic positron emission tomography (PET). In this context, protein functionalisation requires the formation of a covalent bond that must be achieved without compromising the biological properties of the mAb. Photochemistry offers new synthetic routes toward protein conjugates like 89 Zr-mAbs but to harness the potential of light-induced conjugation reactions new photoactivatable reagents are required. Herein, we introduce two photoactivatable DFO-derivatives functionalised with an aryl azide (ArN3 ) for use in light-activated conjugation and radiosynthesis of 89 Zr-mAbs. Incorporation of a tris-polyethylene glycol (PEG)3 linker between DFO and the ArN3 group furnished water-soluble chelates that were used in the one-pot, photoradiosynthesis of different 89 Zr-radiolabelled protein conjugates with radiochemical yields up to 72.9±1.9 %. Notably, the DFO-PEG3 chelates can be readily synthesised in accordance with Good Laboratory Practice (GLP), which will facilitate clinical trials with photoradiolabelled 89 Zr-mAbs.

An intercomparison exercise of good laboratory practices for nano-aerosol size measurements by mobility spectrometers

An intercomparison campaign on nanoparticle size measurement was organized in the frame of the French nanoMetrology club. The aim of this study is to make an inventory of the metrological capabilities of all measurement techniques in France involved in the “nano” size range, including the SMPS (scanning mobility particle sizer) concerning aerosol metrology. For this study, four samples have been proposed, namely (1) a SiO2 colloidal suspension (FD304) consisting of a monomodal population, (2) two samples consisting of two nanoparticle populations of SiO2 having proportions to be determined, and (3) a TiO2 colloidal suspension. Ten SMPS associated with five participants around a common experimental setup were performed in link with a control SMPS to have simultaneous measurements with a same instrument in each laboratory in parallel with the SMPS used by each partner. This article presents SMPS results of this study associated with the description of the experimental setup and the sample preparation protocol with an identified schedule and comparison with SEM measurements. The present paper does not focus on the actual capability of the tested mobility spectrometers, but aims to highlight the good laboratory practices using their own but common resources in terms of aerosol generation and measurement setups.

(Z)-1-Chloro-2,3,3,3-tetrafluoropropene (2017).

(Z)-1-Chloro-2,3,3,3-tetrafluoropropene (HCFO-1224yd(Z)) is a colorless gas used as a single substance or in a mixture with other substances for refrigeration. The 4-h rat inhalation LC50 values from two studies were reported to be >20,180 ppm and >213,100 ppm. HCFO-1224yd(Z) is not expected to undergo significant metabolism. The no-observed-effect level of HCFO-1224yd(Z) for cardiac sensitization (in dogs) was 75,000 ppm. In a 5-day repeat inhalation study in rats, the only observation noted was repetitive movement of the mouth/jaws in some animals in the 50,000-ppm exposure group for 1-2 days during the first 3 exposure days. The toxicological significance of this observation was unknown; therefore, the study no-observed-adverse-effect level (NOAEL) was established at 50,000 ppm. In a good laboratory practice (GLP)-compliant, 4-week inhalation study in rats, there were no test substance-related adverse effects noted at any exposure concentration. The study NOAEL was established at 40,000 ppm. In a GLP-compliant inhalation developmental toxicity study, female rats were exposed for 6 h/day from gestation day 6 through 19. There were no test substance-related adverse effects on either the maternal or fetal rats at any exposure concentration. The NOAEL for developmental effects was established at 20,000 ppm. There are no chronic toxicity or carcinogenicity studies available. HCFO-1224yd(Z) gave mixed results in in vitro genotoxicity assays but was negative in an in vivo micronucleus assay. The NOAEL of 40,000 ppm for HCFO-1224yd(Z) from the 4-week, GLP-compliant inhalation study in rats was used at the point of departure (POD) for workplace environmental exposure level (WEEL) value development. This POD was adjusted to account for interindividual variability, duration of exposure, and database limitations. The resulting 8-h time-weighted average WEEL value of 1000 ppm is expected to provide a significant margin of safety against any potential adverse health effects in workers exposed to HCFO-1224yd(Z).

False-Positive Mycobacterium tuberculosis Detection: Ways to Prevent Cross-Contamination.

The gold standard method for diagnosis of tuberculosis is the isolation of Mycobacterium tuberculosis through culture, but there is a probability of cross-contamination in simultaneous cultures of samples causing false-positives. This can result in delayed treatment of the underlying disease and drug side effects. In this paper, we reviewed studies on false-positive cultures of M. tuberculosis. Rate of occurrence, effective factors, and extent of false-positives were analyzed. Ways to identify and reduce the false-positives and management of them are critical for all laboratories. In most cases, false-positive is occurring in cases with only one positive culture but negative direct smear. The three most crucial factors in this regard are inappropriate technician function, contamination of reagents, and aerosol production. Thus, to reduce false-positives, good laboratory practice, as well as use of whole-genome sequencing or genotyping of all positive culture samples with a robust, extra pure method and rapid response, are essential for minimizing the rate of false-positives. Indeed, molecular approaches and epidemiological surveillance can provide a valuable tool besides culture to identify possible false positives.

Opinion on Current Use of Non-Blinded Versus Blinded Histopathologic Evaluation in Animal Toxicity Studies.

The Society of Toxicologic Pathology (STP) explored current institutional practices for selecting between non-blinded versus blinded histopathologic evaluation during Good Laboratory Practice (GLP)-compliant, regulatory-type animal toxicity studies using a multi-question survey and STP-wide discussion (held at the 2019 STP annual meeting). Survey responses were received from 107 individuals representing 83 institutions that collectively employ 589 toxicologic pathologists. Most responses came from industry (N = 46, mainly biopharmaceutical or contract research organizations) and consultants (N = 24). For GLP-compliant animal toxicity studies, histopathologic evaluation usually involves initial (primary) non-blinded analysis, with post hoc informal blinded re-examination at the study pathologist's discretion to confirm subtle findings or establish thresholds. Initial blinded histopathologic evaluation sometimes is chosen by study pathologists to test formal hypotheses and/or by sponsors to address non-pathologist expectations about histopathology data objectivity. Current practice is that a blinded histopathologic evaluation is documented only if formal blinding (ie, using slides with coded labels) is employed, using simple statements without detailed methodology in the study protocol (or an amendment) and/or pathology report. Blinding is not an appropriate strategy for the initial histopathologic evaluation performed during pathology peer reviews of GLP-compliant animal toxicity studies. [Box: see text].

Evaluating Associations Between Nonclinical Cardiovascular Functional Endpoints and Repeat-dose Cardiovascular Toxicity in the Beagle Dog: A Cross-company Initiative.

Integrating nonclinical in vitro, in silico, and in vivo datasets holistically can improve hazard characterization and risk assessment. In pharmaceutical development, cardiovascular liabilities are a leading cause of compound attrition. Prior to clinical studies, functional cardiovascular data are generated in single-dose safety pharmacology telemetry studies, with structural pathology data obtained from repeat-dose toxicology studies with limited concurrent functional endpoints, eg, electrocardiogram via jacketed telemetry. Relationships between datasets remain largely undetermined. To address this gap, a cross-pharma collaboration collated functional and structural data from 135 compounds. Retrospective functional data were collected from good laboratory practice conscious dog safety pharmacology studies: effects defined as hemodynamic blood pressure or heart rate changes. Morphologic pathology findings (mainly degeneration, vacuolation, inflammation) from related toxicology studies in the dog (3-91 days repeat-dosing) were reviewed, harmonized, and location categorized: cardiac muscle (myocardium, epicardium, endocardium, unspecified), atrioventricular/aortic valves, blood vessels. The prevalence of cardiovascular histopathology changes was 11.1% of compounds, with 53% recording a functional blood pressure or heart rate change. Correlations were assessed using the Mantel-Haenszel Chi-square trend test, identifying statistically significant associations between cardiac muscle pathology and (1) decreased blood pressure, (2) increased heart rate, and between cardiovascular vessel pathology and increased heart rate. Negative predictive values were high, suggesting few compounds cause repeat-dose cardiovascular structural change in the absence of functional effects in single-dose safety pharmacology studies. Therefore, observed functional changes could prompt moving (sub)chronic toxicology studies forward, to identify cardiovascular liabilities earlier in development, and reduce late-stage attrition.