Laboratory Practice Research Articles

Reduce energy consumption in your laboratory – switch ultra-low temperature freezers from – 80 °C to –70 °C. A pilot study on short term storage of plasma samples for coagulation testing

It is common practice in laboratories to store biological samples in ultra-low temperature (ULT) freezers. There is growing interest in raising the temperature of ULT freezers in order to save energy and reduce expenses, as energy conservation becomes increasingly important and sustainable laboratory practices gain popularity. In our laboratory, plasma samples are stored for three months for diagnostic purposes. We therefore took the opportunity to investigate the effect of two different storage temperatures (−70 °C vs −80 °C), on activated partial thromboplastin time (APTT), factor VIII (FVIII), international normalized ratio (INR) and factor VII (FVII) measurements on paired plasma samples collected from 26 individuals after three months of storage. Automated coagulation analysers CS-5100 and ACL TOP were used to perform the tests. We found no consistent difference between the two storage temperatures for any of the four coagulation parameters (all p-values > 0.05). We conclude that the temperature of ULT freezers used to store plasma samples for APTT, FVIII, INR, and FVII measurements can be safely increased from −80 to −70 °C without affecting the stability of the samples.

Diagnosis of avian salmonellosis using PCR kits of different manufacturers

Relevance. The problem in timely detection of this infection is that poultry contaminated with salmonellae may not show clinical signs of the disease. Polymerase chain reaction in the diagnosis of avian salmonellosis is designed for direct detection and identification of pathogenic agents, especially in cases where their detection is difficult by other diagnostic methods. Advantages of PCR tests allow to successfully use this method in veterinary laboratory practice. The aim of this work is to conduct comparative tests of diagnostic PCR kits of different manufacturers in the diagnosis of avian salmonellosis.Methods. To conduct comparative studies of pathological material for the presence of the DNA of Salmonella spp., Salmonella enteritidis and Salmonella typhimurium by molecular biological method, three commercial kits of domestic and imported production for isolation and detection of Salmonella DNA by PCR were used. Determination of the possibility of using membrane glass fiber materials, in the form of cards for taking dry samples of pathogenic material, for subsequent PCR was carried out in comparative tests with parallel studies of biomaterial delivered to the laboratory in frozen form.Results. Comparative studies of pathological material for the presence of Salmonella spp. DNA using three commercial kits for the isolation and detection of Salmonella DNA by polymerase chain reaction, imported and domestic production were carried out. It was found that when comparing the results obtained using kits of different manufacturers for the presence of Salmonella spp. DNA, comparable results were observed. The study was performed within the framework of the state assignment No. FGUG-2022-0009 of (FSC VIEV).

Bacillus subtilis ZB423: 90-Day repeat dose oral (gavage) toxicity study in Wistar rats.

The novel genetically modified probiotic Bacillus subtilis ZB423 was assessed in a 90-day repeated-dose oral toxicity study adhering to Good Laboratory Practice (GLP) and Organization for Economic Cooperation and Development (OECD) guidelines. Spray-dried spores at a concentration of 1.1E12 CFU/g were administered at doses of 130, 260, and 519 mg/kg body weight/day correlating to 1.43 × 1011, 2.86 × 1011, and 5.71 × 1011 CFU/kg/day, respectively, by oral gavage to Wistar rats for a period of 90 consecutive days. Results showed no toxicologically relevant findings for B. subtilis ZB423 from measured parameters. The no observed adverse effect level (NOAEL) of B. subtilis ZB423 is 519 mg/kg body weight/day corresponding to 5.71 × 1011 CFU/kg/day for lyophilized B. subtilis ZB423 spores under the test conditions employed.

Practical Guidance for Clinical Microbiology Laboratories: Updated guidance for processing respiratory tract samples from people with cystic fibrosis.

SUMMARYThis guidance presents recommendations for clinical microbiology laboratories for processing respiratory samples from people with cystic fibrosis (pwCF). Appropriate processing of respiratory samples is crucial to detect bacterial and fungal pathogens, guide treatment, monitor the epidemiology of cystic fibrosis (CF) pathogens, and assess therapeutic interventions. Thanks to CF transmembrane conductance regulator modulator therapy, the health of pwCF has improved, but as a result, fewer pwCF spontaneously expectorate sputum. Thus, the collection of sputum samples has decreased, while the collection of other types of respiratory samples such as oropharyngeal and bronchoalveolar lavage samples has increased. To optimize the detection of microorganisms, including Pseudomonas aeruginosa, Staphylococcus aureus, Haemophilus influenzae, and Burkholderia cepacia complex; other less common non-lactose fermenting Gram-negative bacilli, e.g., Stenotrophomonas maltophilia, Inquilinus, Achromobacter, Ralstonia, and Pandoraea species; and yeasts and filamentous fungi, non-selective and selective culture media are recommended for all types of respiratory samples, including samples obtained from pwCF after lung transplantation. There are no consensus recommendations for laboratory practices to detect, characterize, and report small colony variants (SCVs) of S. aureus, although studies are ongoing to address the potential clinical impact of SCVs. Accurate identification of less common Gram-negative bacilli, e.g., S. maltophilia, Inquilinus, Achromobacter, Ralstonia, and Pandoraea species, as well as yeasts and filamentous fungi, is recommended to understand their epidemiology and clinical importance in pwCF. However, conventional biochemical tests and automated platforms may not accurately identify CF pathogens. MALDI-TOF MS provides excellent genus-level identification, but databases may lack representation of CF pathogens to the species-level. Thus, DNA sequence analysis should be routinely available to laboratories for selected clinical circumstances. Antimicrobial susceptibility testing (AST) is not recommended for every routine surveillance culture obtained from pwCF, although selective AST may be helpful, e.g., for unusual pathogens or exacerbations unresponsive to initial therapy. While this guidance reflects current care paradigms for pwCF, recommendations will continue to evolve as CF research expands the evidence base for laboratory practices.

Lipo-pam™ adjuvanted herpes zoster vaccine induces potent gE-specific cellular and humoral immune responses

Herpes zoster (HZ), also known as shingles, is caused by the reactivation of latent varicella-zoster virus (VZV). Decreased VZV-specific T-cell immune responses significantly contribute to the development of HZ. Shingrix is a recombinant zoster vaccine that is currently used to prevent HZ. However, Shingrix has high reactogenicity and pain at the injection site due to QS21, one of the adjuvant components. In this study, we developed a new herpes zoster vaccine formulation called CVI-VZV-001, containing gE protein and a novel liposome-based adjuvant Lipo-pam™, which consists of two TLR agonists. We evaluated the immunogenicity of CVI-VZV-001 in mouse and rabbit models. CVI-VZV-001 elicited robust gE-specific T-cell immune responses and gE-specific antibody production. Specifically, CVI-VZV-001 induced polyfunctional CD4+ T cell populations that secrete multiple cytokines. Furthermore, CVI-VZV-001 sustained the gE-specific immune responses for up to six months after immunization. To ensure CVI-VZV-001’s safety for further development, we conducted a good laboratory practice (GLP) toxicity test, which confirmed that CVI-VZV-001 is safe for use. At present, CVI-VZV-001 is undergoing phase I clinical trials. This study suggests that CVI-VZV-001 can be a potent candidate for the HZ vaccine with high immunogenicity and safety.

Pengenaan Pajak Terhadap Objek Penelitian Sains yang di Impor Perspektif Maslahah Mursalah

The imposition of import taxes is certainly very high for individuals even though the enforcement limit is at the limit of USD1,500. Even so, the imposition of this tax is also inseparable from scientific research goods such as chemical reaction goods. Chemical reaction goods are goods that produce new chemical compounds such as uranium, cobalt and mercury or laboratory practice goods. Observatory practice items are items that are used as scientific research aids such as star telescopes, electron microscopes, or medical equipment. In terms of benefits, this economic activity is also seen as not meeting the criteria for benefits. This research focuses on the discussion of the Legal Rules for the Imposition of Import Goods Tax According to the Minister of Finance Regulation No. PMK 199/PMK.10/2019 and the imposition of taxes on imported scientific research objects from the perspective of maslahah mursalah. This study uses a normative method with a normative juridical research type. The results of the study prove that the imposition of taxes on imported scientific research objects cannot be said to meet individual benefits because there are parties who consider that the tax is too high to be very detrimental personally. Although the import goods tax is beneficial for the benefit of the wider public, namely for state revenue and infrastructure maintenance.

Potential roles for artificial intelligence in clinical microbiology from improved diagnostic accuracy to solving the staffing crisis.

This review summarizes the current and potential uses of artificial intelligence (AI) in the current state of clinical microbiology with a focus on replacement of labor-intensive tasks. A search was conducted on PubMed using the key terms clinical microbiology and artificial intelligence. Studies were reviewed for relevance to clinical microbiology, current diagnostic techniques, and potential advantages of AI in routine microbiology workflows. Numerous studies highlight potential labor, as well as diagnostic accuracy, benefits to the implementation of AI for slide-based and macroscopic digital image analyses. These range from Gram stain interpretation to categorization and quantitation of culture growth. Artificial intelligence applications in clinical microbiology significantly enhance diagnostic accuracy and efficiency, offering promising solutions to labor-intensive tasks and staffing shortages. More research efforts and US Food and Drug Administration clearance are still required to fully incorporate these AI applications into routine clinical laboratory practices.

The relevance of engineering and technical solutions implemented in the branch of IBCh RAS, and modern problems of vivariums and breeding facility of laboratory animals

The article discusses the relevance of engineering and technical solutions that were implemented in the Branch of Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry of Russian Academy of Sciences (IBH RAS) in the 1980s when designing a laboratory building with a vivarium and a nursery of laboratory animals free of pathogens (SPF). Such laboratory animals are used when performing preclinical studies according to the international standard of good laboratory practice (GLP).

‘New worlds open up’: A qualitative study on Wonder Lab practices in Dutch paediatric palliative care with parents, healthcare professionals, and students

Within paediatric palliative care, it is essential for families and providers to have open, equal, and trusting relationships. In practice, however, building relationships can be challenging. Investing in better understanding the differences in each other's frames of reference and underlying values seems important. Wonder Lab practices provide a space to explore these differences by focusing together on life phenomena in curious and Socratic ways. Wonder Labs were organised with parents, healthcare professionals, and students involved in Dutch paediatric palliative care. The aim of this study was to develop an understanding of how participants experienced participating in Wonder Labs. We conducted twenty in-depth interviews with Wonder Lab participants and used inductive thematic analysis for data interpretation. Five themes were identified: Slowing down, Appreciating stories, Becoming vulnerable, Opening up and diving in, and Reframing perspectives. Participating in Wonder Labs allowed mothers, healthcare professionals, and students to contribute to deepening experiences and gain an expanded understanding of what is at play in caring for children with life-limiting and life-threatening conditions. Through working in pluralised groups, frames of reference and understandings complemented each other and could change. Participants often adopted a more open attitude towards others involved in care after participating and adapted day-to-day practices. Deliberating within paediatric palliative care on sensitive issues and their underlying personal and professional beliefs and values must be part of working together, without specific care situations being the catalyst. This may foster the mutual understanding needed in searching for quality of life, death, and bereavement.

Linking One Health to 3R. Culture of Care as a bridge towards Russell and Burch's highest goal: Replacement

Animal testing has long been ethically controversial, initially due to religious and cultural beliefs and increasingly because of the obvious suffering inflicted. The 3Rs principles are now a central part of laboratory practice and have prompted discussions about adding further Rs to take account of societal values. However, this expansion can lead to contradictions, such as the emphasis on reproducibility, which implies the continued use of animals and thus conflicts with the original highest goal of the 3Rs, replacement. At the same time, the Culture of Care approach associated with the 3Rs promotes a change in the human-animal relationship but has not significantly challenged the use of animals in research. The One Health approach, which recognizes the connection between human, animal and environmental health, is also often criticized for prioritizing human health over animal welfare. This article aims to show how the One Health approach can be consistently extended to interdisciplinarity and intersectionality to include animals as stakeholders in the debate. It will also discuss the potential of the Culture of Care approach to transfer the implications of a consistently conceived One Health approach to the field of animal experimentation, in order to bring the complete renunciation of the use of sentient beings back into the focus of 3R research

High Prevalence of Multistep Algorithms in Diagnostic Clostridioides difficile Laboratory Testing.

Laboratory testing practices for diagnosis of Clostridioides difficile infection (CDI) have evolved in response to published guidelines, availability of highly sensitive nucleic acid amplification tests (NAATs), perceived problems with the specificity of NAATs, and CDI reporting requirements. To assess the current state of laboratory practice for diagnostic CDI testing. An optional 8-item supplemental questionnaire was distributed in December 2019 to the 1374 laboratories participating in the College of American Pathologists C difficile Detection (CDF) proficiency testing program challenge CDF-C. Of 1374 CDF-C participants, 1160 (84.4%) responded, predominantly representing laboratories based in the United States (1077 of 1160; 92.8%). The majority reported using a multistep testing algorithm (684 of 1159; 59.0%). Initial testing with a glutamate dehydrogenase and toxin A/B combination test followed by NAAT for discrepant results was the most common testing method (360 of 1146; 31.4%). NAAT alone (299 of 1146; 26.1%) was next, then NAAT followed by an assay that included toxin A/B enzyme immunoassay if NAAT is positive (258 of 1146; 22.5%). Only 5.4% (62 of 1146) reported using toxin A/B immunoassay alone. Most respondents (1093 of 1131; 96.6%) reported rejecting CDI tests on formed stool, but rejection of CDI testing in pediatric patients was uncommon (211 of 1131; 18.7%). Rejection of CDI testing in patients using laxatives was reported more often by US-based respondents (379 of 1054 [36.0%] versus 9 of 77 [11.7%], P < .001). Multistep algorithms for CDI diagnosis are widely used in line with published recommendations. Most respondents reported rejection of formed stool for CDI testing, but few reported rejection of testing in infants and patients taking laxatives, suggesting these may be areas of opportunity for laboratories to pursue in improving CDI testing practices.

Experiences of early career researchers: Influences on the design and reporting of animal experiments, and the practical and emotional support needed to enhance best practice methods.

While best practice methodology in animal research aims to address reproducibility and translational issues, awareness and implementation remains low. Preclinical systematic reviews have highlighted many flaws, including issues with internal validity and reporting. With early career researchers (ECRs) heavily involved in all aspects of animal experiments, it is crucial we understand what shapes their research practices. Semi-structured interviews were conducted with 13 ECRs, including research masters, PhD and postdoctoral academics. Data were collected and analysed concurrently using constant comparison techniques and an iterative approach. Findings revealed low-level awareness of best practice recommendations but a desire to engage in dedicated workshops on designing and reporting animal experiments. Current laboratory practices and previous literature were main influences on research practice, more than institutional training. An unexpected finding was the discovery of ethical and emotional dilemmas ECRs faced when working with animals. This highlights the need for a multifaceted approach to better support junior researchers, both emotionally and practically, to encourage responsible science.

Variation in liver function testing and the effect of pyridoxal-5-phosphate on ALT, AST and FIB-4 results.

As one of the most requested profiles of blood tests, there is a need for standardization among liver function tests (LFT). Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) are key markers of hepatocellular injury. ALT and AST are used to calculate a Fibrosis-4 (FIB-4) score for assessing liver fibrosis. Despite recommendations by the International Federation of Clinical Chemistry (IFCC) to include pyridoxal-5-phosphate in ALT and AST assay methodologies, most laboratories continue to omit this. Data from the UK NEQAS for Clinical Chemistry Scheme, Distribution 1160 (November 2023), was reviewed to investigate variation in practice regarding liver blood tests in relation to ALT, AST and FIB-4. In addition, a series of questions audited laboratory practice in relation to liver enzymes. Wide variation was seen in LFT profiles offered by laboratories, with 32 different combinations of tests used. The IFCC-recommended methods for ALT and AST are used by one-third of laboratories and give significantly higher results than non-IFCC methods. Laboratories using IFCC methods also reported significantly higher FIB-4 scores. Reference ranges and cut-offs for these tests also varied, and did not account for method-related differences in results. The lack of standardization of LFTs can have a significant impact on patient care. The difference in results for ALT, AST and FIB-4 in laboratories not using IFCC-recommended methods may lead to misdiagnosis. This issue should be addressed by laboratories using methods including pyridoxal-5-phosphate. Until then, method-related reference ranges and cut-offs for ALT, AST and FIB-4 are required.

Pelatihan Pembuatan Beton bagi Siswa SMK N 3 Semarang

&lt;div align="justify"&gt;&lt;p&gt;&lt;em&gt;Concrete is a solid mixture that hardens, consisting of components such as cement, sand, gravel, and water that are mixed. Civil engineering construction is related to concrete, including the construction of roads, buildings, dams, houses, and irrigation channels. Considering the importance of knowledge related to concrete, students of SMK N 3 Semarang who are majoring in construction engineering and housing need to be well informed. Many vocational high school students are still unaware of the proper process of making concrete that meets the desired quality standards. Students of SMKN 3 Semarang, who are expected to be able to work immediately after completing their studies, need to be equipped with knowledge about the concrete-making process. The purpose of this activity is to improve the knowledge of SMKN 3 Semarang students regarding the quality of concrete materials, the making of K-225 quality concrete, and the value of the concrete slump test. The PkM team from Universitas Semarang conducted training on making concrete for SMK N 3 Semarang students using discussion methods and laboratory practice. As a result of this training, the majority of students understand the process of making concrete in terms of material quality, K-225 concrete mix design, and slump test testing.&lt;/em&gt;&lt;/p&gt;&lt;/div&gt;

Integration of Ethnoscience Approach in Physics Learning Based on Laboratory Practice: A Systematic Literature Review

Integration of Ethnoscience Approach in Physics Learning Based on Laboratory Practice: A Systematic Literature Review

MOLECULAR ANALYSIS OF THE MCR-1 GENE IN PSEUDOMONAS AERUGINOSA AND ACINETOBACTER BAUMANII STRAINS

Objective: The emergence of antibiotic resistance in Pseudomonas aeruginosa and Acinetobacter baumanii isolates poses serious risks to public health. Our study aimed to investigate the presence of colistin resistance and mcr-1 gene positivity in these isolates. Material and Method: Culture, biochemical tests, antibiotic susceptibility tests, and molecular tests were used to isolate and identify P. aeruginosa and A. baumanii strains. Result and Discussion: A sum of 156 clinical isolates of Pseudomonas aeruginosa (n = 89) and Acinetobacter baumannii (n = 67) were obtained using the 550 clinical samples collected in one year from the largest hospital in Van, Turkey. The results of antibiotic susceptibility tests showed that approximately 82.8% of P. aeruginosa and 94.6% of A. baumannii strains were multidrug-resistant (MDR). Colistin resistance was detected in 11.23% (10/89) of P. aeruginosa isolates and 11.94% (8/67) of A. baumannii isolates using agar dilution and microdilution methods. Out of the 18 colistin-resistant isolates, the mcr-1 gene was detected in three P. aeruginosa and two A. baumannii strains. The detection of plasmid-mediated colistin resistance in P. aeruginosa and A. baumannii is of great concern due to the high potential for colistin resistance to spread in clinical settings. Understanding the unique circumstances of worldwide colistin resistance can be facilitated by promoting the creation of quick processes for identifying colistin resistance profiles and putting them into practice in hospital laboratories. Colistin and carbapenem treatment are two effective ways to treat emerging resistant super-microbes and slow down the emergence of resistance.

Consistency of alerts generated by, and implementation of, the NHS England acute kidney injury detection algorithm in English laboratories.

National Health Services (NHS) England mandates that an acute kidney injury (AKI) detection algorithmbe embedded in laboratories. We evaluated the implementation of the algorithm and the consistency of alerts submitted to the United Kingdom Renal Registry (UKRR). Code was developed to simulate the syntax of the AKI detection algorithm, executed on data from local laboratories submitted to the UKRR, including alerts and serum creatinine (SCr) results spanning 15months before and after the alert submission. Acute kidney injury alerts were categorized into stages 0/1/2/3. Inter-rater agreement (Gwet's AC1) was used to compare local and centrally derived alerts at individual laboratory and commercial laboratory information management system (LIMS) levels, penalizing extreme disagreements. The analysis included 9,096,667 SCr results from 29 labs (475,634 patients; median age 72years, 47% female) between algorithm activation and data extraction (September 30, 2020). Laboratories and the central simulation generated 1,579,633 and 1,646,850 non-zero AKI alerts, respectively. Agreement was high within known laboratory information management system providers (0.97-0.98) but varied across individual laboratories (overall range 0.17-0.98, 0.17-0.23 in three). Agreement tended to be lower (Gwet's AC1 0.88) with the highest baseline SCr quartile (median 164μmol/L). Overall, alerts submitted to the UKRR are a valid source of AKI surveillance but there are concerns about inconsistent laboratory practices, incomplete adoption of the NHSE algorithm code, alert suppression, and variable interpretation of guidelines. Future efforts should audit and support laboratories with low agreement rates, and explore reasons for lower agreement in individuals with pre-existing CKD.

The use of vacuum fine needle aspiration biopsy of small formations in the diagnosis of thyroid cancer

Introduction. Diagnosis and treatment of lesional pathology of the thyroid are important problems in Russia and worldwide. Currently, fine-needle aspiration (FNA) is the main technique of differential diagnosis of benign and malignant tumors of the thyroid. In 2023, clinical guidelines of the European Thyroid Association (ETA) on thyroid lesion treatment were published noting possible minimally invasive treatment of benign and malignant tumors of the thyroid requiring repeat cytologic verification. Despite the proven effectiveness of FNA, its informativity and specificity vary between 60 and 100 %. Additionally, this method is associated with high risk of false negative and false positive results. Therefore, there is a need to improve FNA and minimize its disadvantages.Aim. To evaluate the capabilities of vacuum FNA of small lesions using multiparameter ultrasound in diagnosis of thyroid cancer.Materials and methods. At the scientific and practical problem laboratory Diagnostic Studies and Minimally Invasive Technologies of the Smolensk State Medical University, a device for vacuum FNA was developed. Between 2023 and 2024, patients with thyroid lesions of size &lt;10 mm underwent 25 vacuum FNAs. Correlation analysis with 51 FNAs performed between 2020 and 2023 was performed.Results. A decrease in the number of nondiagnostic (category I per the Bethesda system, 2017) and undetermined (category III per the Bethesda system, 2017) results was observed. Comparison of preoperative cytologic conclusions (category V per the Bethesda system, 2017) with the results of postoperative histological examination in patients of the control group showed benign lesions in 2 (50 %) cases which correspond to false negative results. Vacuum FNA did not show this discrepancy. These results demonstrate a decrease in the probability of false negative results and effectiveness of vacuum FNA.Conclusion. Use of vacuum FNA expedites ultrasound-guided biopsy for small thyroid lesions. Due to high negative pressure (up to –0.8 bar), single-step retrieval of cytologic material is performed which leads to a decrease in the number of nondiagnostic and undetermined cytologic conclusions by 11.7 and 1.9 %, respectively.

27. Somatic genomic testing and variant curation practices in Australian and New Zealand diagnostic testing laboratories

27. Somatic genomic testing and variant curation practices in Australian and New Zealand diagnostic testing laboratories

Baccalaureate and practical nursing students’ experience with academic accommodation: A qualitative descriptive study

BackgroundBased on the National Educational Association of Disabled Students report, there are numerous gaps in supporting students with disabilities. AimWe examined undergraduate students’ experiences with academic accommodation in the nursing program. Presently, there is minimal research that examines the transition of nursing students with academic accommodation needs in higher educational institutions in the classroom, lab, and clinical environment in Canada. MethodA qualitative descriptive design was used to elicit nursing students' experience with academic accommodation in classroom, lab, and clinical practice within higher education. ResultsNine in-depth semi-structured interviews revealed the challenges and barriers nursing students encountered in their academic journey. Three emergent themes were identified: Students’ experience of their academic accommodation journey, the academic accommodation process through the learning accommodation centre, and recommendations from students’ perspectives. ConclusionThe findings from our study have implications relevant in supporting nursing students, understanding students’ academic accommodation experiences and processes, and informing institutional policies within associated degree nursing programs.