Validation of Golden Gate Assemblies Using Highly Multiplexed Nanopore Amplicon Sequencing.

Golden Gate cloning has revolutionized synthetic biology. Its concept of modular, highly characterized libraries of parts that can be combined into higher order assemblies allows engineering principles to be applied to biological systems. The basic parts, typically stored in Level 0 plasmids, are sequence validated by the method of choice and can be combined into higher order assemblies on demand. Higher order assemblies are typically transcriptional units, and multiple transcriptional units can be assembled into multi-gene constructs. Higher order Golden Gate assembly based on defined and validated parts usually does not introduce sequence changes. Therefore, simple validation of the assemblies, e.g., by colony polymerase chain reaction (PCR) or restriction digest pattern analysis is sufficient. However, in many experimental setups, researchers do not use defined parts, but rather part libraries, resulting in assemblies of high combinatorial complexity where sequencing again becomes mandatory. Here, we present a detailed protocol for the use of a highly multiplexed dual barcode amplicon sequencing using the Nanopore sequencing platform for in-house sequence validation. The workflow, called DuBA.flow, is a start-to-finish procedure that provides all necessary steps from a single colony to the final easy-to-interpret sequencing report.

Metrological traceable calibration of organic carbon and elemental carbon based on laboratory-generated reference materials.

Carbonaceous aerosol, including organic carbon (OC) and elemental carbon (EC), has significant influence on human health, air quality and climate change. Accurate measurement of carbonaceous aerosol is essential to reduce the uncertainty of radiative forcing estimation and source apportionment. The accurate separation of OC and EC is controversial due to the charring of OC. Therefore, the development of reference materials (RM) for the validation of OC/EC separation is an important basis for further study. Previous RMs were mainly based on ambient air sampling, which could not provide traceability of OC and EC concentration. To develop traceable RMs with known OC/EC contents, our study applied an improved aerosol generation and mixing technique, providing uniform deposition of particles on quartz filters. To generate OC aerosol with similar pyrolytic property of ambient aerosol, both water soluble organic carbon (WSOC) and water insoluble organic carbon (WIOC) were used, and amorphous carbon was selected for EC surrogate. The RMs were analyzed using different protocols. The homogeneity within the filter was validated, reaching below 2%. The long-term stability of RMs has been validated with RSD ranged from 1.7%-3.2%. Good correlation was observed between nominal concentration of RMs with measured concentration by two protocols, while the difference of EC concentration was within 20%. The results indicated that the newly developed RMs were acceptable for the calibration of OC and EC, which could improve the accuracy of carbonaceous aerosol measurement. Moreover, the laboratory-generated EC-RMs could be suitable for the calibration of equivalent BC concentration by Aethalometers.

Postoperative laboratory testing in the era of outpatient total joint arthroplasty: Targeted patient selection and associated cost savings.

Background: With the advent of outpatient total joint arthroplasty (TJA), the days of routinely drawing postoperative labs (complete blood counts [CBCs] and metabolic panels [CMPs/BMPs]) to monitor for complications are behind us. However, there does exist a subset of at-risk patients that may benefit from diligent postoperative monitoring, though the circumstances under which labs should be ordered remains unclear and subject to surgeon discretion. A systematic review of the literature was therefore conducted to evaluate the utility of postoperative laboratory testing, approaches to targeted patient selection and associated cost-savings. Methods: The PubMed, MEDLINE, EBSCOhost, and Google Scholar electronic databases were searched on August 17, 2023, to identify all studies published since January 1, 2000, that evaluated the role of postoperative lab testing in TJA. (PROSPERO study protocol registration: CRD42023437334). Articles were included if a full-text English manuscript was available and the study assessed the utility of routine postoperative labs in TJA. 19 studies were included comprising 34,166 procedures. The mean Methodological index for Nonrandomized Studies score was 18.2 ± 1.5. Results: Abnormal postoperative lab results were common and infrequently required clinical intervention. Among several identified risk factors for patients that may benefit from postoperative laboratory monitoring, preoperative lab values proved excellent discriminators of transfusion requirement and metabolite-associated intervention. Selective testing demonstrated the ability to generate substantial cost-savings. Conclusion: Routine postoperative laboratory testing offers little clinical utility and produces unnecessary expenditures. Preoperative lab values offer the greatest predictive utility for postoperative transfusion requirement and metabolite-associated clinical intervention, with a preoperative hemoglobin threshold of 111.5 g/L offering an area under the curve (AUC) of 0.93 for predicting postoperative transfusion. Further investigations are needed for metabolic panel predictive models and should incorporate preoperative lab values. The refinement of such models can enable targeted patient selection to avoid unnecessary labs and generate substantial cost savings without compromising patient safety.

Occurrence and predictors of laboratory abnormalities during outpatient parenteral antimicrobial therapy - A multicenter cohort study to inform laboratory test monitoring.

OBJECTIVES: Evidence on the optimal frequency of laboratory testing during outpatient parenteral antimicrobial therapy (OPAT) is lacking. Therefore, we investigated how often and when laboratory abnormalities occur during OPAT and which factors are associated with these abnormalities. METHODS: We performed a multicenter cohort study in four Dutch hospitals among adult patients receiving OPAT and collected routinely obtained laboratory test results. Incidence and incidence rates were calculated for various laboratory abnormalities. Survival analysis was performed to visualize the time to the first occurrence of laboratory abnormalities and Poisson regression analysis to compare the number of abnormalities in the first and second 30 OPAT days among patients receiving OPAT for ≥60 days. Predictors were identified using a multivariable Cox proportional hazard regression model. RESULTS: 45.1% of 1152 included patients developed laboratory abnormalities, but only 2% led to OPAT discontinuation. Hepatotoxicity was most common (33.9 events/1000 OPAT days), with a time-dependent decrease in the occurrence of the first hepatotoxic event, while hypokalemia was rare (1.7 events/1000 OPAT days). In the subgroup of patients receiving ≥60 days of OPAT, nephrotoxicity was more common in days 31-60. We observed partly toxicity-specific associations between antibiotic type, concomitant medication, baseline laboratory values, patient characteristics, and the occurrence of laboratory abnormalities. CONCLUSIONS: While laboratory abnormalities are frequently observed during OPAT, they rarely lead to discontinuation of OPAT. Specific patient, treatment and laboratory characteristics were associated with the occurrence of laboratory abnormalities. Based on our results, we recommend a more personalized laboratory monitoring policy with less blood sampling.

FI-lab in the emergency department and adverse outcomes among acutely hospitalized older adults.

BACKGROUND: The emergency department is treating a growing number of older patients with frailty, which has been linked to poorer outcomes. Urgency is generally emphasized in the emergency department based on indicators such as triage scores and early warning scores for decision-making. However, this approach may not be sufficient for frail older people. The Frailty Index-laboratory (FI-lab) has been used as a simple assessment tool for frailty, but it may also reflect disease severity and predict adverse outcomes in the emergency care setting. Therefore, we aimed to evaluate the association between FI-lab in the emergency room and adverse outcomes during hospitalization through comparison with assessments using triage and early warning scores. METHODS: This was a retrospective cohort study conducted in a tertiary hospital. The study included patients aged 65 years or older who were admitted to the general internal medicine ward after being initially evaluated in the emergency department. FI-lab was calculated using 24 laboratory parameters from blood tests. The National Early Warning Score (NEWS), the Japan Triage and Acuity Scale (JTAS), and the modified JTAS were also used as prognostic indicators, and their association with adverse outcomes was compared with that of FI-lab. RESULTS: In total, 872 patients (mean age, 80.9 years; male, 52.6 %) were analyzed. Patients who died during hospitalization had a higher FI-lab than those who survived. In multiple regression analysis, FI-lab, NEWS, and the modified JTAS were significantly associated with in-hospital death and prolonged length of hospital stay. In contrast, none of these indices were associated with in-hospital falls. The FI-lab was independently associated with the likelihood of discharge to home. CONCLUSIONS: FI-lab evaluated in the emergency department reflected the severity of illness in acutely hospitalized older adults, similarly to NEWS and JTAS, and was a useful indicator for predicting adverse outcomes. These results may indicate the value of FI-lab for older adults in the acute care setting.

Comparative Characteristic of the Methods of Isolation of Oocytes and Spermatozoa in Laboratory Animals (Review).

Over the past decade, there has been an increasing trend in the use of assisted reproductive technologies, which have significantly expanded the opportunities to overcome the problem of infertility. However, the problem of increasing the effectiveness of in vitro fertilization remains open. Isolation of germ cells from animals is a necessary process for various experimental studies. Animal germ cells can be used in experiments to study physical, chemical, genetic, immunological, and microbiological factors affecting reproduction efficiency and for the development of techniques that increase the effectiveness of in vitro fertilization. All of the above determines the relevance of studying existing methods of oocyte and sperm isolation for experimental in vitro studies. Here we discuss the existing methods of sperm and oocyte isolation from animals and their advantages and disadvantages, and also substantiate priority methods for use.

Identification of the genetic background of laboratory rats through amplicon-based next-generation sequencing for single-nucleotide polymorphism genotyping.

BACKGROUND: Laboratory rats, as model animals, have been extensively used in the fields of life science and medicine. It is crucial to routinely monitor the genetic background of laboratory rats. The conventional approach relies on gel electrophoresis and capillary electrophoresis (CE) technologies. However, the experimental and data analysis procedures for both of these methods are time consuming and costly. RESULTS: We established a single-nucleotide polymorphism (SNP) typing scheme using multiplex polymerase chain reaction (PCR) and next-generation sequencing (NGS) to address the genetic background ambiguity in laboratory rats. This methodology involved three rounds of PCR and two rounds of magnetic bead selection to improve the quality of the sequencing data. We simultaneously analysed 100 laboratory rats (including rats of 5 inbred strains and 2 in-house closed colonies), and the sequencing depth varied from an average of 108.25 to 5189.89, with sample uniformity ranging from 82.5 to 97.5%. A total of 98.9% of the amplicons were successfully genotyped (≥ 30 reads). Genetic background analysis revealed that all 38 experimental rats from the 5 inbred strains were successfully identified (without a heterozygous allele). For the 2 in-house closed colonies, the average heterozygosity (0.162 and 0.169) deviated from the typical range of 0.5-0.7, indicating a departure from the ideal heterozygosity level. Additionally, we employed multiplex PCR-CE to validate the NGS-based method, which yielded consistent results for all the rat strains. These results demonstrated that this approach significantly improves efficiency, saves time, reduces costs and ensures accuracy. CONCLUSION: By utilizing NGS technology, our developed method leverages SNP genotyping for genetic background identification in laboratory rats, demonstrating advantages in terms of labour efficiency and cost-effectiveness, thereby rendering it well suited for projects involving extensive sample cohorts.

Challenges with communication of critical laboratory results in a resource-limited setting in South Africa.

Background: Critical laboratory results are test results suggesting a patient is in immediate danger unless treatment is administered promptly. There is a paucity of studies in sub-Saharan Africa on clinicians' utility of these results and affected patients' outcomes. In our resource-limited setting in South Africa, we rely on telephonic communication to convey critical results. Objective: The aim of this study was to determine the average time for clinicians to acknowledge these results on the laboratory information system and to determine the outcome of affected patients. Methods: A retrospective descriptive audit at Tygerberg Academic Hospital was conducted between 01 October 2021 and 31 March 2022. Critical results and the time of acknowledgement by clinicians on the laboratory information system were obtained from inpatients and outpatients. One hundred and twenty inpatient critical results were randomly selected for a folder review to determine patient outcome. Results: Overall, 2514 critical results were reported, and 63 results were excluded. The remaining 2451 results were obtained from 1346 patients. The majority (94.5%) of results were obtained from inpatients, where 1681 (68.6%) were acknowledged within 24 h. The folder audit of 120 patients determined that 40 (33.3%) patients demised. In 82 (68.3%) patients, communication of a critical result did not alter clinical management. Conclusion: Critical laboratory results are crucial to patient care. This study found that approximately one-third of critical laboratory results were not used within 24 h. Engaging clinicians in current practice and implementing a means of improved communication of critical results is required. What this study adds: The study adds to the evidence of challenges experienced with communicating critical results to clinicians which could impact patient care. This is especially true in resource-limited settings; clinicians need to be made aware of the importance of these results, and communication modes need to be improved.

Investigation into the linearity of the Roche c 702 carbamazepine assay.

BACKGROUND: Carbamazepine is an anticonvulsant drug which is monitored in patients due to toxic side effects. At Manchester University NHS Foundation Trust (MFT) carbamazepine is measured using Roche's Kinetic Interaction of Microparticles in Solution (KIMS) method on the c 702 platform. The assay has an upper limit of linearity of 20 mg/L. Samples with concentrations above this limit should be identified and manually diluted. However, a poor EQA return from UK NEQAS for Tox and TDM Distribution 456 has highlighted an issue with the Roche KIMS assay. Sample A of the distribution had a carbamazepine concentration of 36 mg/L but was underreported by several Roche users. This indicated that the assay was not consistently identifying high concentration samples which required a dilution. METHOD: In this investigation, fresh frozen plasma was spiked with carbamazepine concentrations ranging from 15 to 40 mg/L. The spiked samples and EQA material were analysed at two clinical laboratories using the Roche KIMS assay. RESULTS: Samples spiked with concentrations 20-30 mg/L were not consistently identified for dilution by the analyser. This was observed at both hospital sites. Spike samples and EQA with concentrations >30 mg/l were correctly identified at both sites. CONCLUSION: The manual dilution policy has been changed at MFT so all samples with a carbamazepine level ≥15 mg/L will be manually diluted. The problem was reported to Roche who are investigating the issue further. We would suggest that other laboratories look at validating their dilution protocols.

Suggested guide to using lactate gap as a surrogate marker in the diagnosis of ethylene glycol overdose.

BBACKGROUND: Ethylene glycol (EG) poisoning, if not diagnosed rapidly, can lead to poor patient outcomes. Gas Chromatography (GC) is primarily used for EG quantitation which is rarely available, and the turn-around time may be prolonged. Most lactate results from point-of care (POCT) methods are falsely elevated in EG poisoning compared with automated chemistry analyser results. In combination the lactate gap (POCT-Automated chemistry) can be used as surrogate marker in just about all laboratories to indicate likely EG toxicity and guide treatment. CASE REPORT: A male presented by ambulance to hospital with severe agitation requiring mechanical ventilation to facilitate ongoing management. Venous blood gas analysis confirmed a high anion gap metabolic acidosis (HAGMA) with an elevated lactate. The lactate and osmolarity measured in the laboratory showed a normal lactate and high osmolarity, giving a large osmolar gap. The patient was immediately commenced on renal replacement therapy for presumed EG poisoning to minimize kidney injury, and the treatment continued for 19 hours. A very high EG concentration was confirmed by GC the next day. CONCLUSION: An elevated lactate gap along with a HAGMA and osmolar gap can provide rapid surrogate laboratory data indicating EG poisoning enabling timely treatment and better patient outcomes.

Clustering Based on Laboratory Data in Patients With Heart Failure Admitted to the Intensive Care Unit.

BACKGROUND: Heart failure (HF) is a common condition that imposes a significant burden on healthcare systems. We aimed to identify subgroups of patients with heart failure admitted to the ICU using routinely measured laboratory biomarkers. METHODS: A large dataset (N = 1176) of patients with heart failure admitted to the ICU at the Beth Israel Deaconess Medical Center in Boston, USA, between June 1, 2001, and October 31, 2012, was analyzed. We clustered patients to identify laboratory phenotypes. Cluster profiling was then performed to characterize each cluster, using a binary logistic model. RESULTS: Two distinct clusters of patients were identified (N = 679 and 497). There was a significant difference in the mortality rate between Clusters 1 and 2 (50 [7.4%] vs. 109 [21.9%], respectively, p < 0.001). Patients in the Cluster 2 were significantly older (mean [SD] age = 72.35 [14.40] and 76.37 [11.61] years, p < 0.001) with a higher percentage of chronic kidney disease (167 [24.6%] vs. 262 [52.7%], respectively, p < 0.001). The logistic model was significant (Log-likelihood ratio p < 0.001, pseudo R2 = 0.746) with an area under the curve of 0.905. The odds ratio for leucocyte count, mean corpuscular volume (MCV), red blood cell (RBC) distribution width, hematocrit (HcT), lactic acid, blood urea nitrogen (BUN), serum potassium, magnesium, and sodium were significant (all p < 0.05). CONCLUSION: Laboratory data revealed two phenotypes of ICU-admitted patients with heart failure. The two phenotypes are of prognostic importance in terms of mortality rate. They can be differentiated using blood cell count, kidney function status, and serum electrolyte concentrations.

Prevalence, impact and associated factors of abnormal preoperative investigation result in patients undergoing surgery in Dilla University referral hospital: cross-sectional study.

Background: As one domain of preoperative assessment, preoperative investigations are often ordered to evaluate patient's medical condition for risk stratification and assessing patient status to undergoing surgery. Despite the fact that laboratory testing can assist in ensuring the best possible preoperative condition, routine screening examinations have a number of drawbacks. Although there are evidence-based recommendations for which investigations should be done, the tradition of routine preoperative testing is still prevalent and clinical practice with abnormal results detected varies. Method: Institution-based cross-sectional study design was conducted from 1 November to January at Dilla University Referral Hospital. Data was collected from complete pre-anaesthesia check-up sheets, investigations already done. It was collected at the individual level by using, closed-ended self-guided questionnaire. The collected data was entered, cleaned, edited and checked using SPSS version 26 for data processing and analysis. Logistic regression was performed to examine the impacts of abnormal preoperative investigation results and summarised by using tables and figures. An Adjusted odds ratio with 95% CI was computed to determine the level of significance. Result: Data of 208 patients (65.9 female) with mean±standard deviation age 30.83±15.340 years and 22.59±2.99 BMI were analysed. Patients were mostly American Society of Anaesthesiologists I and II underwent National Institute of Clinical and Health Excellence Grade 2 surgeries and surgical shape class 3. Totally, 178 (44.5%) test results were abnormal. CBC is the most detected abnormal result. Only 15 (3.75%) abnormalities had an impact in terms of delay, further investigations, and surgical technique. Comorbidity (AOR 7.982, 95% CI, P=0.041), medication history (AOR 1.463, 95% CI, P=0.013), ASA physical status II (AOR 3.287, 95% CI, P=0.029) and history of smoking (AOR 1.577, 95% CI, P=0.049) were factors which was significantly associated with abnormal preoperative investigation result. Conclusion: Only 0.6% of all tests had a significant impact in terms of changing perioperative anaesthetic management. The significant impact of abnormal investigation result noticed was delayed surgery.

Diagnostics for detection and surveillance of priority epidemic-prone diseases in Africa: an assessment of testing capacity and laboratory strengthening needs.

In 2023, Africa experienced 180 public health emergencies, of which 90% were infectious diseases and 75% were related to zoonotic diseases. Testing capacity for epidemic-prone diseases is essential to enable rapid and accurate identification of causative agents, and for action to prevent disease spread. Moreover, testing is pivotal in monitoring disease transmission, evaluating public health interventions and informing targeted resource allocation during outbreaks. An online, self-assessment survey was conducted in African Union Member States to identify major challenges in testing for epidemic-prone diseases. The survey assessed current capacity for diagnosing priority epidemic-prone diseases at different laboratory levels. It explored challenges in establishing and maintaining testing capacity to improve outbreak response and mitigate public health impact. Survey data analysed diagnostic capacity for priority infectious diseases, diagnostic technologies in use, existing surveillance programmes and challenges limiting diagnostic capacity, by country. The survey result from 15 Member States who responded to the survey, showed high variability in testing capacity and technologies across countries and diverse factors limiting testing capacity for certain priority diseases like dengue and Crimean-Congo haemorrhagic fever. At the same time diagnostic capacity is better for coronavirus disease 2019 (COVID-19), polio, and measles due to previous investments. Unfortunately, many countries are not utilizing multiplex testing, despite its potential to improve diagnostic access. The challenges of limited laboratory capacity for testing future outbreaks are indeed significant. Recent disease outbreaks in Africa have underscored the urgent need to strengthen diagnostic capacity and introduce cost-effective technologies. Small sample sizes and differing disease prioritisation within each country limited the analysis. These findings suggest the benefits of evaluating laboratory testing capacity for epidemic-prone diseases and highlight the importance of effectively addressing challenges to detect diseases and prevent future pandemics.

Biological Reference Ranges for Healthy Indian Blood Donors: Experience of a Tertiary Care Center Vis-à-Vis International Literature.

INTRODUCTION: Complete blood count is the most common, basic test requisitioned in hematology. The normal reference ranges of hematological parameters are required owing to variable socioeconomic, environmental, and genetic factors in populations. The current study determines the reference ranges of the healthy Indian donor population of a high socioeconomic group. METHODS: The study was conducted in the Department of Transfusion Medicine at a tertiary care hospital in India and included 4098 individuals, aged 18-65 years coming for voluntary blood donation from July 2021 to October 2022. Blood samples were collected in K2EDTA, analyzed on the Sysmex XN-31 hematology analyzer, and using statistical tools, the normal reference ranges were calculated. RESULTS: The reference ranges for hemoglobin (HB) (137-185 g/L), WBC (5.1-1.7 × 109/L), platelet count (115.6-370.0 × 109/L) were noted. No statistically significant changes were observed in different age groups. There were gender-wise differences noted in nearly all parameters. The HB and hematocrit (HCT) range was slightly higher in other Indian and other Asian populations with comparable values with the Chinese, Korean populations, and Western populations; RBC parameters were overall comparable with minor differences; the WBC count was higher than the other Indian and Asian populations particularly the upper limit of lymphocyte and monocyte; and the range of platelet counts had a comparable upper limit with all populations and had the lowest lower value in males in our study, which was comparable to only the Chinese population. CONCLUSIONS: It is concluded that reference ranges of common parameters were calculated with minor changes noted in all hematological parameters on comparing with other Indian, Asian population, and Western data.

Data flow in clinical laboratories: could metadata and peridata bridge the gap to new AI-based applications?

In the last decades, clinical laboratories have significantly advanced their technological capabilities, through the use of interconnected systems and advanced software. Laboratory Information Systems (LIS), introduced in the 1970s, have transformed into sophisticated information technology (IT) components that integrate with various digital tools, enhancing data retrieval and exchange. However, the current capabilities of LIS are not sufficient to rapidly save the extensive data, generated during the total testing process (TTP), beyond just test results. This opinion paper discusses qualitative types of TTP data, proposing how to divide laboratory-generated information into two categories, namely metadata and peridata. Being both metadata and peridata information derived from the testing process, it is proposed that the first is useful to describe the characteristics of data, while the second is for interpretation of test results. Together with standardizing preanalytical coding, the subdivision of laboratory-generated information into metadata or peridata might enhance ML studies, also by facilitating the adherence of laboratory-derived data to the Findability, Accessibility, Interoperability, and Reusability (FAIR) principles. Finally, integrating metadata and peridata into LIS can improve data usability, support clinical utility, and advance AI model development in healthcare, emphasizing the need for standardized data management practices.

Effects of acute smoking abstinence among people with schizophrenia: A systematic review and meta-analysis of laboratory studies.

BACKGROUND: Smoking rates in schizophrenia are exceptionally high; however, cessation rates remain low with limited research on effective interventions. A critical component of intervention development is identifying the effects of abstinence that are most salient and therefore may contribute to lapse and relapse. OBJECTIVES: We conducted a systematic review and meta-analysis of controlled laboratory studies investigating acute smoking abstinence effects among people with schizophrenia and schizoaffective disorder. METHODS: This review is reported according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) statement. OVID (MEDLINE, EMBASE, PsycINFO) and PubMed databases were searched from inception until November 2023. RESULTS: We identified (k = 16) articles meeting inclusion criteria; all assessed smoking abstinence (ranging 2-120 h). Acute abstinence resulted in large increases in reward-oriented craving and moderate increases in relief-oriented craving; these effects were greater in studies with longer abstinence duration (high certainty). We also observed significant increases in negative affect and global withdrawal symptoms, as well as memory disruption (moderate certainty). Qualitative synthesis suggests restlessness, irritability, anxiety, and visuospatial working memory may be additionally impacted. Findings with respect to negative symptoms and movement were mixed. CONCLUSIONS: Reward-oriented craving may constitute a key target of smoking cessation interventions for people with schizophrenia. In addition, identification of pharmacological and psychosocial interventions that address abstinence-induced changes in relief-oriented craving, memory, negative affect, restlessness, irritability, and anxiety may strengthen treatment outcomes.

Interactive Notebooks Improve Students' Understanding of Developmental Neurobiology, Attitudes Toward Research, and Experimental Design Competency in a Lecture-Based Neuroscience Course.

Recent efforts to engage postsecondary science, technology, engineering, and mathematics (STEM) students in the rigors of discovery-driven inquiry have centered on the integration of course-based undergraduate research experiences (CUREs) within the biology curricula. While this method of laboratory education is demonstrated to improve students' content knowledge, motivations, affect, and persistence in STEM, CUREs may present as cost- and/or resource-prohibitive. Likewise, not all lecture courses have a concomitant laboratory requirement. With these caveats in mind, we developed the NeuroNotebook intervention, which provided students enrolled in a standalone Developmental Neurobiology course with an immersive, semester-long "dry-lab" experience incorporating many of the same elements as a CURE (e.g., collaboration, use of experimental design skills, troubleshooting, and science communication). Quantitative and qualitative assessment of this intervention revealed positive pre-/post-semester gains in students' content knowledge, attitudes toward the research process, and development of science process skills. Collectively, these data suggest that interventions such as the NeuroNotebook can be an effective alternative to a "wet-lab" experience.

Evaluation of chlorine dioxide exposure in an Australian gnotobiotic mouse research facility.

Exposure to chlorine dioxide by staff working in a gnotobiotic mouse facility at an Australian research institute was measured to determine whether current controls were sufficient to ensure their exposure remains below the current Australian workplace exposure standard. A combination of workplace surveys, interviews with workers, and personal sampling was undertaken to understand the workplace, identify higher-risk tasks, and measure the concentration of chlorine dioxide in the air where the workers conduct routine tasks involving the use of a chlorine dioxide-based disinfectant. Personal sampling utilized the validated Occupational Safety and Health Administration (OSHA) method ID-202, with minor alterations. The tasks identified as being associated with higher airborne exposure to chlorine dioxide were the use of an atomizer to fill isolator ports with aerosolized disinfectant and the use of a disinfectant dunk tank to submerge and surface decontaminate objects. The current work practices in the gnotobiotic facility were found to be compliant with the current 8-hr time-weighted average (TWA) limit of 0.1 ppm (0.28 mg/m3) but were not compliant with the 15-min short-term exposure limit (STEL) of 0.3 ppm (0.83 mg/m3). Improvements in exposure controls, such as implementing the use of a fume cupboard (hood) or other local ventilation when activating the disinfectant solution and improving the utilization of respiratory protective equipment, are therefore required to meet the STEL, but it is recommended that such improvements are also aimed at meeting the proposed Peak limitation of 0.1 ppm that is expected to soon be adopted by Safe Work Australia, replacing the current TWA-8hr and STEL exposure standards.

Minimizing unnecessary proBNP blood tests: an evaluation of a pop-up form implementation in general practice.

OBJECTIVE: To evaluate the impact of a compulsory pop-up form on the ordering pattern of proBNP blood tests by general practitioners in the Capital Region of Denmark. DESIGN: A follow-up study comparing the average number of proBNP tests ordered before and after the implementation of an intervention. SETTING AND SUBJECTS: From 2016 to 2021, the average number of proBNP tests increased by over 300%. In March 2022, a compulsory pop-up form was introduced in the electronic request system (WebReq), requiring general practitioners to select one of three indications for ordering proBNP, as recommended by the Danish Society of Cardiology. The study included 528 general practitioners in the Capital Region of Denmark, with data available from January 2020 to 2023, encompassing 83,576 proBNP results from 56,645 patients. MAIN OUTCOME MEASURE: The average number of proBNP tests ordered per month and the median level of proBNP before and after the intervention. RESULTS: Following the intervention, the average number of proBNP tests per month decreased by 60% over a 22-month follow-up period. The highest reduction was seen among general practitioners who previously ordered the most tests (≥3 per 1000 biochemical tests). In this group, the median proBNP level increased from 12.1 pmol/L before the intervention to 13.5 pmol/L after the intervention (p < 0.0001). CONCLUSIONS: This study demonstrates a significant decrease in the number of proBNP requests from general practitioners in the Capital Region of Denmark after the introduction of a pop-up form in the requisition system containing the current guidelines.

Assessing the physiological effects of microplastics on cultured mussels in the Mediterranean Sea.

Microplastics (MPs) pollution has gained attention due to its ecological threats and potential economic impacts. Yet significant knowledge gaps remain in understanding MPs effects on marine organisms' physiology. This study quantifies the physiological impacts of MPs on farmed mussels (Mytilus galloprovincialis) across various locations in the Mediterranean Sea by combining a laboratory experiment with a Dynamic Energy Budget (DEB) model. Mussels' clearance rates (CR) were measured under different conditions of microplastics and suspended sediment. The DEB model, driven by satellite data and an MPs distribution model, was validated with literature growth and CR data, supporting further the data extracted from the conducted experiment. Results indicate that while the physiological impacts are minimal in most areas, important reductions in CR (8-25%) were estimated in regions like the Gulf of Napoli, leading to reduced growth (6-16%) and reduced reproductive output (7-19%). In addition to microplastic concentrations, seasonal and spatial variations of food availability and suspended inorganic matter importantly control the impacts, with mussels in oligotrophic environments (such as the Gulf of Napoli) showing higher vulnerability to MPs compared to those in more eutrophic locations. This study underscores the utility of bioenergetics models, such as DEB, in evaluating the ecological risks of microplastics and suggests their broader application in MPs research.