Laboratory Preparation Research Articles

Laboratory synthesis and preparation of thermo-responsive polymeric micelle and hydrogel for resveratrol delivery and release

Laboratory synthesis and preparation of thermo-responsive polymeric micelle and hydrogel for resveratrol delivery and release

Recent Trends in Laboratory Preparation and Industrial Production Methods on Cocrystals

Recent Trends in Laboratory Preparation and Industrial Production Methods on Cocrystals

Navigating the depths of refuse-derived fuel in Canada: From heterogeneity to insightful analysis

ABSTRACT Refuse-derived fuel (RDF) presents inherent heterogeneity, encompassing diverse physical and chemical compositions. This study aims to comprehensively investigate the thermogravimetric characteristics of key RDF fractions and compare them with RDF compositions from Canada and other countries. A representative RDF sample was obtained using three ASTM standard procedures, including the quartering technique, manual sorting, and laboratory preparation. Five major RDF fractions – cardboard (46%), mixed papers (17%), mixed plastics (19%), other organics (3%), and fines (13%) – were manually separated and subjected to cryogenic grinding for characterization analysis. Thermogravimetric characterization at 20°C/min in a nitrogen atmosphere, along with proximate/ultimate analysis and heating value measurements, revealed significant variability in decomposition behavior. DTG analysis showed that LDPE exhibited the highest thermal stability, which peaks at 483.6°C, whereas cardboard and mixed paper underwent single-step decomposition, peaks at 361.4°C, and 359.3°C, respectively. In contrast, mixed plastics, other organics, fines, and raw RDF displayed complex, multi-step decomposition behaviors, underscoring the heterogeneous nature of RDF and informing thermal processing optimization. The Canadian RDF sample showed notably higher cardboard 45% and fines content 13% compared to other studies from different countries averaged 25% cardboard, and 8% fines. Additionally, sorting a 2 kg representative sample required 5 man-hours per kilogram, highlighting the labor-intensive nature of the process.

EXTL3 and NPC1 are mammalian host factors for Autographa californica multiple nucleopolyhedrovirus infection

Baculovirus is an obligate parasitic virus of the phylum Arthropoda. Baculovirus including Autographa californica multiple nucleopolyhedrovirus (AcMNPV) has been widely used in the laboratory and industrial preparation of proteins or protein complexes. Due to its large packaging capacity and non-replicative and non-integrative natures in mammals, baculovirus has been proposed as a gene therapy vector for transgene delivery. However, the mechanism of baculovirus transduction in mammalian cells has not been fully illustrated. Here, we employed a cell surface protein-focused CRISPR screen to identify host dependency factors for baculovirus transduction in mammalian cells. The screening experiment uncovered a series of baculovirus host factors in human cells, including exostosin-like glycosyltransferase 3 (EXTL3) and NPC intracellular cholesterol transporter 1 (NPC1). Further investigation illustrated that EXTL3 affected baculovirus attachment and entry by participating in heparan sulfate biosynthesis. In addition, NPC1 promoted baculovirus transduction by mediating membrane fusion and endosomal escape. Moreover, in vivo, baculovirus transduction in Npc1−/+ mice showed that disruption of Npc1 gene significantly reduced baculovirus transduction in mouse liver. In summary, our study revealed the functions of EXTL3 and NPC1 in baculovirus attachment, entry, and endosomal escape in mammalian cells, which is useful for understanding baculovirus transduction in human cells.

An interlaboratory proficiency test using metagenomic sequencing as a diagnostic tool for the detection of RNA viruses in swine fecal material.

Metagenomic shotgun sequencing (mNGS) is a generic molecular diagnostic method, involving laboratory preparation of samples, sequencing, bioinformatic analysis of millions of short sequences, and interpretation of the results. In this paper, we investigated the performance of mNGS on the detection of porcine astroviruses, a model for RNA viruses in a pig fecal material, among six European veterinary and public health laboratories. We showed that different methods for data generation affect mNGS performance among participants and that the selection of reference genomes is crucial for read classification. Follow-up investigation remains a critical factor for the diagnostic interpretation of mNGS results. The paper contributes to potential improvements of mNGS as a diagnostic tool in clinical settings.

Technical note: Optimizing the in situ cosmogenic 36Cl extraction and measurement workflow for geologic applications

Abstract. In situ cosmogenic 36Cl analysis by accelerator mass spectrometry (AMS) is routinely employed to date Quaternary surfaces and assess rates of landscape evolution. However, standard laboratory preparation procedures for 36Cl dating require the addition of large amounts of isotopically enriched chlorine spike solution; these solutions are expensive and increasingly difficult to acquire from commercial sources. In addition, the typical workflow for 36Cl dating involves measuring both 35Cl/37Cl and 36Cl/Cl concurrently on the high-energy (post-accelerator) end of the AMS system, but 35Cl/37Cl determinations using this technique can be complicated by isotope fractionation and system memory during measurement. The traditional workflow also does not provide 36Cl extraction laboratories with the data needed to calculate native Cl concentrations in advance of 36Cl/Cl measurements. In light of these concerns, we present an improved workflow for extracting and measuring chlorine in geologic materials. Our initial step is to characterize 35Cl/37Cl on sample aliquots of up to ∼1 g prepared in Ag(Cl, Br) matrices, which greatly reduces the amount of isotopically enriched spike solution required to measure native Cl content in each sample. To avoid potential issues with isotope fractionation through the accelerator, 35Cl/37Cl is measured on the low-energy, pre-accelerator end of the AMS line. Then, for 36Cl/Cl measurements, we extract Cl as AgCl or Ag(Cl, Br) in analytical batches with a consistent total Cl load across all samples; this step is intended to minimize source memory effects during 36Cl/Cl measurements and allows the preparation of AMS standards that are customized to match known Cl contents in the samples. To assess the efficacy of this extraction and measurement workflow, we compare chlorine isotope ratio measurements on seven geologic samples prepared using standard procedures and the updated workflow. Measurements of 35Cl/37Cl and 36Cl/Cl are consistent between the two workflows, and 35Cl/37Cl values measured using our methods have considerably higher precision than those measured following standard protocols. The chemical preparation and measurement workflow presented here (1) reduces the amount of isotopically enriched chlorine spike used per rock sample by up to 95 %; (2) identifies rocks with high native Cl concentrations, which may be lower priority for 36Cl surface exposure dating, at an early stage of analysis; and (3) allows laboratory users to maintain control over the total chlorine content within and across analytical batches. These methods can be incorporated into existing laboratory and AMS protocols for 36Cl analyses and will increase the accessibility of 36Cl dating for geologic applications.

Spatially refined salinity hazard analysis in gypsum‐affected irrigated soils

AbstractThe global extent of salt‐affected agricultural land, 20% of which is deemed gypsiferous, results in billions of dollars of annual economic loss, a serious problem deserving of attention. However, the analysis of gypsiferous saline soils, such as in the irrigated Lower Arkansas River Valley (LARV) of Colorado, can result in an inflated estimation of soil salinity when using the traditional soil saturated paste extract electrical conductivity (ECe), leading to inaccurate crop yield loss predictions and misguided decisions for remediation. Sparingly soluble gypsum (CaSO4 2H2O) in these soils dissolves more readily during laboratory preparation of saturated paste extracts because of excess soil water dilution coupled with sample disturbance. We present a pragmatic linear‐regression approach to correct for this phenomenon, calibrated using two adapted methods for correcting ECe on an individual sample basis. The novel approach used electrical conductivity of pore water samples from saline fields to evaluate the accuracy of the correction methods. The approach was applied on soil samples from two surface‐irrigated, saline fields in the LARV, which were mapped using electromagnetic induction data and analysis of covariance linear regression, calibrated for ECe and ECe corrected for excess gypsum dissoultion (ECeg). Average ECeg values are as much as 26% lower than uncorrected ECe in gypsum‐biased portions of the fields. Estimation of corn salinity hazard in these gypsum‐affected areas using ECeg in lieu of ECe in a traditional yield response function generated mean relative yield values that are higher by up to 13 percentage points. We discuss lessons learned and suggest enhancements to the techniques.

Laboratory Preparation for Digital Medicine in Healthcare 4.0: An Investigation Into the Awareness and Applications of Big Data and Artificial Intelligence.

Healthcare 4.0. refers to the integration of advanced technologies, such as artificial intelligence (AI) and big data analysis, into the healthcare sector. Recognizing the impact of Healthcare 4.0 technologies in laboratory medicine (LM), we seek to assess the overall awareness and implementation of Healthcare 4.0 among members of the Korean Society for Laboratory Medicine (KSLM). A web-based survey was conducted using an anonymous questionnaire. The survey comprised 36 questions covering demographic information (seven questions), big data (10 questions), and AI (19 questions). In total, 182 (17.9%) of 1,017 KSLM members participated in the survey. Thirty-two percent of respondents considered AI to be the most important technology in LM in the era of Healthcare 4.0, closely followed by 31% who favored big data. Approximately 80% of respondents were familiar with big data but had not conducted research using it, and 71% were willing to participate in future big data research conducted by the KSLM. Respondents viewed AI as the most valuable tool in molecular genetics within various divisions. More than half of the respondents were open to the notion of using AI as assistance rather than a complete replacement for their roles. This survey highlighted KSLM members' awareness of the potential applications and implications of big data and AI. We emphasize the complexity of AI integration in healthcare, citing technical and ethical challenges leading to diverse opinions on its impact on employment and training. This highlights the need for a holistic approach to adopting new technologies.

Effect of different forms of calcium sulfate on the properties of cement

Abstract In our conditions, mainly flue gas desulfurization gypsum (FGD gypsum) is used as a cement setting regulator. When cement is ground, it decomposes into hemihydrate or even anhydrite as the temperature rises. The ground cement is placed in a storage silo where the increased temperature is compounded by increased pressure and the cement properties are affected by the moisture released and by the change in the phase composition of the setting regulator. Under laboratory conditions, hemihydrate and anhydrite II and III were prepared from one type of FGD gypsum. Their purity was monitored by X-ray diffraction analysis and differential thermal analysis. The prepared calcium sulfates were used in different proportions as setting regulators in the laboratory preparation of Portland cements. Technological properties such as strength and solidification were then determined for cements. The hydration process was monitored by the calorimetric method.

Reidentification and determination of drug susceptibility of Corynebacterium diphtheriae and Corynebacterium ulcerans strains isolated from clinical samples in 2023 in Poland.

Despite the fact that the last cases of fully-symptomatic diphtheria were recorded in Poland in 1996 and 2000, infections caused by non-toxin-producing strains of Corynebacterium still occur. According to the epidemiological reports from ECDC in the second half of 2022, there was an increase in the number of diphtheria cases in European Union countries. As a result, the current issue becomes the appropriate preparation of microbiological laboratories for the diagnosis of Corynebacterium microorganisms. Reidentification of diphtheria bacilli isolated from clinical samples and to assess the drug susceptibility of C. diphtheriae strains isolated in Poland. The subject of the research were 18 strains isolated from clinical samples in Poland in 2023. Microbiological and genetic methods were used for the reidentification of the strains. Drug susceptibility was assessed using the disk diffusion method, following the new EUCAST recommendations effective from 2023. It was confirmed that all examined strains belonged to the genus Corynebacterium. It was de-monstrated that C. diphtheriae strains proved to be susceptible to increased exposure to benzylpenicillin and cefotaxime. Results obtained using ciprofloxacin allowed categorizing the strains into the intermediate susceptibility category WZE, except for one strain which was resistant to this antibiotic. All tested bacterial strains were susceptible to erythromycin. The C. ulcerans strain exhibited a similar antibiotic resistance profile to penicillin, cefotaxime, and ciprofloxa-cin, with additional detection of resistance to clindamycin. The toxigenicity of the tested strains was excluded. Based on epidemiological data regarding the emergence of new cases of infections caused by Corynebacterium strains, it is advisable to prepare theoretically and practically laboratories for diagnostics to detect potentially toxigenic diphtheria bacilli. Effective methods for the microbiological diagnosis of diphtheria bacilli are available. It is recommended to monitor the susceptibility to antimicrobial agents in all C. diphtheriae isolates.

Characterization and preparation of food-derived peptides on improving osteoporosis: A review

Osteoporosis is a systemic bone disease characterized by reduced bone mass and deterioration of the microstructure of bone tissue, leading to an increased risk of fragility fractures and affecting human health worldwide. Food-derived peptides are widely used in functional foods due to their low toxicity, ease of digestion and absorption, and potential to improve osteoporosis. This review summarized and discussed methods of diagnosing osteoporosis, treatment approaches, specific peptides as alternatives to conventional drugs, and the laboratory preparation and identification methods of peptides. It was found that peptides interacting with RGD (arginine-glycine-aspartic acid)-binding active sites in integrin could alleviate osteoporosis, analyzed the interaction sites between these osteogenic peptides and integrin, and further discussed their effects on improving osteoporosis. These may provide new insights for rapid screening of osteogenic peptides, and provide a theoretical basis for their application in bone materials and functional foods.

Design of the 1650-1700nm U band doped fiber laser

Increasing the power of thulium-doped fiber lasers is an important direction for the future development of fiber lasers. Thulium ions exhibit excellent optical properties in the U-band, making them valuable for research. This paper has established a modular design process for fiber lasers, investigating the energy level system of Thulium ions, electron transition processes, power propagation equations, pump power threshold, laser output power, and Bragg grating reflectance for specific wavelength photons. The reflectance of the front and rear cavity mirrors and Bragg grating was also redesigned. Finally, by establishing a physical model and employing numerical simulations, the relationship between laser onset power and pump light input power was determined: when the pump light input power reaches 10.1W, a population inversion occurs, and the fiber laser begins to generate laser output power. The laser output power linearly increases with the increase in pump light input power. When the pump light input power reaches 100W, the laser output power reaches its maximum value of 14.17W. This provides valuable data support for the laboratory preparation of 1650-1700nm thulium-doped fiber lasers.

Reduction in the Input of Microplastics into the Aquatic Environment via Wastewater Treatment Plants in Germany

Microplastic (MP) has emerged as a significant environmental challenge due to increased plastic production and its widespread presence in the environment. This study aimed to assess MP concentrations throughout the treatment process at nine wastewater treatment plants (WWTPs) in Germany, from influent to effluent. A customized sampling technique was employed, followed by field and laboratory preparation and the quantification of polymers (PE, PP, PS, PMMA, and PET) using TED-GCMS. MP concentrations decreased progressively in the WWTPs, with influent concentrations ranging from 2.5 to 13.6 mg/L. Effluent concentrations in the conventional WWTPs ranged from 0.001 to 0.051 mg/L, while advanced treatment via filtration yielded concentrations below the limit of quantification at 0.005 mg/L. All tested of the WWTPs demonstrated an over 99% removal efficiency for microplastics. Despite effective retention by the WWTPs, a critical evaluation of the results is necessary. There is a need to optimize existing technologies and enhance the standardization of sampling, processing, and measurement methods, as well as intensify efforts towards creating preventive measures to reduce plastic emissions.

Probing the interaction of Paenibacillus larvae bacteriophage as a biological agent to control the american foulbrood disease in honeybee

American foulbrood (AFB) is a harmful honeybee diseaseprimarily caused by Paenibacillus larvae. The study aims to isolate and identify the AFB causative agent P. larvae and their specific phages to use as a new biological method for AFB disease control. Eight apiaries were inspected for AFB infections. Symptoms of diseased brood comb, were odd brood cells with soft brown decayed brood amongst healthy brood, were identified in the field and demonstrated the prevalence of AFB in every apiary. Three P. larvae isolates were identified using traditional techniques usinga 452-bp PCR amplicon specific to the bacterial 16SrRNA gene andwas compared between Paenibacillus isolates. Additionally, specific phages of P. larvae strains were applied to examine their efficiency in reducing the infection rate under the apiary condition. The infection rate was reduced to approximately 94.6 to 100 % through the application of a phage mixture, as opposed to 20 to 85.7 % when each phage was administered individually or 78.6 to 88.9 % when antibiotic treatment was implemented. Histological studies on phage-treated bee larvae revealed some cells regaining normal shape, with prominent nuclei and microvilli. The gastrointestinal tract showed normal longitudinal and circular muscles, unlike bee larvae treated with bacterial strains with abnormal and destroyed tissues, as shown by the basement membrane surrounding the mid-gut epithelium. Phage techniques exhibited promise in resolving the issue of AFB in honeybees due to their ease of application, comparatively lower cost, and practicality for beekeepers in terms of laboratory preparation.

Experiences of Using a Real Compound Microscope and a Virtual Compound Microscope in Understanding Plant Tissues by First-year Life Science Students at a Tertiary Institution in South Africa

During the COVID-19 outbreak, the majority of tertiary institutions preferred virtual laboratories to face-to-face laboratories which posed a challenge to many students. The objective of the study was to examine students’ experiences at a higher institution using both a physical and virtual microscope. This qualitative study involved ten randomly chosen first-year Life Science students. They had to prepare and view onion cells under a real microscope, while the second assignment required them to observe onion cell preparation in a virtual laboratory. Students completed a questionnaire containing open-ended questions. The results were analyzed using themes, and the study found that pupils prefer physical microscopes to virtual microscopes. This is because students interact with the equipment in the laboratory, which encourages intellectual interests, knowledge sharing, and the development of social connections. Students can manipulate actual objects, manually focus microscopes, and interact in real time with microscopic worlds. This hands-on expertise is critical for instructing students and researchers on laboratory practices. Real microscopes are versatile pieces of equipment that can be utilized for a wide range of applications, from basic instruction to advanced research. Although virtual microscopes have advanced in terms of image quality, real microscopes provide better resolution and clarity. The study will make a substantial addition to university laboratory pedagogies and expose students to scientific investigations, given the majority of students come from rural schools with limited laboratory equipment such as microscopes. Keywords: Physical Microscope, Virtual Microscope, Experiences, Intellectual Interest, Social Interactions

An apparatus and procedure for studying discounting of real outcomes of money and aversive sound.

We developed and examined a laboratory preparation with adult humans that pits shorter term avoidance over longer term positive reinforcement and may serve as a useful laboratory functional analogue of problematic behavior. Participants were exposed to choices between (1) avoiding an aversive sound and acquiring no money or (2) listening to an aversive sound for a set duration and then receiving money. The first choice, avoiding an aversive sound and acquiring no money, was conceptualized as immediate negative reinforcement and no positive reinforcement, whereas the latter choice, listening to an aversive sound for a set duration and then receiving money, was conceptualized as a potential positive punisher paired with a larger later positive reinforcer. We manipulated the duration of the sound and the magnitude of money to identify the point at which individual participants' choices changed from avoiding the sound to choosing the sound plus money. As the sound duration increased, the choice of listening to the sound and receiving money decreased. Similar functions were observed with two different monetary magnitudes. The model has potential applicability to real-world problems such as smoking, addiction, gambling, anxiety disorders, and other impulse control disorders.

Zeolite-templated carbons process with recycled materials and characterisation for energy storage applications

Quantity and quality are the two most important challenges for applying zeolite-templated carbons (ZTCs) into the industry. To provide more information on the mass production in the large-scale facility, depreciation of the quasi-crystal graphite carbon, utilisation of recycled acetylene and zeolite, and online quick quality control for the production of ZTCs, three types of reactors were applied under similar experimental conditions to investigate the ZTC production rate and the generation of the quasi-crystal graphite carbon on the uppermost layer of ZTC. The best specific surface area and carbon yield efficiency from the generation of ZTC were obtained using a fluidised-bed reactor (FBR). The occurrence of the quasi-crystal graphite carbon also disappeared using a FBR. The fluidised-bed-particle kinetic model was also established to demonstrate the feasibility of accomplishing the yield of 0.5 kg per day in the pilot-scale FBR experiments. The utilisation of recycled acetylene could reach 85% and the recovered rate of zeolite was around 60%. An on-site standard operating procedure (SOP) for QA/QC was also proposed for ZTC preparation and characterisation, which prominently shortened the production time for the laboratory preparation of ZTC and achieved superior hydrogen storage and electrochemical capacity.

Impact of varied rates of NPK fertilizer on the microbial diversity of Jatropha zeyheri tea leaves under greenhouse conditions

Fertilization of Nitrogen (N), phosphorus (P) and potassium (K) is an effective strategy for improving tea yield and quality. Even though, NPK fertilization enhances tea yield and quality, addressing soil deficiencies to boost growth, but also impacts soil microbes, influencing nutrient cycling and yield. Therefore, the objective of this study was to assess the effect of different NPK fertilizer application rates on microbial composition of J. zeyheri tea under greenhouse conditions. Six treatments constituting NPK fertilizer rates (2:3:2 ratio) @ 0, 2, 4, 8, 16 and 32 g were arranged in a randomized complete block design, with five replications. 130 days after initiating treatments, leaves were harvested and oven-dried for 72 h at a temperature of 60°C. After laboratory preparations, microorganisms on tea leaves were determined by the principle most probable number using the Tempo reader (BioMerieux, Rodolphe, Durham, United States) instrument. Treatments had a highly significant effect (P ≤ 0.01) on Bacillus cereus contributing 39% in total treatment variation (TTV), whereas increasing NPK fertilizer rates had significant effects (P ≤ 0.05) on Enterobacteriaceae contributing 58% in TTV. Increasing rates of NPK fertilizer did not influence total coliforms contained in J. zeyheri tea leaf tissues. Bacillus cereus and Enterobacteriaceae over increasing NPK fertilizer rates exhibited positive quadratic relations, with density dependent growth pattern. Fertilizer requirements for Bacillus cereus and Enterobacteriaceae under greenhouse conditions were optimized at 1.75 g fertilizer/plant. In conclusion, Bacillus cereus and Enterobacteriaceae were affected by increasing NPK fertilizer rates.

On the value of advanced information about delayed rewards

In a variety of laboratory preparations, several animal species prefer signaled over unsignaled outcomes. Here we examine whether pigeons prefer options that signal the delay to reward over options that do not and how this preference changes with the ratio of the delays. We offered pigeons repeated choices between two alternatives leading to a short or a long delay to reward. For one alternative (informative), the short and long delays were reliably signaled by different stimuli (e.g., SS for short delays, SL for long delays). For the other (non-informative), the delays were not reliably signaled by the stimuli presented (S1 and S2). Across conditions, we varied the durations of the short and long delays, hence their ratio, while keeping the average delay to reward constant. Pigeons preferred the informative over the non-informative option and this preference became stronger as the ratio of the long to the short delay increased. A modified version of the Δ–Σ hypothesis (González et al., J Exp Anal Behav 113(3):591–608. https://doi.org/10.1002/jeab.595, 2020a) incorporating a contrast-like process between the immediacies to reward signaled by each stimulus accounted well for our findings. Functionally, we argue that a preference for signaled delays hinges on the potential instrumental advantage typically conveyed by information.

Bonded lingual spur to intercept thumb sucking habit: A clinical pearl

Thumb sucking is usually treated with fixed and removable palatal cribs and lingual spurs. Bonded lingual spurs were envisioned and designed based on the principles of conventional Orthodontic spurs. Bonded lingual spur has some apparent advantages over banded lingual spur such as no need of molar banding, small size, low cost, superior esthetics, less laboratory preparation, easy insertion and reduced clinical time. This case pearl describes an innovative method of fabrication of bonded lingual spur.