Product Safety Research Articles

Analytical control of residues of the herbicide trifluralin in the assessment of the food safety

Introduction. Trifluralin is a systemic herbicide of the chemical class of dinitroaniline derivatives. During monitoring studies, trifluralin was detected in carrots produced in the Russian Federation. Since trifluralin-based pesticides are not approved for the use in the Russian Federation, residual amounts of the substance are not typical for this crop. To check the contamination of carrots with a pesticide not typical for this crop, confirmatory qualitative and quantitative studies were carried out. Purpose of the work. Increasing the efficiency of identification and reliability of quantitative results when monitoring food products for safety when performing confirmatory analytical studies using the example of determining trifluralin in carrots to assess the safety of food products intended for the consumer. Materials and methods. Gas chromatography coupled with tandem mass spectrometry (GC-MS/MS) was used to identify and quantify trifluralin. Samples were prepared for analysis using the QuEChERS method. Identification was carried out by multiple reaction monitoring (MRM) using a triple quadrupole mass spectrometer using two to four MRM transitions (m/z): 306,1→264,0; 264,0→206,0; 264,0→188,0; 264,0→160,1. Matrix-matched calibration was used for quantitation. Results. The effectiveness of screening studies when monitoring food products using the GC-MS/MS method can be significantly elevated by increasing the number of MRM transitions (at least 3), controlling the reference retention time of the substance, and maintaining the ratio of confirmatory ions. To obtain a reliable quantification of trifluralin content in carrots, the use of a matrix-matched calibration is recommended. Limitation. In the study, only carrots were considered as food products. Conclusion. Confirmation of test results is especially important when determining pesticide residues that are not normally found in a given matrix, or when it is suspected that the maximum permissible level may be exceeded. When conducting a screening study and detecting a food contaminant, contamination of the product can only be reported in advance. Next, a confirmatory analysis is required using a validated quantitative method, including an appropriate calibration procedure. In quantitative analysis, the presence of matrix sample components can cause problems due to sample suppression/enhancement phenomena. Matrix-aware calibration has been shown to be effective in compensating for matrix effects.

Arsenic, cadmium, and chromium concentrations in contrasting phosphate fertilizers and their bioaccumulation by crops: Towards a green label?

Potentially toxic elements such as arsenic (As), cadmium (Cd), and chromium (Cr) are severely regulated in fertilizers and deserve continuous investigation. Phosphate-derived Cd has been a stepping-stone toward achieving sustainable and safe worldwide food production, especially after a new regulation aiming for reduced limits of Cd in P fertilizers (EU, 2019/1009). Three pot experiments were conducted to assess the variability of As, Cd, and Cr concentrations - with a particular focus on Cd - from monoammonium phosphates (MAP 1, MAP 2, and MAP 3 from different geographic origins) and their accumulation in limed and unlimed soils, and contrasting crops, representing staple food and significant sources of these elements for humans (i.e., potato, tobacco, and rice). A diverse array of sensitive techniques for trace elements determination were used to reveal the highest level of Cd of MAP 3 (20.71 mg kg−1 MAP), which loaded the highest amounts of this element to the soil matrix and solution, plant shoots, and xylem sap, contrasting with results for MAP 1 (0.87 mg kg−1 MAP), which has almost ten times less Cd than that required for low-Cd labeling of P fertilizers (≤20 mg Cd kg−1 P2O5). MAP 3 also had the highest Cr concentration (139.3 mg kg−1 MAP). Among crops, rice accumulated 16-fold less Cd than potato plants. Liming decreased Cd in tobacco and potato shoots up to 35%. Moreover, reductions of about 20% were also observed for Cd accumulation in tubers and sap. Conversely, Cd from MAP 3 was always much more accumulated in soil solution, achieving up to 20 μg L−1, while values < 5 μg L−1 (i.e., a groundwater limit) were obtained from MAP 1. Our findings may be used as a reference in developing green labels for fertilizers in scenarios where Cd accumulation represents a potential risk for soil and human health.

Analysis and Optimization of the Effectiveness of Production Safety Standardization Construction Based on Set-Pair Analysis

Analysis and Optimization of the Effectiveness of Production Safety Standardization Construction Based on Set-Pair Analysis

A Systematic Review of Nano-Encapsulation for Improving the Bioavailability of Dietary Supplements and Nutraceuticals

Nanoencapsulation is a growing technique in the field of nutraceuticals and dietary supplements, offering several advantages over non-encapsulated forms. This systematic review assesses the impact of nanoencapsulation on key factors such as bioavailability, stability, tolerance, and encapsulation efficiency. The selected studies demonstrate that nanoencapsulation enhances the protection and controlled release of bioactive compounds, leading to improved efficacy and therapeutic potential. Furthermore, it increases the bioavailability of nutrients and active ingredients while maintaining their stability under different conditions. The review also highlights the positive effects on product safety and consumer tolerance, making nanoencapsulation a promising strategy for optimizing the formulation of dietary supplements. These findings suggest that nanoencapsulation could play a critical role in advancing the effectiveness of nutraceutical products. Keywords: Nanoencapsulation, dietary supplements, nutraceuticals, efficacy, bioavailability, stability

The Influence of Market Orientation on Marketing Performance of Small and Medium Enterprises in Lamongan District

This research aims to analyze the influence of market orientation on innovation capability and marketing performance, as well as examine the mediating role of innovation capability in the relationship between market orientation and marketing performance. The results of this quantitative research show that market orientation contributes to marketing performance and the formation of innovation capabilities, and that innovation capabilities of SMEs are reflected in attractive and safe products. This research contributes to an understanding the dynamics of market orientation, innovation, and marketing performance in SMEs in the context of Lamongan Regency. Managerial implications and policy recommendations are outlined based on research findings, with the hope of providing guidance for SMEs and related parties on improving marketing performance. These findings support the theory that market orientation can improve marketing performance with innovation capabilities. The practical implication of this research is that SMEs can strengthen their market orientation to increase product innovation. Keywords: market orientation, innovation capability, marketing performance

Dysphagia and Muscle Weakness Secondary to Botulinum Toxin Type A Treatment of Cervical Dystonia: A Drug Class Analysis of Prescribing Information.

The first-line management of cervical dystonia (CD) symptoms is intramuscular injection of botulinum toxin type A (BoNTA). However, a comparison of safety among BoNTAs is difficult because, per regulatory authorities, units of BoNTA activity are not interchangeable. Dysphagia and muscle weakness are widely considered two key adverse events to monitor closely in the treatment of CD. This integrated analysis compared the safety of BoNTAs approved for CD in the US by evaluating relationships between the incidence of dysphagia and muscle weakness in prescribing information and the core neurotoxin content. Coefficients The coefficients of determination (R2) and trendlines were estimated via regression-based lines of best fit. Adverse drug reaction (ADR) rates were strongly correlated with core neurotoxin amounts for conventional BoNTAs (slope coefficients: dysphagia = 0.048, R2 = 0.74; muscle weakness = 0.096, R2 = 0.82). The published ADR rates at approved doses for conventional BoNTAs were higher compared with DaxibotulinumtoxinA (DAXI; DAXXIFY®, Revance Therapeutics, Inc., Nashville, TN, USA) by core neurotoxin content. The use of a core neurotoxin amount was found to be an effective method for comparing the safety of BoNTA products. Current clinical trials suggest that DAXI, a novel BoNTA formulation, provides a potentially wider safety margin compared with other approved BoNTAs for CD. The lower amount of core neurotoxin administered at approved doses compared with conventional BoNTAs may explain low on-target ADRs like muscle weakness, whereas reduced diffusion from the injection site is thought to be responsible for low off-target ADRs like dysphagia.

Exploring the synergistic benefits of biochar and gibberellic acid in alleviating cadmium toxicity

Cadmium (Cd) toxicity significantly threatens agricultural productivity and food safety. Developing effective strategies to enhance plant tolerance to Cd stress is essential. This study investigates the synergistic effects of biochar (BC) and gibberellic acid (GA3) on mitigating Cd toxicity in maize (Zea mays), focusing on their impact on oxidative stress markers and antioxidant enzyme activities. Soil samples were collected from the Cholistan Institute of Desert Studies (CIDS) and analyzed for trace metal ions and other properties. Biochar was produced from fruit and vegetable waste, washed, washed, deashed, and mixed with 10 ppm GA3. FH-1036 hybrid maize seeds were sterilized and planted in pots containing soil with varying Cd levels (0, 8, and 16 mg Cd/kg soil). Twelve treatments were established, including control, GA3, BC, and their combinations under different Cd stress levels. Plants were irrigated to maintain 60% field capacity and harvested at the V10 growth stage. Hydrogen peroxide (H2O2) contents and activities of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and ascorbate peroxidase (APX) were measured in roots, stems, and leaves. Statistical analysis was performed using OriginPro 2021, with ANOVA and Fisher’s LSD test used to determine significant differences. GA3 and BC treatments significantly reduced H2O2 levels in maize roots, stems and leaves under Cd stress. The combined treatment of GA3 + BC showed the most significant reduction in H2O2 levels across all plant parts, reducing root H2O2 by 50%, stem H2O2 by 55%, and leaf H2O2 by 53% under severe Cd stress (16 mg Cd/kg). SOD activity increased under non-stress conditions but decreased under Cd stress, with the highest activity observed in the combined treatment. POD activity followed a similar pattern, with GA3 + BC treatment resulting in the most significant increases under non-stress conditions and the least reductions under Cd stress. CAT activity showed substantial increases with GA3 + BC treatment, particularly under severe Cd stress, with a notable rise over the control. APX activity also exhibited enhancements with GA3 and BC treatments, especially in the combined treatment under various Cd stress levels. This study highlights the potential of combined BC and GA3 treatments in improving Cd stress tolerance in maize. Future research should focus on field trials and the long-term impacts of these treatments on crop productivity and soil health.

Estudo das dificuldades encontradas por laboratórios brasileiros na implementação de métodos alternativos ao uso de animais: avaliação de irritação ocular e o teste de Permeabilidade e Opacidade da Córnea Bovina (BCOP)

Introduction: Ensuring the safety of products causing ocular irritation is crucial. The Bovine Corneal Opacity and Permeability (BCOP) test is an accepted alternative method for ocular irritation assessment in regulatory contexts. Objective: This study aimed to identify the main challenges and technical limitations faced by professionals when implementing alternative methods, particularly for ocular irritation assessment. Method: A qualitative research approach was employed using questionnaires (n = 22 respondents) and interviews (n = 7), followed by discourse analysis. Results: Findings reveal that 71% of alternative method practitioners encounter significant difficulties, primarily due to high validation/implementation costs (62%), driven by consumable expenses. Limited access to training (62%) and importation of reagents and equipment (43%) were also noted. Key BCOP limitations include restricted access to bovine eyeballs (50%), less precise risk classification (43%), and substantial eyeball waste (29%) due to damage. Interviews supported these conclusions, highlighting challenges such as travel to slaughterhouses, insufficient technical details in the OECD Guide, the need for additional support materials, and a lack of experienced professionals. These results underscore the need for expanded training opportunities for test implementation. The high cost of imported materials indicates greater validation expenses in Brazil than in other countries. Conclusions: Addressing these challenges requires an open innovation environment, fostering collaboration among the Triple Helix (Companies, Scientific and Technological Institutions, and Government).

Safety and tolerability of a Muse cell-based product in neonatal hypoxic-ischemic encephalopathy with therapeutic hypothermia (SHIELD trial).

Hypoxic-ischemic encephalopathy (HIE), associated with high mortality and neurological sequelae, lacks established treatment except therapeutic hypothermia. Clinical-grade multilineage-differentiating stress-enduring (Muse) cells (CL2020) demonstrated safety and efficacy in nonclinical HIE rat models, thereby leading to an investigator-initiated clinical trial to evaluate CL2020 safety and tolerability in neonatal HIE as a single-center open-label dose-escalation study with 9 neonates with moderate-to-severe HIE who received therapeutic hypothermia. Each patient received a single intravenous injection of CL2020 cells between 5 and 14 days of age. The low-dose (3 patients) and high-dose (6 patients) groups received 1.5 × 106 and 1.5 × 107 cells/dose, respectively. The occurrence of any adverse event within 12 weeks following CL2020 administration was the primary endpoint of this trial. No significant changes in physiological signs including heart rate, blood pressure, and oxygen saturation were observed during or after administration. The only adverse event that may be related to cell administration was a mild γ-glutamyltransferase level elevation in one neonate, which spontaneously resolved without any treatment. All patients enrolled in the trial survived, and normal developmental quotients (≥ 85) in all 3 domains of the Kyoto Scale of Psychological Development 2001 were observed in 67% of the patients in this trial. CL2020 administration was demonstrated to be safe and tolerable for neonates with HIE. Considering the small number of patients, a randomized controlled confirmatory study is warranted to verify these preliminary findings and evaluate the efficacy of this therapy.

Development of an exposure assessment industrial model for Bacillus cereus in rice matrix containing insect chitosan.

An exposure assessment model for industrial use has been developed by using kinetic data from inactivation and growth of Bacillus cereus spores. It can provide a valuable tool for estimating the concentration of B. cereus after a storage period of 24 h at a specified temperature (20 °C) and for an estimation of the percentage of contaminated portions according to the input data of the model. This model considers a rice-derived product that has undergone a standard cooking process at 95 °C for 20 min. According to the results, the presence of chitosan affects the final microbial load after storage, potentially serving as an additional control measure in the event of cold chain abuse or break. Chitosan's antimicrobial properties likely play a role in reducing microbial growth during storage, thereby contributing to enhanced food safety. In practical terms, this suggests that incorporating chitosan into food products, especially those susceptible to microbial contamination like rice derivatives, could help mitigate risks associated with temperature abuse or cold chain disruptions. By acting as a protective barrier against microbial proliferation, chitosan offers a preventive measure to maintain product quality and safety throughout the supply chain. Considering two scenarios, 104 or 107 as initial contamination the model estimated that the 55 and 100% of portions would be respectively contaminated, according to a Performance Criteria of 4 log reductions.

Expression of Genes Related to Meat Productivity, Metabolic and Morphological Significance of Broiler Chickens with the Use of Nutritional Phytochemicals.

The study aimed to analyze gene expression linked to skeletal muscle growth and lipid metabolism in broiler chickens fed with plant extracts. Five groups of chickens were formed: four experimental groups and one control group. The diets of the experimental groups were supplemented with different plant extracts: chicory, St. John's wort, maral root, and creeping thyme, whereas the control group received feed without phytobiotic compounds. Weekly weighings were conducted (n = 36). The chickens were slaughtered at day 26 for tissue sampling of four birds from each group. Gene expression (MYOG, MSTN, FASN) related to muscle growth and fatty acid synthesis was analyzed using the β-actin ACTB gene as a reference. Blood samples were taken at day 35 for biochemical analysis and anatomical dissection was performed. The study revealed that using plant extracts from chicory, thyme, and maral root increased MYOG gene activity by 4.21, 7.45, and 8.93 times, respectively. T. serpyllum extract boosted the MSTN gene by 10.93 times, impacting muscle growth regulation. FASN gene expression for fatty acid synthesis increased significantly by 18.22-184.12 times with plant extracts. The best results regarding meat productivity of chickens were obtained when using R. carthamoides extract. The results of the study will serve as a basis for further development of a phytocomposition designed to increase the meat productivity of broiler chickens in the production of environmentally safe poultry products.

Influence of roof confined water drainage on stress distribution in coal seam

Coal and gas outbursts, along with rock bursts, are common dynamic disasters in coal mining, threatening the safety and green production of coal mines. The distribution of coal seam stress is an external factor contributing to coal mine power disasters. The drainage of confined water in coal seam roof is the primary factor of coal seam stress changes. Using theoretical analysis and numerical simulations, the influence of roof confined water on coal seam stress by looking at the law of stress and stress conservation transfer in the drainage layer is investigated. Our results show that: (1) after drainage, there is an increase in the local stress within the coal seam, with a stress concentration factor of 1.35. The influence range of theoretically calculated stress is consistent with the measured results. (2) the maximum value of drainage boundary stress correlated positively with the angle of the water-rich abnormal area, with a linear fitting of R2 = 0.98. These findings hold significant theoretical and practical implications for preventing and controlling dynamic disasters during the mining of water-rich coal seams.

Optimizing Construction Management and Workflow Integration through Autonomous Robotics for Enhanced Productivity Safety and Precision on Modern Construction Sites

The construction industry is experiencing a technological transformation with the integration of robotics, fundamentally altering traditional construction management and process design. This review explores the role of construction robots in enhancing productivity, safety, and efficiency on construction sites. It examines the various types of robots used, such as bricklaying and welding robots, and analyzes their functions and real-world applications. The paper also addresses the design of workflows and processes around robots, highlighting the challenges and strategies involved in adapting construction projects to a robot-centric approach. Additionally, the impact of robotics on construction management, including shifts in roles, cost efficiency, and collaboration between humans and robots, is thoroughly explored. The review concludes by identifying future trends, challenges, and opportunities in the field of construction robotics, along with relevant policy and regulatory considerations. This paper aims to provide insights into how construction management must evolve to successfully integrate robots, leading to a more efficient, cost effective, and safe construction environment.

The Development ofanAssessment Tool forSuccessful Product Development Based On DFMA andAHP Approach: Automotive Industry Case Study

Successful product development is critical in the automotive industry, offering benefits such as optimisedmanufacturing, reduced costs, and faster market entry. Despite available tools, the complexity of design resources makes the process challenging. This study aimed to develop an assessment tool for evaluating successful product design in the automotive sector, starting with identifying success criteria from reputable research. Expert Choice software was employed to analysethe significance and weightage of the identified criteria. As a result, there are 19 criteria were prioritisedacross four key categories. The requirements such as product design, cost-effectiveness, safety, and environment. Notably, the most crucial criteria identified were recyclable materials (0.139), end-of-lifecycle considerations (0.118), and sustainable manufacturing (0.108984), while the least important were standardisation(0.0155), cabin ergonomics (0.022), and fire safety (0.0158). The resulting assessment tool, validated by four industry and academic experts, showed promise in helping practitioners self-evaluate successful product design within the automotive industry.

Comparative analyses of bacterial contamination and microbiome of broiler carcasses in wet market and industrial processing environments

The slaughtering environment is crucial for the food hygiene and safety of poultry products. Despite the global dominance of industrial processing, live bird slaughtering in wet markets persists due to cultural, religious, and economic reasons. This study aims to reveal the correlation between hygiene scales in wet markets and bacterial contamination levels on broiler carcasses, with a particular focus on pathobiont transmission risks and microbiome characteristics. Wet markets were categorized based on home-made ratings and the Hygiene and Biosecurity Assessment Tool (HABT). The study assessed total aerobic bacterial (TAB) levels, food spoilage and hygiene indicators (Pseudomonas and E. coli), foodborne pathogen Salmonella, and the microbiome of broiler carcasses, intestinal contents, and slaughtering facilities. Comparative analyses were conducted between market and industrial processing environments. TAB levels on broiler carcasses showed a significant negative correlation with hygiene scores, indicating that both HABT and home-made rating tools effectively assess and improve processing hygiene. Industrial processing consistently reduced bacterial contamination compared to wet markets. Although Salmonella spp. prevalence was lower in market-processed carcasses, the study identified significant cross-transmission of pathobionts and variations in bacterial composition with hygiene improvements. Notably, the microbiome analysis revealed overlaps in amplicon sequence variants (ASVs) between carcasses and contamination vectors, highlighting pathobiont transmission risks. The present study confirmed the scales of hygiene standards among wet markets reflect bacterial contamination on broiler carcasses. Enhancing slaughter practices to meet industrial hygiene standards is essential for reducing the transmission of foodborne pathogens and pathobionts, and improving food safety.

Structural Analysis of Hyaluronic Acid Fillers Using Nuclear Magnetic Resonance: Implications for Quality Control and Clinical Performance.

Potential disruptions in the biocompatibility of hyaluronic acid (HA) fillers can arise with mono-linked 1,4-butanediol diglycidyl ether (BDDE) or unreacted (pendant) 1,4-butanediol di-(propan-2,3-diolyl) ether. Assessing the filler's degree of modification involves evaluating improperly cross-linked BDDE. This study analyzed commercially available HA fillers using nuclear magnetic resonance (NMR), focusing on key parameters, such as the degree of modification (MoD), the cross-linker ratio (CrR), and the degree of cross-linking. We assessed thirteen commercially available HA fillers using NMR. The samples were placed in an NMR instrument, and each sample was analyzed for 26 h, including MoD and CrR assessments. MoD 1H ranged from 17.065% to 2.239%, MoD 13C ranged from 12.567% to 1.947%, and CrR 13C ranged from 0.394 to 0.014. Significant distinctions were observed in the CrR 13C values when the MoD values of the products were similar. This study underscores the importance of considering the MoD and the CrR together to ensure optimal cross-linking and minimize the risks associated with residual BDDE impurities. Utilizing NMR for HA gel characterization provides valuable insights regarding product quality control, safety assessments, and clinical performance evaluations for esthetic interventions, contributing to filler product improvements. Further studies correlating NMR findings with real-world outcomes are essential for ensuring safety and efficacy.

MRNA vaccines: A powerful tool in cancer treatment

As an emerging immunotherapy, cancer vaccine is a method of introducing tumor antigens, genes encoding tumor antigens, immune cells and molecules into the body to activate specific immune responses to kill tumors. In April 2010, the world's first related drug, previz, was approved for marketing in United States. The drug extracts peripheral blood mononuclear cells from patients' autoimmune cells, activates them in vitro and then infuses them back into the treatment of asymptomatic or mildly symptomatic hormone-refractory advanced prostate cancer. mRNA vaccines have attracted much attention because of their high efficacy, specificity, versatility, ability to develop rapidly on a large-scale, low-cost production potential and safety. The mRNA vaccines and nano-agent delivery technology are considered to have great potential in cancer immunotherapy. In recent years, developments in the design and delivery methods of mRNA vaccines have accelerated the development and use of mRNA vaccinations against cancer in clinical settings. This review introduces optimization strategies of mRNA vaccinations against cancer, including coding and non-coding region optimization, and emphasizes mRNA vaccine delivery systems. Multiple nanoparticle vectors have currently been developed to improve mRNA stability and administration efficiency. At the same time, we discuss the applications of cancer therapy with mRNA vaccinations, its limitations and future challenges.

Spatial distribution and risk assessment of heavy metal pollution from enterprises in China

Heavy metal pollution (HMP) directly affects the safety of agricultural products, thereby impacting human health. Industrial emissions, as the main source of soil HMP in China, require in-depth research on their pollution risks. Based on national heavy metal (HM) enterprise data, this paper analyzes the spatial distribution characteristics of key enterprises involved in the HMP across the country. It constructs the risk assessment index system of enterprise HMP based on the “source-pathway-receptor” (SPR) process of the HMP, evaluates and partitions the risk of the HMP from enterprises nationwide. The results show that: (1) Enterprises and pollutant discharge outlets are mainly distributed in the eastern and southeastern coastal regions. Jiangxi, Yunnan, Guangdong, and Hunan Province are the main distribution regions of smelting enterprises, with the most types of HM pollutants. The hazard of pollution sources shows a spatial distribution pattern of higher risk in the southwest and north, and lower risk in the central region. Counties with high-risk pollution sources are mainly distributed in Yunnan, Hunan, Guangdong, Inner Mongolia, and Jiangxi Province. (2) The hazard of pollutant transmission pathways shows a spatial distribution pattern of higher risk in the southeast and lower risk in the central region. About 31.5 % of counties are at extremely high risk, mainly distributed in the southeastern coastal regions of Guangdong, Jiangsu, Zhejiang, Jiangxi, Shandong, and Fujian Province. (3) The vulnerability of the receptors shows significant clustering characteristics in the northeast and central regions. About 3.3 % of counties have a receptor vulnerability level of “extremely high,” mainly distributed in Inner Mongolia, Jilin, Heilongjiang, Liaoning Province in the northeast, as well as Hubei and Jiangsu Province. (4) About 1.55 % of counties nationwide have a comprehensive risk level of the HMP classified as “extremely high,” mainly distributed in Guangdong Province and Inner Mongolia. Additionally, some counties in Yunnan, Hunan, Jiangsu, Jiangxi, and Zhejiang Province have a risk of exceeding pollution standards, requiring further preventive measures to reduce pollution risks in the future. This paper can provide a scientific basis for the prevention and control (P&C) of the HMP in China.

Prediction and Visualization of Total Volatile Basic Nitrogen in Yellow Croaker (Larimichthys polyactis) Using Shortwave Infrared Hyperspectral Imaging.

In the present investigation, we have devised a hyperspectral imaging (HSI) apparatus to assess the chemical characteristics and freshness of the yellow croaker (Larimichthys polyactis) throughout its storage period. This system operates within the shortwave infrared spectrum, specifically ranging from 900 to 1700 nm. A variety of spectral pre-processing techniques, including standard normal variate (SNV), multiple scatter correction, and Savitzky-Golay (SG) derivatives, were employed to augment the predictive accuracy of total volatile basic nitrogen (TVB-N)-which serves as a critical freshness parameter. Among the assessed methodologies, SG-1 pre-processing demonstrated superior predictive accuracy (Rp2 = 0.8166). Furthermore, this investigation visualized freshness indicators as concentration images to elucidate the spatial distribution of TVB-N across the samples. These results indicate that HSI, in conjunction with chemometric analysis, constitutes an efficacious instrument for the surveillance of quality and safety in yellow croakers during its storage phase. Moreover, this methodology guarantees the freshness and safety of seafood products within the aquatic food sector.

Quantification of Adeno-Associated Viral Genomes in Purified Vector Samples by Digital Droplet Polymerase Chain Reaction.

Adeno-associated virus (AAV) is a non-pathogenic virus used as a delivery vehicle to transfer therapeutic genes into patients. Accurate quantification of AAV genome copy number in vector preparations is essential for bioprocess optimization and dosage calculation in both preclinical and clinical studies of AAV-based gene therapy products. Currently, a consensus protocol for AAV viral genome titration is lacking. Here, we present a digital droplet PCR (dd_PCR) protocol for the quantification of viral genomes in purified vector samples. Samples are treated with DNase I to eliminate unpackaged contaminant DNA. DNase-treated samples are then mixed with an appropriate primer-probe set (designed according to the target AAV genome) and PCR reagents, and then loaded into a droplet generator. The prepared droplets are transferred into a PCR plate, where PCR amplification is performed and analyzed. The viral genome titer is calculated based on the concentration (copies/µL), accounting for sample dilutions. A successful measurement shows a clear separation of the positive and negative droplet clouds, has at least 10,000 accepted droplets, shows a value between 10 copies/µL and 10,000 copies/µL, and has a coefficient of variation (CV) between repeats lower than 20%. Reliable viral genome titration will aid in the development of safe and effective AAV-based gene therapy products.