2023 White Paper on Recent Issues in Bioanalysis: ISR for ADA Assays, the Rise of dPCR vs qPCR, International Reference Standards for Vaccine Assays, Anti-AAV TAb Post-Dose Assessment, NanoString Validation, ELISpot as Gold Standard (Part 3 - Recommendations on Gene Therapy, Cell Therapy, Vaccines Immunogenicity & Technologies; Biotherapeutics Immunogenicity & Risk Assessment; ADA/NAb Assay/Reporting Harmonization).

The 17th Workshop on Recent Issues in Bioanalysis (17th WRIB) took place in Orlando, FL, USA on June 19-23, 2023. Over 1000 professionals representing pharma/biotech companies, CROs, and multiple regulatory agencies convened to actively discuss the most current topics of interest in bioanalysis. The 17th WRIB included 3 Main Workshops and 7 Specialized Workshops that together spanned 1 week to allow an exhaustive and thorough coverage of all major issues in bioanalysis of biomarkers, immunogenicity, gene therapy, cell therapy and vaccines. Moreover, in-depth workshops on "EU IVDR 2017/746 Implementation and impact for the Global Biomarker Community: How to Comply with these NEW Regulations" and on "US FDA/OSIS Remote Regulatory Assessments (RRAs)" were the special features of the 17th edition. As in previous years, WRIB continued to gather a wide diversity of international, industry opinion leaders and regulatory authority experts working on both small and large molecules as well as gene, cell therapies and vaccines to facilitate sharing and discussions focused on improving quality, increasing regulatory compliance, and achieving scientific excellence on bioanalytical issues. This 2023 White Paper encompasses recommendations emerging from the extensive discussions held during the workshop and is aimed to provide the bioanalytical community with key information and practical solutions on topics and issues addressed, in an effort to enable advances in scientific excellence, improved quality and better regulatory compliance. Due to its length, the 2023 edition of this comprehensive White Paper has been divided into three parts for editorial reasons. This publication (Part 3) covers the recommendations on Gene Therapy, Cell therapy, Vaccines and Biotherapeutics Immunogenicity. Part 1A (Mass Spectrometry Assays and Regulated Bioanalysis/BMV), P1B (Regulatory Inputs) and Part 2 (Biomarkers, IVD/CDx, LBA and Cell-Based Assays) are published in volume 16 of Bioanalysis, issues 8 and 9 (2024), respectively.

[Source Analysis and Health Risk Assessment of Heavy Metals in the Groundwater of Shijiazhuang, a Typical City in North China Plain].

Increasing attention has been paid to the heavy metal pollution in groundwater. The source analysis and risk assessment of heavy metals will provide data and method support for the targeted control of heavy metal pollution in groundwater. In this study, 20 sampling sites were selected in Shijiazhuang City. The APCS-MLR model and health risk model were applied to analyze and evaluate the pollution sources and health risks of 10 types of heavy metals in the groundwater of Shijiazhuang. The results showed that ① the mean concentration of heavy metals in groundwater followed the order of Fe>Zn>Mn>Cu>Al>Pb>Cr>As>Cd>Hg, and the mean ρ(Fe) and ρ(Pb) were 260.3 µg·L-1 and 10.01 µg·L-1, respectively. According to the results of the single factor and Nemerow index, Pb, Fe, and Cd primarily contributed to the heavy metal pollution in the groundwater. ② The concentration of heavy metals ranged from 47.30 to 2560 µg·L-1. In terms of spatial distribution, the highest concentration appeared at S3 (2560 µg·L-1), whereas the lowest concentration was at S9 (47.30 µg·L-1). ③ Source analysis results showed that industrial and agricultural activities, transportation emission, and geological background were the major heavy metal sources, among which the contribution of industrial and agricultural activities was the highest (47.83%). ④ The industrial-agricultural activities posed a potential threat to adults (HI>1); however, the non-cancer and the cancer risks of other sources for both adults and children were at an acceptable level (HI<1) and potential threat level, respectively; industrial-agricultural activities were the major source of non-cancer (adults:52.46%, children:52.45%) and cancer risks (adults:65.22%, children:65.69%), among which Cd and As showed high cancer risk. Therefore, to ensure the safety of the groundwater environment, strictly controlling the pollution sources and further strengthening the risk control of heavy metal pollution in groundwater are necessary.

[Spatial-temporal Distribution and Risk Assessment of Quinolones Antibiotics in Soil of Shijiazhuang City].

Due to their importance in human medicine, quinolones (QNs), as a typical class of antibiotics, are considered to be the "highest priority critically important antimicrobials" by the World Health Organization (WHO). In order to clarify the spatial-temporal variation and risk of QNs in soil, 18 representative topsoil samples were respectively collected in September 2020 (autumn) and June 2021 (summer). The contents of QNs antibiotics in soil samples were determined using high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS), and the ecological risk and resistance risk were calculated using the risk quotient method (RQ). The results showed that:① the average content of QNs decreased from autumn to summer (the average contents of QNs were 94.88 µg·kg-1in autumn and 44.46 µg·kg-1 in summer); the highest values appeared in the middle area. ② The average proportion of silt was without change, whereas the average proportion of clay and sand was increased and decreased, respectively; the average contents of total phosphorus (TP), ammonia nitrogen (NH4+-N), and nitrate nitrogen (NO3--N) also decreased. ③ The content of QNs was significantly correlated with soil particle size, nitrite nitrogen (NO2--N), and nitrate nitrogen (NO3--N) (P<0.05). ④ The combined ecological risk of QNs showed high risk level (RQsum>1), whereas the combined resistance risk of QNs showed medium risk level (0.1

[Spatial-temporal Evolution and Heterogeneity of Carbon Emissions at County-level in China].

Counties are the key spatial units in achieving the reduction and control of carbon emissions. It is of great significance to study and reveal the spatial-temporal evolution characteristics and influencing mechanism of carbon emissions for realizing the "carbon peak and carbon neutral" goal. In this study, the spatial-temporal evolution and heterogeneity of carbon emissions at the county level in China from 2000 to 2017 were analyzed by using mathematical statistics and panel data regression modeling, and the influencing mechanism was explored. The results showed that: ① from 2000 to 2017, the annual growth rate of carbon emissions was 7.12%, which experienced the three stages of "sharp rise, slow rise, and high fluctuation" and finally stabilized at approximately 90×108 t. At the county scale, there was a significant positive spatial autocorrelation. ② The general panel regression model showed that GDP, construction land area, population, per capita GDP, and per capita deposit balance of financial institutions were significantly correlated with carbon emissions, and the former three had the strongest promoting effect on carbon emissions. ③ The goodness of fit of the geographically and temporally weighted regression model was high, and the direction and intensity of the other impact factors changed greatly in spatial-temporal characteristics, except that GDP showed a stable promoting effect globally. The results showed that carbon emission levels and main influencing factors varied among counties in China. This study revealed the heterogeneity of carbon emissions at the county level, which is helpful to optimize the spatial-temporal implementation path of the "dual carbon" target.

Research on Carbon Emission Efficiency Space Relations and Network Structure of the Yellow River Basin City Cluster.

The Yellow River Basin serves as China's primary ecological barrier and economic belt. The achievement of the Yellow River Basin's "double carbon" objective is crucial to China's green and low-carbon development. This study examines the spatial link and network structure of city cluster carbon emission efficiency in the Yellow River Basin, as well as the complexity of the network structure. It focuses not only on the density and centrality of the carbon emission efficiency network from the standpoint of city clusters, but also on the excellent cities and concentration of the city cluster 's internal carbon emission efficiency network. The results show that: (1) The carbon emission efficiency of the Yellow River Basin has been dramatically improved, and the gap between city clusters is narrowing. However, gradient differentiation characteristics between city clusters show the Matthew effect. (2) The distribution of carbon emission efficiency in the Yellow River Basin is unbalanced, roughly showing a decreasing trend from east to west. Lower-level efficiency cities have played a significant role in the evolution of carbon emissions efficiency space. (3) The strength of the carbon emission efficiency network structure in the Yellow River Basin gradually transitions from weakly correlated dominant to weakly and averagely correlated dominant. Among them, the Shandong Peninsula city cluster has the most significant number of connected nodes in the carbon emission efficiency network. In contrast, the emission efficiency network density of the seven city clusters shows different changing trends. Finally, this study suggests recommendations to improve carbon emission efficiency by adopting differentiated governance measures from the perspective of local adaptation and using positive spatial spillover effects.

Predicting the Water Rebound Effect in China under the Shared Socioeconomic Pathways.

The rebound effect exists widely in the fields of energy, irrigation, and other resource utilizations. Previous studies have predicted the evolution of different resource utilizations under the shared socioeconomic pathways (SSPs), but it is still unclear whether total water use has a rebound effect. This study uses the SSPs as the basic prediction framework and evaluates the water resources and economic status of the provinces in China using the hydro-economic (HE) classification method. Then, combined with the SSPs scenario setting parameters, the conditional convergence model and the method recommended by the Food and Agriculture Organization of the United Nations (FAO) are used to simulate the changes in water use efficiency of the different provinces in China under different scenarios. Based on the future GDP forecast data of China's provinces, combined with the forecast of water use efficiency changes, the total water use changes in China's 31 provinces under different pathways from 2016 to 2030 are calculated. Among them, the future GDP data is predicted based on the Cobb-Douglas production function and SSPs scenario settings. Using a comprehensive evaluation of the evolution of the efficiency and the total amount, this study reveals whether there is a rebound effect. The results showed that with the continuous growth in the water use efficiency, the total water use had a "U" type trend, which indicated that there was a rebound effect in the total water use of China under the different SSPs. Based on this information, this study proposes some suggestions for irrigation water-saving technologies and policies.

Integrin αvß5 as a biomarker for the assessment of non-small cell lung cancer metastasis and overall survival.

BACKGROUND AND AIM: There is growing interest in how integrins play a role in cancer disease biology and what applications these may have in anti-cancer therapeutic development. This study investigates integrin αvß5 expression in non-small cell lung carcinoma (NSCLC) and its correlations with clinical information. METHODS: Tumor human tissue microarrays of 147 radically operated Chinese NSCLC patients were retrieved from the pathology archive, and sections were autoimmune stained along with positive and negative controls. Integrin αvß5 (P1F6) mouse monoclonal antibody were validated by both Western blotting and immunoreactivity on the same set of cell pellets, according to a precedent of expression levels in cell flow cytometry. The immunoreactivity of all patients' cases against integrin αvß5 antibody was graded in a semi-quantitative manner by two pathologists blind to any patient data. RESULTS: Overall survival significantly correlated to integrin αvß5 immunoreactivity by Cox regression analysis with P = 0.032. When applying Kaplan-Meier to analyze the comparison between positive and negative expression on lymph node metastasis patients, P = 0.082. Therefore, integrin αvß5 expression emerges as a significant prognostic factor for NSCLC. In total, 39 of 147 (26.5%) showed an integrin αvß5 immunoreactivity positive, as it ranged from 6.1% in squamous cell carcinoma to 19.7% in adenocarcinoma. This expression correlated significantly with histological subtypes (P = 0.007), tumor node metastasis classification of malignant tumors (P = 0.027) and lymph node metastases (P = 0.006). CONCLUSION: This study support the hypothesis that integrin αvß5 is a prognostic biomarker and correlates to metastasis with therapeutic development potential in NSCLC.

MIOTIC study: a prospective, multicenter, randomized study to evaluate the long-term efficacy of mobile phone-based Internet of Things in the management of patients with stable COPD.

Chronic obstructive pulmonary disease (COPD) is a common disease that leads to huge economic and social burden. Efficient and effective management of stable COPD is essential to improve quality of life and reduce medical expenditure. The Internet of Things (IoT), a recent breakthrough in communication technology, seems promising in improving health care delivery, but its potential strengths in COPD management remain poorly understood. We have developed a mobile phone-based IoT (mIoT) platform and initiated a randomized, multicenter, controlled trial entitled the 'MIOTIC study' to investigate the influence of mIoT among stable COPD patients. In the MIOTIC study, at least 600 patients with stable GOLD group C or D COPD and with a history of at least two moderate-to-severe exacerbations within the previous year will be randomly allocated to the control group, which receives routine follow-up, or the intervention group, which receives mIoT management. Endpoints of the study include (1) frequency and severity of acute exacerbation; (2) symptomatic evaluation; (3) pre- and post-bronchodilator forced expiratory volume in 1 second (FEV1) and FEV1/forced vital capacity (FVC) measurement; (4) exercise capacity; and (5) direct medical cost per year. Results from this study should provide direct evidence for the suitability of mIoT in stable COPD patient management.

Spiritual health, clinical practice stress, depressive tendency and health-promoting behaviours among nursing students.

AIM: This paper is a report of an exploration of the association of spiritual health with clinical practice stress, depressive tendency and health-promoting behaviours among nursing students. BACKGROUND: Several studies in western countries have demonstrated an association between spirituality and health. Spirituality-related research in eastern countries, however, is still in its infancy. METHODS: A cross-sectional design was adopted and structured questionnaires were used for data collection. We adopted the Probability Proportional to Size cluster sampling method to recruit nursing students in senior grades. Data were collected in 2005 using the Spiritual Health Scale, Perceived Clinical Practice Stress Scale, Beck Depression Inventory-II and Health Promotion Behaviours Scale. RESULTS: A total of 1276 nursing students with an average age of 20.1 years (sd = 1.6 years) participated in the study. Spiritual health was negatively associated with clinical practice stress (r = -0.211, P < 0.001) and depressive tendency (r = -0.324, P < 0.001) and positively associated with health-promoting behaviours (r = 0.611, P < 0.001). Using hierarchical regression analysis to control for demographic factors, spiritual health was found to be an important predictive factor for clinical practice stress, depressive tendency and health-promoting behaviours. CONCLUSION: These results are consistent with research findings from western countries. Educators should develop strategies to address nursing students' spiritual health. This may help nursing students to manage their stress, to reduce depressive symptoms and to enhance health-promoting behaviours.