Integrative Inducer Intervention and Transcriptomic Analyses Reveal the Metabolism of Paralytic Shellfish Toxins in Azumapecten farreri.

Paralytic shellfish toxins (PSTs) are widely distributed neurotoxins, and the PST metabolic detoxification mechanism in bivalves has received increasing attention. To reveal the effect of phase I (cytochrome P450)-II (GST)-III (ABC transport) metabolic systems on the PST metabolism in Azumapecten farreri, this study amplified stress on the target systems using rifampicin, dl-α-tocopherol, and colchicine; measured PST levels; and conducted transcriptomic analyses. The highest toxin content reached 1623.48 µg STX eq/kg in the hepatopancreas and only 8.8% of that in the gills. Inducer intervention significantly decreased hepatopancreatic PST accumulation. The proportional reductions in the rifampicin-, dl-α-tocopherol-, and colchicine-induced groups were 55.3%, 50.4%, and 36.1%, respectively. Transcriptome analysis showed that 11 modules were significantly correlated with PST metabolism (six positive/five negative), with phase I CYP450 and phase II glutathione metabolism significantly enriched in negatively correlated pathways. Twenty-three phase I-II-III core genes were further validated using qRT-PCR and correlated with PST metabolism, revealing that CYP46A1, CYP4F6, GSTM1, and ABCF2 were significantly correlated, while CYP4F11 and ABCB1 were indirectly correlated. In conclusion, phase I-II-III detoxification enzyme systems jointly participate in the metabolic detoxification of PSTs in A. farreri. This study provides key data support to profoundly elucidate the PST metabolic detoxification mechanism in bivalves.

Contamination Status and Risk Assessment of Paralytic Shellfish Toxins in Shellfish along the Coastal Areas of China.

Paralytic shellfish toxins (PSTs) are widely distributed in shellfish along the coast of China, causing a serious threat to consumer health; however, there is still a lack of large-scale systematic investigations and risk assessments. Herein, 641 shellfish samples were collected from March to November 2020, and the PSTs' toxicity was detected via liquid chromatography-tandem mass spectrometry. Furthermore, the contamination status and potential dietary risks of PSTs were discussed. PSTs were detected in 241 shellfish samples with a detection rate of 37.60%. The average PST toxicities in mussels and ark shells were considerably higher than those in other shellfish. The PSTs mainly included N-sulfonylcarbamoyl toxins (class C) and carbamoyl toxins (class GTX), and the highest PST toxicity was 546.09 µg STX eq. kg-1. The PST toxicity in spring was significantly higher than those in summer and autumn (p < 0.05). Hebei Province had the highest average PST toxicity in spring. An acute exposure assessment showed that consumers in Hebei Province had a higher dietary risk, with mussels posing a significantly higher dietary risk to consumers. This research provides reference for the green and sustainable development of the shellfish industry and the establishment of a shellfish toxin prevention and control system.

Physiological Effects of Oxidative Stress Caused by Saxitoxin in the Nematode Caenorhabditis elegans.

Saxitoxin (STX) causes high toxicity by blocking voltage-gated sodium channels, and it poses a major threat to marine ecosystems and human health worldwide. Our work evaluated the neurotoxicity and chronic toxicology of STX to Caenorhabditis elegans by an analysis of lifespan, brood size, growth ability, reactive oxygen species (ROS) and adenosine triphosphate (ATP) levels, and the overexpression of green fluorescent protein (GFP). After exposure to a series of concentrations of STX for 24 h, worms showed paralysis symptoms and fully recovered within 6 h; less than 5% of worms died at the highest concentration of 1000 ng/mL for first larval stage (L1) worms and 10,000 ng/mL for fourth larval stage (L4) worms. Declines in lifespan, productivity, and body size of C. elegans were observed under the stress of 1, 10, and 100 ng/mL STX, and the lifespan was shorter than that in controls. With STX exposure, the productivity declined by 32-49%; the body size, including body length and body area, declined by 13-18% and 25-27%, respectively. The levels of ROS exhibited a gradual increase over time, accompanied by a positive concentration effect of STX resulting in 1.14-1.86 times higher levels compared to the control group in L4 worms. Conversely, no statistically significant differences were observed between L1 worms. Finally, after exposure to STX for 48 h, ATP levels and GFP expression in C. elegans showed a significant dose-dependent increase. Our study reports the first evidence that STX is not lethal but imposes substantial oxidative stress on C. elegans, with a dose-responsive relationship. Our results indicated that C. elegans is an ideal model to further study the mechanisms underlying the fitness of organisms under the stress caused by paralytic shellfish toxins including STX.

Variations in the toxicity and condition index of five bivalve species throughout a red tide event caused by Alexandrium catenella: A field study.

Harmful red tides in China have caused paralytic shellfish toxins (PSTs) pollution and led to severe socioeconomic effects in shellfish aquaculture. Although shellfish can survive harmful algal blooms, the effects on their Condition Index (CI) have been underestimated. This study sought to evaluate the effects of the profiles and levels of paralytic shellfish toxins on variations in the CI in bivalves under natural blooming conditions. We observed clear soft tissue lesions to varying degrees except in Mytilus galloprovincialis after toxin exposure. Among the five species of shellfish exposed in situ, only M. galloprovincialis accumulated PSTs content above the maximum permitted level (800 µg STX di-HCl eq./kg). The highest toxin content in all sample tissues was observed in Patinopecten yessoensis. Significant interspecies differences in PSTs accumulation among the five bivalve species were observed in the hepatopancreas. A total of nine PSTs components and four new C-11 hydroxyl metabolites (so-called M-toxins) toxins were detected, and detoxification diversity was observed among bivalves. We observed a higher proportion of M-toxin in early stages, and the proportions changed only slightly over time in M. galloprovincialis and Magallana gigas, thus accounting for the significantly higher metabolism rate. Notably, the CI in M. gigas and Argopecten irradians was positively correlated with lowest toxin accumulation of PSTs content, but significantly inhibited. In conclusion, our results revealed a significant inhibitory effect on the CI in shellfish, in a species specific manner, with distinct levels of inhibition correlated with different toxin metabolites. Our study revealed the toxin content of different bivalves exposed to a natural red tide environment and the consequent effects on growth, thus building a foundation for research on the mechanisms underlying the effects of PSTs on growth. These data establish the ecological and economic significance of the effects of harmful algal blooms on bivalves.

Mechanistic insights into inter-chain disulfide bond reduction of IgG1 and IgG4 antibodies.

Therapeutic monoclonal antibodies (mAbs), primarily immunoglobin G1 (IgG1) and IgG4 with an engineered CPPC motif in its hinge region, are predominant biologics. Inter-chain disulfide bonds of IgG mAbs are crucial to maintaining IgG integrity. Inter-chain disulfide bond-reduced low molecular weight (LMW) is considered as one of quality attributes of IgG drug substance and is observed in drug substance manufacturing. In this study, we demonstrate that IgG1 and IgG4 are susceptible to the reducing agent TCEP differently and they follow different pathways to form LMWs. Our study shows that IgG1 is more sensitive to TCEP than IgG4. Both therapeutic IgG1 and human blood plasma IgG1 follow a heavy-heavy-light chain (HHL) pathway, featured with HHL and HH as intermediate species. Human blood plasma IgG4 with a CPSC motif in its hinge region follows heavy-light chain (HL) pathway, featured with HL as the intermediate species. However, therapeutic IgG4 follows a hybrid pathway with the HL pathway as the primary and the HHL pathway as the secondary. These experimental observations are further explained using solvent accessibility of inter-chain disulfide bonds obtained from computational modeling and molecular dynamics simulations. Findings from this study provide mechanistic insights of LMW formation of IgG1 and IgG4, which suggest selection of IgG1 or IgG4 for bispecific antibodies and cysteine-based antibody-drug conjugates. KEY POINTS: • Experimentally discovered preferable disulfide bond reduction pathways between IgG1 and IgG4 antibodies, driven by the different solvent accessibilities of these disulfide bonds. • Computationally explained the solvent accessibility aided by molecular dynamics simulations. • Provided insights in developing robust biologics process and designing bispecific antibodies and cysteine-based antibody-drug conjugates.

Echinacoside alleviates osteoarthritis in rats by activating the Nrf2-HO-1 signaling pathway.

OBJECTIVE: Osteoarthritis (OA) is a progressive disease characterized by degeneration of cartilage and echinacoside (Ech) has anti-inflammatory and antioxidant effects in various human diseases. This study aimed to reveal the effect and potential mechanism of Ech on OA. MATERIALS AND METHODS: The in vitro OA model was established by rat chondrocytes treated with IL-1ß, and the in vivo OA model was established by anterior cruciate ligament transaction. The effect of Ech on the viability, inflammatory response, extracellular matrix (ECM) degradation, and oxidative stress of IL-1ß-treated rat chondrocytes were evaluated by Cell Counting Kit-8 assay, enzyme-linked immunosorbent assay, quantitative real-time PCR, Western blot, and immunofluorescence assay. Meanwhile, the mechanism of Ech was assessed using Western blot, Cell Counting Kit-8 assay, enzyme-linked immunosorbent assay, and immunofluorescence analysis. Moreover, the function of Ech in vivo was analyzed in rat models of OA. RESULTS: Functionally, Ech enhanced the viability of rat chondrocytes, repressed the inflammatory response and ECM degradation of rat chondrocytes induced by IL-1ß with restrained oxidative stress. Mechanically, Ech repressed IL-1ß-induced chondrocyte injury by activating the Nrf2/HO-1 signaling pathway. Meanwhile, Ech alleviated the degree of articular cartilage injury in rats and exerted protective effects on the rat model of OA in vivo. DISCUSSION AND CONCLUSIONS: Ech alleviated OA in rats by activating the Nrf2-HO-1 signaling pathway.

The development of genus-specific and species-specific real-time PCR assays for the authentication of Patagonian toothfish and Antarctic toothfish in commercial seafood products.

BACKGROUND: The substitution or mislabeling of toothfish is an issue of significant concern for seafood authorities; it also reduces the effectiveness of marine conservation and management programs for its over-exploitation and illegal trafficking, boosting the need for identification methods. RESULTS: Two species-specific real-time polymerase chain reaction (PCR) assays for the identification of Patagonian toothfish (Dissostichus eleginoides) and Antarctic toothfish (Dissostichus mawsoni) and a genus-specific real-time PCR assay for Dissostichus spp. identification were developed based on fragments of the 16S rRNA and COI (cytochrome c oxidase subunit I) genes. These methods were confirmed to be rapid, simple, and sensitive (absolute sensitivity of 0.0002 ng µL-1 and relative sensitivity of 0.1 g kg-1 with good specificity). These methods can be applied to processed and commercial fish products. CONCLUSIONS: These approaches can be beneficial for protecting both consumers and producers from economic fraud and might also help protect toothfish from over-exploitation as well as combat illegal, unreported, and unregulated (IUU) fisheries. © 2021 Society of Chemical Industry.

Antibody disulfide bond reduction and recovery during biopharmaceutical process development-A review.

Antibody disulfide bond reduction has been a challenging issue in monoclonal antibody manufacturing. It could lead to a decrease of product purity and failure to meet the targeted product profile and/or specifications. More importantly, disulfide bond reduction could also impact drug safety and efficacy. Scientists across the industry have been examining the root causes and developing mitigation strategies to address the challenge. In recent years, with the development of high titer mammalian cell culture processes to meet the rapidly growing demand for antibody biopharmaceuticals, disulfide bond reduction has been observed more frequently. Thus, it is necessary to continue evolving the disulfide reduction mitigation strategies and developing novel approaches to maintain high product quality. Additionally, in recent years as more complex molecules (such as bispecific and trispecific antibodies) emerge, the molecular heterogeneity due to incomplete formation of the interchain disulfide bonds becomes a more imperative challenging issue. Given the disulfide reduction challenges that biotech industry is facing, in this review, we provide a comprehensive scientific summary of the root cause analysis of disulfide reduction during process development of antibody therapeutics, mitigation strategies and its potential remediated recovery based on published papers. First, this paper intends to highlight different aspects of the root cause for disulfide reduction. Secondly, to provide a broader understanding of the disulfide bond reduction in downstream process, this paper discusses disulfide bond reduction impact on product stability, associated analytical methods for disulfide bond reduction detection and characterization, process control strategies as well as their manufacturing implementation. In addition, brief perspectives on the development of future mitigation strategies are also reviewed, including platform alignment, mitigation strategy application for the emerging new modalities such as bispecific and trispecific antibodies as well as using machine learning to identify molecule susceptibility of disulfide bond reduction. The data in this review are originated from the published papers.

Exertional rhabdomyolysis in newly enrolled cadets of a military academy.

INTRODUCTION/AIMS: Exertional rhabdomyolysis (ER) often occurs during prolonged intense exercise in hot environments, posing a threat to the health of military personnel. In this study we aimed to investigate possible risk factors for ER and provide further empirical data for prevention and clinical treatment strategies. METHODS: A retrospective investigation of 116 concurrent ER cases was conducted. Conditional logistic regression analyses were performed to assess the association between each potential risk (or protective) factor and ER. The clinical characteristics of the 71 hospitalized patients were analyzed descriptively. RESULTS: After screening, the following variables significantly increased the risk of ER: shorter length of service (recruits; odds ratios [OR], 7.49; 95% confidence interval [CI], 2.58-21.75); higher body mass index (BMI; OR, 1.14, 95% CI, 1.03-1.26); lack of physical exercise in the last half year (less than once per month; OR, 3.20; 95% CI, 1.08-9.44); and previous heat injury (OR, 2.94; 95% CI, 1.26-6.89). Frequent fruit consumption (OR, 0.57; 95% CI, 0.33-0.99), active hydration habit (OR, 0.37; 95% CI, 0.20-0.67), water replenishment of more than 2 L on the training day (OR, 0.15; 95% CI, 0.05-0.45), and water replenishment of at least 500 mL within 1 hour before training (OR, 0.33; 95% CI, 0.12-0.88) significantly decreased the risk of ER. Of the 71 hospitalized patients, 41 (57.7%) were diagnosed with hypokalemia on admission. DISCUSSION: In military training, emphasis should be placed on incremental adaptation training before more intense training, and close attention should be given to overweight and previously sedentary recruits. Fluid replenishment before exercise, increased fruit intake, and proper potassium supplementation may help prevent ER.

Simultaneous determination of triazine herbicides and their metabolites in shellfish by HPLC-MS/MS combined with Q/E-Orbitrap HRMS.

Triazine herbicides are used extensively in agriculture and aquaculture worldwide because of their broad effectiveness in weed control. However, after they are discharged into the sea, they seriously contaminate aquatic ecosystems and threaten aquatic organisms, especially shellfish. Currently, there are no established methods for the detection and confirmation of triazine herbicides and their metabolites in biological matrixes. Hence, the food safety of aquatic products cannot be accurately evaluated, which creates a technical barrier against international aquatic product trade. In this study, for the first time, a method was developed for the analysis and confirmation of seven triazine herbicides and 13 metabolites in shellfish, based on alkaline acetonitrile extraction and neutral Al2O3 cartridge purification coupled with internal standard calibration. Specifically, quantitative and qualitative analysis was conducted using high-performance liquid chromatography-triple quadrupole mass spectrometry (HPLC-MS/MS), and accurate identification was carried out by quadrupole orbitrap high-resolution mass spectrometry (Q/E Orbitrap HRMS). The results showed that target analytes demonstrated good linearity within the corresponding range (R2 > 0.995). The limit of detection and limit of quantitation of the proposed method were 0.1 and 0.3 µg/kg, respectively. The average recoveries of analytes were between 70.0% and 120% when spiked at three levels with blank oyster (Crassostrea gigas) as the matrix, and the relative standard deviations (RSDs) were all less than 12% (n=6). The proposed method was successfully applied for the detection of triazine herbicide residues in oyster samples during actual breeding, and the presence of DIP, HP, DEHA, and other metabolites in positive samples was confirmed by Q/E Orbitrap HRMS. This method exhibits high accuracy, high sensitivity, and good reproducibility. It has promising application prospects in the field of hazard analysis and the positive identification of aquatic products.

Characterization of a broad-spectrum endolysin LysSP1 encoded by a Salmonella bacteriophage.

Foodborne pathogens have caused many public health incidents and heavy economic burden. Endolysins have been proven to have efficient bactericidal activity against pathogens with low incidence of resistance. In this study, the recombinant endolysin LysSP1 encoded by Salmonella Typhimurium lytic bacteriophage SLMP1 was obtained by prokaryotic expression, and its characteristics were analyzed. Ethylenediaminetetraacetic acid (EDTA) can be used as the outer membrane permeabilizer to increase the bactericidal activity of LysSP1. Under the synergism of 5 mmol/L EDTA, LysSP1 exhibited a strong bactericidal activity against Salmonella Typhimurium ATCC14028. LysSP1 was stable at 4°C for 7 days and at -20°C for 180 days. LysSP1 remained the optimal activity at 40°C and was efficiently active at alkaline condition (pH 8.0-10.0). Divalent metal ions could not enhance the bactericidal activity of LysSP1 and even caused the significant reduction of bactericidal activity. LysSP1 not only could lyse Salmonella, but also could lyse other Gram-negative strains and Gram-positive strains. These results indicated that LysSP1 is a broad-spectrum endolysin and has potential as an antimicrobial agent against Salmonella and other foodborne pathogens. KEY POINTS: • Recombinant endolysin LysSP1 can be prepared by prokaryotic expression. • LysSP1 has stable nature and strong bactericidal activity on Salmonella Typhimurium with EDTA. • LysSP1 has a broad range of hosts including Gram-negative bacteria and Gram-positive bacteria.

Effects of single and combined exposure to BDE-47 and PFOA on distribution, bioaccumulation, and toxicity in blue mussel (Mytilus galloprovincialis).

The contamination of 2,2',4,4'-Tetrabrmodiphenyl ether (BDE-47) and perfluorooctanoic acid (PFOA) has drawn a worldwide attention over the risks in ecological and food safety. In this work, blue mussel (Mytilus galloprpvincialis) was employed to investigate the combined effects of BDE-47 (10 ng mL-1) and PFOA (100 ng mL-1) on tissue distribution, accumulation, elimination, and toxicity. Results suggested that BDE-47 and PFOA accumulated mostly in digestive gland, followed by gills and gonad, and M. galloprovincialis displayed higher accumulation capacity to BDE-47 than PFOA. Co-exposure treatment reduced the accumulation of BDE-47, and enhanced the accumulation of PFOA. Furthermore, biochemical and histopathological tests revealed that the aggravated toxicity in co-exposure groups was mainly attributed to the oxidative stress and damage of tissue structure. This work could be helpful to get a better understanding of the combined behaviors and cumulative risks of BDE-47 and PFOA in marine ecosystem.

Integrated analysis of physiological, transcriptomics and metabolomics provides insights into detoxication disruption of PFOA exposure in Mytilus edulis.

Perfluorooctanoic acid (PFOA), a persistent environmental contaminant, resists environmental degradation and bioaccumulates in food chains. Lots of literatures have proved that PFOA exposure could disrupt detoxifying function in a variety of organisms, however, it still remained poorly known about this in mollusk. Here, we examined physiological, transcriptomic, and metabolomic responses to PFOA in Mytilus edulis, a model organism frequently used in studies of aquatic pollution. We aimed to characterize PFOA-induced stress responses and detoxification mechanisms. PFOA exposure significantly altered antioxidant enzyme activity levels and the abundances of lipid peroxidation products. In addition, transcriptomic analysis indicated that several genes associated with oxidative stress and detoxication were differentially expressed after PFOA exposure. In combination, transcriptomic and metabolomic analyses showed that PFOA exposure disturbed several metabolic processes in M. edulis, including the lipid metabolism, amino acid metabolism, and carbohydrate metabolism etc. Molecular examination and enzymes assay of PFOA-exposed M. edulis after a 7-day depuration period still did not recover to control levels. The Pathway enrichment analysis proved that several pathways related to detoxification, such as c-Jun N-terminal kinase (JNK) and p38-dependent mitogen-activated protein kinase (MAPK) pathway, Peroxisome proliferator-activated receptor γ (PPARγ) pathway etc, were obviously affected. The present work verifies firstly PFOA disruption to molluscan detoxification and identifies the key pathways to understand the molecular mechanisms thereof. This study provides new insights into the detoxication disruption invoked in response to PFOA exposure in M. edulis.

Implications of cell division cycle associated 4 on the Wilm's tumor cells viability via AKT/mTOR signaling pathway.

OBJECTIVE: The aim of present report was to elucidate the effect of cell division cycle associated 4 (CDCA4) on the proliferation and apoptosis of Wilm's tumor cells, and to further evaluate its underlying mechanism. METHODS: The expression profiles of CDCA4 and clinical information of Wilm's tumor patients were obtained from public Therapeutically Applicable Research to Generate Effective Treatments (TARGET) database portal. Real-time qPCR and western blot analyses were utilized to determine the expression levels of CDCA4. Gain- and loss-of-function of CDCA4 assays were conducted with transfection technology to investigate the biological role of CDCA4 in Wilm's tumor cells. Cell counting kit 8 and flow cytometer assays were employed to examine the effect of CDCA4 on the cells proliferation and apoptosis. Protein expression levels of indicated markers in each group of Wilm's tumor cells were measured by western blot. RESULTS: The transcriptional expression of CDCA4 was drastically upregulated in Wilm's tumor tissues according to the public TARGET database and in Wilm's tumor cells. The cells viability was remarkably reduced whereas the cells apoptosis was increased in CDCA4-knockdown group compared with negative control group. However, CDCA4-overexpression group promoted the cells proliferation and suppressed the cells apoptosis. Furthermore, the protein expression levels of p-AKT, p-mTOR, and Cyclin D1 were significantly reduced after depletion of CDCA4, whereas overexpression of CDCA4 dramatically elevated these markers' expression levels. CONCLUSIONS: CDCA4 is highly expressed in Wilm's tumor and promoted the proliferation whereas inhibited the apoptosis of Wilm's tumor cells through activating the AKT/mTOR signaling pathway.

Development of duplex real-time polymerase chain reaction for simultaneous detection of oilfish- and escolar-derived components.

BACKGROUND: Oilfish (Ruvettus pretiosus) and escolar (Lepidocybium flavobrunneum) are often used to adulterate high-value aquatic products, causing serious economic losses to consumers, and even keriorrhea in severe cases. It was difficult to identify them by morphological features for these two fish were processed into steak or fillet. The purpose of this study, therefore, is to develop an accurate and efficient method for detecting the oilfish- and escolar-derived components. RESULTS: By comparing the mitochondrial 16S ribosomal RNA gene sequences of oilfish, escolar, and 16 varieties susceptible to adulteration, specific primers/probes were designed, and a duplex real-time polymerase chain reaction (PCR) method was established to detect oilfish- and escolar-derived components. The specificity and sensitivity of the method were analyzed, and the method was used to analyze 70 commercial samples to evalutate its applicability to actual samples in the market. The method developed was highly specific, without any cross-reaction on the other 16 species, with a limit of detection (LOD) for DNA of 0.0002 ng µL-1 for R. pretiosus and 0.002 ng µL-1 for L. flavobrunneum. In addition, the LOD for mixed muscle tissues was 0.1 g kg-1 . Oilfish- and escolar-derived components were detected in 12 of the 70 commercial samples, a result that is consistent with the classic DNA barcoding test results. CONCLUSION: The duplex real-time PCR method developed can be used to detect oilfish and escolar specifically, sensitively and accurately; it provides a good technical support for the identification of authentic aquatic products. © 2020 Society of Chemical Industry.

Kinetic modeling of Chinese hamster ovary cell culture: factors and principles.

As a host for therapeutic protein expression, Chinese hamster ovary (CHO) cells are widely utilized in the mainstream biopharmaceutical industry. Cell culture process development plays an important role in transitioning laboratory research to manufacturing. Among different mathematic tools, kinetic modeling is commonly achieved through analyzing cell culture data to design process parameters, optimize media, and scale up bioreactors. In this review, we examine key factors for upstream process development, and summarize currently used kinetic modeling strategies. In addition, two original examples of kinetic modeling application optimizing cell culture performance are presented. A comprehensive understanding is provided for the kinetic modeling and its applications in cell culture process development.

Online/at-line measurement, analysis and control of product titer and critical product quality attributes (CQAs) during process development.

Process analytical technology (PAT) has been defined by the Food and Drug Administration as a system for designing, analyzing, and controlling manufacturing through timely measurements to ensure final product quality. Based on quality-by-design (QbD) principles, real-time or near-real-time data monitoring is essential for timely control of critical quality attributes (CQAs) to keep the process in a state of control. To facilitate next-generation continuous bioprocessing, deployment of PAT tools for real-time monitoring is integral for process understanding and control. Real-time monitoring and control of CQAs are essential to keep the process within the design space and align with the guiding principles of QbD. The contents of this manuscript are pertinent to the online/at-line monitoring of upstream titer and downstream product quality with timely process control. We demonstrated that an ultra-performance liquid chromatography (UPLC) system interfaced with a UPLC-process sample manager (UPLC-PSM) can be utilized to measure titer and CQAs directly from bioreactors and downstream unit operations, respectively. We established online titer measurements from fed-batch and perfusion-based alternating tangential flow bioreactors as well as product quality assessments of downstream operations for real-time peak collection. This integrated, fully automated system for online data monitoring with feedback control is designed to achieve desired product quality.

Nontarget Screening and Toxicity Evaluation of Diol Esters of Okadaic Acid and Dinophysistoxins Reveal Intraspecies Difference of Prorocentrum lima.

High-resolution mass spectrometry (HRMS) analysis with the assistance of molecular networking was used to investigate intracellular toxin profiles of five Prorocentrum lima (P. lima) strains sampled from the north Yellow Sea and South China Sea. Mice were used as a model species for testing the acute toxicity of intracellular okadaic acid (OA) and dinophysistoxins (DTXs) in free and esterified states. Results showed that OA and DTX1 esterified derivatives were detected in all P. lima samples, accounting for 55%-96% of total toxins in five strains. A total of 24 esters and 1 stereoisomer of DTX1 (35S DTX1) were identified based on molecular networking and MS data analysis, 15 esters of which have been reported first. All P. lima strains displayed specific toxin profiles, and preliminary analysis suggested that toxin profiles of the five P. lima strains might be region-related. Moreover, acute toxicity in mice suggested higher toxicity of esters compared with free toxins, which highlights the importance and urgency of attention to esterified toxins in P. lima.

Assessing the proxy response bias of EQ-5D-3 L in general population: a study based on a large-scale representative household health survey using propensity score matching.

BACKGROUND: Proxy respondent-someone who assists the intended respondent or responds on their behalf-are widely applied in the measurement of health-related quality of life (HRQL). However, proxies may not provide the same responses as the intended respondents, which may bias the findings. OBJECTIVES: To determine whether the use of proxies is related to socio-demographic characteristics of the intended respondent, and to assess the possible proxy response bias of Chinese version of EQ-5D-3 L in general population. METHODS: A cross-sectional study based on a provincially representative sample from 2013 National Health Service Survey (NHSS) in Shaanxi, China was performed. HRQL was measured by Chinese version of EQ-5D-3 L. Propensity score matching (PSM) was used to get matched pairs of self-reports and proxy-reports. Before and after PSM, univariate logistic and linear models including the indicator of proxy response as the only independent variable, were employed to assess the possible proxy response bias of the dimensional and overall health status of EQ-5D-3 L respectively. RESULTS: 19.9% of the responses involved a proxy. Before PSM, the proxy-report group was younger in age and reported less unhealthy lifestyle, lower prevalence of disease, and less hospitalization than the self-report group. After PSM, it showed that the proxy-report group was statistically more likely to report health problem on each dimension of EQ-5D-3 L, with odds ratios larger than one comparing with self-report group. The means of EQ-5D-3 L index and EQ VAS of proxy-report group were 0.022 and 0.834 lower than self-report group. CONCLUSIONS: Significantly negative proxy response bias was found in Chinese EQ-5D-3 L in general population, and the magnitude of the bias was larger in physical dimensions than psychological dimensions after using PSM to control confounders.

Prediction for cardiovascular diseases based on laboratory data: An analysis of random forest model.

BACKGROUND: To establish a prediction model for cardiovascular diseases (CVD) in the general population based on random forests. METHODS: A retrospective study involving 498 subjects was conducted in Xi'an Medical University between 2011 and 2018. The random forest algorithm was used to screen out the variables that greatly affected the CVD prediction and to establish a prediction model. The important variables were included in the multifactorial logistic regression analysis. The area under the curve (AUC) was compared between logistic regression model and random forest model. RESULTS: The random forest model revealed the variables, including the age, body mass index (BMI), fasting blood glucose (FBG), diastolic blood pressure (DBP), triglyceride (TG), systolic blood pressure (SBP), total cholesterol (TC), waist circumference, and high-density lipoprotein-cholesterol (HDL-C), were more significant for CVD prediction; the AUC was 0.802 in CVD prediction. Multifactorial logistic regression analysis indicated that the risk factors for CVD included the age [odds ratio (OR): 1.14, 95% confidence intervals (CI): 1.10-1.17, P < .001], BMI (OR: 1.13, 95% CI: 1.06-1.20, P < .001), TG (OR: 1.11, 95% CI: 1.02-1.22, P = .023), and DBP (OR: 1.04, 95% CI: 1.02-1.06, P = .001); the AUC was 0.843 in CVD prediction. The established logistic regression prediction model was Logit P = Log[P/(1 - P)] = -11.47 + 0.13 × age + 0.12 × BMI + 0.11 × TG + 0.04 × DBP; P = 1/[1 + exp(-Logit P)]. People were prone to develop CVD at the time of P > .51. CONCLUSIONS: A prediction model for CVD is developed in the general population based on random forests, which provides a simple tool for the early prediction of CVD.