Learning-Based Non-Intrusive Electric Load Monitoring for Smart Energy Management.

State-of-the-art smart cities have been calling for economic but efficient energy management over a large-scale network, especially for the electric power system. It is a critical issue to monitor, analyze, and control electric loads of all users in the system. In this study, a non-intrusive load monitoring method was designed for smart power management using computer vision techniques popular in artificial intelligence. First of all, one-dimensional current signals are mapped onto two-dimensional color feature images using signal transforms (including the wavelet transform and discrete Fourier transform) and Gramian Angular Field (GAF) methods. Second, a deep neural network with multi-scale feature extraction and attention mechanism is proposed to recognize all electrical loads from the color feature images. Third, a cloud-based approach was designed for the non-intrusive monitoring of all users, thereby saving energy costs during power system control. Experimental results on both public and private datasets demonstrate that the method achieves superior performances compared to its peers, and thus supports efficient energy management over a large-scale Internet of Things network.

Impact of enzymatic reduction on bivalent bispecific antibody fragmentation and loss of product purity upon reoxidation.

Antibody disulfide bond (DSB) reduction during manufacturing processes is a widely observed phenomenon attributed to host cell reductases present in harvest cell culture fluid. Enzyme-induced antibody reduction leads to product fragments and aggregates that increase the impurity burden on the purification process. The impact of reduction on bivalent bispecific antibodies (BisAbs), which are increasingly entering the clinic, has yet to be investigated. We focused on the reduction and reoxidation properties of a homologous library of bivalent BisAb formats that possess additional single-chain Fv (scFv) fragments with engineered DSBs. Despite all BisAbs having similar susceptibilities to enzymatic reduction, fragmentation pathways were dependent on the scFv-fusion site. Reduced molecules were allowed to reoxidize with and without low pH viral inactivation treatment. Both reoxidation studies demonstrated that multiple, complex BisAb species formed as a result of DSB mispairing. Furthermore, aggregate levels increased for all molecules when no low pH treatment was applied. Combined, our results show that complex DSB mispairing occurs during downstream processes while aggregate formation is dependent on sample treatment. These results are applicable to other novel monoclonal antibody-like formats containing engineered DSBs, thus highlighting the need to prevent reduction of novel protein therapeutics to avoid diminished product quality during manufacturing.

Characterization and quantification of succinimide using peptide mapping under low-pH conditions and hydrophobic interaction chromatography.

Characterization of asparagine deamidation and aspartic acid isomerization is an important aspect of biotherapeutic protein analysis due to the potential negative effect of these modifications on drug efficacy and stability. Succinimide has long been known to be an intermediate product of asparagine deamidation and aspartic acid isomerization, but despite the key role of succinimide in these reactions, its analysis remains challenging due to its instability. We have developed a paradigm in which two interlinked analytical methods are used to develop an optimized approach to analyze succinimide. In the first method, low-pH protein digestion is used for detailed characterization of succinimide with peptide mapping. At low pH, succinimide is stable and can be analyzed with accurate mass measurements and tandem mass spectrometry to confirm its identity and localize its modification site. These results are then used to establish a hydrophobic interaction chromatography (HIC)-based method that can be used for release and stability studies. In this method, unmodified protein, deamidated products, and succinimide are well separated and quantified. Good correlation was obtained between the data from low-pH protein digestion-based peptide mapping and the HIC-based method. Method qualification showed that the HIC-based method is robust, accurate, and precise and has excellent linearity.

Quantification of a recombinant antigen in an immuno-stimulatory whole yeast cell-based therapeutic vaccine.

Therapeutic vaccines represent an emerging class of immune-modulatory treatments for cancer, infections, and chronic diseases. One such vaccine was designed as an immune stimulator of the T cell response against HBV antigens to eliminate HBV infected cells and offer a therapeutic avenue to treat patients suffering from chronic hepatitis B infection. Whole deactivated Saccharomyces cerevisiae cells expressing a recombinant fusion of HBV X, S and Core antigens elicit T cell responses in mice and activate human T cells linked with viral clearance. As the therapeutic efficacy of the yeast-based vaccine relies on the production of the recombinant antigen, analytical methods designed to accurately and precisely quantitate the fusion protein in the midst of all the yeast proteins are necessary. We report the development and characterization of western blot, quantitative ELISA and mass spectrometry based orthogonal methods to support the assessment of manufacturing consistency.

A cloud model-based approach for water quality assessment.

Water quality assessment entails essentially a multi-criteria decision-making process accounting for qualitative and quantitative uncertainties and their transformation. Considering uncertainties of randomness and fuzziness in water quality evaluation, a cloud model-based assessment approach is proposed. The cognitive cloud model, derived from information science, can realize the transformation between qualitative concept and quantitative data, based on probability and statistics and fuzzy set theory. When applying the cloud model to practical assessment, three technical issues are considered before the development of a complete cloud model-based approach: (1) bilateral boundary formula with nonlinear boundary regression for parameter estimation, (2) hybrid entropy-analytic hierarchy process technique for calculation of weights, and (3) mean of repeated simulations for determining the degree of final certainty. The cloud model-based approach is tested by evaluating the eutrophication status of 12 typical lakes and reservoirs in China and comparing with other four methods, which are Scoring Index method, Variable Fuzzy Sets method, Hybrid Fuzzy and Optimal model, and Neural Networks method. The proposed approach yields information concerning membership for each water quality status which leads to the final status. The approach is found to be representative of other alternative methods and accurate.

A multidimension cloud model-based approach for water quality assessment.

Lakes are vitally important, because they perform a multitude of functions, such as water supply, recreation, fishing, and habitat. However, eutrophication limits the ability of lakes to perform these functions. In order to reduce eutrophication, the first step is its evaluation. The process of evaluation entails randomness and fuzziness which must therefore be incorporated. This study proposes an eutrophication evaluation method, named Multidimension Normal Cloud Model (MNCM). The model regards each evaluation factor as a one-dimension attribute of MNCM, chooses reasonable parameters and determines the weights of evaluation factors by entropy. Thus, all factors of MNCM belonging to each eutrophication level are generated and the final eutrophication level is determined by the certainty degree. MNCM is then used to evaluate eutrophication of 12 typical lakes and reservoirs in China and its results are compared with those of the reference method, one-dimension normal cloud model, related weighted nutrition state index method, scoring method, and fuzzy comprehensive evaluation method. Results of MNCM are found to be consistent with the actual water status; hence, MNCM can be an effective evaluation tool. With respect to the former one-dimension normal cloud model, parameters of MNCM are improved without increasing its complexity. MNCM can directly determine the eutrophication level according to the degree of certainty and can determine the final degree of eutrophication; thus, it is more consistent with the complexity of water eutrophication evaluation.

Analysis of temporal and spatial trends of hydro-climatic variables in the Wei River Basin.

The Wei River is the largest tributary of the Yellow River in China. The relationship between runoff and precipitation in the Wei River Basin has been changed due to the changing climate and increasingly intensified human activities. In this paper, we determine abrupt changes in hydro-climatic variables and identify the main driving factors for the changes in the Wei River Basin. The nature of the changes is analysed based on data collected at twenty-one weather stations and five hydrological stations in the period of 1960-2010. The sequential Mann-Kendall test analysis is used to capture temporal trends and abrupt changes in the five sub-catchments of the Wei River Basin. A non-parametric trend test at the basin scale for annual data shows a decreasing trend of precipitation and runoff over the past fifty-one years. The temperature exhibits an increase trend in the entire period. The potential evaporation was calculated based on the Penman-Monteith equation, presenting an increasing trend of evaporation since 1990. The stations with a significant decreasing trend in annual runoff mainly are located in the west of the Wei River primarily interfered by human activities. Regression analysis indicates that human activity was possibly the main cause of the decline of runoff after 1970.

A genome-wide CRISPR screen identifies USP1 as a novel regulator of the mammalian circadian clock.

The circadian clock is generated by a molecular timekeeping mechanism coordinating daily oscillations of physiology and behaviors in mammals. In the mammalian circadian clockwork, basic helix-loop-helix ARNT-like protein 1 (BMAL1) is a core circadian component whose defects lead to circadian disruption and elicit behavioral arrhythmicity. To identify previously unknown regulators for circadian clocks, we searched for genes influencing BMAL1 protein level by using a CRISPR/Cas9-based genome-wide knockout library. As a result, we found that the deubiquitinase ubiquitin carboxyl-terminal hydrolase 1 (USP1) positively affects BMAL1 protein abundance. Overexpression of wild-type USP1, but not a deubiquitinase-inactive mutant USP1, upregulated BMAL1 protein level, whereas genetic ablation of USP1 downregulated BMAL1 protein level in U2OS cells. Furthermore, treatment with USP1 inhibitors led to significant downregulation of BMAL1 protein in U2OS cells as well as mouse tissues. Subsequently, genetic ablation or pharmacological inhibition of USP1 resulted in reduced mRNA levels of a panel of clock genes and disrupted circadian rhythms in U2OS cells. Mechanistically, USP1 was able to de-ubiquitinate BMAL1 and inhibit the proteasomal degradation of BMAL1. Interestingly, the expression of Usp1 was much higher than the other two deubiquitinases of BMAL1 (Usp2 and Usp9X) in the mouse heart, implying a tissue-specific function of USP1 in the regulation of BMAL1 stability. Our work thus identifies deubiquitinase USP1 as a previously unknown regulator of the mammalian circadian clock and highlights the potential of genome-wide CRISPR screens in the identification of regulators for the circadian clock.

Characterization of structures and molecular interactions of RNA and lipid carriers using atomic force microscopy.

Ribonucleic acid (RNA) and lipid are essential biomolecules in many biological processes, and hold a great prospect for biomedical applications, such as gene therapy, vaccines and therapeutic drug delivery. The characterization of morphology and intra-/inter-molecular interactions of RNA and lipid molecules is critical for understanding their functioning mechanisms. Atomic force microscopy (AFM) is a sophisticated technique for characterizing biomolecules featured by its piconewton force sensitivity, sub-nanometer spatial resolution, and flexible operation conditions in both air and liquid. The goal of this review is to highlight the representative and outstanding discoveries of the characterization of RNA and lipid molecules through morphology identification, physicochemical property determination and intermolecular force measurements by AFM. The first section introduces the AFM imaging of RNA molecules to obtain high-resolution morphologies and nanostructures in air and liquid, followed by the discussion of employing AFM force spectroscopy in understanding the nanomechanical properties and intra-/inter-molecular interactions of RNA molecules, including RNA-RNA and RNA-biomolecule interactions. The second section focuses on the studies of lipid and RNA encapsulated in lipid carrier (RNA-lipid) by AFM as well as the sample preparation and factors influencing the morphology and structure of lipid/RNA-lipid complexes. Particularly, the nanomechanical properties of lipid and RNA-lipid characterized by nanomechanical imaging and force measurements are discussed. The future perspectives and remaining challenges on the characterization of RNA and lipid offered by the versatile AFM techniques are also discussed. This review provides useful insights on the characterization of RNA and lipids nanostructures along with their molecular interactions, and also enlightens the application of AFM techniques in investigating a broad variety of biomolecules.

FMRP binds Per1 mRNA and downregulates its protein expression in mice.

FMRP, an RNA-binding protein, has previously shown to be involved in regulation of circadian rhythms in flies and mice. However, the molecular mechanism remains elusive. Here we demonstrate that core circadian component Per1 mRNA was a target of FMRP and the association leads to reduced PER1 expression. In Fmr1 KO mice, the oscillation of PER1 protein expression was significantly affected in a temporal and tissue-dependent pattern when compared to WT mice. Our work thus identified Per1 mRNA as a novel target of FMRP and suggested a potential role of FMRP in regulation of circadian function.

Cry1Δ11 mutation induces ADHD-like symptoms through hyperactive dopamine D1 receptor signaling.

Attention-deficit hyperactivity disorder (ADHD) is a highly heritable neurodevelopmental disorder that affects approximately 5.3% of children and approximately 2.5% of adults. There is an intimate relationship between ADHD and sleep disturbance. Specifically, individuals carry a mutation in the core circadian gene CRY1 (c. 1657 + 3A > C), which results in the deletion of exon 11 expression in the CRY1 protein (CRY1Δ11), causing them to exhibit typical ADHD symptoms. However, the underlying mechanism is still elusive. In this study, we demonstrate that Cry1Δ11 (c. 1717 + 3A > C) mice showed ADHD-like symptoms, including hyperactivity, impulsivity, and deficits in learning and memory. A hyperactive cAMP signaling pathway was found in the nucleus accumbens (NAc) of Cry1Δ11 mice. We further demonstrated that upregulated c-Fos was mainly localized in dopamine D1 receptor-expressing medium spiny neurons (DRD1-MSNs) in the NAc. Neuronal excitability of DRD1-MSNs in the NAc of Cry1Δ11 mice was significantly higher than that of WT controls. Mechanistically, the CRY1Δ11 protein, in contrast to the WT CRY1 protein, failed to interact with the Gαs protein and inhibit DRD1 signaling. Finally, the DRD1 antagonist SCH23390 normalized most ADHD-like symptoms in Cry1Δ11 mice. Thus, our results reveal hyperactive DRD1 signaling as an underlying mechanism and therapeutic target for ADHD induced by the highly prevalent CRY1Δ11 mutation.

CMC Regulatory Considerations for Antibody-Drug Conjugates.

Antibody-drug conjugates unite the specificity and long circulation time of an antibody with the toxicity of a chemical cytostatic or otherwise active drug using appropriate chemical linkers to reduce systemic toxicity and increase therapeutic index. This combination of a large biological molecule and a small molecule creates an increase in complexity. Multiple production processes are required to produce the native antibody, the drug and the linker, followed by conjugation of afore mentioned entities to form the final antibody-drug conjugate. The connected processes further increase the number of points of control, resulting in necessity of additional specifications and intensified analytical characterization. By combining scientific understanding of the production processes with risk-based approaches, quality can be demonstrated at those points where control is required and redundant comparability studies, specifications or product characterization are avoided. Over the product development lifecycle, this will allow process qualification to focus on those areas critical to quality and prevent redundant studies. The structure of the module 3 common technical document for an ADC needs to reflect each of the production processes and the combined overall approach to quality. Historically, regulatory authorities have provided varied expectations on its structure. This paper provides an overview of essential information to be included and shows that multiple approaches work as long as adequate cross-referencing is included.

Multiplex Bioanalytical Methods for Comprehensive Characterization and Quantification of the Unique Complementarity-Determining-Region Deamidation of MEDI7247, an Anti-ASCT2 Pyrrolobenzodiazepine Antibody-Drug Conjugate.

Deamidation, a common post-translational modification, may impact multiple physiochemical properties of a therapeutic protein. MEDI7247, a pyrrolobenzodiazepine (PBD) antibody-drug conjugate (ADC), contains a unique deamidation site, N102, located within the complementarity-determining region (CDR), impacting the affinity of MEDI7247 to its target. Therefore, it was necessary to monitor MEDI7247 deamidation status in vivo. Due to the low dose, a sensitive absolute quantification method using immunocapture coupled with liquid chromatography-tandem mass spectrometry (LBA-LC-MS/MS) was developed and qualified. We characterized the isomerization via Electron-Activated Dissociation (EAD), revealing that deamidation resulted in iso-aspartic acid. The absolute quantification of deamidation requires careful assay optimization in order not to perturb the balance of the deamidated and nondeamidated forms. Moreover, the selection of capture reagents essential for the correct quantitative assessment of deamidation was evaluated. The final assay was qualified with 50 ng/mL LLOQ for ADC for total and nondeamidated antibody quantification, with qualitative monitoring of the deamidated antibody. The impact of deamidation on the pharmacokinetic characteristics of MEDI7247 from clinical trial NCT03106428 was analyzed, revealing a gradual reduction in the nondeamidated form of MEDI7247 in vivo. Careful quantitative biotransformation analyses of complex biotherapeutic conjugates help us understand changes in product PTMs after administration, thus providing a more complete view of in vivo pharmacology.

Atypical Asparagine Deamidation of NW Motif Significantly Attenuates the Biological Activities of an Antibody Drug Conjugate.

Asparagine deamidation is a post-translational modification (PTM) that converts asparagine residues into iso-aspartate and/or aspartate. Non-enzymatic asparagine deamidation is observed frequently during the manufacturing, processing, and/or storage of biotherapeutic proteins. Depending on the site of deamidation, this PTM can significantly impact the therapeutic's potency, stability, and/or immunogenicity. Thus, deamidation is routinely monitored as a potential critical quality attribute. The initial evaluation of an asparagine's potential to deamidate begins with identifying sequence liabilities, in which the n + 1 amino acid is of particular interest. NW is one motif that occurs frequently within the complementarity-determining region (CDR) of therapeutic antibodies, but according to the published literature, has a very low risk of deamidating. Here we report an unusual case of this NW motif readily deamidating within the CDR of an antibody drug conjugate (ADC), which greatly impacts the ADC's biological activities. Furthermore, this NW motif solely deamidates into iso-aspartate, rather than the typical mixture of iso-aspartate and aspartate. Interestingly, biological activities are more severely impacted by the conversion of asparagine into iso-aspartate via deamidation than by conversion into aspartate via mutagenesis. Here, we detail the discovery of this unusual NW deamidation occurrence, characterize its impact on biological activities, and utilize structural data and modeling to explain why conversion to iso-aspartate is favored and impacts biological activities more severely.

A randomized controlled trial: evaluation of efficiency and safety of a novel surgical guide in the extraction of deeply impacted supernumerary teeth in the anterior maxilla.

Background: Preoperative X-ray and cone-beam computed tomography (CBCT) are helpful for locating supernumerary teeth, but the images cannot be transferred to the operation. To design a novel surgical guide plate for intraoperative navigation, we transfer the patient's oral CBCT and gypsum model scan data to a computer for analysis. In our study, we evaluate the efficiency and safety of a novel surgical guide plate for the extraction of deeply impacted supernumerary teeth (DIMSNT) in the anterior maxilla. Methods: Forty patients treated at the Department of School & Hospital of Stomatology, Wenzhou Medical University from March 2019 to December 2020 with DIMSNT (type II/III according to Liu et al.) in the anterior maxilla were randomly divided into 2 groups (20 patients for each group) for the extraction. For group I, a novel surgical guide was selected using CBCT and gypsum model scan. In contrast, for group II who underwent freehand surgery, only the CBCT data was used. The evaluation of operation time, complications, satisfaction score, and the number of cases that underwent extraction immediately after removing the bone were performed to assess the efficiency and safety of this novel surgical plate. Results: All patients completed the surgery successfully. The guides for group I had a good application effect. Group I's operation time (23.35±5.39 min) was shorter than group II (29.60±9.76 min) (P=0.0194). The average pain degree of group I (1.8±1.08) was significantly less than group II (2.82±1.68) (P<0.05). The average swelling score of group I (34) was significantly less than group II (44.7). Patient satisfaction was significantly higher in group I (8.95±1.05) than in group II (7.90±1.51) (P=0.0152). Conclusions: The novel surgical guide assisted with DIMSNT extraction have been effective in improving the quality of the surgery, patient satisfaction, and reduce its difficulty and duration. We can construct a surgical guide plate to guide the incision and osteotomy in DIMSNT surgery through the data analysis of DIMSNT on computer, which has a broad application prospect for clinical use. Trial registration: Chinese Clinical Trial Registry ChiCTR2100054523.

Characterization of N-Terminal Glutamate Cyclization in Monoclonal Antibody and Bispecific Antibody Using Charge Heterogeneity Assays and Hydrophobic Interaction Chromatography.

N-terminal glutamate (E) cyclization to form pyroglutamate (pE) generates charge heterogeneities for mAbs and proteins. Thus far, pE formation rate in lyophilized formulation as compared to in liquid formulation has not been reported. Impact of pE on antibody biological activity has only been predicted or assessed using stressed samples that may contain other confounding degradations besides pE. Additionally, application of hydrophobic interaction chromatography (HIC) to separate pE has not been reported. In our study, N-terminal E cyclization was identified as the major degradation pathway in lyophilized formulation at elevated temperature for both monoclonal antibody (mAb-A) and IgG-like bispecific antibody (bsAb-A). pE was enriched in salt-gradient ion exchange chromatography (IEC) as pre-peak and in HIC as post-peak for both mAb-A and bsAb-A. Structure-function studies with pE-enriched IEC and HIC fractions confirmed that pE did not affect binding activities for mAb-A and bsAb-A. In vitro incubation of bsAb-A in serum and PBS revealed that the serum matrix may play a role in pE conversion in human serum, in contrast to the chemical reaction mechanism reported. These techniques can help in characterization of N-terminal E-to-pE cyclization and quality attribute severity assessment during therapeutic protein product development.

The impact of land use and rainfall patterns on the soil loss of the hillslope.

In order to discuss the effect of rainfall patterns and land use types on soil erosion, the experiment is carried out under natural rainfall events on different kinds of runoff plots in Zhangjiachong watershed. Based on the observed data of 44 individual rainfall events including moderate, heavy and storm rainfall, the differences of erosion modulus among hedgerows plots, terrace plots, and slope plots under different rainfall patterns are analyzed. And the effects of hedgerow and terrace patterns on control of soil loss are revealed by RUSLE. Wilcoxon signed rank test is applied to analyze the significant difference of erosion modulus in different plots and the coefficient of variation is used to compare the characteristics of erosion modulus under different rainfall patterns. The results show that the soil erosion modulus of earth banked terrace has the highest value and the lowest soil erosion modulus occurs in the slope land with hedgerows. The coefficients of variation for soil erosion modulus under heavy and storm rainfall are larger than that of moderate rainfall. Hedgerow pattern can effectively control soil erosion under moderate and heavy rainfall while the effect of hedgerow is considerably weakened under storm rainfall. Earth banked terraces own the highest erosion modulus followed by slope land and stone dike terraces.

Enhanced remediation efficiency of Cr(VI)-contaminated heterogeneous aquifers: Improved sweeping efficiency using shear-thinning fluids.

Low permeability zones (LPZs) are typically bypassed when remedial reagents are injected into heterogeneous aquifers, which hinders the in situ remediation. Although shear-thinning polymers have emerged as promising tools to meet this challenge, their applicability in complex remedial systems remains unconfirmed. We investigated the sweeping efficiencies of calcium polysulfide (CPS) into Cr(VI)-contaminated LPZs using xanthan gum (XG) as the model shear-thinning polymer. Firstly, the compatibility of XG-CPS fluids and their reduction capacities toward Cr(VI) were demonstrated based on batch experiments. The removal rates of Cr(VI) exceeded 85% in the presence of 250-2000 mg/L of XG. Besides, XG-CPS fluids exhibited a greater impact on the permeability decrease of transmissive zones than that of LPZs as confirmed by sand column experiments. Furthermore, the sweeping efficiencies in LPZs during XG-CPS flooding were investigated by multiple sand tank experiments. The sweeping rate in LPZs (rs) in Cr(VI)-contaminated aquifer (1.68 × 10-3/min) was found to be approximately 11% higher than that of uncontaminated system, and two possible reasons behind this phenomenon were proposed. The spatial distribution profiles of Cr under different XG-CPS flooding conditions were depicted based on 20 representative samples. The results indicated that all Cr(VI) in LPZs can be effectively removed either by displacement or immobilization as Cr(III). The percentages of displaced Cr(VI) and immobilized Cr(III) were calculated to be 65%-75% and 25-35%, respectively. This work demonstrates the applicability of XG-CPS fluids as remedial materials for Cr(VI)-contaminated heterogeneous aquifers and provides novel insights into the role of Cr(VI) in in situ remediation using shear-thinning polymers.

Efficaciousness of dexmedetomidine in children undergoing cleft lip and palate repair: a systematic review and meta-analysis.

OBJECTIVE: To systematically assess the efficacy and safety of dexmedetomidine as an anaesthesia adjuvant for cleft lip and palate (CLP) repair in children. DESIGN: Systematic review and meta-analysis. DATA SOURCES: PubMed, Embase, Cochrane, China National Knowledge Infrastructure (CNKI), China Science and Technology Journal Database (VIP) and Wanfang (up to October 2020). Studies in languages other than English and Chinese were excluded. ELIGIBILITY CRITERIA FOR SELECTING STUDIES: Randomised controlled trials (RCTs) evaluating the impact of dexmedetomidine on emergence agitation (EA), the need for postoperative rescue analgesics, postoperative nausea and vomiting (PONV), and other adverse events in paediatric patients during CLP repair. DATA EXTRACTION AND SYNTHESIS: The quality of evidence was assessed by using the Cochrane Review Methods and the Grading of Recommendations Assessment, Development and Evaluation approach. Data were screened, extracted and assessed by two independent authors. Outcomes were reported as a risk ratio (RR) with a 95% CI. A random-effect model was used when heterogeneity was detected. RESULTS: Thirteen studies including 1040 children met the inclusion criteria. The incidence of EA was significantly decreased in the dexmedetomidine group (RR, 0.19; 95% CI 0.10 to 0.36; p<0.00001; I2=56%) as compared with the control group. Paediatric patients receiving dexmedetomidine had lower postoperative analgesic requirements (RR, 0.27; 95% CI 0.10 to 0.73; p=0.01; I2=84%) and a lower incidence of respiratory adverse events (RR, 0.49; 95% CI 0.31 to 0.78; p=0.003; I2=0%). There were no significant differences in the risk of PONV and cardiovascular adverse events. CONCLUSIONS: There was a lack of high-quality studies in this field. Perioperative administration of dexmedetomidine reduced the need for postoperative rescue analgesics and the incidence of EA in children without side effects undergoing CLP repair. However, further verification with larger samples and higher-quality RCTs is needed.

Development of a mechanism of action-reflective, dual target cell-based reporter bioassay for a bispecific monoclonal antibody targeting human CTLA-4 and PD-1.

T-cell-mediated immunotherapy has generated much excitement after the success of therapeutic biologics targeting immune checkpoint molecules. Bispecific antibodies (BsAbs) that recognize two antigen targets are a fast-growing class of biologics offering promising clinical benefits for cancer immunotherapy. Due to the complexity of the molecule structure and the potential mechanism of action (MOA) that involves more than one signaling pathway, it is critical to develop appropriate bioassays for measuring potency and characterizing the biological properties of BsAbs. Here, we present a dual target, cell-based reporter bioassay for a BsAb that binds human CTLA-4 and PD-1 and targets two subsequent signaling pathways that negatively regulate T-cell activation. This reporter bioassay is capable of measuring the potency of both antigen target arms in one assay, which would not be achievable using two single target bioassays. This dual target reporter bioassay demonstrates good performance characteristics suitable for lot release, stability testing, critical quality attribute assessment, and biological properties characterization of the CTLA-4/PD-1 BsAb. Furthermore, this assay can capture the synergistic effect of anti-CTLA-4 and anti-PD-1 activity of the BsAb. Compared to single target assays, this dual target bioassay could better reflect the potential MOA of BsAbs and could be used for evaluation of other bispecific biologics, as well as antibody combination therapies.