Identification of a Fully Human Antibody VH Domain Targeting Anaplastic Lymphoma Kinase (ALK) with Applications in ALK-Positive Solid Tumor Immunotherapy.

The anaplastic lymphoma kinase (ALK, CD247) is a potential target for antibody-based therapy. However, no antibody-based therapeutics targeting ALK have entered clinical trials, necessitating the development of novel antibodies with unique therapeutic merits. Single-domain antibodies (sdAb) bear therapeutic advantages compared to the full-length antibody including deeper tumor penetration, cost-effective production and fast washout from normal tissues. In this study, we identified a human immunoglobulin heavy chain variable domain (VH domain) (VH20) from an in-house phage library. VH20 exhibits good developability and high specificity with no off-target binding to ~6000 human membrane proteins. VH20 efficiently bound to the glycine-rich region of ALK with an EC50 of 0.4 nM and a KD of 6.54 nM. Both VH20-based bispecific T cell engager (TCE) and chimeric antigen receptor T cells (CAR Ts) exhibited potent cytolytic activity to ALK-expressing tumor cells in an ALK-dependent manner. VH20 CAR Ts specifically secreted proinflammatory cytokines including IL-2, TNFα and IFNγ after incubation with ALK-positive cells. To our knowledge, this is the first reported human single-domain antibody against ALK. Our in vitro characterization data indicate that VH20 could be a promising ALK-targeting sdAb with potential applications in ALK-expressing tumors, including neuroblastoma (NBL) and non-small cell lung cancer.

Mechanism and Origins of Enantioselectivity in the Nickel-catalyzed Asymmetric Synthesis of Silicon-Stereogenic Benzosiloles.

As important π-skeletons, benzosiloles often possess unique electronic and optical properties and have been widely used in semiconductor materials. Therefore, great attention has been drawn to the area of developing novel synthetic methods for various benzosiloles. However, the synthesis of enantioenriched silicon-stereogenic benzosiloles is still at an early stage and remains to be explored. Herein, we performed systematic density functional theory studies on the recently reported nickel-catalyzed asymmetric synthesis of silicon-stereogenic benosiloles, which was enabled by an enantioselective desymmetrization of (2-alkenyl)aryl-substituted silacyclobutanes. Our computational study shows that the reaction mechanism involves ligand exchange, oxidative addition, alkene insertion, and hydrogen-transfer coupled reductive-demetalation steps. The proposed transmetalation and ß-hydride elimination mechanism was not found, which might be due to the unfavorable ring strain of the multicyclic intermediates. The novel hydrogen-transfer coupled reductive-demetalation mechanism was shown to be reasonable for the generation of the silicon-stereogenic benzosilole. Noncovalent interactions (including C-H···π and hydrogen bonding) in the rate-determining alkene insertion transition state account for the origins of the enantioselectivity. Our computational study sheds light on the detailed reaction mechanism and also provides insights for the development of novel approaches for synthesis of high-value silicon-stereogenic compounds.

A Survey on Federated Recommendation Systems.

Federated learning has recently been applied to recommendation systems to protect user privacy. In federated learning settings, recommendation systems can train recommendation models by collecting the intermediate parameters instead of the real user data, which greatly enhances user privacy. In addition, federated recommendation systems (FedRSs) can cooperate with other data platforms to improve recommendation performance while meeting the regulation and privacy constraints. However, FedRSs face many new challenges such as privacy, security, heterogeneity, and communication costs. While significant research has been conducted in these areas, gaps in the surveying literature still exist. In this article, we: 1) summarize some common privacy mechanisms used in FedRSs and discuss the advantages and limitations of each mechanism; 2) review several novel attacks and defenses against security; 3) summarize some approaches to address heterogeneity and communication costs problems; 4) introduce some realistic applications and public benchmark datasets for FedRSs; and 5) present some prospective research directions in the future. This article can guide researchers and practitioners understand the research progress in these areas.

Association between CT-based adipose variables, preoperative blood biochemical indicators and pathological T stage of clear cell renal cell carcinoma.

Background: Clear cell renal cell carcinoma (ccRCC) is corelated with tumor-associated material (TAM), coagulation system and adipocyte tissue, but the relationships between them have been inconsistent. Our study aimed to explore the cut-off intervals of variables that are non-linearly related to ccRCC pathological T stage for providing clues to understand these discrepancies, and to effectively preoperative risk stratification. Methods: This retrospective analysis included 218 ccRCC patients with a clear pathological T stage between January 1st, 2014, and November 30th, 2021. The patients were categorized into two cohorts based on their pathological T stage: low T stage (T1 and T2) and high T stage (T3 and T4). Abdominal and perirenal fat variables were measured based on preoperative CT images. Blood biochemical indexes from the last time before surgery were also collected. The generalized sum model was used to identify cut-off intervals for nonlinear variables. Results: In specific intervals, fibrinogen levels (FIB) (2.63-4.06 g/L) and platelet (PLT) counts (>200.34 × 109/L) were significantly positively correlated with T stage, while PLT counts (<200.34 × 109/L) were significantly negatively correlated with T stage. Additionally, tumor-associated material exhibited varying degrees of positive correlation with T stage at different cut-off intervals (cut-off value: 90.556 U/mL). Conclusion: Preoperative PLT, FIB and TAM are nonlinearly related to pathological T stage. This study is the first to provide specific cut-off intervals for preoperative variables that are nonlinearly related to ccRCC T stage. These intervals can aid in the risk stratification of ccRCC patients before surgery, allowing for developing a more personalized treatment planning.

RNA-seq analysis highlights DNA replication and DNA repair associated with early-onset hearing loss in the cochlea of DBA/2J mice.

AIMS: Age-related hearing loss (ARHL) is a significant health concern, and DBA/2J (D2) and C57BL/6 (B6) mouse strains serve as valuable models for its study. B6 mice, harboring a homozygous ahl allele in Cdh23, manifest high-frequency hearing loss at 3 months. In contrast, D2 mice, carrying the R109H variant of the Fascin-2 gene (Fscn2), experience early-onset hearing loss by 3 weeks. Yet, the underlying molecular mechanisms driving early-onset hearing loss in D2 mice remain elusive. This study aimed to identify novel genes and regulatory pathways as therapeutic targets for early deafness. MAIN METHODS: This study employs RNA-sequencing (RNA-seq) to analyze cochlear mRNA expression at two different ages in D2 and B6 mice, respectively. The differentially expressed genes (DEGs) are uniquely associated with D2 mice by Venn diagram analysis. A protein-protein interaction (PPI) network is further constructed, followed by module analysis utilizing MCODE. Enrichment analysis of GO and KEGG pathways revealed biological functions and molecular pathways. The PPI network and VarElect analysis are conducted for genes within these pathways, facilitating the identification of pivotal genes based on scoring criteria. Subsequently, five genes are meticulously selected and validated through qRT-PCR. KEY FINDINGS: Notably, 1181 DEGs are uniquely associated with D2 mice by Venn diagram analysis. GO and KEGG pathway enrichment analyses shed light on distinctive pathways in D2 mice, encompassing DNA replication, mismatch repair, base excision repair, and nucleotide excision repair, which are associated with apoptosis. Five genes involved in these pathways were finally selected and validated by qRT-PCR. Their down-regulation with age is consistent with RNA-seq result. SIGNIFICANCE: Our study underscores the potential implication of down-regulated genes associated with DNA replication and DNA damage repair in the early-onset hearing loss observed in D2 mice.

Assessment of Novel Mesothelin-Specific Human Antibody Domain VH-Fc Fusion Proteins-Based PET Agents.

Mesothelin (MSLN) is a tumor-associated antigen found in a variety of cancers and is a target for imaging and therapeutic applications in MSLN-expressing tumors. We have developed high affinity anti-MSLN human VH domain antibodies, providing alternative targeting vectors to conventional IgG antibodies that are associated with long-circulating half-lives and poor penetration of tumors, limiting antitumor activity in clinical trials. Based on two newly identified anti-MSLN VH binders (3C9, 2A10), we generated VH-Fc fusion proteins and modified them for zirconium-89 radiolabeling to create anti-MSLN VH-Fc PET tracers. The focus of this study was to assess the ability of PET-imaging to compare the in vivo performance of anti-MSLN VH-Fc fusion proteins (2A10, 3C9) targeting different epitopes of MSLN vs IgG1 (m912; a clinical benchmark antibody with an overlapped epitope as 2A10) for PET imaging in a mouse model of colorectal cancer (CRC). The anti-MSLN VH-Fc fusion proteins were successfully modified and radiolabeled with zirconium-89. The resulting MSLN-targeted PET-imaging agents demonstrated specific uptake in the MSLN-expressing HCT116 tumors. The in vivo performance of the MSLN-targeted PET-imaging agents utilizing VH-Fc showed more rapid and greater accumulation and deeper penetration within the tumor than the full-length IgG1 m912-based PET-imaging agent. Furthermore, PET imaging allowed us to compare the pharmacokinetics of epitope-specific VH domain-based PET tracers. Overall, these data are encouraging for the incorporation of PET imaging to assess modified VH domain structures to develop novel anti-MSLN VH domain-based therapeutics in MSLN-positive cancers as well as their companion PET imaging agents.

Evaluation of the relations between reproduction-related pituitary and ovarian hormones and abdominal fat area-related variables determined with computed tomography in overweight or obese women who have undergone bariatric surgery: a cross-sectional study.

Background: An understanding of the associations between midregion fat depots and systemic hormone levels will be crucial for developing health-promotion messages aimed at overweight or obese women. However, related research in this area is rare. The present study was performed to identify and quantify fat-related reproduction pituitary and ovarian hormones in overweight or obese women. Methods: A total of 250 eligible overweight or obese women scheduled to undergo laparoscopic sleeve gastrectomy (LSG) from a single center were retrospectively included in this study. Computed tomography (CT) images at the level of the umbilicus were selected, and abdominal fat areas were measured and calculated. The reproduction-related pituitary and ovarian hormones were also measured. The correlations among the parameters were examined using Spearman correlation test. Multiple linear regression analysis was performed after log and ß-transformation of the hormone levels and fat area-related variables. Results: Positive correlations were detected for prolactin (PRL) with total fat area (TFA) [ß=0.045; P=0.029; 95% confidence interval (CI): 0.004-0.085] and subcutaneous fat area (SFA) (ß=0.066; P=0.023; 95% CI: 0.009-0.123), whereas estradiol showed a negative correlation with visceral fat area (VFA) (ß=-0.056, P=0.005; 95% CI: -0.096 to -0.017) and relative VFA (rVFA) (ß=-0.068; P=0.001; 95% CI: -0.109 to -0.027) and a positive correlation with SFA (ß=0.036; P=0.042; 95% CI: 0.001-0.071). Progesterone (PROG) was negatively correlated with both VFA (ß=-0.037; P=0.002; 95% CI: -0.061 to -0.013) and rVFA (ß=-0.039; P=0.002; 95% CI: -0.063 to -0.014). The final results revealed that TFA was increased by 3.1% and SFA was increased by 4.7% with a doubling of PRL concentration; VFA was reduced by 2.5% and rVFA was reduced by 2.6% with a doubling of PROG concentration; and VFA was reduced by 3.8%, rVFA was reduced by 4.6%, and SFA was increased by 2.5% with a doubling of estradiol concentration. Conclusions: There exist certain associations between some reproduction-related pituitary and ovarian hormones and fat areas. Our findings provide new insights into the associations between midregion fat depots and systemic hormone levels in overweight or obese women.

Catalytic Enantioselective Biltz Synthesis.

The Biltz synthesis establishes straightforward access to 5,5-disubstituted (thio)hydantoins by combining a 1,2-diketone and a (thio)urea. Its appealing features include inherent atom and step economy together with the potential to generate structurally diverse products. However, control of the stereochemistry of this reaction has proven to be a daunting challenge. Herein, we describe the first example of enantioselective catalytic Biltz synthesis which affords more than 40 thiohydantoins with high stereo- and regio-control, irrespective of the symmetry of thiourea structure. A one pot synthesis of corresponding hydantoins is also documented. Remarkably, experimental studies and DFT calculations establish the reaction pathway and origin of stereoselectivity.

Prediction of Fuhrman nuclear grade for clear cell renal carcinoma by a multi-information fusion model that incorporates CT-based features of tumor and serum tumor associated material.

PURPOSE: Prediction of Fuhrman nuclear grade is crucial for making informed herapeutic decisions in clear cell renal cell carcinoma (ccRCC). The current study aimed to develop a multi-information fusion model utilizing computed tomography (CT)-based features of tumors and preoperative biochemical parameters to predict the Fuhrman nuclear grade of ccRCC in a non-invasive manner. METHODS: 218 ccRCC patients confirmed by histopathology were retrospectively analyzed. Univariate and multivariate logistic regression analyses were performed to identify independent predictors and establish a model for predicting the Fuhrman grade in ccRCC. The predictive performance of the model was evaluated using receiver operating characteristic (ROC) curves, calibration, the 10-fold cross-validation method, bootstrapping, the Hosmer-Lemeshow test, and decision curve analysis (DCA). RESULTS: R.E.N.A.L. Nephrometry Score (RNS) and serum tumor associated material (TAM) were identified as independent predictors for Fuhrman grade of ccRCC through multivariate logistic regression. The areas under the ROC curve (AUC) for the multi-information fusion model composed of the above two factors was 0.810, higher than that of the RNS (AUC 0.694) or TAM (AUC 0.764) alone. The calibration curve and Hosmer-Lemeshow test showed the integrated model had a good fitting degree. The 10-fold cross-validation method (AUC 0.806) and bootstrap test (AUC 0.811) showed the good stability of the model. DCA demonstrated that the model had superior clinical utility. CONCLUSION: A multi-information fusion model based on CT features of tumor and routine biochemical indicators, can predict the Fuhrman grade of ccRCC using a non-invasive approach. This model holds promise for assisting clinicians in devising personalized management strategies.

Shortest path core-selection incentive for federated learning platform with medical applications.

As the main technology to solve data islands and mine data value, federated learning has been widely researched and applied, and more and more federated learning platforms are emerging. The federated learning platform organizes users, devices and data to collaborate in a crowdsourcing manner and complete specific federated learning tasks. This paper designs the shortest path core-selection incentive mechanism by combining the VCG auction mechanism and the core concept of cooperative games. This mechanism solves the problems of overpayment, false-name attack, and deviation from the core of the VCG mechanism, and saves the expenditure of the federated learning task issuer. It adopts the VCG-nearest principle in the core selection, so that the federated learning task participants get rewards as close as possible to the outcome of VCG mechanism. This mechanism can guarantee four economic attributes: incentive compatibility, individual rationality, alliance rationality, and maximization of social efficiency. Medical experimental results illustrate the effectiveness of the mechanism.

Preclinical assessment of a novel human antibody VH domain targeting mesothelin as an antibody-drug conjugate.

Mesothelin (MSLN) has been a validated tumor-associated antigen target for several solid tumors for over a decade, making it an attractive option for therapeutic interventions. Novel antibodies with high affinity and better therapeutic properties are needed. In the current study, we have isolated and characterized a novel heavy chain variable (VH) domain 3C9 from a large-size human immunoglobulin VH domain library. 3C9 exhibited high affinity (KD [dissociation constant] <3 nM) and binding specificity in a membrane proteome array (MPA). In a mouse xenograft model, 3C9 fused to human IgG1 Fc was detected at tumor sites as early as 8 h post-infusion and remained at the site for over 10 days. Furthermore, 3C9 fused to a human Fc domain drug conjugate effectively inhibited MSLN-positive tumor growth in a mouse xenograft model. The X-ray crystal structure of full-length MSLN in complex with 3C9 reveals interaction of the 3C9 domains with two distinctive residue patches on the MSLN surface. This newly discovered VH antibody domain has a high potential as a therapeutic candidate for MSLN-expressing cancers.

Development of an efficient method for selection of stable cell pools for protein expression and surface display with Expi293F cells.

Compare with transient expression, stable cell lines generally have higher productivity and better quality for protein expression. However, selection of stable cell line is time-consuming and laborious. Here we describe an optimized selection method to achieve high-efficient stable cell pools with Expi293F suspension cells. This method only takes 2-3 weeks to generate stable cell pools with 2- to 10-fold higher productivity than transient gene expression (TGE). In fed-batch culture with Yeastolate, >1 g/L yield was achieved with our KTN0239-IgG stable cell pool in shaker flasks. This method can be also applied to efficiently display proteins on the cell surface.

Feasibility of 3.0 T balanced fast field echo non-contrast-enhanced whole-heart coronary magnetic resonance angiography.

Background: Coronary artery disease (CAD) is one of the most common diseases seriously harmful to human health caused by atherosclerosis. Besides coronary computed tomography angiography (CCTA) and invasive coronary angiography (ICA), coronary magnetic resonance angiography (CMRA) has become an alternative examination. The purpose of this study was to prospectively evaluate the feasibility of 3.0 T free-breathing whole-heart non-contrast-enhanced coronary magnetic resonance angiography (NCE-CMRA). Methods: After Institutional Review Board approval, the NCE-CMRA data sets of 29 patients acquired successfully at 3.0 T were evaluated independently by two blinded readers for visualization and image quality of coronary arteries using the subjective quality grade. The acquisition times were recorded in the meantime. A part of the patients had undergone CCTA, we represented stenosis by scores and used the Kappa to evaluate the consistency between CCTA and NCE-CMRA. Results: Six patients did not get diagnostic image quality because of severe artifacts. The image quality score assessed by both radiologists is 3.2±0.7, which means the NCE-CMRA can show the coronary arteries excellently. The main vessels of the coronary artery on NCE-CMRA images are considered reliably assessable. The acquisition time of NCE-CMRA, is 8.8±1.2 min. The Kappa of CCTA and NCE-CMRA on detecting stenosis is 0.842 (P<0.001). Conclusions: The NCE-CMRA results in reliable image quality and visualization parameters of coronary arteries within a short scan time. The NCE-CMRA and CCTA have a good agreement for detecting stenosis.

Human Action Recognition From Various Data Modalities: A Review.

Human Action Recognition (HAR) aims to understand human behavior and assign a label to each action. It has a wide range of applications, and therefore has been attracting increasing attention in the field of computer vision. Human actions can be represented using various data modalities, such as RGB, skeleton, depth, infrared, point cloud, event stream, audio, acceleration, radar, and WiFi signal, which encode different sources of useful yet distinct information and have various advantages depending on the application scenarios. Consequently, lots of existing works have attempted to investigate different types of approaches for HAR using various modalities. In this article, we present a comprehensive survey of recent progress in deep learning methods for HAR based on the type of input data modality. Specifically, we review the current mainstream deep learning methods for single data modalities and multiple data modalities, including the fusion-based and the co-learning-based frameworks. We also present comparative results on several benchmark datasets for HAR, together with insightful observations and inspiring future research directions.