The hook effect is best explained by how the analyte signal generated from the assay is compromised due to either antibody excess or antigen excess. Reporting false negatives can also impact clinical decisions that may have adverse effects on the patient. The clinical impact of the hook effect will lead to reporting either inaccurately low or false-negative results. Six different patient pools were tested using immunoassay screening methods and also the LC-MS/MS confirmation method. For the immunoassay screening method, 50 ​μL of each patient sample is pooled together for every 100 patient samples. If there is a sample with the hook effect, it would give a very low or a negative result when the pool is tested neat and it would give a positive result when the pool is tested in a 1∶100 dilution. The drugs tested included Cannabinoids, Benzodiazepines, Amphetamines, Ecstasy, EDP, Opiates, Heroin, Cocaine, Fentanyl, Oxycodone and a combined method for Buprenorphine/Norbuprenorphine. Patient Pool number 6 showed a positive Buprenorphine and Norbuprenorphine that was traced back to the sample with the hook effect based on the suggested protocol. The random and different patient pools were prepared by using a Gilson GX241 liquid handler. The neat result for Pool number 6 showed the immunoassay for Buprenorpone and Norpuprenorphine was “Not Detected” at 2.1 ​μg/L while the LC-MS/MS result was 2,780 ​μg/L (cut-off <10 ​μg/L). However, the result for the 1 in 100 dilutions of the pool for the immunoassay was “Detected” at 37 ​μg/L without multiplying by the dilution factor. The result for the LC-MS/MS was 2,970 ​μg/L. The suggested protocol is practical and cost-effective to avoid the clinical impact of reporting either inaccurately low or false-negative results. Also, can be used for testing when suspected samples with the hook effect are investigated.

Abstract

Objective

Multiple reaction monitoring (MRM), is a highly sensitive targeted mass spectrometry technique that is used to validate discovery phase mass spectrometry data. Through this simple technique, relative protein expressions can be acquired without using commercially available antibodies.

Methods

Our previous work showed Gipie silencing of UM-HACC-2A (adenoid cystic carcinoma) cells resulted in upregulation of a plethora of immune proteins in immune cells in 3D ACC - immune co-culture model.

Results

MRM enabled determining relative protein expression of significantly upregulated immune proteins due to silencing of Gipie. Granzyme A, CD48, granzyme B, HLA class I, antigen B, HLA class I, antigen A, galectin 1, vimentin, endoplasmic reticulum chaperone BiP, and dipeptidyl peptidase 1 differential expressions were validated. Whereas coactosin-like protein expressions were found non-significant among control and Gipie silenced ACC.

Conclusions

This manuscript presents the validation data of previous discovery phase mass spectrometry and also accentuate the use of MRM in translational or preclinical studies in the field of biomedical sciences.

Hemophilia is a hemorrhagic disease caused by a lack of clotting factors due to genetic mutations, and is a representative of rare diseases. The prevalence rate of hemophilia is about 2.73/100,000–3.09/100,000 in China, and the absolute number of patients with hemophilia is large given a huge population base. As a part of the Healthy China initiative, prevention and treatment of hemophilia has attracted much attention in the country. With comprehensive promotion of the construction of the grading diagnosis and treatment system for hemophilia, China requires hospitals applying for comprehensive management centers and diagnosis and treatment centers for hemophilia to be able to independently carry out screening and confirming tests related to hemophilia diagnosis. Identifying the type of pathogenic mutation through genetic diagnosis is of great value for assessing the risk of clinical bleeding in hemophilia patients, studying the induction mechanism of clotting factor inhibitors, conducting gene therapy and promoting eugenics. The mutation types of F8 gene are similar in Chinese patients to international data reports. The F9 gene mutations are mainly single-base point mutations, with no mutation hotspots found in Chinese patients. Historically in China, the treatment of hemophilia has gone through insufficient on-demand treatment and exploration of low-dose prophylaxis, and currently, it is moving towards high-dose prophylaxis with better efficacy and individualized prophylaxis, achieving more ideal treatment effects, reducing patient inconvenience and discomfort, and avoiding high treatment costs as much as possible. Based on related registration data and literature, this report comprehensively summarizes the progress and points out the shortcomings in clinical research and medical care as regards hemophilia in China, in the hope of helping further improve the diagnosis and treatment of hemophilia in the country.

Aluminum (Al) exhibits excellent electrical conductivity, mechanical ductility, and good chemical compatibility with high-ionic-conductivity electrolytes. This makes it more suitable as an anode material for all-solid-state lithium batteries (ASSLBs) compared to the overly reactive metallic lithium anode and the mechanically weak silicon anode. This study finds that the pre-lithiated Al anode demonstrates outstanding interfacial stability with the Li₆PS₅Cl (LPSCl) electrolyte, maintaining stable cycling for over 1200 h under conditions of deep charge–discharge. This paper combines the pre-lithiated Al anode with a high-nickel cathode, LiNi_0.8Co_0.1Mn_0.1O₂, paired with the highly ionic conductive LPSCl electrolyte, to design an ASSLB with high energy density and stability. Using anode pre-lithiation techniques, along with dual-reinforcement technology between the electrolyte and the cathode active material, the ASSLB achieves stable cycling for 1000 cycles at a 0.2C rate, with a capacity retention rate of up to 82.2%. At a critical negative-to-positive ratio of 1.1, the battery’s specific energy reaches up to 375 Wh kg^-1, and it maintains over 85.9% of its capacity after 100 charge–discharge cycles. This work provides a new approach and an excellent solution for developing low-cost, high-stability all-solid-state batteries.

Hygroscopic hydrogel is a promising evaporative-cooling material for high-power passive daytime cooling with water self-regeneration. However, undesired solar and environmental heating makes it a challenge to maintain sub-ambient daytime cooling. While different strategies have been developed to mitigate heat gains, they inevitably sacrifice the evaporation and water regeneration due to highly coupled thermal and vapor transport. Here, an anisotropic synergistically performed insulation-radiation-evaporation (ASPIRE) cooler is developed by leveraging a dual-alignment structure both internal and external to the hydrogel for coordinated thermal and water transport. The ASPIRE cooler achieves an impressive average sub-ambient cooling temperature of ~ 8.2 °C and a remarkable peak cooling power of 311 W m^-2 under direct sunlight. Further examining the cooling mechanism reveals that the ASPIRE cooler reduces the solar and environmental heat gains without comprising the evaporation. Moreover, self-sustained multi-day cooling is possible with water self-regeneration at night under both clear and cloudy days. The synergistic design provides new insights toward high-power, sustainable, and all-weather passive cooling applications.

The paper published in SCIENCE CHINA Chemistry systematically summarizes the research progress and innovative strategies for improving the performance of NaₓTMO₂ cathode materials through interface regulation engineering in recent years. This work reveals the enhancement mechanisms of interface engineering, such as inorganic/organic coatings, heterogeneous interface phase designing, and surface doping. The phase evolution behaviors, ion-transfer kinetics, and electrochemical properties of NaₓTMO₂ resulted from interfacial modulations are concluded in depth.

Researchers at Xi'an Jiaotong University have made a significant advancement in the development of silicon anodes for lithium-ion batteries (LIBs), introducing a novel binder that dramatically improves the stability and performance of silicon-based electrodes. This breakthrough addresses one of the most critical challenges in silicon anode technology: the unstable solid electrolyte interphase (SEI) layer, which has long hindered the commercialization of high-capacity silicon anodes.

A study in Science China Life Sciences reveals how hybrid fish maintain fertility despite genomic divergence. By analyzing structural variations, gene expression, and chromatin accessibility in testicular cells, researchers identified conserved allelic and testis-specific genes, with rnf212b as a key regulator of reproductive traits.

The β cell proliferation capacity determines its mass and islet function, thus playing a pivotal role in maintaining glucose homeostasis. A study published in Science Bulletin Journal reported a pancreatic endogenous ribonuclease 4 (Rnase4), which governs β cell number and islet size under both physiological and pathological conditions. This factor is selectively expressed in β cells and acts in a cell-autonomous manner to drive β cell proliferation through the Axl-PI3K/Akt/FoxO1/Pdx1 signaling pathway. Absence of Rnase4 diminished β cell population, leading to systemic insulin deficiency; exogenous supplementation of this factor could rescue β cell loss and islet dysfunction induced by genetic, chemical, or inflammatory challenges. The findings highlight a key β-cell self-derived factor in regulating its proliferation and body glucose metabolism.