Labeling Procedure Research Articles

Fully automated 18F-fluorination of N-succinimidyl-4-[18F]fluorobenzoate ([18F]SFB) for indirect labelling of nanobodies

N-succinimidyl-4-[18F]fluorobenzoate ([18F]SFB), a widely used labeling agent to introduce the 4-[18F]fluorobenzoyl-prosthetic group, is normally obtained in three consecutive steps from [18F]fluoride ion. Here, we describe an efficient one-step labeling procedure of [18F]SFB starting from a tin precursor. This method circumvents volatile radioactive side-products and simplifies automatization. [18F]SFB was obtained after HPLC purification in a yield of 42 + 4% and a radiochemical purity (RCP) > 99% (n = 6). In addition, we investigate the automation of the coupling of [18F]SFB to a nanobody (cAbBcII10, targeting β-lactamase enzyme) and purification by size exclusion chromatography (PD-10 desalting column) to remove unconjugated reagent. Production and use of [18F]SFB were implemented on a radiosynthesis unit (Neptis®). The fully automated radiosynthesis process including purification and formulation required 160 min of synthesis time. [18F]SFB-labeled nanobody was obtained in a yield of 21 + 2% (activity yield 12 + 1% non-decay corrected) and a radiochemical purity (RCP) of > 95% (n = 3). This approach simplifies [18F]SFB synthesis to one-step, enhances the yield in comparison to the previous report and enables the production of radiolabeled nanobody on the same synthesis module.

Assessing kernel processing score of harvested corn silage in real-time using image analysis and machine learning

Whole Plant Corn Silage (WPCS), produced by Self-Propelled Forage Harvesters (SPFH), is among the most common dietary ingredients for dairy cows. Kernel processor settings at harvest can impact WPCS quality and milk production. Having the capability to monitor machine performance and WPCS quality during harvest would allow farmers, nutritionists, and custom harvesters to optimize feed quality as the WPCS enters storage. A real-time quality assessment system was developed - a device to acquire images of chopped and processed silage moving through the spout of a SPFH and an image analysis algorithm to estimate the Kernel Processing Score (KPS) given a set of images. The system represents automation of the quality assessment process and could be incorporated in a real-time machine parameter tuning system. An initial experiment of corn particle detection in high-resolution images of stationary WPCS resulted in good detection performance (precision of 0.7381, recall of 0.9624, and intersection-over-union of 0.7104) but exposed inadequacies in the image labeling procedure and was not able to accurately estimate KPS. An improved camera system was designed and consisted of specialized exposure components to capture images at 121 frames per second. This system was able to acquire pictures silage moving at a rate of 36.9 m s−1. The KPS estimation was done offline and occurred in two steps: whole undamaged kernels were counted in a set of 1,000 images using machine learning; KPS was then estimated using a linear regression model previously obtained with the comparison between mechanical sieving KPS and number of kernels per image (r(38) = 0.967, p < 0.0001). A comparison between the estimated score to mechanically sieved KPS showed high correlation (r(38) = 0.977, p < 0.0001) and standard error of 2.71 %. Considering the computing power of high-end GPUs, the method is restrained by its current image acquisition rate, that results in one estimate in <10 s.

Mechanism of titanium electrochemical oxidation via isotopic labeling, high resolution ion depth profiling, and impedance spectroscopy

The electrochemical oxidation mechanism of titanium between 0 and 10 V vs saturated calomel reference electrode (SCE) was examined for ultra-thin Ti films sputtered onto Si(001) substrates and exposed in-situ to H218O, and then anodized in D216O. The effects of this isotopic labeling procedure were studied using medium energy ion scattering (MEIS) and nuclear reaction profiling (NRP). Both MEIS and NRP results are consistent in showing that the titanium oxide layer is composed of two distinct regions, Ti16O2/Ti18O2/Ti/Si(001) for the entire range of the formation voltages (0–10 V vs SCE). The outermost region consists entirely of 16O, and the 18O region is always adjacent to the Ti metal. The two distinct structures observed can be consistent with both the point defect model (PDM) or high-field model (HFM), assuming the mobility of titanium cations is much higher compared to the mobility of oxygen ions. No Ti or 18O loss into the electrolyte during anodization is detected. Linear growth rate is observed in 0–10 V range vs SCE with experimental anodization ratio of 24.5±0.6 Å V−1. Mott–Schottky (MS) analyses show positive slopes, indicating formation of an n-type TiO2 semiconductor, with O vacancies (or Ti interstitials) as major charge carriers in the 0–10 V range. Charge carrier densities, ND = (0.8-5.0 )×1021 cm−3 were calculated from Mott–Schottky analysis and were well within the range of results reported in the literature. We observe derease in the charge carrier densities above ∼4 V vs SCE, that can be connected to defects annihilation or minor modification in the structure of the growing TiO2 film.

Clinically Applicable Homogeneous Assay for Serological Diagnosis of Alpha-Fetoprotein by Impact Electrochemistry.

Tumor protein quantification with high specificity, sensitivity, and efficiency is of great significance to enable early diagnosis and effective treatment. The existing methods for protein analysis usually suffer from high cost, time-consuming operation, and insufficient sensitivity, making them not clinically friendly. In this work, a label-free homogeneous sensor based on the nano-impact electroanalytic (NIE) technique was proposed for the detection of tumor protein marker alpha-fetoprotein (AFP). The detection principle is based on the recovery of current of single PtNP catalyzed hydrazine oxidation due to the release of the pre-adsorbed passivating aptamers on PtNPs from the competition of the stronger binding between the specific interaction of the AFP aptamer and AFP. The intrinsic one-by-one analytical ability of NIE allows highly sensitive detection, which can be further improved by reducing the reaction/incubation volume. Meanwhile, the current sensor avoids a laborious labeling procedure as well as the separation and washing steps due to the in situ characteristic of NIE. Accordingly, the current sensor enables efficient, highly sensitive, and specific AFP analysis. More importantly, the reliable detection of AFP in diluted real sera from hepatocellular carcinoma (HCC) patients is successfully achieved, indicating that the impact electrochemistry-based sensing platform has great potential to be applied in point-of-care devices for HCC liquid biopsy.

Biomimetic 3D Recognition with 2D Flexible Nanoarchitectures for Ultrasensitive and Visual Extracellular Vesicle Detection.

Despite increasing recognition of extracellular vesicles being important circulating biomarkers in disease diagnosis and prognosis, current strategies for extracellular vesicle detection remain limited due to the compromised sample purification and extensive labeling procedures in complex body fluids. Here, we developed a 2D magnetic platform that greatly improves capture efficiency and readily realizes visible signal conversion for extracellular vesicle detection. The technology, termed high-affinity recognition and visual extracellular vesicle testing (HARVEST), leverages 2D flexible Fe3O4-MoS2 nanostructures to recognize extracellular vesicles through multidentate affinity binding and feasible magnetic separation, thus enhancing the extracellular vesicle capture performance with both yield and separation time, affording high sensitivity with the detection limit of 20 extracellular vesicle particles/μL. Through integration with lipid labeling chemistry and the fluorescence visualization system, the platform enables rapid and visible detection. The number of extracellular vesicles can be feasibly determined by smart mobile phones, readily adapted for point-of-care diagnosis. When clinically evaluated, the strategy accurately differentiates melanoma samples from the normal cohort with an AUC of 0.98, demonstrating the efficient extracellular vesicle detection strategy with 2D flexible platforms for cancer diagnosis.

99m Tc]Tc-HYNIC-EcgDf21: A defensin short analogue with potential application in infection foci imaging.

Opportunistic infections are a problem of great relevance in public health and the precise detection and localization of infection in the early stages of the disease is of great importance for patient management as well as cost containment. Our proposal seeks to contribute to developing a new agent that meets the needs of diagnosis and follow-up of fungal and bacterial infections, focused on the design of a radiotracer with the potential for recognition of hidden infection foci. Defensins are plant antimicrobial peptides that not only show activity against plant pathogens but also against human ones. A short analogue of EcgDf1 defensin, EcgDf21d (NH2 -ERFTGGHCRGFRRRCFCTKHC-COOH), was labelled through the formation of a 99m Tc-HYNIC complex which was assessed for physicochemical and biological behaviour both in vitro and in vivo. The [99m Tc]Tc-HYNIC-EcgDf21 labelling procedure rendered a single product with remarkably high RCP and stability in the labelling milieu. The Log p value indicated that [99m Tc]Tc-HYNIC-EcgDf21 has a hydrophilic behaviour, confirmed by the biodistribution profiles. The optimal uptake value was obtained for Candida albicans infection model reaching a lesion/muscle ratio of 3, this correlates with in vitro binding studies, and the lesion can be definitely observed in the scintigraphic images.

Introducing ISAP and MATSS: Mental stress induced speech utterance procedure and obtained dataset

Mental stress persisting for long can cause severe health issues. There are various approaches available in the literature for investigating stress through speech utterances. The available procedure to obtain speech under stress dataset requires the speakers to undergo the actual stress situations in a real environment with limited control or inducing stress with a mental task in a lab environment. These approaches either suffer from ethical issues or unreliable labeling of the obtained speech samples. In this paper, we attempt to overcome these limitations with Induced mental Stress based speech production And labeling Procedure (ISAP), for obtaining speech utterances under mental stress along with labeling the samples simultaneously. The proposed ISAP can be incorporated by future studies as per their need to create a speech under stress dataset. We also present the obtained dataset, the baseline experiments, and classification results with various machine learning models. A total of 1260 speech utterances are obtained, with ISAP able to induce stress in 54.4% of the cases. The accuracy of the SVM classifier in recognizing three stress classes, namely, No Stress, Low Stress, and High Stress is found to be 57.1%.

Large-scale group decision-making to facilitate inter-rater reliability of human-factors analysis for the railway system

The interdependency analysis of human factors usually needs to identify and label factors from accident/incident reports using a binary framework. The labeling procedure involves enormous knowledge and requires a large group of raters, but the raters’ opinions usually highly contradict each other. Thus, inter-rater reliability should be implemented to improve the labeling consistency. However, the inter-rater reliability coefficient is commonly computed on a category-by-category basis, which is inefficient especially when there are many raters. To overcome this deficiency, a feedback-autonomy-based consensus model is proposed to determine the inter-rater reliability of human-factors labeling results involving a large group of raters. The proposed model can manage various non-cooperative behaviors and provide feedback adjustment suggestions to raters for the labeling procedure. Notably, the autonomy mechanism allows raters to freely choose and adjust labels referring to the reference opinions without causing over-adjustment issue. The inter-rater reliability and the interdependency results are performed on 279 accident/incident reports. We apply a commercial tool to obtain the reference results, which are consistent with the unanimous group opinions. Besides, other comparative studies clarify the advantages of the proposed consensus model. The proposed model is convenient for reducing conflicts among raters that usually exist in human-factors analysis.

An interpretable unsupervised Bayesian network model for fault detection and diagnosis

Process monitoring is a critical activity in manufacturing industries. A wide variety of data-driven approaches have been developed and employed for fault detection and fault diagnosis. Analyzing the existing process monitoring schemes, prediction accuracy of the process status is usually the primary focus while the explanation (diagnosis) of a detected fault is relegated to a secondary role. In this paper, an interpretable unsupervised machine learning model based on Bayesian Networks (BN) is proposed to be the fundamental model supporting the process monitoring scheme. The proposed methodology is aligned with the recent efforts of eXplanatory Artificial Intelligence (XAI) for knowledge induction and decision making, now brought to the scope of advanced process monitoring. A BN is capable of combining data-driven induction with existing domain knowledge about the process and to display the underlying causal interactions of a process system in an easily interpretable graphical form. The proposed fault detection scheme consists of two levels of monitoring. In the first level, a global index is computed and monitored to detect any deviation from normal operation conditions. In the second level, two local indices are proposed to examine the fine structure of the fault, once it is signaled at the first level. These local indices support the diagnosis of the fault, and are based on the individual unconditional and conditional distributions of the monitored variables. A new labeling procedure is also proposed to narrow down the search and identify the fault type. Unlike many existing diagnosis methods that require access to faulty data (supervised diagnosis methods), the proposed diagnosis methodology belongs to the class that only requires data under normal conditions (unsupervised diagnosis methods). The effectiveness of the proposed monitoring scheme is demonstrated and validated through simulated datasets and an industrial dataset from semiconductor manufacturing.

Aptamer recognition-promoted specific intercalation of iridium complexes in G-quadruplex DNA for label-free and enzyme-free phosphorescence analysis of kanamycin

In consideration of relevance of antibiotic with food security, it is extremely desirable to propose sensitive and credible methods for antibiotic screening. Nevertheless, most of known approaches are developed based on fluorescence technique, which suffered from the interferences of background fluorescence and autoluminescence, and tedious labeling procedures, ascribing to the deficiency of high-performance and multifunctional dyes. Herein, we developed a novel iridium (III) complex (Ir-QAU)-based aptamer-promoted phosphorescence sensor for label-free, enzyme-free and highly sensitive detection of target antibiotic (kanamycin, Kan) based on target-switched hybridizing chain reaction (HCR). Ir-QAU was elaborately devised to present a signal-on response to G-quadruplex (G4) DNA against other DNAs due to its specific intercalation in G4 DNA and subsequent restriction of intra-molecular rotation. The recognition of H1 by Kan promoted the formation of Kan@H1 complexes, which hybridized with H2 and H3 via toehold-mediated hybridization reaction, subsequently switching HCR to produce large numbers of G4 DNA. Compared to Kan absence, abundant Ir-QAU was locked in G4 DNA to yield a significantly increased luminescence, which switches the luminescence analysis process of Kan with a limit of detection down to 0.38 pM. Furthermore, the Ir-QAU-based sensor was triumphantly applied to detect Kan in milk sample. We anticipate this work will disclose a new way to development of high-efficiency and practical luminescence sensor, and show a great potential for antibiotic-related food security.

An ultrasensitive electrochemical self-signal circulating tumor DNA recognition strategy employing black phosphorous nanosheets assembled with flavin adenine dinucleotide

A self-signal electrochemical sensing platform was constructed for direct recognition of circulating tumor DNA (ctDNA) employing black phosphorous nanosheets (BPNS) assembled with flavin adenine dinucleotide (FAD) prepared by ultrasonication approach. FAD provided a highly efficient and stable dispersing medium for acquiring highly dispersed BPNS in aqueous phase. The obtained FAD/BPNS nanocomposite exhibited favorable electrochemical redox activity and was utilized as the interface for the immobilization and hybridization of DNA. The amine-terminated probe ssDNA was covalently assembled onto the FAD/BPNS nanocomposite with plentiful phosphonate groups accompanied by the decrease of the self-signal. The self-redox signal of the nanointerface regenerated after the probe hybridized with the complementary sequence as a result of DNA transformation. Electrochemical response enhanced with complementary DNA concentration from 1.0 × 10-18 to 1.0 × 10-8 mol/L with a detection limit of 2.6 × 10-19 mol/L. The DNA determination platform revealed outstanding sensitivity, specificity and stableness, and was successfully employed in the detection of ctDNA associated with colorectal cancer. The developed biosensing strategy is simple to accomplish and has the potency for the application for diverse morbific gene without sophisticated label procedure.

Differential CFTR-Interactome Proximity Labeling Procedures Identify Enrichment in Multiple SLC Transporters.

Proteins interacting with CFTR and its mutants have been intensively studied using different experimental approaches. These studies provided information on the cellular processes leading to proper protein folding, routing to the plasma membrane, recycling, activation and degradation. Recently, new approaches have been developed based on the proximity labeling of protein partners or proteins in close vicinity and their subsequent identification by mass spectrometry. In this study, we evaluated TurboID- and APEX2-based proximity labeling of WT CFTR and compared the obtained data to those reported in databases. The CFTR-WT interactome was then compared to that of two CFTR (G551D and W1282X) mutants and the structurally unrelated potassium channel KCNK3. The two proximity labeling approaches identified both known and additional CFTR protein partners, including multiple SLC transporters. Proximity labeling approaches provided a more comprehensive picture of the CFTR interactome and improved our knowledge of the CFTR environment.

Assessing Cellular Uptake of Exogenous Coenzyme Q10 into Human Skin Cells by X-ray Fluorescence Imaging.

X-ray fluorescence (XRF) imaging is a highly sensitive non-invasive imaging method for detection of small element quantities in objects, from human-sized scales down to single-cell organelles, using various X-ray beam sizes. Our aim was to investigate the cellular uptake and distribution of Q10, a highly conserved coenzyme with antioxidant and bioenergetic properties. Q10 was labeled with iodine (I2-Q10) and individual primary human skin cells were scanned with nano-focused beams. Distribution of I2-Q10 molecules taken up inside the screened individual skin cells was measured, with a clear correlation between individual Q10 uptake and cell size. Experiments revealed that labeling Q10 with iodine causes no artificial side effects as a result of the labeling procedure itself, and thus is a perfect means of investigating bioavailability and distribution of Q10 in cells. In summary, individual cellular Q10 uptake was demonstrated by XRF, opening the path towards Q10 multi-scale tracking for biodistribution studies.

Gene-dosage effect of Pfkfb3 on monocyte/macrophage biology in atherosclerosis.

Macrophage-rich atherosclerotic arteries are highly active in glycolysis. PFKFB3, a key glycolytic enzyme, has emerged as a potential therapeutic target in atherosclerosis. Small-molecule inhibitors of PFKFB3, such as 3PO and PFK158, have demonstrated efficacy in hampering atherogenesis in preclinical models. However, genetic studies elucidating the role of Pfkfb3 in atherogenesis need to be conducted to validate pharmacological findings and to unveil potential pharmacological side effects. Apoe-/- mice with global heterozygous or myeloid cell-specific Pfkfb3 deficiency were fed a Western diet (WD), after which atherosclerosis development was determined. Monocyte subsets in atherosclerotic mice and patients were examined by flow cytometry. Monocyte infiltration was assayed by a Ly6Chi monocyte-specific latex labelling procedure. In situ efferocytosis was assessed on mouse aortic root sections. Additionally, metabolic status, macrophage motility, efferocytosis, and involved mechanisms were analysed in peritoneal macrophages. Global heterozygous or myeloid cell-specific Pfkfb3 deficiency reduced atherogenesis in Apoe-/- mice. Mechanistic studies showed that PFKFB3 controlled the proliferation and infiltration of proinflammatory monocytes. Moreover, PFKFB3 expression was associated with inflammatory monocyte expansion in patients with atherosclerotic coronary artery disease. Surprisingly, homozygous loss of Pfkfb3 impaired macrophage efferocytosis and exacerbated atherosclerosis in Apoe-/- mice. Mechanistically, PFKFB3-driven glycolysis was shown to be essential for actin polymerization, thus aiding the efferocytotic function of macrophages. Collectively, these findings suggest the existence of a double-edged sword effect of myeloid PFKFB3 on the pathogenesis of atherosclerosis and highlight the need for caution in developing anti-atherosclerotic strategies that target PFKFB3.

Wettability‐Based Cell Sorting: Exploring Label‐Free Isolation Strategy for Mixed Primary Glial Cell Population (Adv. Mater. Interfaces 22/2022)

Wettability-Based Cell Sorting A wettability-controlled substrate can be used to separate multiple cell types without the need for a complex labeling procedure, as shown in article number 2200388 by Eunyoung Park, Jennifer Shin, and co-workers. Mixed glial brain cells cultured on a hydrophobic substrate spontaneously segregate into substrate-adherent microglia (green-yellow) and spheroidal aggregates of astrocyte/oligodendrocyte progenitor cells (pink-orange). The hydrophobic poly(cyclohexyl methacrylate) (pCHMA) thin-film substrate permits the natural segregation of glial cell subtypes with dissimilar adhesion properties.

Live Imaging of Microtubule Dynamics in Glioblastoma Cells Invading the Zebrafish Brain.

With a dismal median survival time in real populations-between 6 to 15 months-glioblastoma (GBM) is the most devastating malignant brain tumor. Treatment failure is mainly due to the invasiveness of GBM cells, which speaks for the need for a better understanding of GBM motile properties. To investigate the molecular mechanism supporting GBM invasion, new physiological models enabling in-depth characterization of protein dynamics during invasion are required. These observations would pave the way to the discovery of novel targets to block tumor infiltration and improve patient outcomes. This paper reports how an orthotopic xenograft of GBM cells in the zebrafish brain permits subcellular intravital live imaging. Focusing on microtubules (MTs), we describe a procedure for MT labeling in GBM cells, microinjecting GBM cells in the transparent brain of 3 days post fertilization (dpf) zebrafish larvae, intravital imaging of MTs in the disseminating xenografts, altering MT dynamics to assess their role during GBM invasion, and analyzing the acquired data.

Microscale Thermophoresis and Molecular Modelling to Explore the Chelating Drug Transportation in the Milk to Infant.

The microscale thermophoresis (MST) technique was utilized to investigate lactoferrin–drug interaction with the iron chelator, deferiprone, using label-free system. MST depends on the intrinsic fluorescence of one interacting partner. The results indicated a significant interaction between lactoferrin and deferiprone. The estimated binding constant for the lactoferrin–deferiprone interaction was 8.9 × 10−6 ± 1.6, SD, which is to be reported for the first time. Such significant binding between lactoferrin and deferiprone may indicate the potentiation of the drug secretion into a lactating mother’s milk. The technique showed a fast and simple approach to study protein–drug interaction while avoiding complicated labeling procedures. Moreover, the binding behavior of deferiprone within the binding sites of lactoferrin was investigated through molecular docking which reflected that deferiprone mediates strong hydrogen bonding with ARG121 and ASP297 in pocket 1 and forms H-bond and ionic interaction with ASN640 and ASP395, respectively, in pocket 2 of lactoferrin. Meanwhile, iron ions provide ionic interaction with deferiprone in both of the pockets. The molecular dynamic simulation further confirmed that the binding of deferiprone with lactoferrin brings conformational changes in lactoferrin that is more energetically stable. It also confirmed that deferiprone causes positive correlation motion in the interacting residues of both pockets, with strong negative correlation motion in the loop regions, and thus changes the dynamics of lactoferrin. The MM-GBSA based binding free energy calculation revealed that deferiprone exhibits ∆G TOTAL of −63,163 kcal/mol in pocket 1 and −63,073 kcal/mol in pocket 2 with complex receptor–ligand difference in pocket 1 and pocket 2 of −117.38 kcal/mol and −111.54 kcal/mol, respectively, which in turn suggests that deferiprone binds more strongly in the pocket 1. The free energy landscape of the lactoferrin–deferiprone complex also showed that this complex remains in a high energy state that confirms the strong binding of deferiprone with the lactoferrin. The current research concluded that iron-chelating drugs (deferiprone) can be transported from the mother to the infant in the milk because of the strong attachment with the lactoferrin active pockets.

Feasibility of 18F-FDG Labelling of Leucocytes in a Centre Without an On-Site Cyclotron and Monitoring of Radiation Dose to Occupational Worker in the Labelling Procedure.

Objectives: Differentiation of infection from sterile inflammation is still a major concern for clinicians. The 18F-WBC positron emission tomography/computed tomography scan has been considered a promising tool for accurate diagnosis of infection owing to its high specificity, but it renders the availability of a medical cyclotron a necessity. The aim of the present study was to determine the feasibility of labeling leukocytes and establish the protocol in a center without the availability of an on-site medical cyclotron. The secondary aim was to monitor radiation doses to occupational workers involved in labeling of leukocytes with 18F-FDG. Materials and Methods: Leukocyte separation was performed and leukocytes were radiolabeled with 18F-FDG in a sterile environment according to the procedure described by Bhattacharya et al. In vitro leukocyte viability was assessed using the trypan dye exclusion technique. Labeling efficiency and yield were also estimated for all radiolabeling procedures. Whole-body and extremity doses received by the personnel involved in the radiolabeling procedure were also estimated using pocket dosimeters. Results: Leukocyte labeling was carried out in 35 runs, during which there were two failed labeling attempts due to clotting of the blood sample. The total time involved in the whole procedure was around 2.5 h. The average labeling efficiency was 78.01% ± 6.99% (range 63.46%-86.54%), cell viability was 98%, and the cell suspension was stable up to 4 h. The mean dose was measured as 17 μSv at the chest level and 32 μSv at the extremity level, per procedure. Conclusions: Labeling of leukocytes with 18F-FDG is possible at a tertiary nuclear medicine setup without the availability of an on-site medical cyclotron, with reasonable labeling efficiency of 78.01% ± 6.99%. In addition, in-house labeling of leukocytes with 18F-FDG is safe and the radiation doses incurred by the personnel during the labeling procedure are well within the occupational dose limits established by the national regulatory authority.

Preclinical studies of automated radiolabeled microcarriers for radiosynovectomy of inflammatory joint disease

The broad implementation of therapeutic loaded nano- and microcarriers has led to significant progress in the treatment of inflammatory joint diseases, especially in the case of radiological synovectomy using β-emitting radioisotope labeled carriers. However, the lack of automated procedures for radiolabeling of these carriers seriously limits their further translation into clinical practice. Here, we develop the automated process of microcarriers radiolabeling for radiosynovectomy of inflammatory joint disease (synovitis in an arthritis rat model). The fabricated polymeric microparticles (MPs) possess high biocompatibility in vitro and in vivo, can be effectively loaded with 188Re therapeutic radionuclide (∼ 97,98%), and exhibit good radiochemical stability (up to 95%). In vivo biodistribution demonstrates the accumulation of intra-articular injected 188Re-MPs in the knee joint (> 96%ID/g for 7 d) without radiation exposure to other healthy organs. It has been shown that the applied radiosynovectomy with 188Re-MPs can significantly reduce knee joint swelling (knee joint with synovitis ∼ 1.8 ± 0.1 cm versus treated knee joint ∼ 1.1 ± 0.1 cm) and leads to the improvement of inflammation in synovium and cartilage recovery, decreasing the levels of inflammatory factors (TNF-α, IL-6, and IL-1b). Although there are very limited clinical trials on radiosynovectomy of inflammatory joint disease, this study is a step towards implementation of the developed automated labeling procedure for the real application of radiosynovectomy with 188Re in clinics.

Wettability‐Based Cell Sorting: Exploring Label‐Free Isolation Strategy for Mixed Primary Glial Cell Population

AbstractGlial brain cells, including astrocytes, oligodendrocytes, and microglia, have received much attention as crucial players in neurological diseases. As a result of their critical roles, numerous in vitro studies are being conducted, necessitating the use of appropriate isolation methods for different glial cell types. The most effective glial isolation at the moment is labeling‐based protocols that require expensive antibodies and equipment. More commonly used label‐free methods are better known for higher accessibility but with compromised accuracy and longer isolation time. In this study, a cost‐effective surface wettability‐based isolation protocol for mixed glial cell population that does not require labeling is established. Specifically, when cultured on a hydrophobized substrate coated with a thin‐film of poly (cyclohexyl methacrylate)(pCHMA), glial cells spontaneously segregate into substrate‐adherent microglia and spheroids of astrocyte/oligodendrocyte progenitor cells in suspension. In comparison to existing label‐free isolation methods, the strategic approach ensures rapid isolation and high purity. Furthermore, this is the first to demonstrate that the hydrophobic property of pCHMA‐coated substrate can be used to sort multiple brain glial cell types based on their differential adhesion properties without the need for complex labeling procedure. The study emphasizes the wettability‐controlled substrate's potential for segregating different cell types of various pathophysiological states.