Commercial Human Serum Research Articles

The novel immunomodulatory biologic LMWF5A for pharmacological attenuation of the \u201ccytokine storm\u201d in COVID-19 patients: a hypothesis

BackgroundA common complication of viral pulmonary infections, such as in the ongoing COVID-19 pandemic, is a phenomenon described as a “cytokine storm”. While poorly defined, this hyperinflammatory response results in diffuse alveolar damage. The low molecular weight fraction of commercial human serum albumin (LMWF5A), a novel biologic in development for osteoarthritis, demonstrates beneficial in vitro immunomodulatory effects complimentary to addressing inflammation, thus, we hypothesize that LMWF5A could improve the clinical outcomes of COVID-19 by attenuating hyperinflammation and the potential development of a cytokine storm.Presentation of the hypothesisA variety of human in vitro immune models indicate that LMWF5A reduces the production of pro-inflammatory cytokines implicated in cytokine storm associated with COVID-19. Furthermore, evidence suggests LMWF5A also promotes the production of mediators required for resolving inflammation and enhances the barrier function of endothelial cultures.Testing the hypothesisA randomized controlled trial, to evaluate the safety and efficacy of nebulized LMWF5A in adults with Acute Respiratory Distress Syndrome (ARDS) secondary to COVID-19 infection, was developed and is currently under review by the Food and Drug Administration.Implications of hypothesisIf successful, this therapy may attenuate the cytokine storm observed in these patients and potentially reduce mortality, increase ventilation free days, improve oxygenation parameters and consequently lessen the burden on patients and the intensive care unit.ConclusionsIn conclusion, in vitro findings suggest that the immunomodulatory effects of LMWF5A make it a viable candidate for treating cytokine storm and restoring homeostasis to the immune response in COVID-19.

Selective Detection of Folic Acid Using 3D Polymeric Structures of 3-Carboxylic Polypyrrole.

The detection of folic acid in biological samples or pharmaceutical products is of great importance due to its implications in the biological functions of the human body, along with the development and growth of the fetus. The deficiency of folic acid can be reversed by the intake of different pharmaceutical formulations or alimentary products fortified with this molecule. The elaboration of sensing platforms represents a continuous work in progress, a task in which the use of conductive polymers modified with different functionalities represents one of the outcoming strategies. The possibility of manipulating their morphology with the use of templates or surfactants represents another advantage. A sensing platform based on carboxylic functionalized polypyrrole was synthesized via the electrochemical approach in the presence of a polymeric surfactant on a graphite-based surface. The sensor was able to detect the folic acid from 2.5 μM to 200 μM with a calculated limited of detection of 0.8 μM. It was employed for the detection of the analyte from commercial human serum and pharmaceutical products with excellent recovery rates. The results were double checked using an optimized spectrophotometric procedure that confirmed furthermore the performances of the sensor related to real samples assessment.

Modification of chlorosulfonated polystyrene substrates for bioanalytical applications.

In this work the modification of polystyrene micro-well plates and their use as bioanalytical platform is described. A wet-chemical procedure was applied for the chlorosulfonation of these polystyrene substrates (PS) resulting in well-controlled and reactive surfaces. This method enabled the production of transparent and stable substrates under ambient conditions. The chlorosulfonyl moieties at the substrate surface were converted under mild conditions into different functional groups. The modification of PS served to increase the hydrophilic properties of the surface and thus, the improvement of interaction with biocompounds. The resulting substrates were characterized by contact angle measurements, X-ray Photoelectron Spectroscopy and colorimetry. PS substrates modified with different functional groups and attachment approaches (covalent link and direct adsorption of the antibodies) were used as the platform for immunoassays and the results compared to a commercial Human Serum Albumin ELISA kit. Aminated surfaces gave better results than those with carboxyl, alkene or epoxy groups and even the commercial kit.

Enhanced Performance of Reagent-Less Carbon Nanodots Based Enzyme Electrochemical Biosensors.

This work reports on the advantages of using carbon nanodots (CNDs) in the development of reagent-less oxidoreductase-based biosensors. Biosensor responses are based on the detection of H2O2, generated in the enzymatic reaction, at 0.4 V. A simple and fast method, consisting of direct adsorption of the bioconjugate, formed by mixing lactate oxidase, glucose oxidase, or uricase with CNDs, is employed to develop the nanostructured biosensors. Peripherical amide groups enriched CNDs are prepared from ethyleneglycol bis-(2-aminoethyl ether)-N,N,N′,N′-tetraacetic acid and tris(hydroxymethyl)aminomethane, and used as precursors. The bioconjugate formed between lactate oxidase and CNDs was chosen as a case study to determine the analytical parameters of the resulting L-lactate biosensor. A linear concentration range of 3.0 to 500 µM, a sensitivity of 4.98 × 10−3 µA·µM−1, and a detection limit of 0.9 µM were obtained for the L-lactate biosensing platform. The reproducibility of the biosensor was found to be 8.6%. The biosensor was applied to the L-lactate quantification in a commercial human serum sample. The standard addition method was employed. L-lactate concentration in the serum extract of 0.9 ± 0.3 mM (n = 3) was calculated. The result agrees well with the one obtained in 0.9 ± 0.2 mM, using a commercial spectrophotometric enzymatic kit.

The presence of S-sulfonated transthyretin in commercial human serum albumin solutions: Potential contribution to neuropathy

BackgroundCommercial solutions of human serum albumin (HSA) are administered to critically ill patients for the treatment of shock, restoration of blood volume, and the acute management of burns. Previously, conflicting results on the effects of HSA administration have been reported varying from a favorable increase in total plasma antioxidant capacity to a higher mortality rate in traumatic brain injury (TBI) patients. These results could be partially explained due to the known heterogeneity of HSA solutions. We report the discovery of S-sulfonated human transthyretin (hTTR) in HSA solutions. MethodsProteomics was performed on commercially available solutions of 5% HSA by LC-MS analysis. The MS charge envelope for hTTR was deconvolved to the uncharged native hTTR parent mass (13,762 Da). The parent mass was then integrated, and relative proportions of the 2 major species of hTTR, native and S-sulfonated hTTR (13,842 Da), were calculated. ResultsThe majority of hTTR found in 5% commercial HSA solutions is in the S-sulfonated form regardless of the age of the HSA solution. S-sulfonation of hTTR at the free cysteine residue in position 10 appears to be the result of a mixed disulfide exchange possibly with S-cysteinylated hTTR or S-cysteinylated HSA. hTTR is a tetramer composed of four identical monomers each containing a reduced cysteine residue in position 10. S-sulfonation of hTTR at this cysteine residue can destabilize the hTTR tetramer, an important step in the formation of TTR-related amyloid fibrils. ConclusionsAdministration of a commercial HSA solution that already contains S-sulfonated hTTR could potentially contribute to the development of amyloid-related/polyneuropathy in the critically ill.

On the Mechanisms of Action of the Low Molecular Weight Fraction of Commercial Human Serum Albumin in Osteoarthritis.

The low molecular weight fraction of commercial human serum albumin (LMWF5A) has been shown to successfully relieve pain and inflammation in severe osteoarthritis of the knee (OAK). LMWF5A contains at least three active components that could account for these anti-inflammatory and analgesic effects.We summarize in vitro experiments in bone marrow–derived mesenchymal stem cells, monocytic cell lines, chondrocytes, peripheral blood mononuclear cells, fibroblast-like synoviocytes, and endothelial cells on the biochemistry of anti-inflammatory changes induced by LMWF5A. We then look at four of the major pathways that cut across cell-type considerations to examine which biochemical reactions are affected by mTOR, COX-2, CD36, and AhR pathways. All three components show anti-inflammatory activities in at least some of the cell types.The in vitro experiments show that the effects of LMWF5A in chondrocytes and bone marrow–derived stem cells in particular, coupled with recent data from previous clinical trials of single and multiple injections of LMWF5A into OAK patients demonstrated improvements in pain, function, and Patient Global Assessment (PGA), as well as high responder rates that could be attributed to the multiple mechanism of action (MOA) pathways are summarized here. In vitro and in vivo data are highly suggestive of LMWF5A being a disease-modifying drug for OAK.

Assessment of total, ligand-induced peroxisome proliferator activated receptor \u03b3 ligand activity in serum

BackgroundHumans are exposed to a complex mixture of environmental chemicals that impact bone and metabolic health, and traditional exposure assessments struggle to capture these exposure scenarios. Peroxisome proliferator activated receptor-gamma (PPARγ) is an essential regulator of metabolic and bone homeostasis, and its inappropriate activation by environmental chemicals can set the stage for adverse health effects. Here, we present the development of the Serum PPARγ Activity Assay (SPAA), a simple and cost-effective method to measure total ligand activity in small volumes of serum.MethodsFirst, we determined essential components of the bioassay. Cos-7 cells were transfected with combinations of expression vectors for human PPARγ and RXRα, the obligate DNA-binding partner of PPARγ, along with PPRE (DR1)-driven luciferase and control eGFP reporter constructs. Transfected cells were treated with rosiglitazone, a synthetic PPARγ ligand and/or LG100268, a synthetic RXR ligand, to characterize the dose response and determine the simplest and most efficacious format. Following optimization of the bioassay, we assessed the cumulative activation of PPARγ by ligands in serum from mice treated with a PPARγ ligand and commercial human serum samples.ResultsCos-7 cells endogenously express sufficient RXR to support efficacious activation of transfected PPARγ. Co-transfection of an RXR expression vector with the PPARγ expression vector did not increase PPRE transcriptional activity induced by rosiglitazone. Treatment with an RXR ligand marginally increased PPRE transcriptional activity in the presence of transfected PPARγ, and co-treatment with an RXR ligand reduced rosiglitazone-induced PPRE transcriptional activity. Therefore, the final bioassay protocol consists of transfecting Cos-7 cells with a PPARγ expression vector along with the reporter vectors, applying rosiglitazone standards and/or 10 μL of serum, and measuring luminescence and fluorescence after a 24 h incubation. Sera from mice dosed with rosiglitazone induced PPRE transcriptional activity in the SPAA in a dose-dependent and PPARγ-dependent manner. Additionally, human serum from commercial sources induced a range of PPRE transcriptional activities in a PPARγ-dependent manner, demonstrating the ability of the bioassay to detect potentially low levels of ligands.ConclusionsThe SPAA can reliably measure total PPRE transcriptional activity in small volumes of serum. This system provides a sensitive, straightforward assay that can be reproduced in any cell culture laboratory.

Facile synthesis of cellulose microfibers supported palladium nanospindles on graphene oxide for selective detection of dopamine in pharmaceutical and biological samples

The cost-effective synthesis of novel functional nanomaterials has received significant attention in the physical and chemical sciences due to their improved surface area, high catalytic activity along with unique morphological features. This paper reports a facile and eco-friendly synthesis of spindle-like palladium nanostructures (PdSPs) on graphene oxide-cellulose microfiber (GO-CMF) composite for the first time. The GO-CMF/PdSPs composite was synthesized by an electrochemical method without the use of additional surfactants and capping agents. The synthesized materials were characterized and confirmed by using transmission electron microscopy, high-resolution scanning electron microscopy, X-ray diffraction spectroscopy, Raman spectroscopy and Fourier-transform infrared spectroscopy. As-synthesized GO-CMF/PdSPs composite modified electrode was used as a selective electrocatalyst for the oxidation of dopamine (DA). The electrochemical redox behaviors of DA were investigated using cyclic voltammetry (CV). The CV results revealed that the GO-CMF/PdSPs composite modified electrode has 10 folds enhanced oxidation current response to DA than GO, PdSPs and GO-CMF modified GCEs. Under optimized conditions, the GO-CMF/PdSPs composite sensor exhibits a linear response to DA in the concentration range from 0.3 to 196.3 μM with the lower detection limit of 23 nM. The nanocomposite electrode also shows promising features towards the reliable and selective detection of DA, which includes high stability, reproducibility and high selectivity towards the commonly interfering species such as ascorbic acid, uric acid, and dihydroxybenzene isomers. The sensor was successfully tested for the real-time detection of DA in the commercial DA injections and human serum samples.

Stepwise frontal affinity chromatography model for drug and protein interaction.

Frontal affinity chromatography is an efficient technique that combines affinity interaction and high-performance liquid chromatography, and frontal analysis has been used in studying the interaction between drugs and proteins. Based on frontal analysis, stepwise frontal analysis has been established. The present study aimed to use the Lineweaver-Burk plot in stepwise frontal analysis by taking the weighted average of time data. Commercial human serum albumin (HSA) and alpha-1-acid glycoprotein (AGP) columns were used as an affinity column. Warfarin and digitoxin were chosen as model drugs for the HSA column, whereas verapamil and tamsulosin were selected as model drugs for the AGP column. The time data obtained by frontal analysis and stepwise frontal analysis were compared, and the results revealed good correlation (r2 = 0.9946-0.9998). Frontal analysis and stepwise frontal analysis were also used to analyze the equilibrium dissociation constants (Kd) of model drugs on the HSA and AGP columns. The Kd values were compared with literature values, which revealed the same order of magnitude. These results illustrate that conversion of the time data is reasonable and feasible. The Lineweaver-Burk plot can be used in the stepwise frontal analysis model to study the characteristics of the interaction between drugs and proteins. Graphical abstract ᅟ.

Enhanced amperometric detection of paracetamol by immobilized cobalt ion on functionalized MWCNTs - Chitosan thin film

In the present study, a nanocomposite of f-MWCNTs-chitosan-Co was prepared by the immobilization of Co(II) on f-MWCNTs-chitosan by a self-assembly method and used for the quantitative determination of paracetamol (PR). The composite was characterized by field emission scanning electron microscopy (FESEM) and energy dispersive x-ray analysis (EDX). The electroactivity of cobalt immobilized on f-MWCNTs-chitosan was assessed during the electro-oxidation of paracetamol. The prepared GCE modified f-MWCNTs/CTS-Co showed strong electrocatalytic activity towards the oxidation of PR. The electrochemical performances were investigated by cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS) and differential pulse voltammetry (DPV). Under favorable experimental conditions, differential pulse voltammetry showed a linear dynamic range between 0.1 and 400 μmol L−1 with a detection limit of 0.01 μmol L−1 for the PR solution. The fabricated sensor exhibited significant selectivity towards PR detection. The fabricated sensor was successfully applied for the determination of PR in commercial tablets and human serum sample.

The impact of oxidative stress on binding of drugs with plasma proteins studied by fluorescence anisotropy methods.

The aim of this study was to investigate the effect of oxidative stress on drug binding affinity, free and bound fraction. The fluorescence anisotropy method was used to examine the interactions of commercial human serum albumin (HSA) and human plasma proteins with ochratoxin A (OTA) or carboxylate form of camptothecin (CPT-C). In-vitro method revealed significant reduction in bound fraction of drugs to HSA oxidized by chloramine T. Oxidative stress was determined by measuring the plasma level of advanced oxidation protein products (AOPP). A significant positive correlation between the AOPP and the plasma free fraction of tested drugs in thirty healthy patients was also found. The oxidative stress monitoring by AOPP is very important for improvement of dosage of drugs highly bound to albumin and with narrow therapeutic index. As a result of severe oxidative stress, the drug pharmacokinetics and therapeutic effects are prone to change. This study highlights the issues of therapeutic drug monitoring and it can explain the behaviour of drugs in pathological situations.

Spontaneous Release of Human Serum Albumin S-Bound Homocysteine in a Thiol-Free Physiological Medium

Elevated plasma homocysteine (Hcy) concentrations independently predict cardiovascular disease. However, the transport of Hcy into pathological tissues such as atherosclerotic plaques, characterized by relatively low local thiol concentrations, is largely unknown. We sought to address if albumin-bound Hcy can be released in a thiol-free medium with or without previous incubation with Hcy and homocysteine thiolactone (HTL). We found that Hcy release was dependent on the baseline amount of albumin-bound Hcy. After 48 h incubation in a thiol- free medium the quantity of albumin-bound Hcy released from commercial human serum albumin (HSA) was 1.15 mmol/mol prot. If HSA was pre-incubated with 50 µmol/L reduced Hcy and then transferred into a free-thiol medium (HSA-S-Hcy), the amount of Hcy released after 48 h increased to 33.5 mmol/mol prot. If HSA was pre-incubated with 5 mmol/L HTL and then with 50 µmol/L of reduced Hcy (HSA-HTL-S-Hcy), the amount of Hcy released increased to 92.8 mmol/mol prot. Hcy release from HSA-HTL-S-Hcy further increased in presence of cysteine (Cys), glutathione (GSH), or Cys + GSH in the medium. Therefore, a significant amount of albumin-bound Hcy is released into thiol-free environments, similar to atherosclerotic plaques, with potential deleterious effects on vascular homeostasis, atherosclerosis and thrombosis.

Commutability of control materials for external quality assessment of serum apolipoprotein A-I measurement.

The aim of the current study was to evaluate the commutability of commercial control materials and human serum pools and to investigate the suitability of the materials for the external quality assessment (EQA) of serum apolipoprotein A-I (apo A-I) measurement. The Clinical and Laboratory Standards Institute (CLSI) EP14-A3 protocol was used for the commutability study. Apo A-I concentrations in two levels of commercial control materials used in EQA program, two fresh-frozen human serum pools (FSPs) and two frozen human serum pools prepared from residual clinical specimens (RSPs) were measured along with 50 individual samples using nine commercial assays. Measurement results of the 50 individual samples obtained with different assays were pairwise analyzed by Deming regression, and 95% prediction intervals (PIs) were calculated. The commutability of the processed materials was evaluated by comparing the measurement results of the materials with the limits of the PIs. The FSP-1 was commutable for all the 36 assay pairs, and FSP-2 was commutable for 30 pairs; RSP-1 and RSP-2 showed commutability for 27/36 and 22/36 assay pairs, respectively, whereas the two EQA materials were commutable only for 4/36 and 5/36 assay pairs, respectively. Non-commutability of the tested EQA materials has been observed among current apo A-I assays. EQA programs need either to take into account the commutability-related biases in the interpretation of the EQA results or to use more commutable materials. Frozen human serum pools were commutable for most of the assays.

Synthesis of neodymium-iron nanoperovskite for sensing applications of an antiallergic drug

A facile route was used for the fabrication of NdFeO$_{3}$ (NF) perovskite. The orthorhombic structure of single phase nanocrystalline NdFeO$_{3}$ perovskite was confirmed by different characterization techniques: XRD, FTIR, TEM, BET, SEM, and EDAX. The nanoperovskite was successfully used as a sensor component with a carbon paste electrode (CpE) for the determination of ketotifen in pharmaceutical formulations. Ketotifen, an antiallergic drug, is utilized for the treatment of asthma attacks, rhinitis, skin allergies, and anaphylaxis. Sodium dodecyl sulfate (SDS) was employed in a surface assembly mode to improve the sensitivity of the sensor response. The proposed sensor, NdFeO$_{3}$ in situ modified carbon paste electrode in the presence of SDS (NFMCpE-SDS), showed a higher current response towards ketotifen oxidation compared to other surfaces. The enhanced signal response is attributed to the catalytic properties of the sensor and the relative increase in surface area. A linear relationship between the oxidation peak current and ketotifen concentration was obtained in the concentration range of 0.8 $\mu $mol L$^{-1}$ to 360 $\mu $mol L$^{-1}$. The surface showed long-term stability of 1 month of operation and the limits of detection and quantification of ketotifen were 2.92 nmol L$^{-1}$ and 9.71 nmol L$^{-1}$, respectively, with high sensitivity of 311 $\mu $A/mmol L$^{-1}$. The composite electrode proved to be selective for drug sensing in the presence of different interfering species and applicable for drug determination in commercial tablets, human serum, and urine.

Elimination of erroneous results in flow cytometry caused by antibody binding to Fc receptors on human monocytes and macrophages.

Nonspecific binding of monoclonal antibodies (mAbs) to Fc-receptors on leukocytes is an important cause of background fluorescence in flow cytometry, and failing to block such nonspecific binding can lead to erroneous results. A major part of previous studies on blocking reagents for flow cytometry have been done in mice, and published results are not completely in agreement. In humans, Fc-receptors are found on most leukocytes, with highest abundance on monocytes/macrophages. Therefore, in the present study our aim was to thoroughly investigate the efficiency of different commonly used blocking reagents regarding inhibition of nonspecific binding of mouse mAbs to human peripheral blood mononuclear cells (MNCs) and monocyte-derived macrophages (MDMs). Monocytes and MDMs showed strong nonspecific binding of IgG1 and IgG2a isotypes, but not IgG2b. In contrast, B-cells, T-cells, and NK-cells did not substantially bind any of the mouse isotype control antibodies evaluated (IgG1, IgG2a, and IgG2b). Importantly, we show that binding of IgG1 and IgG2a to monocytes and MDMs can be eliminated by blocking, either with a commercial Fc-blocking reagent, with mouse or human serum, or with mouse or human IgG in high concentration. Previously, isotype controls have been widely used in flow cytometry assays. However, we show that such controls may be highly unreliable, and we believe they should not be used as gating controls, or to determine background signal. Based on these results, as well as considerations of price and applicability, our recommendation is not to use isotypes as gating controls in flow cytometry, but instead to use 100 μg/mL of purified human IgG as blocking reagent for elimination of nonspecific binding of mouse mAbs to human MNCs and MDMs. © 2016 International Society for Advancement of Cytometry.

A Human Serum-Based Enzyme-Free Continuous Glucose Monitoring Technique Using a Needle-Type Bio-Layer Interference Sensor

The incidence of diabetes is continually increasing, and by 2030, it is expected to have increased by 69% and 20% in underdeveloped and developed countries, respectively. Therefore, glucose sensors are likely to remain in high demand in medical device markets. For the current study, we developed a needle-type bio-layer interference (BLI) sensor that can continuously monitor glucose levels. Using dialysis procedures, we were able to obtain hypoglycemic samples from commercial human serum. These dialysis-derived samples, alongside samples of normal human serum were used to evaluate the utility of the sensor for the detection of the clinical interest range of glucose concentrations (70–200 mg/dL), revealing high system performance for a wide glycemic state range (45–500 mg/dL). Reversibility and reproducibility were also tested over a range of time spans. Combined with existing BLI system technology, this sensor holds great promise for use as a wearable online continuous glucose monitoring system for patients in a hospital setting.

The low molecular weight fraction of commercial human serum albumin induces acetylation of α-tubulin and reduces transcytosis in retinal endothelial cells

It has long been appreciated that the microtubule network plays a critical role in endothelial cell function. Chemical inhibition of tubulin polymerization has been shown to drastically increases endothelial permeability via interactions with the actin cytoskeleton. Conversely, stabilization of microtubules significantly decreases vascular permeability. The purpose of this investigation was to determine if the low molecular weight fraction of commercial 5% human serum albumin (LMWF5A) alters endothelial cell cytoskeletal dynamics and function. To investigate this, human retinal endothelial cells (HREC) were treated with LMWF5A and the acetylation of α-tubulin was determined by immunofluorescent staining and immunoblotting. In addition, permeability assays were performed to evaluate functional changes. We found that HREC treated with LMWF5A exhibit a rapid increase in the amount and distribution of acetylated α-tubulin. This was accompanied by a reduction in macromolecular permeability. Calcium depletion and inhibition of PI3-kinase reduced LMWF5A-induced acetylation while p38 MAPK inhibition potentiated this effect. These findings suggest that LMWF5A mediates changes in the microtubule network and reduces transcytosis in HREC.

A Graphene/Gelatin Composite Material for the Entrapment of Hemoglobin for Bioelectrochemical Sensing Applications

In the present work, a novel graphene (GN) and gelatin (GTN) composite was prepared and used as an immobilization matrix for hemoglobin (Hb). Compared with Hb immobilized on a bare, GN or GTN modified glassy carbon electrode (GCE), a stable and pair of well-defined quasi redox couple was observed at an Hb modified GN/GTN composite GCE at a formal potential of −0.306 V versus Ag|AgCl. The direct electrochemical behavior of Hb was greatly enhanced by the presence of both GTN and GN. A heterogeneous electron transfer rate constant (Ks) was calculated as 3.82 s−1 for Hb immobilized at GN/GTN modified GCE, which indicates the fast direct electron transfer of Hb toward the electrode surface. The biosensor shows a stable and wide linear response for H2O2 in the linear response range from 0.1 μM to 786.6 μM with an analytical sensitivity and limit of detection of 0.48 μAμM−1 cm−2 and 0.04 μM, respectively. The fabricated biosensor holds its high selectivity in the presence of potentially active interfering species and metal ions. The biosensor shows its satisfactory practical ability in the commercial contact lens solution and human serum samples.

The fluorescence detection of glutathione by ∙OH radicals’ elimination with catalyst of MoS2/rGO under full spectrum visible light irradiation

In this study, a new method for the detection of glutathione (GSH) was designed based on the ∙OH radicals’ elimination system due to the reducing ability of GSH for the first time. Fluorescence method with terephthalic acid (TA) as the probe was employed for the quantification of ∙OH radicals’ production and elimination. Experimental conditions of ∙OH radicals’ production were optimized in detail, and ∙OH radicals were found to be efficiently produced by the excellent catalysis performance of MoS2/rGO under full spectrum visible light irradiation. The introduction of GSH make fluorescent intensity decrease due to the elimination of ∙OH radicals. For the present fluorescence based GSH sensor, a wide detection range of 60.0–700.0µM and excellent selectivity have been achieved. Furthermore, it has been successfully employed for the determination of GSH in commercial drug tablets and human serum.

Low Molecular Weight Fraction of Commercial Human Serum Albumin Induces Morphologic and Transcriptional Changes of Bone Marrow-Derived Mesenchymal Stem Cells.

Osteoarthritis (OA) is the most common chronic disease of the joint; however, the therapeutic options for severe OA are limited. The low molecular weight fraction of commercial 5% human serum albumin (LMWF5A) has been shown to have anti-inflammatory properties that are mediated, in part, by a diketopiperazine that is present in the albumin preparation and that was demonstrated to be safe and effective in reducing pain and improving function when administered intra-articularly in a phase III clinical trial. In the present study, bone marrow-derived mesenchymal stem cells (BMMSCs) exposed to LMWF5A exhibited an elongated phenotype with diffuse intracellular F-actin, pronounced migratory leading edges, and filopodia-like projections. In addition, LMWF5A promoted chondrogenic condensation in "micromass" culture, concurrent with the upregulation of collagen 2α1 mRNA. Furthermore, the transcription of the CXCR4-CXCL12 axis was significantly regulated in a manner conducive to migration and homing. Several transcription factors involved in stem cell differentiation were also found to bind oligonucleotide response element probes following exposure to LMWF5A. Finally, a rapid increase in PRAS40 phosphorylation was observed following treatment, potentially resulting in the activation mTORC1. Proteomic analysis of synovial fluid taken from a preliminary set of patients indicated that at 12 weeks following administration of LMWF5A, a microenvironment exists in the knee conducive to stem cell infiltration, self-renewal, and differentiation, in addition to indications of remodeling with a reduction in inflammation. Taken together, these findings imply that LMWF5A treatment may prime stem cells for both mobilization and chondrogenic differentiation, potentially explaining some of the beneficial effects achieved in clinical trials.