Human Serum Concentrations Research Articles

Zinc concentrations in human milk and infant serum during the first six months of lactation

Normal supply of zinc to the newborn via milk is essential for normal development. Using ICP-OES, we analyzed changes in the level of Zn in milk and infant serum in the neonatal period (Day 1 and Day 28 post partum) and at 6 months after delivery, in the cohort of 60 mothers and exclusively breastfed babies. Zn level in the serum showed increase (significant at 6 months) during this period (mg/mL): Day 1: 0.52±0.12; Day 28: 0.59±0.19; 6 months: 0.68±0.28. The concentration of Zn in the milk showed an opposite (decreasing) trend during the follow up: Day 1: 4.70±1.74mg/L; Day 28: 2.65±1.06; 6 months: 0.46±0.36. A significant negative correlation was established between serum and milk [Zn] at day 28 (R=−0.338; p=0.008), whereas a positive correlation was found at 6 months between these parameters (R=0.306; p=0.018). There was no significant correlation between [Zn] in the milk and serum and infants' body mass, mothers' age and mass at delivery. The level of Zn in the milk at 6 months of lactation is not sufficient to meet the recommended values. This implies that in Serbian population, Zn supplementation might be needed in the later phase of lactation.

Non-enzymatic Fluorescent Biosensor for Glucose Sensing Based on ZnO Nanorods

We have developed a non-enzymatic fluorescent biosensor for glucose sensing based on ZnO nanorods. ZnO nanorods of high density, high crystallinity, and good alignment were grown on low-cost industrial copper substrates at low temperature. To grow them directly on the substrates without using a seed layer, we utilized a simple one-step seedless hydrothermal method, which is based on galvanic cell structure. Herein, the glucose-treated ZnO nanorods together with the ultraviolet (UV) irradiation of the sample during the photoluminescent measurement played the role of a catalyst. They decomposed glucose into hydrogen peroxide (H2O2) and gluconic acid, which is similar to the glucose oxidase enzyme (GOx) used in enzymatic sensors. Due to the formation of H2O2, the photoluminescence intensity of the UV emission peak of ZnO nanorods decreased as the glucose concentration increased from 1 mM to 100 mM. In comparison with glucose concentration of a normal human serum, which is in the range of 4.4–6.6 mM, the obtained results show potential of non-enzymatic fluorescent biosensors in medical applications.

Ultrasensitive electrochemical immunosensor for quantitative detection of SCCA using Co3O4@CeO2-Au@Pt nanocomposite as enzyme-mimetic labels

Recently early diagnosis of squamous cell carcinoma antigen (SCCA) as a tumor maker of various cancers has increasingly attracted a lot of attention with heightening of incidence rate of cancer. The SCCA with low concentration in human serum should be diluted before detecting. Thus, an immunoassay with high sensitivity is significant for early detecting SCCA. Therefore, a nonenzymatic sandwich-type electrochemical immunosensor herein was conducted to quantitative detection of squamous cell carcinoma antigen (SCCA). The amino functionalized cobaltosic oxide @ ceric dioxide nanocubes with core-shell morphology were prepared to combine sea-urchin like gold @ platinum nanoparticles (Co3O4@CeO2-Au@Pt), and used as labels to conjugate with secondary antibodies for signal amplification. Due to the synergetic effect, excellent electrochemical property and superior auxiliary catalytic activity of Co3O4@CeO2-Au@Pt, high electrocatalytic current responses toward the reduction of hydrogen peroxide (H2O2) were achieved. Besides, the electrodeposited gold nanoparticles (D-Au NPs) which were modified on glassy carbon electrodes (GCE) were used as antibodies carriers and sensing platforms. With the well cooperation of Co3O4@CeO2-Au@Pt and D-Au NPs, a broad linear range from 100fg/mL to 80ng/mL with a low detection limit of 33 fg/mL for detecting SCCA was achieved. In addition, the immunosensor displayed with good reproducibility, high selectivity and stability. The results are satisfactory when the proposed method has been applied to analyze human serum samples, indicating that the potential application is promising in clinical monitoring of tumor markers.

Anti-apoptotic protein Lifeguard does not act as a tumor marker in breast cancer.

The aim of the present study was to use an enzyme-linked immunosorbent assay (ELISA) to determine the concentrations of Lifeguard (LFG) protein in the serum of 36 patients diagnosed with breast cancer and to compare these values with the concentrations of LFG protein in the serum of 7 healthy volunteers in order to detect a possible association between the expression of LFG in the serum and the degree of malignancy of the disease. Although there is no direct association between the LFG protein concentration in the serum and the degree of malignancy of breast cancer, a statistically significant distribution of the concentration in all investigated samples was observed. This indicated an association between the LFG protein concentration in human serum with a currently unknown factor.

Evaluation of the immunomodulatory effects of 2,3,3,3-tetrafluoro-2-(heptafluoropropoxy)-propanoate in C57BL/6 mice.

2,3,3,3-tetrafluoro-2-(heptafluoropropoxy)-propanoate was designed to replace perfluorooctanoic acid (PFOA), which has been mostly phased out of U.S. production due to environmental persistence, detectable human and wildlife serum concentrations, and reports of systemic toxicity. In rodent models, PFOA exposure suppresses T cell-dependent antibody responses (TDAR) and vaccine responses in exposed humans. To determine replacement compound effects on TDAR and related parameters, male and female C57BL/6 mice were gavaged with 0, 1, 10, or 100 mg/kg/day for 28 days. Mice immunized with antigen on day 24 were evaluated for TDAR and splenic lymphocyte subpopulations five days later. Serum and urine were collected for test compound concentrations and liver peroxisome proliferation was measured. Relative liver weight at 10 and 100 mg/kg and peroxisome proliferation at 100 mg/kg were increased in both sexes. TDAR was suppressed in females at 100 mg/kg. T lymphocyte numbers were increased in males at 100 mg/kg; B lymphocyte numbers were unchanged in both sexes. Females had less serum accumulation and higher clearance than males, and males had higher urine concentrations than females at all times and doses. While this PFOA-replacement compound appears less potent at suppressing TDAR relative to PFOA, it produces detectable changes in parameters affected by PFOA; further studies are necessary to determine its full immunomodulatory profile and potential synergism with other per- and polyfluoroalkyl substances of environmental concern.

A Colorimetric CMOS-Based Platform for Rapid Total Serum Cholesterol Quantification

Elevated cholesterol levels are associated with a greater risk of developing cardiovascular disease and other illnesses, making it a prime candidate for detection on a disposable biosensor for rapid point of care diagnostics. One of the methods to quantify cholesterol levels in human blood serum uses an optically mediated enzyme assay and a bench top spectrophotometer. The bulkiness and power hungry nature of the equipment limits its usage to laboratories. Here, we present a new disposable sensing platform that is based on a complementary metal oxide semiconductor process for total cholesterol quantification in pure blood serum. The platform that we implemented comprises readily mass-manufacturable components that exploit the colorimetric changes of cholesterol oxidase and cholesterol esterase reactions. We have shown that our quantification results are comparable to that obtained by a bench top spectrophotometer. Using the implemented device, we have measured cholesterol concentration in human blood serum as low as 29 $\mu \text{M}$ with a limit of detection at 13 $\mu \text{M}$ , which is approximately 400 times lower than average physiological range, implying that our device also has the potential to be used for applications that require greater sensitivity.

Extended Gate Field-Effect Transistor Biosensors for Point-Of-Care Testing of Uric Acid.

An enzyme-free redox potential sensor using off-chip extended-gate field effect transistor (EGFET) with a ferrocenyl-alkanethiol modified gold electrode has been used to quantify uric acid concentration in human serum and urine. Hexacyanoferrate (II) and (III) ions are used as redox reagent. The potentiometric sensor measures the interface potential on the ferrocene immobilized gold electrode, which is modulated by the redox reaction between uric acid and hexacyanoferrate ions. The device shows a near Nernstian response to uric acid and is highly specific to uric acid in human serum and urine. The interference that comes from glucose, bilirubin, ascorbic acid, and hemoglobin is negligible in the normal concentration range of these interferents. The sensor also exhibits excellent long term reliability and is regenerative. This extended gate field effect transistor based sensor is promising for point-of-care detection of uric acid due to the small size, low cost, and low sample volume consumption.

Polybrominated diphenyl ether, polychlorinated biphenyl, and dechlorane plus concentrations in human serum, and their relationships with thyroid hormones.

Persistent halogenated organic compounds, including polybrominated diphenyl ethers (PBDEs), polychlorinated biphenyls (PCBs), and dechlorane plus (DP), were determined in human serum collected in Weifang and Nanning, China, and the relationships between their concentrations and thyroid hormone concentrations were determined. The total PBDE (Σ10PBDEs), total PCB (Σ14PCBs), and total DP (ΣDP) concentrations were higher in Weifang (medians 220, 4.3, and 2.2 ng/g lipid, respectively) than in Nanning (medians 180, 1.9, and 1.5 ng/g lipid, respectively). The donors’ ages correlated with Σ14PCBs but not with Σ10PBDEs or ΣDP in Weifang. Age positively correlated with Σ10PBDEs but not Σ14PCBs and ΣDP in Nanning. The correlations between the individual PBDE, PCB, and DP congener and thyroid hormone concentrations were analyzed by stepwise multiple linear regression after log2-transforming the variables. Triiodothyronine (T3) concentrations were associated with BDE-99 concentrations and ΣDP in Weifang but with BDE-153 concentrations and Σ10PBDEs in Nanning. A positive correlation was found, for the first time, between anti-DP and free-T3 concentrations in Weifang, but free-T3 concentrations correlated with PCB-118 and PCB-170 concentrations in Nanning. Thyroxine (T4) concentrations correlated with total dioxin-like PCB concentrations in Weifang but with PCB-156 concentrations in Nanning. No significant correlations were found between the analyte and free-T4 concentrations in Weifang, but negative correlations were found between free-T4 and total dioxin-like PCB and BDE-190 concentrations in Nanning. No significant correlations were found between the analyte and thyroid-stimulating hormone (TSH) concentrations in Weifang but negative correlations were found between Σ10PBDEs and the TSH concentrations in Nanning. The correlations between the analyte and thyroid hormone concentrations followed different trends in Weifang and Nanning but were similar to those that have been found in animal and epidemiological studies.

Prediction of electron density and trace element concentrations in human blood serum following radioiodine therapy in differentiated thyroid cancer patients

Abstract The aim of this study is to investigate trace element levels (Cr, Cu, Fe, Mg, Mn, Se, Zn, As, B, Si and Co) in serum samples of patients with papillary thyroid cancer. The analysis of trace elements levels of samples carried out using ICP-OES. Elements in blood samples were determined by ICP-OES after using Hettich Universal Centrifuge and differences between the values of Zn, Mg, Fe and Cu of blood samples were obtained pre-therapy and post-therapy. The concentrations of elements were normalized to percentage. Later, electron densities ( N el ) of blood samples have been determined using ZXCOM program. A venture was made to perform a correlation between electron density and pre-posttreatment. This attempt is quite new in literature. Interestingly, it was generally observed that the electron densities of blood samples posttreatment are higher than pretreatment. Among elements, normalized concentration values of Cu and electron densities were in particular compared with patient type. Also, statistical analysis of blood results was achieved for 11 patients and 11 normal individual as control group for pre-posttreatment. Wilcoxon test which is a non-parametric test was used.

3D-copper oxide and copper oxide/few-layer graphene with screen printed nanosheet assembly for ultrasensitive non-enzymatic glucose sensing

Screen-printed copper oxide (CuO) and CuO/few-layer graphene on graphite electrodes were used to fabricate the ultrasensitive nonenzymatic glucose biosensors. Flower-like CuO and flower-like CuO/few-layer graphene composites were prepared by screen-printing method and characterized by X-ray diffraction (XRD), field-emission scanning electron microscopy (FESEM), and high-resolution transmission electron microscopy (HETEM). On the basis of their cyclic voltammetry (CV) and chronoamperometry results, it was concluded that the addition of graphene to CuO significantly improved the performance of the fabricated glucose sensors, exhibiting high and reproducible sensitivity of 3120 μAmM−1 cm−2 with three linear ranges from 4 μM to 13.5 mM and the detection limit of 4 μM (S/N = 3) in a fast response time of 2 s. In addition, the fabricated sensors could effectively avoid the disturbance by interferents, such as Ascorbic Acid (AA), Uric Acid (UA), and Dopamine (DA). Most importantly, the testing results of real blood serum samples demonstrated that the electrodes were applicable and acceptable for the determination of glucose concentrations in human serum. The efficiencies of two non-enzymatic glucose biosensors for glucose determination were comparable with that of a commercial enzymatic sensor.

All-in-one dual-aptasensor capable of rapidly quantifying carcinoembryonic antigen

Using a dual DNA aptamer (CEA aptamer linked to hemin aptamer), capable of rapidly capturing carcinoembryonic antigen (CEA) and hemin, an all-in-one dual-aptasensor with 1,1′-oxalyldiimidazole (ODI) chemiluminescence detection was developed for the early diagnosis of human cancer. CEA and hemin competitively bound with the dual DNA aptamer while the mixture in a detection cell was incubated for 30min at room temperature. When Amplex Red and H2O2 were added in the detection cell after the incubation, the yield of resorufin formed from the reaction Amplex Red and H2O2 depended on the concentration of HRP-mimicking G-quardruplex DNAzyme formed from the binding interaction between hemin and the dual DNA aptamer. Bright red light was observed with the addition of ODI and H2O2 in the detection cell containing resorufin. Relative CL intensity of all-in-one dual-aptasensor, operated with the competitive reaction of CEA and hemin in the presence of the dual aptamer, was exponentially decreased with the increase of CEA concentration in human serum. The limit of detection (LOD=3σ) of the all-in-one dual-aptasensor which operated with excellent accuracy, precision, and reproducibility was as low as 0.58ng/ml. The good correlation between the easy to use all-in-one dual-aptasensor and conventional enzyme-linked immunosorbent assay (ELISA), operated with time consuming procedures (e.g., long incubations and multiple washings), indicates that the rapid all-in-one dual-aptasensor can be applied as a novel clinical tool for the early diagnosis of breast cancer.

Thyronamines and Derivatives: Physiological Relevance, Pharmacological Actions, and Future Research Directions.

Thyronamines (3-T1AM, T0AM) are endogenous compounds probably derived from L-thyroxine or its intermediate metabolites. Combined activities of intestinal deiodinases and ornithine decarboxylase generate 3-T1AM in vitro. Alternatively, 3-T1AM might be formed by the thyroid gland and secreted into the blood. 3-T1AM and T0AM concentrations have been determined by liquid chromatography-tandem mass spectrometry analysis (LC-MS/MS) in tissues, serum, and cell lines. However, large variations of 3-T1AM concentrations in human serum were reported by LC-MS/MS compared with a monoclonal antibody-based immunoassay. These differences might be caused by strong binding of the highly hydrophobic 3-T1AM to apolipoprotein B100. Pharmacological administration of 3-T1AM results in dose-dependent reversible effects on body temperature, cardiac function, energy metabolism, and neurological functions. The physiological relevance of these actions is unclear, but may occur at tissue concentrations close to the estimated endogenous concentrations of 3-T1AM or its metabolites T0AM or thyroacetic acid (TA1). A number of putative receptors, binding sites, and cellular target molecules mediating actions of the multi-target ligand 3-T1AM have been proposed. Among those are members of the trace amine associated receptor family, the adrenergic receptor ADRα2a, and the thermosensitive transient receptor potential melastatin 8 channel. Preclinical studies employing various animal experimental models are in progress, and more stable receptor-selective agonistic and antagonistic analogues of 3-T1AM are now available for testing. The potent endogenous thyroid hormone-derived biogenic amine 3-T1AM exerts marked cryogenic, metabolic, cardiac and central actions and represents a valuable lead compound linking endocrine, metabolic, and neuroscience research to advance development of new drugs.

Impedimetric immunosensor for the label-free and direct detection of botulinum neurotoxin serotype A using Au nanoparticles/graphene-chitosan composite

In this work, a novel nanocomposite film consisting of the Au nanoparticles/graphene-chitosan has been designed to construct an impedimetric immunosensor for a rapid and sensitive immunoassay of botulinum neurotoxin A (BoNT/A). BoNT/A antibody was immobilized on glassy carbon electrode modified with Au nanoparticles/graphene-chitosan for the signal amplification. The fabrication of immunosensor was extensively characterized by using transmission electron microscopy (TEM), scanning electron microscopy (SEM), atomic force microscopy (AFM), X-ray diffraction (XRD), Fourier transform infrared (FTIR), cyclic voltammetry (CV), and electrochemical impedance spectroscopy (EIS). The impedance changes, due to the specific immuno-interactions at the immunosensor surface that efficiently restricted the electron transfer of redox probe Fe(CN)64−/3− were utilized to detect BoNT/A. The measurements were highly targeted specific and linear with logarithmic BoNT/A concentrations in PBS, milk and human serum across a 0.27–268pgmL−1 range and associated with a detection limit of 0.11pgmL−1.

Metformin Effect on Nontargeted Metabolite Profiles in Patients With Type 2 Diabetes and in Multiple Murine Tissues

Metformin is the first-line oral medication to increase insulin sensitivity in patients with type 2 diabetes (T2D). Our aim was to investigate the pleiotropic effect of metformin using a nontargeted metabolomics approach. We analyzed 353 metabolites in fasting serum samples of the population-based human KORA (Cooperative Health Research in the Region of Augsburg) follow-up survey 4 cohort. To compare T2D patients treated with metformin (mt-T2D, n = 74) and those without antidiabetes medication (ndt-T2D, n = 115), we used multivariable linear regression models in a cross-sectional study. We applied a generalized estimating equation to confirm the initial findings in longitudinal samples of 683 KORA participants. In a translational approach, we used murine plasma, liver, skeletal muscle, and epididymal adipose tissue samples from metformin-treated db/db mice to further corroborate our findings from the human study. We identified two metabolites significantly (P < 1.42E-04) associated with metformin treatment. Citrulline showed lower relative concentrations and an unknown metabolite X-21365 showed higher relative concentrations in human serum when comparing mt-T2D with ndt-T2D. Citrulline was confirmed to be significantly (P < 2.96E-04) decreased at 7-year follow-up in patients who started metformin treatment. In mice, we validated significantly (P < 4.52E-07) lower citrulline values in plasma, skeletal muscle, and adipose tissue of metformin-treated animals but not in their liver. The lowered values of citrulline we observed by using a nontargeted approach most likely resulted from the pleiotropic effect of metformin on the interlocked urea and nitric oxide cycle. The translational data derived from multiple murine tissues corroborated and complemented the findings from the human cohort.

Dose optimization of voriconazole/anidulafungin combination against Aspergillus fumigatus using an in vitro pharmacokinetic/pharmacodynamic model and response surface analysis: clinical implications for azole-resistant aspergillosis.

Combination therapy of voriconazole with an echinocandin is often employed in order to increase the efficacy of voriconazole monotherapy. Four clinical Aspergillus fumigatus isolates with different in vitro susceptibilities to voriconazole (MIC 0.125-2 mg/L) and anidulafungin (MEC 0.008-0.016 mg/L) were tested in an in vitro pharmacokinetic/pharmacodynamic model simulating human serum concentrations of standard dosages of voriconazole and anidulafungin. Fungal growth was assessed using galactomannan production and quantitative PCR. Drug concentrations were determined with bioassays. Pharmacodynamic interactions were assessed using Bliss independence analysis (BI) and Loewe additivity-based canonical mixture response-surface non-linear regression analysis (LA). Probability of target attainment (PTA) was estimated with Monte Carlo analysis for different doses of anidulafungin (25, 50 and 100 mg) and azole resistance rates (5%-25%). Synergy [BI 51% (8%-80%), LA 0.63 (0.38-0.79)] was found at low anidulafungin (fCmax/MEC <10) and voriconazole (fAUC/MIC <10) exposures, whereas antagonism [BI 12% (5%-18%, LA 1.12 (1.04-4.6)] was found at higher drug exposures. The largest increase in PTA was found with 25 mg of anidulafungin and voriconazole MIC distributions with high (>10%) resistance rates. PTAs for isolates with voriconazole MICs of 1, 2 and 4 mg/L was 78%, 12% and 0% with voriconazole monotherapy and 96%-100%, 68%-82% and 9%-20% with combination therapy, respectively. Optimal activity was associated with a voriconazole tCmin/MIC ratio of 1.5 for monotherapy and 0.75 for combination therapy. The present study indicated that the combination of voriconazole with low-dose anidulafungin may increase the efficacy and reduce the cost and potential toxicity of antifungal therapy, particularly against azole-resistant A. fumigatus isolates and in patients with subtherapeutic serum levels. This hypothesis warrants further in vivo verification.

Simultaneous and multiplexed detection of exosome microRNAs using molecular beacons

Simultaneous and multiplexed detection of microRNAs (miRNAs) in a whole exosome is developed, which can be utilized as a PCR-free efficient diagnosis method for various diseases. Exosomes are small extracellular vesicles that contain biomarker miRNAs from parental cells. Because they circulate throughout bodily fluids, exosomal biomarkers offer great advantages for diagnosis in many aspects. In general, PCR-based methods can be used for exosomal miRNA detection but they are laborious, expensive, and time-consuming, which make them unsuitable for high-throughput diagnosis of diseases. Previously, we reported that single miRNA in the exosomes can be detected specifically using an oligonucleotide probe or molecular beacon. Herein, we demonstrate for the first time that multiple miRNAs can be detected simultaneously in exosomes using miRNA-targeting molecular beacons. Exosomes from a breast cancer cell line, MCF-7, were used for the production of exosomes because MCF-7 has a high level of miR-21, miR-375, and miR-27a as target miRNAs. Molecular beacons successfully hybridized with multiple miRNAs in the cancer cell-derived exosomes even in the presence of high human serum concentration. In addition, it is noteworthy that the choice of fluorophores for multiplexing biomarkers in an exosome is crucial because of its small size. The proposed method described in this article is beneficial to high-throughput analysis for disease diagnosis, prognosis, and response to treatment because it is a time-, labor-, and cost-saving technique.

Contribution of Direct and Indirect Exposure to Human Serum Concentrations of Perfluorooctanoic Acid in an Occupationally Exposed Group of Ski Waxers.

The contribution of direct (i.e., uptake of perfluorooctanoic acid (PFOA) itself) and indirect (i.e., uptake of 8:2 fluorotelomer alcohol (FTOH) and metabolism to PFOA) exposure to PFOA serum concentrations was investigated using a dynamic one-compartment pharmacokinetic (PK) model. The PK model was applied to six occupationally exposed ski waxers for whom direct and indirect exposures via inhalation were characterized using multiple measurements with personal air sampling devices. The model was able to predict the diverging individual temporal trends of PFOA in serum with correlation coefficients of 0.82-0.94. For the four technicians with high initial concentrations of PFOA in serum (250-1050 ng/mL), the ongoing occupational exposure (both direct and indirect) was of minor importance and net depuration of PFOA was observed throughout the ski season. An estimated average intrinsic elimination half-life of 2.4 years (1.8-3.1 years accounting for variation between technicians and model uncertainty) was derived for these technicians. The remaining two technicians, who had much lower initial serum concentrations (10-17 ng/mL), were strongly influenced by exposure during the ski season with indirect exposure contributing to 45% of PFOA serum concentrations. On the basis of these model simulations, an average metabolism yield of 0.003 (molar concentration basis; uncertainty range of 0.0006-0.01) was derived for transformation of 8:2 FTOH to PFOA. An uncertainty analysis was performed, and it was determined that the input parameters quantifying the intake of PFOA were mainly responsible for the uncertainty of the metabolism yield and the initial concentration of PFOA in serum was mainly contributing to the uncertainty of estimated serum half-lives.

Defining the Active Fraction of Daptomycin against Methicillin-Resistant Staphylococcus aureus (MRSA) Using a Pharmacokinetic and Pharmacodynamic Approach

Our objective was to study the pharmacodynamics of daptomycin in the presence of varying concentrations of human serum (HS) in vitro to quantify the fraction of daptomycin that is ‘active’. Time kill experiments were performed with daptomycin (0 to 256 mg/L) against two MRSA strains at log-phase growth, in the presence of HS (0%, 10%, 30%, 50%, 70%) combined with Mueller-Hinton broth. Daptomycin ≥ 2 mg/L achieved 99.9% kill within 8 h at all HS concentrations; early killing activity was slightly attenuated at higher HS concentrations. After 1 h, bacterial reduction of USA300 upon exposure to daptomycin 4 mg/L ranged from -3.1 to -0.5 log10CFU/mL in the presence of 0% to 70% HS, respectively. Bactericidal activity was achieved against both strains at daptomycin ≥ 4 mg/L for all fractions of HS exposure. A mechanism-based mathematical model (MBM) was developed to estimate the active daptomycin fraction at each %HS, comprising 3 bacterial subpopulations differing in daptomycin susceptibility. Time-kill data were fit with this MBM with excellent precision (r2 >0.95). The active fraction of daptomycin was estimated to range from 34.6% to 25.2% at HS fractions of 10% to 70%, respectively. Despite the reported low unbound fraction of daptomycin, the impact of protein binding on the activity of daptomycin was modest. The active fraction approach can be utilized to design in vitro experiments and to optimize therapeutic regimens of daptomycin in humans.

Quantum dots assisted laser desorption/ionization mass spectrometric detection of carbohydrates: qualitative and quantitative analysis.

A quantum dots (QDs) assisted laser desorption/ionization mass spectrometric (QDA-LDI-MS) strategy was proposed for qualitative and quantitative analysis of a series of carbohydrates. The adsorption of carbohydrates on the modified surface of different QDs as the matrices depended mainly on the formation of hydrogen bonding, which led to higher MS intensity than those with conventional organic matrix. The effects of QDs concentration and sample preparation method were explored for improving the selective ionization process and the detection sensitivity. The proposed approach offered a new dimension to the application of QDs as matrices for MALDI-MS research of carbohydrates. It could be used for quantitative measurement of glucose concentration in human serum with good performance. The QDs served as a matrix showed the advantages of low background, higher sensitivity, convenient sample preparation and excellent stability under vacuum. The QDs assisted LDI-MS approach has promising application to the analysis of carbohydrates in complex biological samples.

Silver nanoflower–reduced graphene oxide composite based micro-disk electrode for insulin detection in serum

Sensitive and selective determination of protein biomarkers remains a significant challenge due to the existence of various biomarkers in human body at a low concentration level. Therefore, new technologies were incessantly steered to detect tiny biomarkers at a low concentration level, yet, it is difficult to develop reliable, stable and sensitive detection methods for disease diagnostics. Therefore, the present study demonstrates a methodology to detect insulin in serum at low levels based on Ag nanoflower (AgNF) decorated reduced graphene oxide (rGO) modified micro-disk electrode arrays (MDEAs). The morphology of AgNF-rGO composite was characterized by scanning electron microscopy, the structure was analyzed using X-ray diffraction patterns and Raman spectra. The hybrid interface exhibited enhanced electrical conductivity when compared with its individual elements and had improved capturing ability for antibody–antigen binding towards insulin detection. In order to measure quantitatively the insulin concentration in PBS and human serum, the change in impedance (ΔZ) from electrochemical impedance spectroscopy was analyzed for various concentrations of insulin in [Fe(CN)6]3−/4− redox couple. The electrode with adsorbed antibodies showed an increase in ΔZ for the addition of antigen concentrations over a working range of 1–1000ngmL−1. The detection limits were 50 and 70pgmL−1 in PBS and human serum, respectively.