In-depth investigation of altered glycosylation in human haptoglobin associated cancer by mass spectrometry.

Serum haptoglobin (Hp), a highly sialylated biomolecule with four N-glycosylation sites, is a positive acute-phase response glycoprotein that acts as an immunomodulator. Hp has gained considerable attention due to its potential as a signature molecule that exhibits aberrant glycosylation in inflammatory disorders and malignancies. Its glycosylation can be analyzed qualitatively and quantitatively by various methods using mass spectrometry. In this review, we have provided a brief overview of Hp structure and biological function and described mass spectrometry-based techniques for analyzing glycosylation ranging from macroheterogeneity to microheterogeneity of Hp in diseases and cancer. The sugars on haptoglobin can be a sweet bridge to link the potential of cancer-specific biomarkers to clinically relevant applications.

Nanocomposite films as electrochemical sensors for detection of catalase activity.

Cross-linked hydrogel substrates have garnered attention as they simultaneously enable oxidoreductase reactions in a control volume extended to adsorption of redox capacitors for amplification of electrochemical signals. In this study, the effect of catalase immobilization in mold-casted alginate-based thin films (1 mm × 6 mm × 10 mm) containing multi walled carbon nanotubes (MWCNT) coated with chitosan has been studied via amperometry. The amperometric response was measured as a function of peroxide concentration, at a fixed potential of -0.4 V vs. SPCE in phosphate-buffered saline (pH = 7.4). Results indicate substrate detection is not diffusion-limited by the 100 µm thick chitosan layer, if the cationic polyelectrolyte is in contact with the sensing carbon electrode, and the linear detection of the enzyme absent in solution is enabled by immobilization (R 2 = 0.9615). The ferricyanide-mediated biosensor exhibited a sensitivity of 4.55 µA/mM for the optimal formulation at room temperature comparable to other nanomaterial hybrid sensing solution namely amine-functionalized graphene with an average response time of 5 s for the optimal formulation. The suitability of the optimized chitosan-coated alginate slabs nano-environment for co-encapsulation of catalase and carbon nanotubes was confirmed by cyclic voltammetry.

Supervised Learning for Predictive Pore Size Classification of Regenerated Cellulose Membranes Based on Atomic Force Microscopy Measurements.

Nanoporous dialysis membranes made of regenerated cellulose are used as molecular weight cutoff standards in bioseparations. In this study, mesoporous standards with Stokes' radii (50 kDa/2.7 nm, 100 kDa/3.4 nm and 1000 kDa/7.3 nm) and overlapping skewed distributions were characterized using AFM, with the specific aim of generating pore size classifiers for biomimetic membranes using supervised learning. Gamma transformation was used prior to conducting discriminant analysis in terms of the area under the receiver operating curve (AUC) and classification accuracy (Acc). Monte Carlo simulations were run to generate datasets (n = 10) on which logistic regression was conducted using a constant ratio of 80:20 (measurement:algorithm training), followed by algorithm validation by WEKA. The proposed algorithm can classify the 1000 kDa vs. 100 kDa (AUC > 0.8) correctly, but discrimination is weak for the 100 kDa vs. 50 kDa (AUC < 0.7), the latter being attributed to the instrument accuracy errors below 5 nm. As indicated by the results of the cross-validation study, a test size equivalent to 70% (AUCtapping = 0.8341 ± 0.0519, Acctapping = 76.8% ± 5.9%) and 80% (AUCfluid = 0.7614 ± 0.0314, Acctfluid = 76.2% ± 1.0%) of the training sets for the tapping and fluid modes are needed for correct classification, resulting in predicted reduction of scan times.

Sustainable, Alginate-Based Sensor for Detection of Escherichia coli in Human Breast Milk.

There are no existing affordable diagnostics for sensitive, rapid, and on-site detection of pathogens in milk. To this end, an on-site colorimetric-based sustainable assay has been developed and optimized using an L16 (54) Taguchi design to obtain results in hours without PCR amplification. To determine the level of Escherichia coli (E. coli) contamination, after induction with 150 µL of breast milk, the B-Per bacterial protein extraction kit was added to a solution containing an alginate-based microcapsule assay. Within this 3 mm spherical novel sensor design, X-Gal (5-Bromo-4-Chloro-3-Indolyl ß-d-Galactopyranoside) was entrapped at a concentration of 2 mg/mL. The outward diffusing X-Gal was cleaved by ß-galactosidase from E. coli and dimerized in the solution to yield a blue color after incubation at 40 °C. Color intensity was correlated with the level of E. coli contamination using a categorical scale. After an 8 h incubation period, a continuous imaging scale based on intensity normalization was used to determine a binary lower limit of detection (LOD), which corresponded to 102 colony forming unit per mL (CFU/mL) and above. The cost of the overall assay was estimated to be $0.81 per sample, well under the $3 benchmark for state-of-the-art immune-based test kits for pathogen detection in biofluids. Considering the reported binary LOD cutoff of 102 CFU/mL and above, this proposed hydrogel-based assay is suited to meet global requirements for screening breast milk or milk for pathogenic organisms of 104 CFU/mL, with a percentage of false positives to be determined in future efforts.

Molecular Engineering for Enhanced Charge Transfer in Thin-Film Photoanode.

We developed three types of dithieno[3,2-b;2',3'-d]thiophene (DTT)-based organic sensitizers for high-performance thin photoactive TiO2 films and investigated the simple but powerful molecular engineering of different types of bonding between the triarylamine electron donor and the conjugated DTT π-bridge by the introduction of single, double, and triple bonds. As a result, with only 1.3 µm transparent and 2.5-µm TiO2 scattering layers, the triple-bond sensitizer (T-DAHTDTT) shows the highest power conversion efficiency (η = 8.4%; VOC = 0.73 V, JSC = 15.4 mA·cm-2, and FF = 0.75) in an iodine electrolyte system under one solar illumination (AM 1.5, 1000 W·m-2), followed by the single-bond sensitizer (S-DAHTDTT) (η = 7.6%) and the double-bond sensitizer (D-DAHTDTT) (η = 6.4%). We suggest that the superior performance of T-DAHTDTT comes from enhanced intramolecular charge transfer (ICT) induced by the triple bond. Consequently, T-DAHTDTT exhibits the most active photoelectron injection and charge transport on a TiO2 film during operation, which leads to the highest photocurrent density among the systems studied. We analyzed these correlations mainly in terms of charge injection efficiency, level of photocharge storage, and charge-transport kinetics. This study suggests that the molecular engineering of a triple bond between the electron donor and the π-bridge of a sensitizer increases the performance of dye-sensitized solar cell (DSC) with a thin photoactive film by enhancing not only JSC through improved ICT but also VOC through the evenly distributed sensitizer surface coverage.

Sex differences in the association between obesity and albuminuria among Korean adults: a cross-sectional study using the Korea National Health and Nutrition Examination Survey data.

BACKGROUND: The association between obesity and albuminuria in the general population remains unclear. We aimed to identify the association between obesity and albuminuria as well as sex differences regarding the associations using several obesity indices, including waist circumference (WC), body mass index (BMI), and waist-to-height ratio (WHR). METHODS: This study included 3841 subjects (1730 males and 2111 females; age 20-80 years) who participated in the Fifth Korea National Health and Nutrition Examination Survey conducted in 2011. Subjects with hypertension, diabetes, renal failure, or a malignant tumor and those who were pregnant or menstruating were excluded. Albuminuria was defined as a urinary albumin-to-creatinine ratio ≥30 mg/g. Anthropometric parameters were categorized into sex-specific quartiles. Logistic regression models were used to assess the associations between each anthropometric parameter and albuminuria. RESULTS: All of the obesity indices of the fourth quartile group of females showed a twofold higher risk for albuminuria than the second quartile group, and it was persistently significant after adjusting for age, smoking, and physical activity. After further adjustment for high blood pressure and impaired fasting glucose and triglyceride levels, WC and BMI of the fourth quartile group of females still showed a significantly higher risk for albuminuria than the second quartile group (odds ratios 1.96 and 2.24; 95 % confidence intervals 1.03-3.74 and 1.15-4.37). None of the associations between albuminuria and the obesity indices were significant in males. CONCLUSION: Higher WC and BMI were significantly associated with the risk of albuminuria among females, but not males.

Rapid, Affordable, and Point-of-Care Water Monitoring Via a Microfluidic DNA Sensor and a Mobile Interface for Global Health.

Contaminated water is a serious concern in many developing countries with severe health consequences particularly for children. Current methods for monitoring waterborne pathogens are often time consuming, expensive, and labor intensive, making them not suitable for these regions. Electrochemical detection in a microfluidic platform offers many advantages such as portability, minimal use of instrumentation, and easy integration with electronics. In many parts of the world, however, the required equipment for pathogen detection through electrochemical sensors is either not available or insufficiently portable, and operators may not be trained to use these sensors and interpret results, ultimately preventing its wide adoption. Counterintuitively, these same regions often have an extensive mobile phone infrastructure, suggesting the possibility of integrating electrochemical detection of bacterial pathogens with a mobile platform. Toward a solution to water quality interventions, we demonstrate a microfluidic electrochemical sensor combined with a mobile interface that detects the sequences from bacterial pathogens, suitable for rapid, affordable, and point-of-care water monitoring. We employ the transduction of DNA hybridization into a readily detectable electric signal by means of a conformational change of DNA stem-loop structure. Using this platform, we successfully demonstrate the detection of as low as 100 nM E. coli sequences and the automatic interpretation and mapping of the detection results via a mobile application.

Simultaneous sorting of multiple bacterial targets using integrated dielectrophoretic-magnetic activated cell sorter.

The ability to rapidly and accurately sort multiple types of biological targets-such as molecules, viruses, bacteria or mammalian cells-from complex sample mixtures is an essential component for a wide range of diagnostic and therapeutic strategies. However, most current selection methods for cell separation are either limited with regard to throughput, as is the case for Fluorescence Assisted Cell Sorting (FACS), or else only allow binary separation of targets that have been labeled via a single parameter, such as Magnetic Activated Cell Sorting (MACS). We report here the integrated Dielectrophoretic-Magnetic Activated Cell Sorter (iDMACS), an integrated platform that combines two different force fields in a single microfluidic device for highly efficient multi-target separation. We describe the underlying physics and design of the iDMACS device and demonstrate approximately 900-fold enrichment of multiple bacterial target cell types with over 95% purity after a single round of separation.

Multitarget magnetic activated cell sorter.

Magnetic selection allows high-throughput sorting of target cells based on surface markers, and it is extensively used in biotechnology for a wide range of applications from in vitro diagnostics to cell-based therapies. However, existing methods can only perform separation based on a single parameter (i.e., the presence or absence of magnetization), and therefore, the simultaneous sorting of multiple targets at high levels of purity, recovery, and throughput remains a challenge. In this work, we present an alternative system, the multitarget magnetic activated cell sorter (MT-MACS), which makes use of microfluidics technology to achieve simultaneous spatially-addressable sorting of multiple target cell types in a continuous-flow manner. We used the MT-MACS device to purify 2 types of target cells, which had been labeled via target-specific affinity reagents with 2 different magnetic tags with distinct saturation magnetization and size. The device was engineered so that the combined effects of the hydrodynamic force produced from the laminar flow and the magnetophoretic force produced from patterned ferromagnetic structures within the microchannel result in the selective purification of the differentially labeled target cells into multiple independent outlets. We demonstrate here the capability to simultaneously sort multiple magnetic tags with >90% purity and >5,000-fold enrichment and multiple bacterial cell types with >90% purity and >500-fold enrichment at a throughput of 10(9) cells per hour.

Multitarget dielectrophoresis activated cell sorter.

The ability to rapidly and efficiently isolate specific viruses, bacteria, or mammalian cells from complex mixtures lies at the heart of biomedical applications ranging from in vitro diagnostics to cell transplantation therapies. Unfortunately, many current selection methods for cell separation, such as magnetic activated cell sorting (MACS), only allow the binary separation of target cells that have been labeled via a single parameter (e.g., magnetization). This limitation makes it challenging to simultaneously enrich multiple, distinct target cell types from a multicomponent sample. We describe here a novel approach to specifically label multiple cell types with unique synthetic dielectrophoretic tags that modulate the complex permittivities of the labeled cells, allowing them to be sorted with high purity using the multitarget dielectrophoresis activated cell sorter (MT-DACS) chip. Here we describe the underlying physics and design of the MT-DACS microfluidic device and demonstrate approximately 1000-fold enrichment of multiple bacterial target cell types in a single-pass separation.

Selection of mammalian cells based on their cell-cycle phase using dielectrophoresis.

An effective, noninvasive means of selecting cells based on their phase within the cell cycle is an important capability for biological research. Current methods of producing synchronous cell populations, however, tend to disrupt the natural physiology of the cell or suffer from low synchronization yields. In this work, we report a microfluidic device that utilizes the dielectrophoresis phenomenon to synchronize cells by exploiting the relationship between the cell's volume and its phase in the cell cycle. The dielectrophoresis activated cell synchronizer (DACSync) device accepts an asynchronous mixture of cells at the inlet, fractionates the cell populations according to the cell-cycle phase (G(1)/S and G(2)/M), and elutes them through different outlets. The device is gentle and efficient; it utilizes electric fields that are 1-2 orders of magnitude below those used in electroporation and enriches asynchronous tumor cells in the G(1) phase to 96% in one round of sorting, in a continuous flow manner at a throughput of 2 x 10(5) cells per hour per microchannel. This work illustrates the feasibility of using laminar flow and electrokinetic forces for the efficient, noninvasive separation of living cells.