Biochemical Analysis Systems Research Articles

Function evaluation of the autoanalyser internal quality control of the Kehua Polaris c2000 automatic biochemical analyser

Objective: The present study aims to (1) evaluate the autoanalyser quality control (QC) module of the Kehua Polaris c2000 automatic modularised biochemical analyser and (2) verify the impact of the analyser’s automatic implementation of internal QC (IQC) testing at a set time point on the results of IQC and turnaround time (TAT). Methods: For 5 consecutive days, three different methods were used to conduct IQC. Method 1: Internal QC was carried out at 8:00 every day. Method 2: The QC products were placed in the instrument calibration QC plate in the afternoon, and the instrument was set to automatically measure the products at 7:00 the next day. Method 3: The QC products were placed in the instrument calibration QC plate, and the instrument was set up for automatic measurement at 7:30 every day. All three methods were compared and evaluated. The effect of IQC on the TAT was monitored using method 2. Results: There were no statistical differences in the IQC results between methods 2 and 1. However, there were statistical differences in some items between the results of methods 3 and 1; thus, the IQC results of method 2 can be adopted. The implementation of method 2 for IQC can help achieve a significant TAT-saving of 35.23 min during the automatic retest process after the IQC indicates an out-of-control situation. This time reduction is highly valuable for improving efficiency and streamlining the testing workflow. Conclusion: Using the autoanalyser QC module of the Kehua Polaris c2000 automatic modular biochemical analysis system to perform IQC has no impact on the IQC test results and can save TAT, as well as automatically correct most out-of-control occurrences.

Droplet Transportation on Liquid-Infused Asymmetrically Structured Surfaces by Mechanical Oscillation and Viscosity Control.

The transportation of droplets on solid surfaces has received significant attention owing to its importance in biochemical analysis and microfluidics. In this study, we propose a novel strategy for controlling droplet motion by combining an asymmetric structure and infused lubricating oil on a vibrating substrate. The transportation of droplets with volumes ranging from 10 to 90 μL was realized, and the movement speed could be adjusted from 1.45 to 10.87 mm/s. Typical droplet manipulations, including droplet transportation along a long trajectory and selective movement of multiple droplets, were successfully demonstrated. Through experimental exploration and theoretical analysis, we showed that the adjustment of droplet transport velocity involves an intricate interaction among the Ohnesorge number, droplet volume, and input amplitude. It can potentially be used for the more complex manipulation of liquid droplets in microfluidic and biochemical analysis systems.

Microbial machinery dealing diverse aromatic compounds: Decoded from pelagic sediment ecogenomics in the gulfs of Kathiawar Peninsula and Arabian Sea

Aromatic hydrocarbons are persistent pollutants in aquatic systems as endocrine disruptors, significantly impacting natural ecosystems and human health. Microbes perform as natural bioremediators to remove and regulate aromatic hydrocarbons in the marine ecosystem. The present study focuses upon the comparative diversity and abundance of various hydrocarbon-degrading enzymes and their pathways from deep sediments along the Gulf of Kathiawar Peninsula and Arabian Sea, India. The elucidation of large number of degradation pathways in the study area under the presence of a wide range of pollutants whose fate needs to be addressed. Sediment core samples were collected, and the whole microbiome was sequenced. Analysis of the predicted ORFs (open reading frames) against the AromaDeg database revealed 2946 aromatic hydrocarbon-degrading enzyme sequences. Statistical analysis portrayed that the Gulfs were more diverse in degradation pathways compared to the open sea, with the Gulf of Kutch being more prosperous and more diverse than the Gulf of Cambay. The vast majority of the annotated ORFs belonged to groups of dioxygenases that included catechol, gentisate, and benzene dioxygenases, along with Rieske (2Fe–2S) and vicinal oxygen chelate (VOC) family proteins. From the sampling sites, only 960 of the total predicted genes were given taxonomic annotations, which mention the presence of many under-explored marine microorganism-derived hydrocarbon degrading genes and pathways. Through the present study, we tried to unveil the array of catabolic pathways of aromatic hydrocarbon degradation and genes from a marine ecosystem that upholds economic and ecological significance in India. Thus, this study provides vast opportunities and strategies for microbial resource recovery in marine ecosystems, which can be investigated to explore aromatic hydrocarbon degradation and their potential mechanisms under various oxic or anoxic environments. Future studies should focus on aromatic hydrocarbon degradation by considering degradation pathways, biochemical analysis, enzymatic, metabolic, and genetic systems, and regulations.

A Novel Detachable, Reusable, and Versatile Acoustic Tweezer Manipulation Platform for Biochemical Analysis and Detection Systems.

Multifunctional, integrated, and reusable operating platforms are highly sought after in biochemical analysis and detection systems. In this study, we demonstrated a novel detachable, reusable acoustic tweezer manipulation platform that is flexible and versatile. The free interchangeability of different detachable microchannel devices on the acoustic tweezer platform was achieved by adding a waveguide layer (glass) and a coupling layer (polydimethylsiloxane (PDMS) polymer film). We designed and demonstrated the detachable multifunctional acoustic tweezer platform with three cell manipulation capabilities. In Demo I, the detachable acoustic tweezer platform is demonstrated to have the capability for parallel processing and enrichment of the sample. In Demo II, the detachable acoustic tweezer platform with capability for precise cell alignment is demonstrated. In Demo III, it was demonstrated that the detachable acoustic tweezer platform has the capability for the separation and purification of cells. Through experiments, our acoustic tweezer platform has good acoustic retention ability, reusability, and stability. More capabilities can be expanded in the future. It provides a simple, economical, and multifunctional reusable operating platform solution for biochemical analysis and detection systems.

An integrated microfluidic system for multi-target biochemical analysis of a single drop of blood

Herein, we described an integrated microfluidic system for multi-target biochemical analysis of micro-volumes of blood samples. This system mainly consists of a microfluid handling module, an integrated microwell array chip, a temperature control module and a spectral detection module. A novel microfluidic plasma extraction approach was developed by coupling the filter membrane-based plasma separation technique with the microfluidic liquid-handling technique. With this approach, quantitative extraction of trace blood with volumes as low as a drop (30 μL) can be automatically realized. Multiple operations in multi-target biochemical analysis are achieved including micro-volume blood collecting, quantitative plasma extraction, plasma dilution, plasma distribution, transferring, biochemical reaction and absorption spectroscopy detection. This system was applied to the multi-target biochemical analysis of glucose, cholesterol and total protein in blood samples and could achieve the determination of the target analytes of a 30-μL blood sample within 11 min. The experimental results are consistent with those obtained by a commercial biochemical analyzer.

Experimental study on micro-grinding and flow characteristics of quartz glass with micro-channel

As an important part of the microfluidic system, the microfluidic channel is a key factor to improve the microfluidic performance. The glass microfluidic channel has huge application potential in the fields of medical diagnosis, biochemical analysis and microelectromechanical systems. The micro-channel was ground on the surface of quartz glass by micro-grinding machining. The micro-ground micro-channel with different depths ranged from 80 μm to 200 μm were obtained. The influences of the machining depth and liquid flow rate on flow characteristics of micro-channel were investigated. The experimental results show that when the flow velocity was less than 2.5 m/s, the flow characteristics of the micro-channel first decreased and then increased with the increasing machining depth. When the flow velocity was greater than 2.5 m/s, it decreased as the increased depth.

Research advances in clinical biochemical analysis systems based on microfluidic driving and control technique

Microfluidic techniques have the features of high throughput, low consumption, as well as ease of miniaturization, integration, and automation, thus emerging as a feasible solution to cost-effective clinical biochemical analysis. Various clinical biochemical analysis systems based on the microfluidic technique, with different driving and control methods, are introduced in this article, in addition to commercially available microfluidic biochemical analysis instruments. The characteristics of these systems are comprehensively reviewed.

Scheduling Problem of Biochemical Analyzer and Exploration of Neural Network-Greedy Algorithm

The mathematical model of biochemical analysis system was established based on neural network-greedy algorithm. The optimal task scheduling sequence was solved by neural network algorithm. At the same time, the local optimization was obtained by combining greedy algorithm. In this way, the task scheduling problem in biochemical analyzer was transformed into a mathematical problem, and the mathematical model of scheduling algorithm was established. On the platform of MATLAB, eight groups of simulation tests were carried out on the same task scheduling problem using the neural network-greedy scheduling algorithm and the traditional fixedperiod scheduling algorithm. The task-time Gantt charts of the two algorithms were compared under different scheduling orders. The results showed that the average speed of the neural network-greedy algorithm was improved by 31% compared with that of the fixed-period scheduling algorithm. The mathematical model of biochemical analysis system on scheduling problem established by neural network-greedy scheduling algorithm has high efficiency compared with the traditional fixed-period scheduling algorithm.

Cell-Derived Plasma Membrane Vesicles are Permeable to Hydrophilic Macromolecules

Giant Plasma Membrane Vesicles (GPMVs) are a widely used model system for biochemical and biophysical analysis of the isolated mammalian plasma membrane (PM). A core advantage of these vesicles is that they maintain the native lipid and protein diversity of the plasma membrane while affording the experimental flexibility of synthetic giant vesicles. In addition to fundamental investigations of PM structure and composition, GPMVs have been used to evaluate the binding of proteins and small molecules to cell-derived membranes, and the permeation of drug-like molecules through them.

Correlation between intestinal mucosal permeability and prognosis in patients with liver cirrhosis

Objective: To investigate the correlation between changes in intestinal mucosal permeability and prognosis of patients with liver cirrhosis. Methods: Data of 89 cases with liver cirrhosis who were hospitalized in the Hepatology Department of Shanxi Provincial Hospital of Traditional Chinese Medicine from January 2017 to August 2017 were collected as the liver cirrhosis experimental group, and 40 healthy subjects were randomly selected as the healthy control group. JY-DLT, the Intestinal Mucosal Barrier Biochemical Index Analysis System was used to measure the levels of serum diamine oxidase (DAO), D-lactic acid, and endotoxin (ETX) in two groups to evaluate intestinal mucosal barrier function. Spearman's rank correlation test was used to evaluate the correlation between liver cirrhosis prognosis and intestinal mucosal permeability. The results of the two groups were compared by Mann-Whitney H test of two independent samples. One-way Anova was used for intergroup comparison. The pairwise comparison between groups was performed using the LSD or SNK test. Results: The level of ETX in patients with decompensated cirrhosis was significantly higher than that in the compensated phase, and the difference was statistically significant (P < 0.05). The levels of DAO, D-lactic acid and ETX in the liver cirrhosis group were significantly higher than those in the healthy control group, and the differences were statistically significant (P < 0.01). The plasma levels of DAO, D-lactic acid and ETX in the Child-Pugh grade groups of patients with liver cirrhosis were significantly higher than those in the healthy control group, and the differences were statistically significant (P < 0.05). The results of intergroup comparison showed that there were statistically significant differences in DAO, D-lactic acid and ETX levels between Child-Pugh grade A and grade B groups (t = -4.255, 2.527, -2.179, P < 0.05). Furthermore, there were statistically significant differences in the levels of D-lactic acid and ETX between the Child-Pugh grade A and grade C groups (t = -2.693, -4.248, P < 0.01).The plasma levels of DAO, D-lactic acid and ETX levels were positively correlated (r = 0.205, 0.372, 0.342, P < 0.01). D-lactic acid and ETX levels were positively correlated with CTP score, Forns' index, RPR index, APRI score, FIB-4 index and FibroScan score(P < 0.01). Conclusion: The three indices (plasma DAO, D-lactic acid, and ETX) can accurately detect the changes in intestinal mucosal permeability. Moreover, the higher index of intestinal mucosal permeability causes the more severe degree of liver cirrhosis and the correlation between the intestinal mucosal permeability and the prognosis score of liver cirrhosis provides a reference for a new evaluation system and new ideas for the treatment of liver cirrhosis.

High resolution measurement of refractive index with resistance to temperature crosstalk through an all fiber MZI-PMF structure

A simple cascaded structure composed of all-fiber Mach-Zehnder interferometer (MZI) and polarization-maintaining fiber (PMF) is proposed and experimentally demonstrated for simultaneous refractive index (RI) and temperature measurement. By splicing a section of thin no-core fiber (TNCF) between two conventional no-core fibers (NCFs) with large lateral offset, a part of the light in the MZI structure can interact directly with the external environment to obtain a high RI sensitivity. A Panda-type PMF is cascaded with the MZI to eliminate temperature crosstalk in the RI measurement. The experimental results show that the RI sensitivity of MZI is as high as −13862 nm/RIU, and the temperature sensitivities of MZI and PMF are −0.4 nm/℃ and 1.47 nm/℃, respectively. With the help of a sensitivity matrix, the change of RI and temperature can finally be demodulated. The RI and temperature resolutions of the cascaded structure can achieve 1.83 × 10−6 RIU and 0.014 ℃ respectively, to the best of our knowledge, which are the highest among all fiber structures without special materials coating or filling for simultaneous measurement. This structure presents outstanding advantages of easy fabrication, low cost and less sample consumption, which has great potential for integrated biochemical analysis systems.

Bio-Protocol Watermarking on Digital Microfluidic Biochips

Advancements in digital microfluidic biochip (DMFB) technologies are paving the way for low-cost and automated platforms for implementing bio-protocols. However, the deployment of DMFBs outside of controlled settings will make them vulnerable to intellectual property (IP) theft. Bio-protocol development requires large investments for cross-domain innovations in biochemical analysis, microfluidics, and cyberphysical systems. We propose a watermarking technique for bio-protocol IP protection-a first in microfluidics-that hierarchically embeds a secret signature across these domains. Such a signature can be exclusively attributed to the owner (like a hash). The proposed solution takes into account the inherent variability in domain-specific parameters such as mixing ratio, sensor calibration, and incubation time. We describe watermarking techniques of varying complexities for different bio-protocol steps. These include watermarking for bio-protocol synthesis parameters and the cyberphysical systems control path parameters. A watermarking scheme based on integer linear programming is proposed for the sample-preparation step of a bio-protocol. The practicality of our solution is demonstrated through case studies involving an immunoassay and several mixing ratios required in the sample-preparation process of bio-protocols. The effectiveness of this approach is evaluated through various security metrics: proof of ownership score, the probability of successful tampering of the watermark, and the probability of coincidence. We also analyze the integrity of the watermark against various possible attacks: brute force search, the insertion of a new watermark, and the watermarking of more parameters.

Piezoelectric peristaltic micropump integrated on a microfluidic chip

Micropump is extremely useful in miniature biochemical analysis systems for its abilities of manipulating and transporting minute liquid. We developed a novel peristaltic micropump to pump fluids in microfluidic chip in this article. The micropump consists of three piezoelectric actuators, and part of a microfluidic chip as a compressible microchannel. The fluid in the microchannel is transported owning to a series of impacting actions on the microchannel by the piezoelectric actuators. The microchannel is integrated fabricated with the microfluidic chip, which ensures a tiny dead volume less than 300 nL. Being beneficial from the plug-and-play design of the microfluidic chip, there is no cross-contamination for the reagent has no contact with the reusable actuators. Calibration shows the micropump can transport liquid at a high resolution of 8.4 nL/s, a maximum flow rate of 102 nL/s, and backpressure of up to 2.0 K Pa. A demonstration of mixing liquids on a chip was carried out using two micropumps, indicating its great potential for manipulating microamount reagent in biochemical application.

Comparative phosphoproteomic analysis reveals signaling networks regulating monopolar and bipolar cytokinesis

The successful completion of cytokinesis requires the coordinated activities of diverse cellular components including membranes, cytoskeletal elements and chromosomes that together form partly redundant pathways, depending on the cell type. The biochemical analysis of this process is challenging due to its dynamic and rapid nature. Here, we systematically compared monopolar and bipolar cytokinesis and demonstrated that monopolar cytokinesis is a good surrogate for cytokinesis and it is a well-suited system for global biochemical analysis in mammalian cells. Based on this, we established a phosphoproteomic signature of cytokinesis. More than 10,000 phosphorylation sites were systematically monitored; around 800 of those were up-regulated during cytokinesis. Reconstructing the kinase-substrate interaction network revealed 31 potentially active kinases during cytokinesis. The kinase-substrate network connects proteins between cytoskeleton, membrane and cell cycle machinery. We also found consensus motifs of phosphorylation sites that can serve as biochemical markers specific to cytokinesis. Beyond the kinase-substrate network, our reconstructed signaling network suggests that combination of sumoylation and phosphorylation may regulate monopolar cytokinesis specific signaling pathways. Our analysis provides a systematic approach to the comparison of different cytokinesis types to reveal alternative ways and a global overview, in which conserved genes work together and organize chromatin and cytoplasm during cytokinesis.

Paper-based bipolar electrode-electrochemiluminescence (BPE-ECL) device with battery energy supply and smartphone read-out: A handheld ECL system for biochemical analysis at the point-of-care level

In this work, a handheld paper-based bipolar electrode-electrochemiluminescence (P-BPE-ECL) system has been proposed. In this system, a rechargeable battery is used for power supply, while a smartphone is applied for read-out of ECL signal. For the P-BPE-ECL, the carbon ink-based BPE and driving electrodes are screen-printed on the paper, and the wax-screen-printing is employed to fabricate microfluidic channels on electrode-patterned paper. Moreover, the luminol/H2O2-based ECL reaction is applied to demonstrate the quantitative ability of the P-BPE-ECL system. Under optimized conditions, H2O2 can be determined over the range of 5–5000μM, with a detection limit of 1.75μM, and relative standard deviations (RSDs) of 7.59%, 3.82% and 4.93% for 10, 100 and 1000μM H2O2 (n=5). Thus, this system has an acceptable sensitivity, dynamic range, stability and reproducibility. Finally, the applicability of the P-BPE-ECL system is demonstrated for detection of glucose in phosphate buffer solution (PBS) and artificial urine (AU) samples, with the detection limits of 0.017mM and 0.030mM, respectively. Moreover, the P-BPE-ECL system has the ability to perform high-throughput detection of glucose. Therefore, the developed system can provide a new platform for many applications such as point-of-care testing, health diagnosis and environmental monitoring.

Differences in Hematological Characteristics, Including Cholesterol and Apolipoprotein B and E, between Hepatitis B Virus and Hepatitis C Virus Patients in Korea

Hepatitis B virus (HBV) and hepatitis C virus (HCV) chronically cause hepatitis, liver cirrhosis, and hepatocellular carcinoma, and biomarkers related to liver damage are elevated in HBV and HCV patients. However, comparisons of biomarkers between HBV and HCV patients have not previously been reported. The aim of this study was to investigate differences in hematological biomarker in the sera of HBV and HCV patients and to find a key biomarker to differentiate between HBV and HCV infections. HBV (n=115) and HCV (n=128) samples (serum and whole blood) were collected and tested using a biochemical analysis system. The obtained data were analyzed with SPSS 18.0 statistical software. The mean age of the HCV group (60.3±14.1) was much higher than that of the HBV group (51.1±12.4). Male and female rates were 71.3% and 28.7% in the HBV group and 53.9% and 46.1% in the HCV group, respectively (p = 0.005). AST, ALT, and TG values were higher in the HCV group than in the HBV group. Although γ-GTP and LDL levels were higher in the HBV group than in the HCV group, apoB and apoE levels were much higher in HCV group than in HBV group (p < 0.001). There were no significant differences in the other hematological biomarkers between the HBV and HCV groups. In conclusion, HBV rates were higher in male patients, and HCV rates were higher in older patients. In particular, apoE and apoB were more highly expressed in HCV patients, and they might be key markers to differentiate HCV infection.

Deep Proteomic Evaluation of Primary and Cell Line Motoneuron Disease Models Delineates Major Differences in Neuronal Characteristics

The fatal neurodegenerative disorders amyotrophic lateral sclerosis and spinal muscular atrophy are, respectively, the most common motoneuron disease and genetic cause of infant death. Various in vitro model systems have been established to investigate motoneuron disease mechanisms, in particular immortalized cell lines and primary neurons. Using quantitative mass-spectrometry-based proteomics, we compared the proteomes of primary motoneurons to motoneuron-like cell lines NSC-34 and N2a, as well as to non-neuronal control cells, at a depth of 10,000 proteins. We used this resource to evaluate the suitability of murine in vitro model systems for cell biological and biochemical analysis of motoneuron disease mechanisms. Individual protein and pathway analysis indicated substantial differences between motoneuron-like cell lines and primary motoneurons, especially for proteins involved in differentiation, cytoskeleton, and receptor signaling, whereas common metabolic pathways were more similar. The proteins associated with amyotrophic lateral sclerosis also showed distinct differences between cell lines and primary motoneurons, providing a molecular basis for understanding fundamental alterations between cell lines and neurons with respect to neuronal pathways with relevance for disease mechanisms. Our study provides a proteomics resource for motoneuron research and presents a paradigm of how mass-spectrometry-based proteomics can be used to evaluate disease model systems.

Research of Laser Induced Fluorescence Detection Techniques Based on Microfluidic Devices

Laser induced fluorescence (LIF) detection technique is one of the most sensitive detection method in micro total analytical systems (μTAS) as its good monochromaticity, strong collimation and high optical density. It is widely used in biochemical analysis systems and the detection limit could reach 10-9-10-12mol/L. Due to the important role in μTAS, different optical structure arrangements of LIF detection techniques were continuously developed. We reviewed the recent progress of the optical structure arrangement and the applications of the LIF detection techniques.

Molecular Architecture of the Bacterial Flagellar Motor in Cells

The flagellum is one of the most sophisticated self-assembling molecular machines in bacteria. Powered by the proton-motive force, the flagellum rapidly rotates in either a clockwise or counterclockwise direction, which ultimately controls bacterial motility and behavior. Escherichia coli and Salmonella enterica have served as important model systems for extensive genetic, biochemical, and structural analysis of the flagellum, providing unparalleled insights into its structure, function, and gene regulation. Despite these advances, our understanding of flagellar assembly and rotational mechanisms remains incomplete, in part because of the limited structural information available regarding the intact rotor–stator complex and secretion apparatus. Cryo-electron tomography (cryo-ET) has become a valuable imaging technique capable of visualizing the intact flagellar motor in cells at molecular resolution. Because the resolution that can be achieved by cryo-ET with large bacteria (such as E. coli and S. enterica) is limited, analysis of small-diameter bacteria (including Borrelia burgdorferi and Campylobacter jejuni) can provide additional insights into the in situ structure of the flagellar motor and other cellular components. This review is focused on the application of cryo-ET, in combination with genetic and biophysical approaches, to the study of flagellar structures and its potential for improving the understanding of rotor–stator interactions, the rotational switching mechanism, and the secretion and assembly of flagellar components.

Clinical application of elevated platelet-activating factor acetylhydrolase in patients with hepatitis B.

BackgroundThe aim of this study was to investigate the variation of platelet-activating factor acetylhydrolase (PAF-AH) in patients with various stages of hepatitis B infection and evaluate the association between PAF-AH activity and chronic severe hepatitis B (CSHB) and mortality in patients with hepatitis B.MethodsSerum PAF-AH activity was measured in 216 patients with hepatitis B and in 152 healthy controls using an automatic biochemical analysis system. Spearman correlation was used to investigate the correlation between PAF-AH activity and other biochemical indicators. The receiver operating characteristic (ROC) curve and multivariable logistic regression analysis were used to evaluate the ability of PAF-AH activity to predict CSHB and mortality in patients with hepatitis B.ResultsThe PAF-AH activities in patients with CSHB (1320 ± 481 U/L) were significantly higher than those in healthy controls and in other hepatitis B groups (all P < 0.01). In patients with hepatitis B, PAF-AH activity correlated with total bilirubin (r = 0.633), total bile acid (r = 0.559), aspartate aminotransferase (r = 0.332), apolipoprotein B (r = 0.348), high-density lipoprotein cholesterol (r = −0.493), and apolipoprotein AI (r = −0.530). The areas under the ROC curves for the ability of PAF-AH activity to predict CSHB and mortality in patients with hepatitis B were 0.881 (95% confidence interval (CI): 0.824–0.937, P < 0.001) and 0.757 (95% CI: 0.677–0.837, P < 0.001), respectively. Multivariate logistic regression analysis showed PAF-AH activity to be an independent factor predicting CSHB with an odds ratio of 1.003 (95% CI: 1.002–1.005, P < 0.001).ConclusionElevated PAF-AH in patients with hepatitis B was significantly associated with liver damage. Thus, serum PAF-AH could be used as a novel indicator for predicting CSHB and mortality in patients with hepatitis B. Further, PAF-AH activity was an independent factor predicting CSHB.