Identification of Apolipoprotein AI as a serum biomarker of chronic kidney disease in liver transplant recipients, using proteomic techniques

Chronic kidney disease (CKD) is a common complication post‐orthotopic liver transplantation (OLT). Development of CKD is detected by monitoring serum urea and creatinine, however disease can occasionally be at an advanced stage before they become abnormal. Therefore, more accurate parameters are required. In order to identify novel biomarkers of CKD, serum was obtained from 47 OLT recipients with CKD (glomerular filtration rate <60 mL/min) and 23 with normal renal function (glomerular filtration rate >90 mL/min). Using the proteomic technique SELDI‐TOF‐MS, three protein biomarkers (55.6 kDa, 9.5 kDa and 11.4 kDa) were identified that, together, could stratify patients into cases or controls with a sensitivity and specificity of 93.6 and 91.3%, respectively. The area under the curve was 0.94. The primary splitter of the groups at 55.6 kDa was an alternative version of a molecule at 27.8 kDa, which was subsequently identified by 1‐D SDS‐PAGE and LC‐ESI‐MS/MS to be Apolipoprotein AI. Protein expression was shown to be reduced in CKD, by both ELISA (p = 0.057) and Western blot analysis (p = 0.003). Apolipoprotein AI is a novel, accurate marker of CKD post‐OLT. It does require further validation in a large, more diverse patient population but could potentially improve detection of CKD.     

Anatomic site‐specific proteomic signatures of gastrointestinal stromal tumors

The gastrointestinal stromal tumor (GIST) is the most common mesenchymal malignancy of the gastrointestinal tract. Its clinical course ranges widely from a curable disorder to a highly malignant disease. Although its clinical and molecular characteristics depend on the anatomic site of origin, the molecular background of GIST arising in different anatomical site has not been studied yet. To investigate the proteomic background of GIST, we examined the proteomic features corresponding to the anatomic site of tumor origin. Comparison of the proteomic profile of gastric (23 cases) and small intestinal (9 cases) GIST by 2‐DE revealed 105 protein spots with significantly different intensity (p <0.01) between the two groups. Mass spectrometric study identified 68 distinct proteins for these 105 protein spots, including cancer‐associated ones such as prohibitin, pigment epithelium‐derived factor, and alpha‐actinin 4. The intensity of 37/105 (35.2%) protein spots was significantly concordant with the corresponding mRNA levels (p <0.01). Although both 2‐D DIGE and microarray experiments showed significant up‐regulation of vimentin expression in small intestinal GIST, Western blotting did not show a significant difference between the two groups. In conclusion, our study demonstrates the proteins specially expressed in GIST depending on their site of origin, as well as the unique advantage offered by use of proteomics to acquire such data. The identified proteins may provide clues to understanding the different characteristics of GIST depending on their site of origin.     

Quantitative proteomic analysis of a paired human liver healthy versus carcinoma cell lines with the same genetic background to identify potential hepatocellular carcinoma markers

To comprehensively measure global changes in protein expression associated with human hepatocellular carcinoma (HCC), comparative proteomic analysis of two cell lines derived from the healthy and carcinoma tissue of a same donor respectively was conducted using quantitative amino acid-coded mass tagging /stable isotope labeling with amino acids in cell culture-based LC-MS/MS approach. Among a total of 501 proteins precisely quantified, the expressions of 128 proteins were significantly altered including 70 proteins up-regulated and 58 down-regulated in HCC cells. According to their previously characterized functions, the differentially expressed proteins were found associated with nine functional categories including glycolysis, stress response, cell communication, cell cycle, apoptosis/death, etc. For example, multiple enzymes involving glycolysis pathway were found differentially regulated in HCC cells, illustrating the critical participation of glycolysis in the HCC transformation. The accuracy of certain differentially expressed proteins identified through the amino acid-coded mass tagging-based quantification was validated in the paired cell lines, and later their pathological correlations were examined in multiple clinical pairs of normal versus tumor tissues from HCC specimen by using a variety of biological approaches including Western blotting and in situ immunoassays. These consistencies suggested that multiple proteins such as HSP27, annexin V, glyceraldehyde-3-phosphate dehydrogenase, nucleolin and elongation factor Tu could be the biomarkers candidates for diagnosis of HCC.     

Expression of tropomyosin alpha 4 chain is increased in esophageal squamous cell carcinoma as evidenced by proteomic profiling by two-dimensional electrophoresis and liquid chromatography-mass spectrometry/mass spectrometry

To identify proteins associated with esophageal carcinogenesis, we performed protein profiling of 16 esophageal squamous cell carcinomas (ESCCs) and paired noncancerous tissues by 2-DE and MS/MS. In cancerous tissues, three spots showed significant up-regulation in the amount of protein, while eight spots were significantly down-regulated. The identities of the spots were determined by PMF with LC-MS/MS and were confirmed by immunoblotting. The up-regulated proteins were tropomyosin alpha 4 chain, transgelin, and pyruvate kinase. The down-regulated proteins were serum albumin precursor, isoforms of annexin A1, tropomyosin beta chain, 14-3-3 protein sigma, and isoforms of serotransferrin precursor. In all 16 cases, up-regulation of the tropomyosin alpha 4 chain was confirmed by immunoblotting. Localization of the tropomyosin alpha 4 chain in ESCC cells and adjacent fibroblasts was confirmed by immunohistochemistry.     

Proteomics analysis of cellular components in lentiviral vector production using Gel-LC-MS/MS

Among the gene therapy vectors developed to date, lentiviral vectors persist in the host and are therefore best suited for long-term gene transfer and gene-replacement therapies. Human immunodeficiency virus (HIV)-1-based lentiviral vector products are currently produced by transient transfection, filtration and ultracentrifugation, and the quality of the resultant vector product is variable. We reason that by identifying host proteins co-produced with viral vectors we may better understand the mechanism of viral vector production, and improve the safety and quality of the resultant products. Our LC-MS/MS studies identified both viral vector proteins and host proteins, including nuclear proteins, elongation factors and chaperone and heat shock proteins (HSP), and confirmed the presence of known HIV-incorporated proteins, e.g. elongation factor-1α, HSP70 and Histone 2A, demonstrating the capability and viability of LC-MS/MS methods for the proteomic analysis of highly complex samples. Evaluation of the functions of the identified components is in progress to understand their implication for product quality and safety. These studies support the development of improved production and characterisation methods and advance the clinical application of lentiviral vector-based products.     

Data‐pulse‐voltage reduction of AC‐PDPs by using metastable‐particle priming

Abstract— It is shown that space charges dominate the build‐up of an address discharge when it is preceded by a priming discharge in less than 32 μsec. When the separation of these discharges (cease period) exceeds 32 μsec, the space charges diffuse away and the metastable particles start playing the role of priming. The priming effect of the metastable particles is not too strong, which is desirable for adopting a data‐pulse‐voltage reduction technique for PDPs. By choosing the length of the cease period to be between 32 and 80 μsec in the Address‐While‐Display drive scheme, the data pulse voltage was reduced to 20 V. This leads to a considerable cost reduction of data driver ICs.     

Wide‐locking range LC‐tank divide‐by‐4 injection‐locked frequency divider using transformer feedback

A wide locking range, injection locked frequency divider (ILFD) circuit topology is explored. The modulus‐four ILFD utilizes a cross‐coupled voltage‐controlled oscillator in conjunction with transformer feedback, parallel‐tuned resonator, and two‐segment, series mixers at the injection point. The transformer feedback and two‐segment mixing circuit topology achieves a locking range of 2.7 GHz (14.1 to 16.8 GHz) at an injection point bias of 0.9 Vdc and 0 dBm injection power. Spectral measurements at the ILFD output demonstrate proper phase‐lock operation and expected phase noise reduction using a high quality signal source at the ILFD input. The ILFD is implemented in the TSMC 0.18 μm 1P6M CMOS process, utilizes a die area of 0.839 × 0.566 mm² and consumes 16.56 mW. © 2015 Wiley Periodicals, Inc. Int J RF and Microwave CAE 25:557–562, 2015.     

Dual‐band rotary standing‐wave voltage‐controlled oscillator

In this article, a dual‐band rotary standing‐wave oscillator (RSWO) is introduced that generates sinusoidal signals by the formation of a standing wave on a ring (closed‐loop)‐distributed composite right/left‐handed (CRL) Inductor‐Capacitor (LC) transmission line network. The LC network consists of four unit cells of CRL LC resonator stacked in series, and two pairs of cross‐coupled transistors are used to compensate for the loss of LC resonator. Varactors are used as the control to switch on/off the high‐ or low‐frequency bands. In the fundamental mode, the RSWO operates at the high‐frequency band. In the harmonic mode, the oscillator provides low‐frequency band outputs. The dual‐band function exploits the multiple oscillation modes of the CRL RSWO. The proposed RSWO has been implemented with the Taiwan Semiconductor Manufacturing Company, Limited (TSMC) 0.18‐μm SiGe BiCMOS technology. It can generate differential signals in the high‐band frequency range of 6.73–8.60 GHz and in the low‐band frequency range of 3.68–3.73 GHz. The die area of the RSWO is 1.123 × 1.123 mm². © 2014 Wiley Periodicals, Inc. Int J RF and Microwave CAE 24:536–543, 2014.     

Low‐cost manufacturing of patterned films with nano‐precision for display applications

Abstract— Much attention has been given to methodologies for patterning optical films with nanoscale precision at the scale and economics required by the flat‐panel‐display industry. By using Fluorocur® mold materials, the PRINT® (Pattern Replication in Nonwetting Template) technology enables low‐cost manufacturing of precise microscale and nanoscale features with single‐nanometer precision from virtually any material.     

A polarizer‐free flexible display using dye‐doped liquid‐crystal gels

Abstract— A reflective‐type polarizer‐free flexible display using a dye‐doped liquid‐crystal (LC) gels is demonstrated. Compared to the conventional guest‐host LC mode, it has high contrast ratio and brightness due to the combining of both scattering and absorption. Such a gel‐like flexible display is bendable and trimable. In this paper, a three‐step switch using distinct dye‐doped LC gels is also demonstrated. The potential applications are e‐paper and decorative displays.     

A proteomic approach combining MS and bioinformatic analysis for the detection and identification of biomarkers of administration of exogenous human growth hormone in humans

An integrated MS-based proteomic approach is described that combines MALDI-MS and LC-MS with artificial neural networks for the identification of protein and peptide biomarkers associated with recombinant human growth hormone (rhGH) administration. Serum from exercised males administered with rhGH or placebo was analysed using ELISA to determine insulin-like growth factor-I concentrations. Diluted serum from rhGH- and placebo-treated subjects was analysed for protein biomarkers by MALDI-MS, whereas LC-MS was used to analyse tryptically digested ACN-depleted serum extracts for peptide biomarkers. Ion intensities and m/z values were used as inputs to artificial neural networks to classify samples into rhGH- and placebo-treated groups. Six protein ions (MALDI-MS) correctly classified 96% of samples into their respective groups, with a sensitivity of 91% (20 of 22 rhGH treated) and specificity of 100% (24 of 24 controls). Six peptide ions (LC-MS) were also identified and correctly classified 93% of samples with a sensitivity of 90% (19 of 21 rhGH treated) and a specificity of 95% (20 of 21 controls). The peptide biomarker ion with the highest significance was sequenced using LC-MS/MS and database searching and found to be associated with leucine-rich α-2-glycoprotein.