Immuno-chromatographic Analysis for HPV-16 and 18 E7 Proteins as a Biomarker of Cervical Cancer Caused by Human Papillomavirus

Among the more than 120 different types of human papillomavirus (HPV), types 16 and 18 have been known to be high risk agents that cause cervical cancer. We examined, in an immuno-chromatographic analysis, the potential of using the early gene product. E7 protein, as a diagnostic marker of cervical cancer caused by HPV. We developed monoclonal antibodies specific to HPV-16 and 18 E7 proteins that were produced from bacterial cells using, gene recombinant technology. For each E7 protein, the optimal antibody pair was selected using the immuno-chromatographic sandwich-type binding system based on the lateral flow through membrane pores. Under these conditions, this rapid testing assay had a detection capability as low as 2 ng/mL of E7 protein. Furthermore, since viral analysis required the host cell to be lysed using chemicals such as detergents, it was possible that the E7 protein was structurally damaged during this process, which would result in a decrease in detection sensitivity. Therefore, we examined the detrimental effects caused by different detergents on the E7 protein using HeLa cells as the host. In these experiments, we found that the damage caused by the detergent, nonylphenylpolyethylene glycol (NP-40), was minimal relative to Triton X-100 commonly used or the cell lysis. Temperature also affected the stability oldie E7 protein, and we found that the E7 protein was stabilized at 4 T. for about 2 h, which was 4 times longer than at room temperature. Finally, a HPV-infected cervical cancer cell line, which was used as a real sample model, was treated using the optimized conditions and the presence of E7 proteins were analyzed by immuno-chromatography. The results of this experiment demonstrated that this rapid test could specifically detect HPV-infected samples.Cho IH, 2009, ANAL CHIM ACTA, V632, P247, DOI 10.1016/j.aca.2008.11.019Liang YJ, 2008, INT J BIOCHEM CELL B, V40, P2431, DOI 10.1016/j.biocel.2008.04.003Ressler S, 2007, CLIN CANCER RES, V13, P7067, DOI 10.1158/1078-0432.CCR-07-1222Jeon JH, 2007, EXP MOL MED, V39, P621Mirecka EA, 2006, PROTEIN EXPRES PURIF, V48, P281, DOI 10.1016/j.pep.2006.04.017Nishimura A, 2006, MICROBES INFECT, V8, P984, DOI 10.1016/j.micinf.2005.10.015Cho JH, 2006, ANAL CHEM, V78, P793, DOI 10.1021/ac051453vHuh KW, 2005, P NATL ACAD SCI USA, V102, P11492, DOI 10.1073/pnas.0505337102Ono T, 2003, J IMMUNOL METHODS, V272, P211DeFilippis RA, 2003, J VIROL, V77, P1551, DOI 10.1128/JVI.77.2.1551-1563.2003Jeon JH, 2002, EXP MOL MED, V34, P496VINCE A, 2002, J CLIN VIROL, V25, P109Chen XJS, 2001, J MOL BIOL, V307, P173Paek SH, 2000, METHODS, V22, P53, DOI 10.1006/meth.2000.1036Walboomers JMM, 1999, J PATHOL, V189, P12Wertlake P, 1999, J REPROD MED, V44, P11JEON S, 1995, P NATL ACAD SCI USA, V92, P1654WILBUR DC, 1994, AM J CLIN PATHOL, V101, P209SCHEFFNER M, 1993, CELL, V75, P495SELVEY LA, 1992, J VIROL METHODS, V37, P119MAY K, 1991, AM J OBSTET GYNECOL, V165, P2000DYSON N, 1989, SCIENCE, V243, P934NELSON PH, 1989, EMBO J, V8, P3905GISSMANN L, 1986, VIRAL ETIOLOGY CERVI, P217CHANG JY, 1985, EUR J BIOCHEM, V151, P217

Electrophysiological Study of Potassium Channel in Parotid Gland Acinar Cell

When the salicary gland acinar cells are stimulated vy secretafofesm its intracellular Ca^(2+) lons are increased and potassium channels are opened. the purpose of this experiment is to study the characteristics of potassium channel and its channel activity induced by Phenylephrine using dissociated aclnar cells from the parotid gland. the parotid and its channelactivity induced by Phenylephrine using dissociated aclnar cells from the parotid gland. The parotid gland acinar cells were collected from Sprague-Dawley rat by trypsln and collafenase treatment. Isolated acinar cells were placed in 100% oxygenated tyrode solution. To record channel activities, mocropipette filled with HEPES-buffered high potassium solution with 1-5 ㏁ resistance were used. Membrane currents were recorded by cell-attached patch clamp method. the channel activities were recorded and analyzed on pCiamp soft ware. Two distinct types of potassium channels in parotid gland acinar cells were observed. The one type which had large conductance about 170pS was voltage dependent. The other type which had small conductance was uoltage independent. The potassium current was activated by phenylephrine (1×10^(-4) M) which is an α-adrenergic agonist at 60 mV holding potential (Vp) by increasing open probability

Biphasic Electrical Pulse by a Micropillar Electrode Array Enhances Maturation and Drug Response of Reprogrammed Cardiac Spheroids

Direct reprogramming is an efficient strategy to produce cardiac lineage cells necessary for cardiac tissue engineering and drug testing for cardiac toxicity. However, functional maturation of reprogrammed cardiomyocytes, which is of great importance for their regenerative potential and drug response, still remains challenging. In this study, we propose a novel electrode platform to promote direct cardiac reprogramming and improve the functionality of reprogrammed cardiac cells. Nonviral cardiac reprogramming was improved via a three-dimensional spheroid culture of chemically induced cardiomyocytes exposed to a small-molecule cocktail. A micropillar electrode array providing biphasic electrical pulses mimicking the heartbeat further enhanced maturation and electrophysiological properties of reprogrammed cardiac spheroids, leading to proper responses and increased sensitivity to drugs. On the basis of our results, we conclude that our device may have a wider application in the generation of functional cardiac cells for regenerative medicine and screening of novel drugs.11Nsciescopu

Effects of exercise-induced beta-hydroxybutyrate on muscle function and cognitive function

Recent studies have shown that exercise improves skeletal muscle and cognitive function by stimulating the secretion of numerous molecules. In particular, previous studies have suggested that exercise-induced beta-hydroxybutyrate (BHB) release might improve skeletal muscle and cognitive function, but to date these studies have been limited to cell and animal models. Therefore, we aimed to determine how an exercise-induced increase in BHB affects skeletal muscle and cognitive function at a cellular level, in an animal model, and in humans. The effects of BHB on skeletal muscle and cognitive function were determined by treating C2C12 and C6 cell lines with BHB, and by measuring the skeletal muscle and serum BHB concentrations in aged mice after endurance or resistance exercise. In addition, serum BHB concentration was measured before and after high-speed band exercise in elderly people, and its relationships with muscle and cognitive function were analyzed. We found that BHB increased cell viability and brain-derived neurotrophic factor expression level in C6 cells, and endurance exercise, but not resistance exercise, increased the muscle BHB concentration in aged mice. Furthermore, the BHB concentration was positively related to skeletal muscle and cognitive function. Exercise did not increase the serum BHB concentration in the elderly people and BHB did not correlate with cognitive function, but after excluding the five people with the highest preexisting serum concentrations of BHB, the BHB concentrations of the remaining participants were increased by exercise, and the concentration showed a tendency toward a positive correlation with cognitive function. Thus, the BHB released by skeletal muscle following endurance exercise may improve muscle and cognitive function in animals and humans. © 2021 The Authors. Physiological Reports published by Wiley Periodicals, Inc. on behalf of The Physiological Society and the American Physiological Society.TRU

Grain-boundary conduction in gadolinia-doped ceria: The effect of SrO addition

This study examined the effect of adding SrO on the grain-boundary conduction of Ce(0.9)Gd(0.1)O(1.95) (gadolinia-doped ceria) containing 500 ppm SiO(2). The apparent grain-boundary resistivity at 300 degrees C decreased drastically from 746.7 to 0.90-1.97 k Omega cm upon doping with >= 1 mol % SrO, while the grain-interior resistivity increased gradually from 3.1 to 11.6 k Omega cm as the SrO concentration was increased up to 5 mol %. Therefore, doping with 1 mol % SrO resulted in the minimum total resistivity. The electron probe X-ray microanalysis and the analysis of the lattice parameters suggest that the 140-500-fold enhancement in the grain-boundary conduction is attributed to the scavenging of the highly resistive siliceous phase by the SrO-containing phase. (C) 2008 The Electrochemical Society

Characteristics of wet and dry weather heavy metal discharges in the Yeongsan Watershed, Korea

A comprehensive water quality monitoring program was conducted in the Yeongsan (YS) River, Korea from 2005 to present to investigate wet and dry weather pollutant discharge in an attempt to establish point and non-point pollution management strategies. As part of this monitoring program, 11 heavy metal species were measured during dry and wet weather conditions in the YS River, where Gwangju City (GJ), a subcatchment of the YS River, was further monitored to clarify the responsibility of different metal species discharged into the mainstream. Monthly grab water samples showed that greater amounts of metals along the YS River were discharged during the wet summer months due largely to storm runoff. In addition, further monitoring results revealed that GJ, a highly urbanized area, was a significant contributor of the heavy metals being discharged into the YS River during both wet and dry weather. The most abundant metal species discharged from GJ were manganese, aluminum and iron with different contributions of wet and dry weather flows to the total discharge load. Wet weather flow was a significant contributor to the annual dissolved metal loads, accounting for 44–93% of the annual load depending on the metal species, with the exception of chromium and cadmium (9% and 27%, respectively). Mostly, metal loads during wet weather were shown to be proportional to the rainfall depth and antecedent dry period. A substantial fraction of metals were also associated with solids, suggesting that sedimentation might be an appropriate management practice for reducing the metal load generated in GJ. Overall, although dissolved metal concentrations in YS River were at an acceptable level for aquatic community protection, continual metal discharge throughout the year was considered to be a potential problem in the long-term due to gradual water quality degradation as well as continuous metal accumulation in the system.clos