Characterization of Recombinant Bovine Sperm Hyaluronidase and Identification of an Important Asn-X-Ser/Thr Motif for Its Activity
-
- Journal of Microbiology and Biotechnology
- /
- v.28 no.9
- /
- pp.1547-1553
- /
- 2018
Hyaluronidases are a family of enzymes that catalyse the breakdown of hyaluronic acid, which is abundant in the extracellular matrix and cumulus oocyte complex. To investigate the activity of recombinant bovine sperm hyaluronidase 1 (SPAM1) and determine the effect of the Asn-X-Ser/Thr motif on its activity, the bovine SPAM1 open reading frame was cloned into the mammalian expression vector pCXN2 and then transfected to the HEK293 cell line. Expression of recombinant bovine hyaluronidase was estimated using a hyaluronidase activity assay with gel electrophoresis. Recombinant hyaluronidase could resolve highly polymeric hyaluronic acid and also caused dispersal of the cumulus cell layer. Comparative analysis with respect to enzyme activity was carried out for the glycosylated and deglycosylated bovine sperm hyaluronidase by N-glycosidase F treatment. Finally, mutagenesis analysis revealed that among the five potential N-linked glycosylation sites, only three contributed to significant inhibition of hyaluronic activity. Recombinant bovine SPAM1 has hyaluronan degradation and cumulus oocyte complex dispersion ability, and the N-linked oligosaccharides are important for enzyme activity, providing a foundation for the commercialization of hyaluronidase.
In this paper, performance of the CPC(complex phase code) which is recently proposed as a practical phase encoding method for phase-code multiplexes holographic memory system is comparatively analyzed with those of the conventional phase codes such as PR(pure random code), RCE(random code with equality), WHM(Walsh Hadamard Matrix). In computer simulation, the size of an address bean is fixed at 32
Ferimzone Z is a fungicide for effectively controlling rice blast. Under light irradiation conditions, it undergoes a rapid conversion to its E-stereoisomer. Given the importance of isomers in risk assessments of residues in crops, an analytical method was developed for individual isomer quantification. A comparative analysis performed using two columns in HPLC-MS/MS demonstrated that the isomers were successfully separated using the Cadenza column. For the brown rice sample preparation, 5 g of the homogenized sample was saturated with 7 mL of water. The sample was then extracted with a 10 mL mixed solvent of acetonitrile and ethyl acetate (1:1, v/v) that contained 0.1% formic acid, and it was subsequently partitioned with magnesium sulfate and sodium chloride. The upper layer was purified using dSPE containing C18 and PSA sorbents. The established method was subjected to method validation, and it showed recovery rates of 90.6-98.8% (RSD ≤ 3.9%) at concentrations of 0.01, 0.1, 2 mg/kg, with a soft matrix effect (%ME) ranging from -3.1% to +6.5%. This method can be employed in monitoring studies of brown rice to determine the conversion ratio from the Z isomers to the E isomers.
This study investigated the problems of remarried wife with alcoholic problem. Specifically, this study regarded the wife's alcoholic problem as the problems of dysfunctional interaction among family members rather than an individual problem. The study had three purposes as follows. First, what were the interactions sustaining the conflict of remarried family? Second, what were the characteristics of remarried family relating to family conflict? Third, what were the interventions of family therapy to resolve the conflicts of remarried family? The total family counseling was 9 sessions, which included individual, couple, and family counseling. The study used constant comparative analysis by using open coding method to find the factors relating interactions sustaining the conflict of remarried family, the characteristics of remarried family relating to family conflict, and the interventions of family therapy to resolve the conflicts of remarried family. Also, the study used Miles and Huberman's matrix and graphical network display to show the interaction of family members. The study found intriguing factors, counteracting factors, relationship level, and relational elements to wife's alcoholic problems among family members. The study revealed husband's rigid family rules, husband's execution of patriarchal authority, husband's pursuit of family identity, and husband's emphasis of union among family members as the characteristics of remarried family. On the other hand, the study found the reinforcement of spousal subsystem, the pursuit of change in communication pattern between husband and wife, the amelioration of rigid family rules, and the acceptance of differences among family members as the intervention of family therapy to resolve the conflicts of remarried family.
The purpose of this paper is to measure the clustering change and analyze empirical results. Additionally, by using k-means, hierarchical, and mixed models on Asian container ports over the period 2006-2015, the study aims to form a cluster comprising Busan, Incheon, and Gwangyang ports. The models consider the number of cranes, depth, birth length, and total area as inputs and container twenty-foot equivalent units(TEU) as output. Following are the main empirical results. First, ranking order according to the increasing ratio during the 10 years analysis shows that the value for average linkage(AL), mixed ward, rule of thumb(RT)& elbow, ward, and mixed AL are 42.04% up, 35.01% up, 30.47%up, and 23.65% up, respectively. Second, according to the RT and elbow models, the three Korean ports can be clustered with Asian ports in the following manner: Busan Port(Hong Kong, Guangzhou, Qingdao, and Singapore), Incheon Port(Tokyo, Nagoya, Osaka, Manila, and Bangkok), and Gwangyang Port(Gungzhou, Ningbo, Qingdao, and Kasiung). Third, optimal clustering numbers are as follows: AL(6), Mixed Ward(5), RT&elbow(4), Ward(5), and Mixed AL(6). Fourth, empirical clustering results match with those of questionnaire-Busan Port(80%), Incheon Port(17%), and Gwangyang Port(50%). The policy implication is that related parties of Korean seaports should introduce port improvement plans like the benchmarking of clustered seaports.
Purpose SPECT/CT was noted for its excellent correction method and qualitative functions based on fusion images in the early stages of dissemination, and interest in and utilization of quantitative functions has been increasing with the recent introduction of companion diagnostic therapy(Theranostics). Unlike PET/CT, various conditions like the type of collimator and detector rotation are a challenging factor for image acquisition and reconstruction methods at absolute quantification of SPECT/CT. Therefore, in this study, We want to find out the effect on the radioactivity concentration estimate by the increase or decrease of the total acquisition time according to the number of projections and the acquisition time per projection among SPECT/CT imaging conditions. Materials and Methods After filling the 9,293 ml cylindrical phantom with sterile water and diluting 99mTc 91.76 MBq, the standard image was taken with a total acquisition time of 600 sec (10 sec/frame × 120 frames, matrix size 128 × 128) and also volume sensitivity and the calibration factor was verified. Based on the standard image, the comparative images were obtained by increasing or decreasing the total acquisition time. namely 60 (-90%), 150 (-75%), 300 (-50%), 450 (-25%), 900 (+50%), and 1200 (+100%) sec. For each image detail, the acquisition time(sec/frame) per projection was set to 1.0, 2.5, 5.0, 7.5, 15.0 and 20.0 sec (fixed number of projections: 120 frame) and the number of projection images was set to 12, 30, 60, 90, 180 and 240 frames(fixed time per projection:10 sec). Based on the coefficients measured through the volume of interest in each acquired image, the percentage of variation about the contrast to noise ratio (CNR) was determined as a qualitative assessment, and the quantitative assessment was conducted through the percentage of variation of the radioactivity concentration estimate. At this time, the relationship between the radioactivity concentration estimate (cps/ml) and the actual radioactivity concentration (Bq/ml) was compared and analyzed using the recovery coefficient (RC_Recovery Coefficients) as an indicator. Results The results [CNR, radioactivity Concentration, RC] by the change in the number of projections for each increase or decrease rate (-90%, -75%, -50%, -25%, +50%, +100%) of total acquisition time are as follows. [-89.5%, +3.90%, 1.04] at -90%, [-77.9%, +2.71%, 1.03] at -75%, [-55.6%, +1.85%, 1.02] at -50%, [-33.6%, +1.37%, 1.01] at -25%, [-33.7%, +0.71%, 1.01] at +50%, [+93.2%, +0.32%, 1.00] at +100%. and also The results [CNR, radioactivity Concentration, RC] by the acquisition time change for each increase or decrease rate (-90%, -75%, -50%, -25%, +50%, +100%) of total acquisition time are as follows. [-89.3%, -3.55%, 0.96] at - 90%, [-73.4%, -0.17%, 1.00] at -75%, [-49.6%, -0.34%, 1.00] at -50%, [-24.9%, 0.03%, 1.00] at -25%, [+49.3%, -0.04%, 1.00] at +50%, [+99.0%, +0.11%, 1.00] at +100%. Conclusion In SPECT/CT, the total coefficient obtained according to the increase or decrease of the total acquisition time and the resulting image quality (CNR) showed a pattern that changed proportionally. On the other hand, quantitative evaluations through absolute quantification showed a change of less than 5% (-3.55 to +3.90%) under all experimental conditions, maintaining quantitative accuracy (RC 0.96 to 1.04). Considering the reduction of the total acquisition time rather than the increasing of the image acquiring time, The reduction in total acquisition time is applicable to quantitative analysis without significant loss and is judged to be clinically effective. This study shows that when increasing or decreasing of total acquisition time, changes in acquisition time per projection have fewer fluctuations that occur in qualitative and quantitative condition changes than the change in the number of projections under the same scanning time conditions.
Purpose: Raynaud scan is divided to flow, blood pool and local-delay image. Usually, we evaluate comparison through blood pool and local-delay image. We will evaluate about usability when comparative observe blood image and local-delay image in Raynaud scan that used
The wall shear stress in the vicinity of end-to end anastomoses under steady flow conditions was measured using a flush-mounted hot-film anemometer(FMHFA) probe. The experimental measurements were in good agreement with numerical results except in flow with low Reynolds numbers. The wall shear stress increased proximal to the anastomosis in flow from the Penrose tubing (simulating an artery) to the PTFE: graft. In flow from the PTFE graft to the Penrose tubing, low wall shear stress was observed distal to the anastomosis. Abnormal distributions of wall shear stress in the vicinity of the anastomosis, resulting from the compliance mismatch between the graft and the host artery, might be an important factor of ANFH formation and the graft failure. The present study suggests a correlation between regions of the low wall shear stress and the development of anastomotic neointimal fibrous hyperplasia(ANPH) in end-to-end anastomoses. 30523 T00401030523 ^x Air pressure decay(APD) rate and ultrafiltration rate(UFR) tests were performed on new and saline rinsed dialyzers as well as those roused in patients several times. C-DAK 4000 (Cordis Dow) and CF IS-11 (Baxter Travenol) reused dialyzers obtained from the dialysis clinic were used in the present study. The new dialyzers exhibited a relatively flat APD, whereas saline rinsed and reused dialyzers showed considerable amount of decay. C-DAH dialyzers had a larger APD(11.70
The wall shear stress in the vicinity of end-to end anastomoses under steady flow conditions was measured using a flush-mounted hot-film anemometer(FMHFA) probe. The experimental measurements were in good agreement with numerical results except in flow with low Reynolds numbers. The wall shear stress increased proximal to the anastomosis in flow from the Penrose tubing (simulating an artery) to the PTFE: graft. In flow from the PTFE graft to the Penrose tubing, low wall shear stress was observed distal to the anastomosis. Abnormal distributions of wall shear stress in the vicinity of the anastomosis, resulting from the compliance mismatch between the graft and the host artery, might be an important factor of ANFH formation and the graft failure. The present study suggests a correlation between regions of the low wall shear stress and the development of anastomotic neointimal fibrous hyperplasia(ANPH) in end-to-end anastomoses. 30523 T00401030523 ^x Air pressure decay(APD) rate and ultrafiltration rate(UFR) tests were performed on new and saline rinsed dialyzers as well as those roused in patients several times. C-DAK 4000 (Cordis Dow) and CF IS-11 (Baxter Travenol) reused dialyzers obtained from the dialysis clinic were used in the present study. The new dialyzers exhibited a relatively flat APD, whereas saline rinsed and reused dialyzers showed considerable amount of decay. C-DAH dialyzers had a larger APD(11.70