BioBrick-based 'Quick Gene Assembly' in vitro.

Because of the technological limitations of de novo DNA synthesis in (i) making constructs containing tandemly repeated DNA sequence units, (ii) making an unbiased DNA library containing DNA fragments with sequence multiplicity in a specific region of target genes, and (iii) replacing DNA fragments, development of efficient and reliable biochemical gene assembly methods is still anticipated. We succeeded in developing a biological standardized genetic parts that are flanked between a common upstream and downstream nucleotide sequences in an appropriate plasmid DNA vector (BioBrick)-based novel assembly method that can be used to assemble genes composed of 25 tandemly repeated BioBricks in the correct format in vitro. We named our new DNA part assembly system: 'Quick Gene Assembly (QGA)'. The time required for finishing a sequential fusion of five BioBricks is less than 24 h. We believe that the QGA method could be one of the best methods for 'gene construction based on engineering principles' at the present time, and is also a method suitable for automation in the near future.

[Basic Evaluation of Microsemi LC-767CRP as POCT Model for Simultaneous Measurement of CBC and CRP].

Microsemi LC-767CRP (LC-767, Horiba, Ltd.) is capable of simultaneous measuring of complete blood count (CBC) including 3-part differentials (3-part Diff.) of white blood cells (WBC) and C-reactive protein (CRP) in 4 minutes. Data obtained using LC-767 were intra-assay-reproducible (n = 10, CV = 0.6-4.0% for CBC, 0.6-2.5% for 3-part Diff. and 2.8-7.7% for CRP). They also showed the good linearity, no definite carry-over and the excellent correlations with routine instruments in our institution. Concerning CRP, the minimal detectable concentration revealed < 0.1 mg/dL, and prozone was observed in the sample containing > 30 mg/dL of CRP. LC-767 showed better correlation with a routine instrument in monocyte percentage than LC-667, probably due to modification of the hemolysis solution ratio and diluent temperature. In conclusion, LC-767 provided accurate CBC and CRP results, and showed improvement in CRP linearity and monocyte percentage compared with LC-667. LC-767, which is equipped with a bar-code reader with easy accessibility to electronic medical record, is suitable as the next-generation point of care testing model in the era of information and network-oriented medicine.

[Experience of Collaborative Research through Department of Medical Instrumental Research and Technology in Kyoto Prefectural University of Medicine].

Both of Kyoto Prefectural University of Medicine which offers high, technical and safe medical treatment and Horiba, Ltd. which has small CBC analyzers in a core product established a joint research institute for development of advanced laboratory test analyzer from January 1, 2012 in Kyoto Prefectural University of Medicine as the "advanced treatment hospital" where the Ministry of Health, Labour and Welfare has got approved. Clinical needs about analyzer and reagent for a laboratory test are being investigated to the emergency medical care unit and the intensive care unit as well as the laboratory test part in the affiliated hospital and many medical departments of the pediatrics, the internal medicine and the surgery. Developing the new analyzer based on high technology, evaluating the performance of them and spreading them to a medical examination and treatment site is our main target.

[Effect of sample volumes and utilized anticoagulants on the measurement of complete blood counts including 3-part differentials using capillary blood].

We collected the different volumes of venous blood(60 microL, 125 microL, 250 microL and 500 microL) into micro sample cups from respective volunteer to compare complete blood count (CBC) among 4 sample volumes, and found that 60 microL of sample volume seemed enough for CBC measurement using Microsemi LC-667 CRP (Horiba Co.). Subsequently, we measured CBC using 60 microL of peripheral blood after combining one of the 3 capillary tubes (heparin coated, EDTA coated and plain) with either EDTA coated or plain micro sample cups to examine the effect of anticoagulants contained into these commercially available maneuvers for capillary blood sampling. When we used the plain micro sample cup, platelet aggregation and false increase of white blood cell(WBC) count were observed irrespective to the combination of capillary tubes. We also tried whether commercially recommended volume (250 microL) of sample could be obtained by either fingertip or earlobe puncture from volunteers, and found that 7 of 16 fingertip and only 1 earlobe punctures could achieve sufficient volume. Whereas, at least 60 microL of sample were available more than 80% of volunteers by both methods, and CBC data obtained from these lesser samples obtained by fingertip puncture showed no statistically significant differences when compared with those of conventional venous samples (2 mL). From these findings, we concluded that at least 60 microL of capillary blood obtained from fingertip then collected into EDTA coated micro sample cup was enough to measure CBC using Microsemi LC-667 CRP.

[Clinical evaluation of arteriosclerosis using high sensitive CRP in multiphasic screening].

In order to identify the chronic inflammation, hsCRP measurement at several time are recommended. The continuous higher value of CRP is observed as the signal of the chronic inflammation. In this case the average of CRP was more than 0.15mg/dl and the standard deviation was less than 100%. In this study, the hsCRP showed the biggest AUC (Aria Under Curve) at acute myocardial infarction (AMI). The blood pressure showed the biggest AUC at stroke. The event rate for AMI of patients who have the continuous higher CRP value is higher than the normal control. The relative risk for AMI is 20 fold. On the other hand, the relative risk for stroke is less than 1 fold. The key factor of the disease was suggested difference between AMI and stroke.