Microproteomics with microfluidic-based cell sorting: Application to 1000 and 100 immune cells.

Ultimately, cell biology seeks to define molecular mechanisms underlying cellular functions. However, heterogeneity within cell populations must be considered for optimal assay design and data interpretation. Although single-cell analyses are desirable for addressing this issue, practical considerations, including assay sensitivity, limit their broad application. Therefore, omics studies on small numbers of cells in defined subpopulations represent a viable alternative for elucidating cell functions at the molecular level. MS-based proteomics allows in-depth proteome exploration, although analyses of small numbers of cells have not been pursued due to loss during the multistep procedure involved. Thus, optimization of the proteomics workflow to facilitate the analysis of rare cells would be useful. Here, we report a microproteomics workflow for limited numbers of immune cells using non-damaging, microfluidic chip-based cell sorting and MS-based proteomics. Samples of 1000 or 100 THP-1 cells were sorted, and after enzymatic digestion, peptide mixtures were subjected to nano-LC-MS analysis. We achieved reasonable proteome coverage from as few as 100-sorted cells, and the data obtained from 1000-sorted cells were as comprehensive as those obtained using 1 µg of whole cell lysate. With further refinement, our approach could be useful for studying cell subpopulations or limited samples, such as clinical specimens.

Impact of combined lipid lowering and blood pressure control on coronary plaque: myocardial ischemia treated by percutaneous coronary intervention and plaque regression by lipid lowering and blood pressure controlling assessed by intravascular ultrasonography (MILLION) study.

The aim of the study was to elucidate the aggressive reduction of both low-density lipoprotein cholesterol (LDL-C) and blood pressure (BP) reduced coronary atherosclerotic plaque volume compared with a standard treatment of LDL-C and BP in Japanese patients with coronary artery disease (CAD). This study is a prospective, randomized, and open-labelled with a blind-endpoint evaluation study. A total of 97 patients (81 men, mean age 62.0 ± 9.6) with CAD undergoing intravascular ultrasonography (IVUS)-guided percutaneous coronary intervention (PCI) were randomized, and 68 patients had IVUS examinations at baseline and at 18-24 months follow-up. Patients were randomly assigned to standard or aggressive strategies targeting LDL-C and a BP of 100 mg/dL and 140/90 mmHg vs. 70 mg/dL and 120/70 mmHg, respectively. The primary endpoint was the percent change in coronary plaque volume. Both standard and aggressive strategies succeeded to achieve target levels of LDL-C and BP; 74.9 ± 14.7 vs. 63.7 ± 11.9 mg/dL (NS) and 124.1 ± 9.4/75.8 ± 7.7 vs. 113.6 ± 9.6/65.8 ± 9.4 mmHg (systolic BP; NS, diastolic BP; p < 0.05), respectively. Both groups showed a significant reduction in the coronary plaque volume of -9.4 ± 10.7% and -8.7 ± 8.6% (NS) in standard and aggressive therapies, respectively. Both standard and aggressive intervention significantly regressed coronary plaque volume by the same degree, suggesting the importance of simultaneous reductions of LDL-C and BP for prevention of CAD.

Impact of combined lipid lowering with blood pressure control on coronary plaque regression: rationale and design of MILLION study.

A line of epidemiological studies suggests that the accumulation of coronary risk factors promotes the progression of coronary atherosclerosis. Recent clinical studies showed that aggressive low-density lipoprotein (LDL) cholesterol-lowering therapy using statins could regress coronary atheroma and reduce major cardiovascular events. Additionally, therapy that controlled amlodipine-based blood pressure reduced major cardiovascular events in patients with hypertension compared with an atenolol-based regimen. An open-label randomized multicenter study is primarily planned to evaluate the changes in coronary atheroma volume using intravascular ultrasonography 18-24 months after intensive lowering of LDL-cholesterol and blood pressure compared with a standard therapy indicated by current guidelines in Japanese patients with coronary artery disease (CAD). The secondary endpoints include changes in serum lipid levels, inflammatory markers, glucose markers and blood pressure. In total, 100 subjects with CAD who are undergoing percutaneous coronary intervention will be tested. The MILLION study will provide new evidence and therapeutic standards for the prevention of CAD in Japanese patients by controlling both LDL-C levels and blood pressure.

Prescription rate of medications potentially contributing to lower urinary tract symptoms and detection of adverse reactions by prescription sequence symmetry analysis.

BACKGROUND: The lower urinary tract symptoms (LUTS) increases with age and can have a significant effect on the quality of life of the patients. Elderly patients, who are often characterized by a decline in physiological functional and polypharmacy, are susceptible to adverse drug reactions to pharmacotherapy. LUTS can also be a side effect of medication. The purpose of this study was to investigate the possible association between the initiation of LUTS-causing drug therapy and the onset of LUTS. METHODS: Drug dispensing data at the individual level were retrieved from the CISA (Platform for Clinical Information Statistical Analysis: http://www.cisa.jp) database. A retrospective study was conducted by reviewing patients with LUTS who were dispensed drugs that increased the risk of LUTS between April 2011 and March 2012. Prescription sequence symmetry analysis (PSSA) was employed to investigate the associations between the dispensing of medicines of LUTS and that of LUTS-causing drugs. RESULTS: LUTS-causing drugs were frequently dispensed to patients with LUTS. The use of medications potentially contributing to LUTS was associated with polypharmacy [number of prescription drugs:12.13 ± 6.78 (user) vs. 5.67 ± 5.24 (nonuser)] but not patient age [ age: (71.38 ± 13.28 (user) vs. 70.45 ± 14.80 (nonuser)]. Significant adverse drug events were observed the use of donepezil, cyclophosphamide, antiparkinson drugs, antidepressant, diazepam, antipsychotic drugs for peptic ulcer, tiotropium bromide, and opioids. CONCLUSIONS: The use of prescription LUTS-causing drugs was correlated with polypharmacy. The adverse drug events associated with LUTS-causing drugs were highly prevalent in elderly patients. To prevent of adverse drug events in patients with LUTS, pharmacists and physicians should regularly review medication lists and reduce the prescribed medicines.

Development of a methodology for the detection of hospital financial outliers using information systems.

Comparison of financial indices helps to illustrate differences in operations and efficiency among similar hospitals. Outlier data tend to influence statistical indices, and so detection of outliers is desirable. Development of a methodology for financial outlier detection using information systems will help to reduce the time and effort required, eliminate the subjective elements in detection of outlier data, and improve the efficiency and quality of analysis. The purpose of this research was to develop such a methodology. Financial outliers were defined based on a case model. An outlier-detection method using the distances between cases in multi-dimensional space is proposed. Experiments using three diagnosis groups indicated successful detection of cases for which the profitability and income structure differed from other cases. Therefore, the method proposed here can be used to detect outliers.

The effects of amino-acid mutations on specific interactions between urokinase-type plasminogen activator and its receptor: Ab initio molecular orbital calculations.

During cancer invasion, the binding of urokinase-type plasminogen activator (uPA) to its receptor (uPAR) on the surface of a cancer cell is considered a trigger for invasion. Here, we present a stable structure of the solvated complex formed between uPA and uPAR (uPA-uPAR) and investigate the specific interactions between uPA and uPAR by ab initio fragment molecular orbital (FMO) calculations. The result indicates that the electrostatic interactions between the charged amino acid residues existing in both uPA and uPAR make a large contribution to the binding between uPA and uPAR. In particular, Lys23, Lys46, Lys98 and Lys61 of uPA are found to have strong attractive interactions with uPAR. To elucidate the effect of these residues on the interactions between uPA and uPAR, we substituted each of them with the uncharged amino acid Leu and investigated the interactions between the mutated uPA and wild-type uPAR. The interaction energies indicate that Lys46 and Lys98, which bind uPA to the rim of the central ligand-binding cavity of uPAR, make greater contributions to the binding between uPA and uPAR than Lys23, which is positioned at the bottom of the ligand-binding cavity of uPAR. The effect of hydrating water molecules located between uPA and uPAR is also investigated to be significant for the specific interactions between uPA and uPAR. These results are expected to be informative for developing new peptide antagonists that block the binding of uPA to uPAR.

Ab initio molecular orbital calculations on specific interactions between urokinase-type plasminogen activator and its receptor.

Cancer invasions and metastases are controlled by various proteases. In particular, the binding of urokinase-type plasminogen activator (uPA) to the uPA receptor (uPAR) existing on the surface of cancer cell is considered to be a trigger for cancer invasions. In the present study, we determined the structure of uPA and uPAR complex in water and investigated the specific interactions between uPA and uPAR by ab initio molecular orbital (MO) calculations based on fragment MO method. The result indicates that the 20-26 amino acid residues of uPA are important for the binding between uPA and uPAR, and that the electrostatic interactions between the charged amino acid residues existing in both uPA and uPAR have large contribution to the binding. The influence of crystal water molecules existing between uPA and uPAR was also investigated to be significant on the specific interactions between uPA and uPAR. These results are expected to be informative for developing new medicines blocking the binding of uPA and uPAR.

A proposal of speculative operation on distributed system for FEM-based ablation simulator.

This study aims to provide physics-based force feedback system on distributed system for simulating invasive operation such as ablation. conventional PC-based VR surgical simulators with haptic interaction are hard to provide sufficient computational resources for the simulation of physics-based soft tissue fracture. For proper presentation of force feedback as real operations, physics-based simulation is inevitable. At the same time finite element method requires huge computational complexites. In this paper, the authors propose server-side speculative operation method on application layer for hiding the calculation latencies. The proposed method would achieve the response acceleration without the decomposition of conventional simulation process. The theoritical estimates of speculation parameters are mentioned.

Organ exclusion simulation with multi-finger haptic interaction for open surgery simulator.

Exclusion is a surgical manipulation of pushing aside organ in open surgery. Recently, training opportunity of surgeon is decreasing due to animal protection and patient's rights. In this study, we propose an organ exclusion training simulator with multi-finger haptic device and stress visualization. The method was applied to a medical application of exclusion which is an important manipulation to make a hidden tissue visible or to enlarge workspace. The system equips FEM-based soft tissue deformation and multi-finger haptic device. Real-time simulation was achieved with a prototype system. Experimental results of training trial suggested the effectiveness of the system and stress visualization for exclusion training. Results of subjective evaluation by surgeons were highly positive as to realism of manipulation and usefulness of the simulator.

A novel approach for training of surgical procedures based on visualization and annotation of behavioural parameters in simulators.

Recording performance during training sessions on simulators is becoming a new standard for assessment of surgical skills and thus a significant part of training. Typical simulator-based training can be assessed using criteria that cover the whole procedure to make distinction between skill levels. Studies so far have rarely addressed the challenge of how to provide better feedback about the user's performance on a surgical simulator. Our approach for surgical training is Annotated Simulation Records (ASRs) and visualization of behavioural parameters of interaction in surgery. This paper briefly outlines a framework for building user-defined skill models and presents initial results. We demonstrate the ASR-based approach in force exertion tasks on elastic objects by utilizing a cardiovascular surgeon's recorded interaction on an aorta palpation simulator.

Transfusion checkup system using mobile terminals.

Kyoto University Hospital introduces a new hospital information system on January 2005. The system introduces ubiquitous information system using mobile terminals for nursing information support and implements a transfusion checkup system, as a single error in the transfusion process may result in a serious accident. The developed system carefully designed to suit real workflow and to make full use of the benefits of mobile terminals increases efficiency and the safety of transfusion.

Electronic clinical path system based on semistructured data model using personal digital assistant for onsite access.

Clinical Paths (Paths) have been introduced by different hospitals for patient care management. An Electronic Clinical Path (ECP) with onsite access provision seems to improve the efficiency of medical staffs because they can share vast medical information about patients at a time and also can reuse accumulated data easily, which is impossible with paper-based Path. Data model is the basis for implementing ECP. However, there is no established model for ECP. The purpose of this study is to introduce a model for ECP and implement an ECP with onsite access system. We introduced a Semistructured Data Model (SSDM) for ECP, and implemented a Web application system based on this model using Personal Digital Assistant (PDA) as inputting device. Our system functioned as expected with wireless LAN, and users handled the data on bedside using PDA. By introducing SSDM, we showed the correspondence between schema of Paths and implementation of ECP.