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Biomedical subjects

Ken Yamazaki

Publications and source records attributed to Ken Yamazaki.

12 recordsLinked to original sources
PubMed2006

Screening for ligands using a generic and high-throughput light-scattering-based assay.

Rapid identification of small molecules that interact with protein targets using a generic screening method greatly facilitates the development of therapeutic agents. The authors describe a novel method for performing homogeneous biophysical assays in a high-throughput format. The use of light scattering as a method to evaluate protein stability during thermal denaturation in a 384-well format yields a robust assay with a low frequency of false positives. This novel method leads to the identification of interacting small molecules without the addition of extraneous fluorescent probes. The analysis and interpretation of data is rapid, with sensitivity for protein stability comparable to differential scanning calorimetry. The authors propose potential uses in drug discovery, structural genomics, and functional genomics as a method to evaluate small-molecule interactions, identify natural cofactors that stabilize target proteins, and identify natural substrates and products for previously uncharacterized protein targets.

Binding Sites↗
PubMed2006

A biomechanical assessment of infra-laminar hooks as an alternative to supra-laminar hooks in thoracolumbar fixation.

STUDY DESIGN: Biomechanical study, using human cadaveric spines, of two types of posterior fixation in the thoracolumbar spine. OBJECTIVES: To compare fixation in the thoracolumbar spine using pedicle screws combined with infra-laminar hooks (at both the cranial and caudal ends of the same vertebra) with fixation using pedicle screws alone. SUMMARY OF BACKGROUND DATA: The standard method of using laminar hooks is to place supra-laminar hooks at the level above the vertebra being fixed by the cranial pedicle screws, and infra-laminar hooks at the level of the vertebra receiving the caudal pedicle screws. However, using this method, a single normal motion segment is killed. In the proposed technique, infra-laminar hooks will be used at the same level as the cranial pedicle screws, thus sparing the superior adjacent segment. METHODS: PEDICLE SCREW LOOSENING TEST: Nine vertebrae (T9-L2) had pedicle screws inserted into both pedicles. On one side, the pedicle screw was connected to a fixation rod, while on the other side the pedicle screw was combined with an infra-laminar hook and connected to a rod. The rod on one side was then pushed with increasing force until loosening of the screw. The same test was repeated on the other rod until loosening of the screw. The reverse test (i.e., pulling until loosening) was done in a similar manner on another nine vertebrae. BIOMECHANICAL STIFFNESS TEST: Eight spines (T12-L2) were biomechanically tested in: axial compression, flexion, extension, left and right lateral bending, and left and right axial torsion. This sequence was applied to: 1) the intact spine; 2) after applying pedicle screws and infra-laminar hooks with rods to the destabilized spine; and 3) after removal of the hooks (with the pedicle screw and rods still inserted). From the load-deformation curves obtained, stiffness values were calculated. RESULTS: PEDICLE SCREW LOOSENING TEST: The pedicle screw loosening tests showed that a pedicle screw combined with an infra-laminar hook offers significantly greater fixation strength as compared to a pedicle screw alone. Biomechanical stiffness test. Fixation by pedicle screws combined with infra-laminar hooks, placed on the vertebra fixed by pedicle screws at both the cranial and caudal ends, offered a stiffer construct as compared to fixation by pedicle screws alone. CONCLUSIONS: This method offers a firm construct without sacrificing a normal motion segment.

Bone Screws↗
PubMed2006

Establishment of perineural invasion models and analysis of gene expression revealed an invariant chain (CD74) as a possible molecule involved in perineural invasion in pancreatic cancer.

PURPOSE: Perineural invasion causes frequent local recurrence even after resection and a poor prognosis for pancreatic cancer. We established perineural invasion models and analyzed the molecular mechanism of perineural invasion in pancreatic cancer. EXPERIMENTAL DESIGN: Seven pancreatic cancer cell lines with or without human peripheral nerves were s.c. implanted in nonobese diabetes/severe combined immunodeficient mice. We compared expression profiles among high and low perineural invasion cell lines by using an oligonucleotide microarray. We examined up-regulation of the invariant chain (CD74) in high perineural invasion cell lines in mRNA and protein levels and surgical cases immunohistochemically. RESULTS: Four of seven pancreatic cancer cell lines (CaPan1, CaPan2, CFPAC, and MPanc96) showed perineural invasion to s.c. transplanted human peripheral nerves. Moreover, CaPan1 and CaPan2 (high perineural invasion group) also resulted in a high frequency of perineural invasion to mouse s.c. peripheral nerves, whereas three pancreatic cancer cell lines HPAFII, AsPC1, and Panc1 (low perineural invasion group) did not show perineural invasion to either human or mouse nerves. We identified 37 up-regulated genes and 12 down-regulated genes in the high perineural invasion group compared with the low perineural invasion group. Among them, CD74 was up-regulated in the high perineural invasion group in mRNA and protein levels. Furthermore, immunohistochemical expression of CD74 in clinical cases revealed its significant overexpression in pancreatic cancer with perineural invasion (P < 0.008). CONCLUSIONS: This is the first report of perineural invasion models using human pancreatic cancer cell lines. In combination with gene expression profiling, it was indicated that CD74 could be a candidate molecule involved in perineural invasion. These models provide new approaches for study of perineural invasion in pancreatic cancer.

Aged↗
PubMed2006

Variations in carotid arterial compliance during the menstrual cycle in young women.

The effect of menstrual cycle phase on arterial elasticity is controversial. In 10 healthy women (20.6+/-1.5 years old, mean+/-s.d.), we investigated the variations in central and peripheral arterial elasticity, blood pressure (carotid and brachial), carotid intima-media thickness (IMT), and serum oestradiol and progesterone concentrations at five points in the menstrual cycle (menstrual, M; follicular, F; ovulatory, O; early luteal, EL; and late luteal, LL). Carotid arterial compliance (simultaneous ultrasound and applanation tonometry) varied cyclically, with significant increases from the values seen in M (0.164+/-0.036 mm2 mmHg-1) and F (0.171+/-0.029 mm2 mmHg-1) to that seen in the O phase (0.184+/-0.029 mm2 mmHg-1). Sharp declines were observed in the EL (0.150+/-0.033 mm2 mmHg-1) and LL phases (0.147+/-0.026 mm2 mmHg-1; F=8.51, P<0.05). Pulse wave velocity in the leg (i.e. peripheral arterial stiffness) did not exhibit any significant changes. Fluctuations in carotid arterial elasticity correlated with the balance between oestradiol and progesterone concentrations. No significant changes were found in carotid and brachial blood pressures, carotid artery lumen diameter, or IMT throughout the menstrual cycle. These data provide evidence that the elastic properties of central, but not peripheral, arteries fluctuate significantly with the phases of the menstrual cycle.

Adolescent↗
PubMed2006

A biomechanical study of 3 different types of sublaminar wire used for constructs in the thoracic spine.

A biomechanical study was carried out on 3 different types of sublaminar wire used in constructs to secure the thoracic spine: stainless steel monofilament wire (steel wire), titanium cable (cable), and ultra-high molecular weight polyethylene tape (tape). Two experiments were carried out. Experiment 1: Thirty-one fresh human thoracic vertebrae classified as osteoporotic (bone mineral density of <0.8 g/cm2) were used. The steel wire, cable, or tape was placed sublaminarly and a tensile force was applied until the steel wire, cable, or tape cut 5 mm through the lamina, and the force at this point was noted. Experiment 2: Seven fresh human thoracic spines (T7-T10) were biomechanically tested as follows: axial compression (250 N), flexion (7.5 Nm), extension (7.5 Nm), left lateral bending (7.5 Nm), right lateral bending (7.5 Nm), left axial torsion (10 Nm), and right axial torsion (10 Nm). This sequence was applied to the intact spine. The spine was then de-stabilized and then restabilized using one or other of the 3 different types of sublaminar wires. The biomechanical testing was then repeated on the restabilized spine and stiffness curves were generated. In the laminar cut-through test, the cut-through force for tape was higher than that for either steel wire or cable. In the biomechanical stiffness testing, there was no significant difference between the 3 different sublaminar wiring constructs in any of the loading modes tested. The results of both experiments suggest that tape is as good, if not better, than steel wire or cable as a sublaminar wiring construct material.

Aged↗
PubMed2004

Unfavorable effects of resistance training on central arterial compliance: a randomized intervention study.

BACKGROUND: Reductions in the compliance of central arteries exert a number of adverse effects on cardiovascular function and disease risk. Endurance training is efficacious in increasing arterial compliance in healthy adults. We determined the effects of resistance training on carotid arterial compliance using the intervention study design. METHODS AND RESULTS: Twenty-eight healthy men 20 to 38 years old were randomly assigned to the intervention group (n=14) and the control group (n=14). Control subjects were instructed not to alter their normal activity levels throughout the study period. Intervention subjects underwent 3 supervised resistance training sessions per week for 4 months and detraining for a subsequent 4 months. The resistance training increased maximal strength in all muscle groups tested (P<0.001). There were no significant differences in baseline arterial compliance and beta-stiffness index between the intervention and control groups. In the intervention group, carotid arterial compliance decreased 19% (P<0.05), and beta-stiffness index increased 21% (P<0.01) after resistance training. These values returned completely to the baseline levels during the detraining period. Arterial compliance did not change in the control group. In both groups, there were no significant changes in brachial and carotid blood pressure, carotid intima-media thickness, lumen diameter, and femoral arterial compliance. Changes in carotid artery compliance were significantly and negatively related to corresponding changes in left ventricular mass index (r=-0.56, P<0.001) and left ventricular hypertrophy index (r=-0.68, P<0.001). CONCLUSIONS: In marked contrast to the beneficial effect of regular aerobic exercise, several months of resistance training "reduces" central arterial compliance in healthy men.

Adult↗
PubMed2003

Overexpression of KIT in chromophobe renal cell carcinoma.

We analysed gene-expression profiles in 15 surgical specimens of conventional, papillary, and chromophobe renal cell carcinomas (RCCs) using high-density oligonucleotide arrays. From about 12,000 genes targeted by the array, 67 were upregulated specifically in each histological type of RCC. The oncogene KIT was one of the genes whose expression was upregulated specifically in chromophobe RCCs. Immunohistochemical analysis demonstrated the KIT gene product on the cell membrane of chromophobe RCC in all cases, although it was not detected in conventional RCCs or non-neoplastic kidneys except for weak staining in the cytoplasm of renal tubules. These results suggest that each histological subtype of RCC has a unique gene-expression profile, and in particular indicates for the first time that KIT could be a useful marker for chromophobe RCC. As overexpression of KIT might be involved in tumor growth, KIT could be a new therapeutic target in this special type of RCC.

Biomarkers, Tumor↗
PubMed2003

Expression profiling in multistage hepatocarcinogenesis: identification of HSP70 as a molecular marker of early hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) associated with chronic liver disease evolves from precancerous lesions and early HCC to a progressed form. Nodule-in-nodule-type HCC (progressed HCC within early HCC) represents the transition from early to progressed HCC and, therefore, is useful in molecular genetic analysis of HCC progression during multistage carcinogenesis. We compared expression profiles among 7 early components and 7 progressed components of nodule-in-nodule-type HCCs and their corresponding noncancerous liver tissues with oligonucleotide array. Of the approximately 12,600 genes that were analyzed, a set of 95 genes provided a molecular signature that distinguished between early HCC components and their noncancerous liver tissues, and a set of 92 genes distinguished between progressed and early HCC components. Of these genes, the most abundantly up-regulated gene in early HCC components (P <.001) was heat-shock protein 70 (HSP70). Real-time quantitative reverse transcription polymerase chain reaction (RT-PCR) confirmed this finding. Further immunohistochemical examination of HSP70 revealed its significant overexpression in early HCC compared with precancerous lesions (P <.001) and in progressed HCC compared with early HCC (P <.001). In conclusion, molecular signatures were clearly different in noncancerous liver tissue as compared with the early and progressed components of nodule-in-nodule-type HCC. Moreover, HSP70 could be a sensitive marker for the differential diagnosis of early HCC from precancerous lesion or noncancerous liver, a difficult distinction for pathologists due to very well differentiated histology with little atypia in early HCC.

Algorithms↗
PubMed2002

pc1 and psc1, zebrafish homologs of Drosophila Polycomb and Posterior sex combs, encode nuclear proteins capable of complex interactions.

Drosophila Polycomb group proteins are thought to form multimeric nuclear complexes that are responsible for stable transmission of repressed states of gene expression during the proliferation of differentiating embryos. In this study, we cloned, sequenced, and characterized two Polycomb group homologs, designated pc1 and psc1, in zebrafish. Amino acid sequence analyses determined that pc1 is a structural homolog of Drosophila Polycomb and that psc1 is a homolog of Drosophila Posterior sex combs. Northern blots and whole-mount in situ hybridization revealed that pc1 and psc1 had overlapping expression patterns at successive stages of embryogenesis. Immunocytochemistry localized both Pc1 and Psc1 protein in blastomere nuclei. Pull-down assays and two-hybrid system deletion analyses showed that these proteins were capable of homotypic and heterotypic interactions and identified the regions required for these interactions. The evidence supports the idea that zebrafish Polycomb group proteins, like those of other species, form nuclear complexes with compositions that may vary in a spatio-temporal manner during development.

Animals↗
PubMed2002

Rescue of hereditary form of dilated cardiomyopathy by rAAV-mediated somatic gene therapy: amelioration of morphological findings, sarcolemmal permeability, cardiac performances, and the prognosis of TO-2 hamsters.

The hereditary form comprises approximately 1/5 of patients with dilated cardiomyopathy (DCM) and is a major cause of advanced heart failure. Medical and socioeconomic settings require novel treatments other than cardiac transplantation. TO-2 strain hamsters with congenital DCM show similar clinical and genetic backgrounds to human cases that have defects in the delta-sarcoglycan (delta-SG) gene. To examine the long-term in vivo supplement of normal delta-SG gene driven by cytomegalovirus promoter, we analyzed the pathophysiologic effects of the transgene expression in TO-2 hearts by using recombinant adeno-associated virus vector. The transgene preserved sarcolemmal permeability detected in situ by mutual exclusivity between cardiomyocytes taking up intravenously administered Evans blue dye and expressing the delta-SG transgene throughout life. The persistent amelioration of sarcolemmal integrity improved wall thickness and the calcification score postmortem. Furthermore, in vivo myocardial contractility and hemodynamics, measured by echocardiography and cardiac catheterization, respectively, were normalized, especially in the diastolic performance. Most importantly, the survival period of the TO-2 hamsters was prolonged after the delta-SG gene transduction, and the animals remained active, exceeding the life expectancy of animals without transduction of the responsible gene. These results provide the first evidence that somatic gene therapy is promising for human DCM treatment, if the rAAV vector can be justified for clinical use.

Animals↗
PubMed2002

Effect of cycling experience and pedal cadence on the near-infrared spectroscopy parameters.

PURPOSE: Previously we demonstrated that the method to reorder near-infrared spectroscopy (NIRS) parameters against crank angle could serve as a useful measure in providing circulatory dynamics and metabolic changes in a working muscle during pedaling exercise. To examine further applicability of this method, we investigated the effects of cycling experience and pedal cadence on the NIRS parameters. METHODS: Noncyclists (NON), triathletes (TRI), and cyclists (CYC) performed pedaling exercises at a work intensity of 75% VO2max while changing pedal cadence (50, 75, 85, and 95 rpm). Physiological and biomechanical responses and NIRS parameters were measured. RESULTS: NIRS measurements determined with the reordered NIRS change demonstrated significant differences depending on the factors. The bottom peak of reordered NIRS changes in muscle blood volume and oxygenation level shifted upward with an increase in pedal cadence in NON but remained unchanged in CYC. The reordered NIRS change demonstrated a temporary increase at the crank angle corresponding to the relaxation phase of the working muscle. This temporary increase was observed even in the highest pedal cadence in CYC. The difference in levels between the peak of the temporary increase and the bottom peak of reordered NIRS change (LPB-diff) for CYC at 85 rpm was significantly larger than that for NON. The results with NIRS parameters corresponded to changes in pedal force and myoelectric activity during pedal thrust. CONCLUSIONS: The bottom peak level of the reordered NIRS changes and LPB-diff determined for blood volume are available to detect noninvasively the differences in circulatory dynamics and metabolic change during pedaling exercises performed at different pedal cadences and also to estimate the difference of physiological and technical developments for endurance cycling in athletes.

Adaptation, Physiological↗