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Biomedical subjects

M Hamada

Publications and source records attributed to M Hamada.

At least 19 recordsLinked to original sources

Different real-time PCR assays could lead to a different result of detection of varicella-zoster virus in facial palsy.

Real-time PCR is a useful tool for rapid detection of viral genomic DNA. However, there are many types of real-time PCR, and this variation may induce different results. The sensitivity of two different real-time PCR assays was evaluated for the detection of the varicella-zoster virus (VZV) genome: LightCycler PCR and TaqMan PCR. Auricular skin cells and saliva were sampled from 201 patients with facial nerve paralysis. A hundred and seventy-one of these patients were diagnosed clinically with Bell's palsy, and the remaining 30 with Ramsay Hunt syndrome. In 30 specimens obtained from Ramsay Hunt syndrome patients, VZV DNA was detected in 26 skin and 3 saliva specimens using the LightCycler PCR, while 28 skin and 9 saliva specimens were positive using the TaqMan PCR. None of the patients with Bell's palsy were positive for VZV by the LightCycler PCR, whereas five of these patients were positive by the TaqMan PCR. The TaqMan PCR assay has a better sensitivity compared to the LightCycler PCR for the detection of VZV genome from patients with facial palsy. Further study is required to develop a more sensitive real-time PCR.

Chickenpox↗

Induction of endonuclease G-mediated apopotosis in human oral squamous cell carcinoma cells by protein kinase C inhibitor safingol.

PKC inhibitor safingol suppressed the growth of human oral squamous cell carcinoma (SCC) cells significantly at concentrations that inhibit PKC isoforms. Safingol inhibited the translocation of PKC following treatment with 12-o-tetradecanoylphorbol 13-acetate (TPA) in PKC alpha-EGFP-transfected cells, but not in PKC beta-EGFP- transfected cells, indicating selective inhibition for PKC alpha in oral SCC cells. Flow cytometric analysis and DNA analysis by agarose gel electrophoresis revealed an increase in the proportion of sub-G(1) cells and DNA fragmentation in safingol-treated cells. Mitochondrial membrane potential was decreased, and cytochrome c was released from mitochondria. However, the safingol-induced cell death was not accompanied by activation of caspase 3 and cleavage of poly (ADP-ribose) polymerase (PARP). The broad-spectrum caspase inhibitor BD-fmk failed to prevent safingol-induced cell death. Another apoptogenic factor endonuclease G, but not apoptosis-inducing factor (AIF), was also released from mitochondria and translocated to the nucleus. These results suggest that PKC alpha inhibitor safingol induces an endonuclease G- mediated apoptosis in a caspase-independent manner.

Apoptosis↗

A very rare anatomic variation of the right ventricular branch: right ventricular descending branch from the anterior interventricular branch.

The anterior interventricular branch of the left coronary artery has the most constant distribution in the human heart and rarely gives off right ventricular branches. Here we report a case with a right ventricular branch which diverged from the anterior interventricular branch and descended on the anterior right ventricular wall parallel to the anterior interventricular sulcus; we termed it the "right ventricular descending branch." This artery gave a collateral artery to the occluded anterior interventricular branch at the apex, and had prevented anterior myocardial infarction. The right ventricular descending branch should be precisely identified in order to perform successful myocardial revascularization procedures such as coronary artery bypass grafting and percutaneous coronary intervention, especially in a patient with anterior interventricular branch occlusion.

Aged↗

Comparison between the cross-sectional area of bone-patellar tendon-bone grafts and multistranded hamstring tendon grafts obtained from the same patients.

This study intraoperatively compared the cross-sectional area (CSA) and graft-tunnel fit of bone-patellar tendon-bone (BTB) and multistranded hamstring tendon (STG) grafts harvested from the same patient. Twenty-two patients with a mean age of 26 years were the subjects of the study. Each BTB graft was harvested from the central third of the patellar tendon. Tripled or quadrupled semitendinosus tendon with or without gracilis tendon was used as a STG graft. CSAs of both grafts in the same patients were intraoperatively measured using a custom-made area micrometer during primary ACL reconstruction and revision surgery or during combined ACL and PCL reconstruction. Graft-tunnel fit was calculated by dividing the CSA of the graft by that of the tibial bone tunnel. The average CSA of STG grafts was significantly greater than that of BTB grafts. The average value of graft-tunnel fit for the STG grafts% was also greater than that of the BTB grafts%. Thus STG grafts have a larger CSA and closer graft-tunnel fit than BTB grafts in a clinical setting.

Adolescent↗

[Emergency coronary artery bypass grafting for acute coronary syndrome with preoperative intraaortic balloon pumping; comparative surgical outcome and long-term results].

With recent technical improvements in catheter interventional therapy, percutaneous coronary intervention (PCI) has now become the treatment of first choice for acute coronary syndrome (ACS). The objective of the present study was to evaluate critically the timing of coronary artery bypass grafting (CABG) for severe ACS with preoperative intraaortic balloon pumping (IABP). Since 1994, a total of 70 patients have gone emergency or urgent CABG for ACS. Of 70 patients, 50 patients required preoperative IABP. There were 22 patients (17 men, 5 women) with acute myocardial infarction (AMI), with a mean age of 67.7 years, and 28 patients (19 men, 9 women) with unstable angina pectoris (UAP), with a mean age of 69.2 years. There was a significant difference, between AMI and UAP, in the prevalence of emergency operation (95.5% vs 25.0%), in preoperative cardiogenic shock (81.8% vs 17.9%), in the level of preoperative CPK-MB (196.7 IU/l vs 2.0 IU/l) and in preoperative ejection fraction (41.8% vs 47.3%). Two patients in AMI required percutaneous cardiopulmonary support (PCPS). Thirteen patients in AMI and 22 patients in UAP presented left main trunk (LMT) disease. Of the 13 LMT patients in AMI, 4 patients were AMI due to acute occlusion in the LMT. The AMI patients received 2.45 distal anastomoses on average, while the UAP patients 3.14 distal anastomoses (p = 0.019). Excluding the mean number of distal anastomoses, there was no difference in the intraoperative technical factors, such as aortic cross clamping duration, cardiopulmonary bypass duration, rate of complete revascularization, between AMI and UAP. There were postoperative significant differences in low cardiac output syndrome (LOS) [45.6% in AMI vs 3.6% in UAP] and in prolongation of mechanical ventilation (59.1% in AMI vs 14.3% in UAP). The hospital mortality was 9.1% (2/22) in AMI, and 3.6% (1/28) in UAP, with no significant difference. Of these 3 patients, 1 patient died from perioperative cerebrovascular accident (CVA), another from LOS, and the other from postoperative mesenteric ischemia, with an overall mortality of 6.0% (3/50). The overall patency rate of the grafts was 100% in AMI and 96.6% in UAP. The 5-year-survival rate excluding in-hospital death was 72.5% in AMI, and 89.6% in UAP. The 5-year-cardiac event-free rate was 77% in AMI and 89.4% in UAP. The overall survival rate, and cardiac event-free rate, at 5 years was 80.8%, and 83.8%, respectively. In conclusion, for ACS cases, especially UAP cases of LMT, in which symptoms, findings of ischemia and hemodynamics are stabilized by medical intervention including IABP; emergency surgery could be avoided immediately after coronary angiography. Recovery in the ischemic myocardium is intended by IABP, and urgent surgery should be performed after sufficient and precise preoperative examinations. An improvement not only in the perioperative but also long-term results can be expected by performing complete revascularizations.

Aged↗

Beneficial effect of replacing of angiotensin-converting enzyme inhibitor with angiotensin II antagonist for heart failure patients.

OBJECTIVE: Angiotensin-converting enzyme (ACE) inhibitors and beta-blockers improve prognosis inpatients with chronic heart failure. Some patients, however, show little response to combined treatment with an ACE inhibitor and a beta-blocker. In addition, the ACE inhibitor cannot completely suppress angiotensin II production. Our objective was to examine whether replacing the ACE inhibitor with an angiotensin II antagonist can improve the condition of patients with chronic heart failure. METHODS: In 11 patients with chronic heart failure treated with an ACE inhibitor and a beta-blocker, who have had severe left ventricular dysfunction or high plasma level of natriuretic peptides, left ventricular dimension, and fractional shortening, plasma atrial and brain natriuretic peptide (ANP and BNP) levels were determined before and 3 months after the change of treatment. RESULTS: After substituting the ACE inhibitor with an angiotensin II antagonist, patients showed New York Heart Association functional class improvement, and significant decrease in left ventricular dimension and BNP. CONCLUSION: In patients with severe chronic heart failure treated with an ACE inhibitor and a beta-blocker, replacing the ACE inhibitor with an angiotensin II antagonist may be effective. However, this has to be confirmed by an adequately powered randomized controlled study.

Adult↗

Correlation between age and vital organ function following deep hypothermic circulatory arrest.

BACKGROUND: We investigated the correlation bet-ween perioperative variables such as patients' age and vital organ function after operation on thoracic aorta using deep hypothermic circulatory arrest (DHCA). METHODS: Ninety-five patients who underwent replacement of thoracic aorta under DHCA for acute or chronic aortic dissection and aortic aneurysm, and survived more than 10 days were divided into group I (age less than 60, n=17), group II (between 60 and 69, n=39), and group III (over 70, n=39). Concomitant procedures such as aortic root replacement and coronary artery bypass grafting were performed in 9, 4, and 1 patients in group I, II, and III, respectively. Postoperative pulmonary, renal, and hepatic function within 10 days were compared. Correlation between other perioperative variables and organ function was also investigated among all 95 patients. RESULTS: Postoperative pulmonary, renal, and hepatic function in group III was not significantly inferior to those in groups I and II. The operation time, and amount of red blood cells and fresh frozen plasma used during operation, were significantly greater in group I, which showed an intimate correlation to significant elevation of hepatic enzymes. CONCLUSIONS: Although lower functional reserve of vital organs in the elderly patients was predicted, they showed an acceptable functional recovery after operation with DHCA. Other perioperative variables such as operation time and blood transfusion showed a negative impact on postoperative hepatic function.

Age Factors↗

UCS15A, a non-kinase inhibitor of Src signal transduction.

Src tyrosine kinase plays key roles in signal transduction following growth factor stimulation and integrin-mediated cell-substrate adhesion. Since src-signal transduction defects are implicated in a multitude of human diseases, we have sought to develop new ways to identify small molecule inhibitors using a yeast-based, activated-src over-expression system. In the present study, we describe the identification of a unique src-signal transduction inhibitor, UCS15A. UCS15A was found to inhibit the src specific tyrosine phosphorylation of numerous proteins in v-src-transformed cells. Two of these phosphoproteins were identified as bona-fide src substrates, cortactin and Sam68. UCS15A differed from conventional src-inhibitors in that it did not inhibit the tyrosine kinase activity of src. In addition, UCS15A appeared to differ from src-destabilizing agents such as herbimycin and radicicol that destabilize src by interfering with Hsp90. Our studies suggest that UCS15A exerted its src-inhibitory effects by a novel mechanism that involved disruption of protein-protein interactions mediated by src. One of the biological consequences of src-inhibition by UCS15A was its ability to inhibit the bone resorption activity of osteoclasts in vitro. These data suggest that UCS15A may inhibit the bone resorption activity of osteoclasts, not by inhibiting src tyrosine kinase activity, but by disrupting the interaction of proteins associated with src, thereby modulating downstream events in the src signal transduction pathway.

3T3 Cells↗

Construction of FLAG and histidine tagging vectors for Schizosaccharomyces pombe.

Schizosaccharomyces pombe is becoming an increasingly popular model system for investigating important cellular processes. To facilitate detection, purification and functional studies of Sz. pombe gene products, we constructed two tagging expression vectors for use in Sz. pombe. These vectors allow proteins to be expressed ectopically as fusion proteins with a FLAG epitope and six histidine residue tags attached to their N-terminus or C-terminus. The function and applicability of these vectors were examined and the results are shown using the N-terminal tagging vector encoding Sfc6p, a subunit of the Sz. pombe RNA polymerase III general transcription factor, TFIIIC.

Amino Acid Sequence↗

Molecular recognition by chiral cationic micellar and micelle-like aggregates in electrokinetic capillary chromatography.

We examined the enantiomer separation with micelles and a micelle-like polymer made with trimethylammonium-terminated surfactants all of whose hydrocarbon chains contain hydrogen bonding valinediamide moieties in electrokinetic chromatography (EKC). The surfactants used were 3-(N-dodecanoyl-L-valylamino)-propyltrimethylammonium bromide (surfactant 1) and 6-(N-nonanoyl-L-valylamino)hexyl-trimethylammonium bromide (surfactant 2); the micelle-like polymer was derived from 3-(N-10-undecenoyl-L-valyl)aminopropyltrimethylammonium bromide (surfactant 3). N-Acylamino acids and their isopropyl esters were separated with enantiomers with the same configuration as the chiral surfactant and which were retained to a greater extent than the counterparts in micelles. The micellar hydrophobic environment, in which amides function as hydrogen bonding sites with solutes, and ceased micellar kinetic association-dissociation with polymerization are discussed.

Amino Acids↗

Biochemical and morphological changes in herniated human intervertebral disc.

The molecular and morphologic features of herniated human intervertebral disc tissues are of particular importance to clarify the pathogenesis. The present study analyzed the biochemical and morphological features of herniated intervertebral disc tissues to determine the constituent factors responsible for intervertebral disc herniation. A total of 32 herniated disc specimens and 4 control disc samples were analyzed. Collagen subunit composition, collagenase activity, lipid peroxidation level, caspase-3 activity, metal levels, morphologic studies, and genetic analysis were performed on herniated disc tissues of chronic (group A) and acute (group B) group and compared with findings of control group. Nick translation analysis in situ revealed apoptotic-positive stained DNA fragments as black-brown spots in herniated disc tissues. The presence of type II collagen in control disc samples and its absence in herniated samples were confirmed immunohistochemically. The increased caspase-3 activity, the apoptotic-positive stained DNA fragments, and the electron microscopic findings suggest enhanced programmed cell death in herniated discs. The significant increase in lipid peroxidation levels and collagenase activity, and the low metal levels suggest the enhancement of cell death signals in herniated discs, caused by oxygen stress. Linkage analysis of herniated disc tissues in Japanese individuals may suggest ethnic variation. These findings may be helpful in understanding the pathogenesis of herniated disc disease.

Adolescent↗

Effect of beta-blockers on circulating levels of inflammatory and anti-inflammatory cytokines in patients with dilated cardiomyopathy.

OBJECTIVES: This study was designed to evaluate the beneficial effect of beta-blockers on circulating cytokine levels in patients with dilated cardiomyopathy (DCM). BACKGROUND: Elevated circulating levels of inflammatory cytokines have been reported in patients with DCM. However, alterations of the levels of inflammatory and anti-inflammatory cytokines in association with beta-blocker therapy are unknown. METHODS: We studied 32 patients with idiopathic DCM who had been treated with digitalis, diuretics and angiotensin-converting enzyme inhibitors. In addition to this combination therapy, beta-blockers were started in all patients. Serum levels of interleukin (IL)-10, tumor necrosis factor-alpha (TNF-alpha) and soluble TNF receptors (sTNF-R1 and R2) were measured at baseline and 12 weeks after the initiation of beta-blocker therapy. We also measured plasma levels of neurohumoral factors, as well as left ventricular (LV) size and function. Ten age-matched subjects with no cardiac disease served as the control group. RESULTS: Baseline levels of IL-10, TNF-alpha and sTNF-R2 were significantly higher in patients with DCM than in control subjects (p < 0.05). There was a significant positive correlation between IL-10 and TNF-alpha levels (r = 0.545, p = 0.029). The TNF-alpha/IL-10 ratio correlated well with plasma epinephrine levels (r = 0.677, p = 0.025), and the level of sTNF-R2 was closely related to LV size. Serum levels of IL-10, TNF-alpha and sTNF-R2 were significantly decreased during beta-blocker therapy (p < 0.005). CONCLUSIONS: Our findings indicate that beta-blockers have an important immunoregulatory role in modifying the dysregulated cytokine network in DCM. This effect of beta-blockers may be partly responsible for the efficacy of therapeutic drugs for heart failure.

Adrenergic beta-Antagonists↗

Single- versus bi-socket anterior cruciate ligament reconstruction using autogenous multiple-stranded hamstring tendons with endoButton femoral fixation: A prospective study.

PURPOSE: This prospective study was conducted to compare the single-socket and the bi-socket anterior cruciate ligament (ACL) reconstruction techniques in terms of outcome. TYPE OF STUDY: Nonrandomized control trial. METHODS: There were 160 consecutive patients with unilateral chronic ACL insufficiency who underwent endoscopic single- or bi-socket ACL reconstruction alternately using multiple-stranded medial hamstring tendon and EndoButton (Smith & Nephew, Andover, MD) femoral fixation. All patients underwent the same postoperative rehabilitation protocol. Of them, 106 patients (57 single, 49 bi) were available for 2-year follow-up. RESULTS: According to the IKDC Knee Ligament Evaluation Form, 23 patients (40%) of the single-socket group were subjectively graded as normal, 30 (53%) as nearly normal, and 4 (7%) as abnormal. Twenty-six patients (53%) of the bi-socket group were graded as normal, 21 (43%) as nearly normal, and 2 (4%) as abnormal (P =.19). The mean side-to-side anterior laxity difference (KT-1000 manual maximum force) was 0.9 +/- 1.8 mm for the single-socket group and 0.7 +/- 1.2 mm for the bi-socket group (P =.44). Fifty-three of 57 patients (93%) in the single-socket group and all patients in the bi-socket group showed anterior laxity differences of +/-3 mm or less (P =.12). There were no differences in thigh muscle strength between the groups. CONCLUSIONS: Both single- and bi-socket ACL reconstruction using autogenous multiple-stranded hamstring tendons with EndoButton fixation provided satisfactory anterior stability, and there were no statistically significant differences in subjective results or measured restored stability between the 2 groups.

Adolescent↗

Allosterically controlled single-chained maxizymes with extremely high and specific activity.

For the treatment of chronic myelogenous leukemia (CML), attempts have been made to design various ribozyme motifs that can specifically recognize and cleave BCR-ABL fusion mRNAs. In the case of L6 BCR-ABL b2a2 mRNA, it is difficult to cleave the abnormal mRNA specifically because the mRNA includes no sequences that can be cleaved efficiently by conventional hammerhead ribozymes near the BCR-ABL junction. We recently succeeded in designing a novel maxizyme, which specifically cleaves BCR-ABL fusion mRNA, as a result of the formation of a dimeric structure [Kuwabara, T.; et al. Mol. Cell 1998, 2, 617-627; Tanabe, T.; et al. Nature 2000, 406, 473-474]. Specifically, we tailored the maxizyme with molecular switching function: the maxizyme splices a cleavable GUC site, but only when it appears within a strand of mRNA that possesses the abnormal splice junction. We demonstrated that this approach is generalizable [Tanabe, T.; et al. Biomacromolecules 2000, 1, 108-117]. All the maxizymes designed in the past functioned as a result of the formation of a dimeric structure. Questions have been asked whether a similar molecular switching might be possible within a single molecule when two monomer units of the maxizyme were connected via a linker sequence. We found that an analogous conformational change could not be induced within a single molecule when two maxizyme units were simply connected via a nonregulatable linker sequence. However, an active conformation was achieved by the introduction of an antisense modulator within the linker sequence that adjusted the overall structure to the correct form. Results of studies in cultured cells suggested that the desired conformational change could indeed be induced within the modified single-chained maxizyme and such a construct caused apoptosis only in leukemic cells with the Philadelphia chromosome.

Allosteric Regulation↗

Parental enforcement of bedtime during childhood modulates preference of Japanese junior high school students for eveningness chronotype.

We examined the effect of home bedtime discipline during childhood on morningness and eveningness (M-E) preference by Japanese junior high school students. M-E was assessed by the M-E Questionnaire (MEQ) of Torsvall and Akerstedt (the higher the score, the greater the preference for morningness), and parental determination of bedtime during childhood was ascertained using an original questionnaire. The average M-E score of adolescents living in urban Kochi City (mean +/- SD; 15.10 +/- 3.42) was significantly lower (P < .01) than the score of those in suburban districts (16.14 +/- 3.44). Overall, 43.1% of the junior high school students in Kochi City compared to 53.0% of the students living in suburban districts had their bedtime decided during childhood by parents (P < .01). In Kochi City, the M-E score for boys (14.62 +/- 3.51) was lower (P < .01) than girls (15.53 +/- 3.28). During childhood, parents decided the bedtime for 49% of the girls compared to 36.6% of the boys (P < .01). Boys whose bedtime was not decided by parents during childhood had a somewhat stronger preference for eveningness (14.20 +/- 3.53) (P < .05) compared to those whose bedtime was decided by parents (15.12 +/- 3.36). The results suggest bedtime discipline at home during childhood has an effect on adolescent chronotype, modulating the extent of shift to evening ness in Japanese junior high school boys in particular.

Adolescent↗