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Biomedical subjects

Osamu Suzuki

Publications and source records attributed to Osamu Suzuki.

At least 19 recordsLinked to original sources

Novel correction methods as alternatives for the six-dimensional correction in CyberKnife treatment.

PURPOSE: During CyberKnife treatment, the 6D correction method is used to correct patient positional errors, including rotational ones. We developed novel correction methods for translating rotational errors into 3D, with the aim of making their correction safer than with 6D correction and as accurate as possible. MATERIALS AND METHODS: These novel correction methods were named the gravity correction and the beam correction method. With the gravity correction method, the beam coordinates after rotation are corrected to match the tumor gravity point with 3D translational components translated by the affine transformation matrix. For beam correction, the beam coordinates are corrected to match the translated tumor target coordinates for each treatment beam. The effectiveness and impact of these methods were demonstrated by means of dose volume histogram (DVH) shift evaluation. For analysis of the treatment data of 10 patients, the treatment beam was rotated in three patterns of rotational degree and corrected with the two methods. The amount of tumor gravity point shift in the rotation was also calculated, and the deterioration of the tumor DVH was studied. RESULTS: In the case of +/-1 degrees , +/-3 degrees , and +/-1 degrees rotation for the X, Y, Z axes, the tumor gravity point of all 10 patients moved around 2.4 mm on average. Tumor DVH was deteriorated worse as the distance between the tumor gravity point and the rotational origin became more distant. With the planned D90, which represents the dose above which 90% of the tumor volume is irradiated set at 100%, the postrotational average D90 dose deteriorated to 96.12% after (+/-1 degrees , +/-3 degrees , and +/-1 degrees ) rotation. The dose was improved to 99.9% (SD +/- 0.41) after the gravity correction, or to 99.87% (SD +/- 0.55) after the beam correction. CONCLUSION: The correction methods developed by us can correct tumor DVH findings to the same degree as with 6D correction and are safer because the movement required for correcting the linac is not rotational but translational only.

Head and Neck Neoplasms↗

Genetic stock identification of chum salmon in the Bering Sea and North Pacific Ocean using mitochondrial DNA microarray.

A newly developed DNA microarray was applied to identify mitochondrial (mt) DNA haplotypes of more than 2200 chum salmon in the Bering Sea and North Pacific Ocean in September 2002 and also 2003, when the majority of maturing fish were migrating toward their natal river. The distribution of haplotypes occurring in Asian and North American fish in the surveyed area was similar in the 2 years. A conditional maximum likelihood method for estimation of stock compositions indicated that the Japanese stocks were distributed mainly in the north central Bering Sea, whereas the Russian stocks were mainly in the western Bering Sea. The North American stocks were abundant in the North Pacific Ocean around the Aleutian Islands. These results indicate that the Asian and North American stocks of chum salmon are nonrandomly distributed in the Bering Sea and the North Pacific Ocean, and further the oligonuleotide DNA microarray developed by us has a high potential for identification of stocks among mixed ocean aggregates of high-seas chum salmon.

Animals↗

Preliminary results of phase I trial of oral uracil/tegafur (UFT), leucovorin plus irinotecan and radiation therapy for patients with locally recurrent rectal cancer.

BACKGROUND: Surgical attempts for locally recurrent rectal cancer often fail due to local re-recurrence and distant metastasis. Preoperative chemoradiation may enhance better local control and survival. The aim of this study was to assess the safety of oral uracil and tegafur (UFT) plus leucovorin (LV), and irinotecan combined with radiation and determine the maximum-tolerated dose (MTD) and dose limiting toxicity (DLT) of the triple drug regimen. PATIENTS AND METHODS: Patients with locally recurrent rectal cancer received escalating doses of irinotecan on days 1, 8, 15, and 22 (starting at 30 mg/m2, with 10 mg increments between consecutive cohorts) and fixed doses of UFT (300 mg/m2) plus LV (75 mg/day) on days 3 to 7, 10 to 14, 17 to 21, and 24 to 28. Radiation was given 5 days per week totaling 40 to 50 Gy (2Gy/day). RESULTS: Six patients were treated at the starting dose, and 2 received the full scheduled chemoradiotherapy. The other 4 patients had grade 3 diarrhea and diarrhea was the DLT. One patient had partial response and he had subsequently radical surgical resection. Median progression free survival for local recurrence was 320 days. CONCLUSION: Irinotecan plus UFT/LV with concomitant radiotherapy in patients with locally recurrent rectal cancer was not feasible due to diarrhea in this setting. Modification of the treatment is needed.

Journal Article↗

Diagnostic accuracy of cardiac markers for myocardial damage after radiofrequency catheter ablation.

AIMS: This study compares serum markers of myocardial damage incurred during radiofrequency catheter ablation (RFCA). METHODS AND RESULTS: Blood was sampled from 34 patients with atrial flutter (n = 16), atrioventricular nodal reentrant tachycardia (AVNRT; n = 13), and Wolff-Parkinson-White syndrome (WPW; n = 5) to measure creatine kinase MB subfraction (CK-MB), human heart-type fatty acid protein (h-FABP), and cardiac troponin T (cTnT) values at baseline and after RFCA. The controls comprised 12 patients without significant elevation of all myocardial markers during electrophysiological study (EPS) without RFCA. h-FABP values did not elevate significantly, whereas CK-MB and cTnT demonstrated significant change after RFCA (P < 0.001). Neither peak h-FABP nor CK-MB correlated with following RFCA parameters: delivery duration, number of RFCA discharges, and cumulative RFCA energy. In contrast, correlations were significant between mean peak values of cTnT and these RFCA parameters (all P < 0.05). The sensitivity (71.6%) and specificity (35.6%) of h-FABP were inferior to those of cTnT (93.3% and 89.8%, respectively). CONCLUSION: h-FABP is an insensitive and less specific marker of myocardial damage in RFCA much along the lines of CK-MB and when compared with the proven accuracy of cTnT.

Adult↗

Analyses of the cDNA and genomic DNA sequences encoding the luteinizing hormone beta-subunit precursor protein in the rabbit.

To examine the molecular basis for efficient induction of superovulation in the rabbit, we determined the cDNA sequences of the luteinizing hormone beta-subunit (LHB) from Japanese White (JW), New Zealand White (NZW), and Dutch-Belted (Dutch) rabbits, and we compared these LHB sequences with those of other mammals. Using 5'- and 3'-rapid amplification of cDNA ends (RACE) with pituitary cDNA libraries, we found that the LHB cDNAs of all three breeds are the same length (523 bp from the 5'-end to the polyA site) and have putative AATAAA polyadenylation signal sequences at nucleotides 504 to 509. Northern blot analysis indicated that the approximately 600-nt mRNA encoding JW LHB is slightly longer than the LHB mRNAs of the other two breeds. The NZW and Dutch rabbit LHB coding sequences are 426 bp long, and their G+C contents are higher (>73%) than those of other mammalian LHBs (<70%). The predicted 141-amino-acid sequences of the JW and NZW LHB proteins are identical, and the Dutch LHB and JW/NZW sequences differ at only two residues. The exon-intron configuration of the NZW LHB gene (three exons and two introns) is similar to that of other mammalian LHB genes, and the sequences of NZW rabbit and other mammalian LHB promoter regions are highly conserved. Phylogenetic analysis of the deduced amino acid sequences of the three rabbit LHB proteins indicated that the rabbit occupies a phylogenetic position between rodents and domestic animals, and is far from humans. The results suggest that LH prepared from rodents or domestic animals, if available, would be a better inducer for superovulation in rabbits than human LH/CG.

Amino Acid Sequence↗

Simultaneous determination of selegiline and desmethylselegiline in human body fluids by headspace solid-phase microextraction and gas chromatography-mass spectrometry.

A method for the simultaneous determination of selegiline and its metabolite, desmethylselegiline, in human whole blood and urine is presented. The method, which combines a fiber-based headspace solid-phase microextraction (SPME) technique with gas chromatography-mass spectrometry (GC-MS), required optimization of various parameters (e.g., salt additives, extraction temperatures, extraction times and the extraction properties of the SPME fiber coatings). Pargyline was used as the internal standard. Extraction efficiencies for both selegiline and desmethylselegiline were 2.0-3.4% for whole blood, and 8.0-13.2% for urine. The regression equations for selegiline and desmethylselegiline extracted from whole blood were linear (r(2)=0.996 and 0.995) within the concentration ranges 0.1-10 and 0.2-20 ng/ml, respectively. For urine, the regression equations for selegiline and desmethylselegiline were linear (r(2)=0.999 and 0.998) within the concentration ranges 0.05-5.0 and 0.1-10 ng/ml, respectively. The limit of detection for selegiline and desmethylselegiline was 0.01-0.05 ng/ml for both samples. The lower and upper limits of quantification for each compound were 0.05-0.2 and 5-20 ng/ml, respectively. Intra- and inter-day coefficients of variation for selegiline and desmethylselegiline in both samples were not greater than 8.7 and 11.7%, respectively. The determination of selegiline and desmethylselegiline concentrations in Parkinson's disease patients undergoing continuous selegiline treatment is presented and is shown to validate the present methodology.

Amphetamines↗

The relationship between sperm morphology and in vitro fertilization ability in mice.

In vitro fertilization (IVF) is widely used in reproduction research, but the sperm of some inbred strains of mice yield low fertilization rates in IVF. To determine the cause of this problem, we examined the effect of epididymal sperm morphology, in particular, tail bending and the presence and type of cytoplasmic droplet (CD), on fertilizability in vitro. Sperm suspensions were obtained from the following five strains: C57BL/6J, BALB/cA, C3H/HeN, DBA/2J, and 129 x 1/SvJ. The sperm were fixed in 10% formalin and three parts of the sperm, namely the head, tail, and CD, were examined. We recorded the proportion of abnormal sperm heads and hairpins at the neck; tails were categorized as straight, proximal bent, or distal bent; and the CDs were categorized as none, light-type, and heavy-type. Based on these parameters, we determined the correlations between sperm morphology and fertilizability in vitro, as judged by IVF using ICR oocytes. The proportion of sperm with a hairpin neck was higher in strain C57BL/6J, while abnormal head morphology occurred significantly more often in strain BALB/cA. The percentage of sperm with a proximal bent tail was highest in strain DBA/2J and lowest in strain 129 x 1/SvJ. A heavy-type CD was observed more frequently in the 129 x 1/SvJ and C57BL/6J strains than in the other three strains in which a light-type CD predominated. The rank order of the fertilization rates was 129 x 1/SvJ < C57BL/6J < C3H/HeN < BALB/cA < DBA/2J. In addition, fertilization rate was positively correlated with a proximal bent tail, but negatively correlated with a heavy-type CD and distal bent tail. This new classification system establishes that the morphological characteristics of epididymal sperm differ among inbred strains of mice and that tail and CD morphology are closely related to fertilization ability in IVF. Thus, our results provide a novel method for assessing the quality of mouse sperm used for IVF.

Animals↗

Elevated extracellular calcium stimulates secretion of bone morphogenetic protein 2 by a macrophage cell line.

It has been suggested that macrophages and multinucleated giant cells are responsible for phagocytosis of resorbable calcium phosphate (CaP) compounds implanted in bone defects. However, function of macrophages around the CaP, if continuously exposed to various concentration of extracellular calcium ions ([Ca(2+)](o)), is still unknown. The present study showed that when resorbable octacalcium phosphate was implanted in mouse calvaria, macrophage-like cells were observed around the implant during bone formation. Then, experiments were designed to investigate whether secretion of bone morphogenetic protein 2 (BMP-2) is enhanced by [Ca(2+)](o) in a macrophage cell line (J774A.1) in vitro. The mRNA expression and the secretion of BMP-2 in J774A.1 cells were significantly increased when incubated in the medium with [Ca(2+)](o) up to 14mM. The promotion of mRNA expression was maintained even when incubated with a small amount of minute CaP crystals. The present results suggest that [Ca(2+)](o) above physiological concentration may stimulate macrophages to induce osteogenic cytokine, such as BMP-2, for bone formation by osteoblast.

Animals↗

Determination of platinum derived from cisplatin in human tissues using electrospray ionization mass spectrometry.

Determination of platinum (Pt) derived from cisplatin in tissues was performed by electrospray ionization mass spectrometry (ESI-MS) using silver (Ag) as the internal standard. Pt and Ag reacted with diethyldithiocarbamate (DDC), and were extracted using isoamylalcohol and acidified with oxalic acid. The compounds were termed Pt(DDC)(3)(+) and Ag(DDC)(2)(+), based on their m/z values exhibiting the highest peaks at m/z 639 and m/z 405, respectively. The limit of detection was 30 pg and the quantitation range was from 100 to 10,000 pg using 5 mg tissue. The present method allowed the determination of Pt in wet-ashed tissue in 10 min.

Antineoplastic Agents↗

Sequence analysis of cDNA encoding rabbit follicle-stimulating hormone beta-subunit precursor protein.

To understand the molecular basis of the rabbit's efficient superovulation, we determined the cDNA sequence of the follicle-stimulating hormone (FSH) beta-subunit precursor protein using a combination of 5'- and 3'-rapid amplification of cDNA ends (RACE) with pituitary cDNA libraries of the Japanese White rabbit and compared it with those of other mammals. RACE experiments detected at least three transcripts for the FSHbeta precursor protein in the libraries. The transcripts had lengths of 457, 1,621, and 1,767 bp, from the 5'-end to the poly(A) site. The shortest and mid-length transcripts had the putative polyadenylation signal sequence AATAAA at nucleotides 436 and 1,601, respectively, whereas the longest form had an ATTAAA sequence at nucleotide 1,745 of the cDNA sequence. These transcripts are likely to be polyadenylation variants of one large transcript because they share the same coding sequence for the precursor protein (130 amino acid residues in length). However, only a few shortest variants seem to be formed because the shortest variants were not detected by Northern blot analysis. Phylogenetic analysis of the deduced amino acid sequence indicates that the rabbit is phylogenetically closer to humans than to the other mammals, suggesting that an FSH preparation from human sources would be superior as a follicle stimulant for the induction of superovulation.

Amino Acid Sequence↗

Bone formation enhanced by implanted octacalcium phosphate involving conversion into Ca-deficient hydroxyapatite.

The present study was designed to investigate whether hydrolysis of synthetic octacalcium phosphate (OCP) into hydroxyapatite affects bone formation. Mouse bone marrow stromal ST-2 cells and primary calvarial osteoblastic cells were cultured on the dishes pre-coated with OCP or its hydrolyzed Ca-deficient hydroxyapatite (OCP hydrolyzate; HL). The capacity of proliferation and differentiation was determined up to day 20. Granules of OCP and HL were implanted into critical-size rat calvaria defects for 4 and 12 weeks, and then bone formation was measured by histomorphometry. Structural changes of incubated and implanted OCP were determined by X-ray diffraction (XRD) and Fourier transform infrared spectroscopy (FTIR). The proliferation of both ST-2 and primary osteoblasts cultured on OCP or HL was initially inhibited, whereas their differentiation to osteoblasts was promoted at last. Implantation of OCP in bone defect more significantly enhanced bone formation than that of HL until 12 weeks. OCP tended to convert to apatite in vitro and in vivo. The conversion of the implanted OCP was ascertained to advance gradually with implantation periods. Taken together, these results suggest that OCP supports appositional bone formation and OCP-apatite conversion may be involved in this stimulatory capacity of OCP.

Animals↗

Surface chemistry and biological responses to synthetic octacalcium phosphate.

Octacalcium phosphate (OCP) has been suggested as a precursor of biological apatite in bone, dentin, and cementum because its existence explains the nonstoichiometry of apatite crystals in their compositions. Synthetic inorganic calcium phosphate compounds have been used clinically to fill bone defects, and sintered hydroxyapatite (HA) and beta-tricalcium phosphate (beta-TCP), bone substitute materials, are known to be osteoconductive, with beta-TCP also being bioresorbable. Nonsintered synthetic OCP has been shown to enhance bone regeneration accompanied by conversion into hydrolyzed apatitic products in situ and biodegradation. The surfaces of the OCP implant and the converted apatite seem to be continuously exposed to biological constituents, such as extracellular matrices, inorganic biominerals, and cellular components. This article reviews the surface reaction of OCP implants and the biological responses, such as experimentally stimulated bone formation on synthetic OCP, the mechanism of OCP hydrolysis into apatite, and the adsorption of biomolecules onto OCP and the converted apatite, of particular interest in reactive bone induction with synthetic OCP implants.

Adsorption↗

Octacalcium phosphate combined with collagen orthotopically enhances bone regeneration.

Octacalcium phosphate (OCP) is resorbable bone regenerative material, but its brittleness makes it difficult to maintain its shape without restraint. We have engineered a scaffold constructed of synthetic OCP and porcine collagen sponge (OCP/Collagen) and investigated whether OCP/Collagen composite could improve bone regeneration. To examine this hypothesis, bone regeneration by the implantation of OCP/Collagen was compared with those by OCP and collagen. Radiographic and histological examination was performed and the percentage of newly formed bone (n-Bone%) in the defect was determined by a histomorphometrical analysis. OCP/Collagen, OCP, or collagen was implanted into the critical-sized defects in rat crania and fixed at 2, 4, or 8 weeks after implantation. OCP/Collagen improved the handling performance than the granules of OCP, and synergistically enhanced the bone regeneration beyond expectation, which were composed of bone nucleation by OCP and cell infiltration by collagen. Histomorphometrical analysis showed that n-Bone% +/- standard error treated with OCP/Collagen (48.4 +/- 5.14) was significantly higher than those with OCP (27.6 +/- 4.04) or collagen (27.4 +/- 5.69) in week 8. The present study suggests that the combination OCP with collagen elicited the synergistic effect for bone regeneration.

Animals↗

Intracavitary brachytherapy for carcinoma of the uterine cervix--comparison of HDR (Ir-192) and MDR (Cs-137).

PURPOSE: To compare the results of high dose rate (HDR) (Ir-192) and medium dose rate (MDR) (Cs-137) intracavitary brachytherapy (ICRT) for carcinoma of the uterine cervix. MATERIALS AND METHODS: Between May 1991 and March 2001, a total of 206 patients with Stage I-IVA previously untreated cervical cancer were treated with ICRT combined with external beam radiotherapy (EBRT). HDR was administered to a total of 135 patients: 22 patients in Stage I, 49 in Stage II, 56 in Stage III, and eight in Stage IVA. MDR was administered to a total of 71 patients: six patients in Stage I, 27 in Stage II, 33 in Stage III, and five in Stage IVA. The MDR at point A was 30 Gy/hour for HDR and 1.7 Gy/hour for MDR treatment, and the corresponding median follow-up periods for survivors were 55 and 68 months. RESULTS: For the HDR group, 5-year cause-specific survival rates were 90%, 78%, 53% and 33% for Stages I, II, III, and IVA, respectively. For the MDR group, the corresponding rates were 100%, 76%, 51%, and 40%. In the HDR group, 19 patients (14%) developed Grade 2 or higher late complications, and, in the MDR group, four patients (6%) did. CONCLUSIONS: There was no statistically significant difference in cause-specific survivals between the results of HDR and MDR brachytherapy for cervical cancer. The incidence of late complications tended to be higher for the HDR group than for the MDR group, but did not show a statistically significant difference (p=0.07).

Adult↗

High-dose-rate brachytherapy combined with long-term hormonal therapy for high-risk prostate cancer: results of a retrospective analysis.

PURPOSE: High-dose-rate (HDR) brachytherapy combined with hormonal therapy (HTx), without the addition of external beam radiation therapy (EBRT) for high-risk prostate cancer was evaluated retrospectively. MATERIALS AND METHODS: Between May 1995 and April 2002, 35 patients with prostate cancer [Stage > or = T2b (UICC 1997) or tumor grading=3 or prostate-specific antigen (PSA) level > or = 20 ng/mL] were treated with HDR brachytherapy combined with HTx. Most patients (74%) had two or more of these factors. All patients received Iridium-192 HDR brachytherapy with a total dose of 54 Gy/9 fractions/5 days (48 Gy/8 fractions/5 days for the first 6 cases) in one implant session. The median neoadjuvant HTx [luteinizing hormone-releasing hormone (LH-RH) agonist and antiandrogen] period was 7 months. The median adjuvant HTx (ATH) (LH-RH agonist) period was 40 months, and median follow-up was 57 months (range, 23-117 months). RESULTS: The 5-year actuarial biochemical control, local control, and disease-free rates were 62%, 96%, and 76% respectively. No patients experienced local and/or regional relapse without distant progression. The 5-year actuarial cause-specific survival and overall survival rates were 89% and 87%, respectively. The acute and late toxicity were moderate and well tolerated. CONCLUSION: HDR brachytherapy plus long-term HTx is at least as effective as conventional EBRT plus long-term HTx.

Aged↗

Transgene insertion pattern analysis using genomic walking in a transgenic mouse line.

A transgene mapping technique (Noguchi et al., Exp. Anim. 53:103-111, 2004) is described that can be used to analyze transgene integration patterns in transgenic mice. The technique was used to reveal that a transgenic mouse line (GM1-sy#116) harbored inverted and direct tandem repeats of both intact and partial pCAGGS-based transgenes in the G2 region of chromosome 1. This complicated concatenation of transgenes may have been caused by simple end-joining of DNA constructs fragmented by exposure to UV transillumination during gel-purification, and by nuclease digestion inside zygote pronuclei. The results suggest that care should be taken to avoid unwanted fragmentation during the preparation of vector constructs.

Animals↗

The regulatory roles of cell surface sialylation and N-glycans in human B cell lymphoma cell adhesion to galectin-1.

Surface sialylation and glycosylation of tumor cells is known to affect various biological phenomena. In the present study, we analyzed the regulatory roles of cell surface sialylation in cell adhesion to galectin-1 in the human diffuse large B cell lymphoma (DLBCL) cell line, HBL-2, and Burkitt's lymphoma cell line, HBL-8. Vibrio cholerae neuraminidase treatment enhanced HBL-2 cell adhesion to galectin-1, suggesting that sialic acid inhibits HBL-2 cell adhesion to galectin-1. The data from employing two different neuraminidases, Vibrio cholerae and Newcastle disease virus neuraminidase, showed that alpha2,6-linked sialic acid plays an important role in the inhibition of HBL-2 cell adhesion to galectin-1. In addition, neuraminidase treatment enhanced the cell adhesion to galectin-1 much more with the highly sialylated HBL-8 3G3 clone than with the hyposialylated HBL-8 3D2 clone. Flow cytometric analysis revealed the expression of partially sialylated L-PHA reactive oligosaccharides on the surfaces of HBL-2 cells. Swainsonine (SW) treatment also enhanced HBL-2 cell adhesion to galectin-1. These data indicate that SW treatment decreased sialylated L-PHA reactive oligosaccharides resulting in cell surface desialylation and leading to enhancement of cell adhesion to galectin-1. In conclusion, alteration of cell surface sialylation or N-glycan expression regulates cell adhesion to galectin-1 in human malignant lymphoma.

Burkitt Lymphoma↗