Biomedical subjects

N Shigematsu

Publications and source records attributed to N Shigematsu.

At least 37 records · Page 2Linked to original sources

PubMed1997

Probing the non-proline cis peptide bond in beta-lactamase from Staphylococcus aureus PC1 by the replacement Asn136 --> Ala.

A non-proline cis peptide is present between Glu166 and Ile167 in the active site of beta-lactamase from Staphylococcus aureus PC1. To examine the role of the interaction between the side chain of Asn136 and the main chain of Glu166, the site-directed mutant N136A was produced. The enzyme shows no measurable hydrolytic activity toward a variety of penicillins or cephalosporins except for the chromogenic cephalosporin, nitrocefin. For nitrocefin, the progress curve exhibits a fast burst with a stoichiometry of 1 mol of degraded substrate per mole of enzyme followed by a slow phase with a hydrolysis rate that is reduced by approximately 700-fold compared with that of the wild-type enzyme. Thus, the mutant enzyme is deacylation defective. Monitoring the hydrolysis of nitrocefin after preincubation with a number of beta-lactam compounds shows that cephalosporins form stable acyl complexes with the enzyme, whereas penicillins do not. The molecular weight of the mutant was determined by electrospray mass spectrometry, and the presence of the stable acyl enzyme adducts with cephaloridine and cefotaxime was confirmed by both electrospray and MALDI mass spectrometry. Therefore, in addition to impairing deacylation, the acylation machinery has been altered compared with the wild-type enzyme to act on cephalosporins and not on penicillins. Urea denaturation and thermal unfolding studies show that the N136A mutant enzyme is less stable than the wild-type enzyme. However, stability against chemical denaturation of the mutant enzyme is enhanced in the presence of cephaloridine beyond the stability of the wild-type protein. This is attributed to accumulation of favorable interactions between the cephaloridine and the protein, which play a role in the folded state and not in the unfolded state.

Alanine↗

PubMed1997

Radiotherapy for centrally recurrent cervical cancer of the vaginal stump following hysterectomy.

PURPOSE: This study was performed to establish the classification and the treatment modality for recurrent cervical cancer of the vaginal stump after hysterectomy. PATIENTS AND METHODS: Ninety patients with centrally recurrent cervical cancer of the vaginal stump following hysterectomy were treated with high-dose-rate intracavitary brachytherapy with or without external irradiation. The intervals between primary surgery and vaginal recurrences varied from 3 months to 36 years. Tumor size of the vaginal stump was determined by bimanual rectovaginal examination at the time of recurrence and was classified into three groups, i.e., small (no palpable tumor), medium (less than 3 cm), and large (3 cm or more). RESULTS: The 10-year survival rates for all patients were 52%. Survival was greatly influenced by the tumor sizes of the vaginal stump. The 10-year survival rates of patients with small, medium, and large size tumors were 72, 48, and 0%, respectively. All patients with large size tumors died within 5 years. Of 90 patients, 75 (83%) were determined by physical examination to be free of tumor on at least one visit within 2 months of the completion of treatment (CR). The remaining 15 patients (17%) had physical findings suggestive of residual tumor (Residual). The overall 10-year survival rate for all patients with CR was 63%, compared with 10% for the patients with Residual (P < 0.0001). The incidences of distant metastases of the patients with or without local failure were 55 and 13%, respectively (P < 0.0001). The patients with local failure had significantly higher incidence of metastases. Most patients with small size tumor were treated with brachytherapy alone, and the survival rates of these patients were not improved by combination with external irradiation. CONCLUSION: These results suggest that tumor size was a significant prognostic factor for recurrent cervical cancer of the vaginal stump. Patients with small size tumors were recommended to be treated with brachytherapy alone.

Adult↗

PubMed1997

Total synthesis of WS9326A, a potent tachykinin antagonist from Streptomyces violaceoniger.

Total synthesis of the cyclic peptide lactone WS9326A, a potent tachykinin antagonist isolated from Streptomyces violaceoniger strain 9326, has been achieved via Cbz-Thr(Boc-allo-Thr-Asn-Ser(Bzl)-(E)delta MeTyr-Leu-D-Phe-OTce, which was cyclized (Phe and allo-Thr) using an active ester method with N-hydroxysuccinimide. Finally the unique N-acyl group, the 2-(1(Z)-pentenyl)cinnamoyl moiety, was introduced onto the amino group in the Thr unit. The key step of the synthesis involves the preparation of the E-isomer of the dehydro-N-methyltyrosine (delta MeTyr) unit. The debenzoxylation reaction of the threo- and erythro-isomer of the beta-benzoxy-N-methyltyrosine derivatives gave exclusively the Z-isomer of Cbz-Thr-delta MeTyr(MOM)-OMe, which was then converted to the desired E-isomer by photochemical isomerization of Cbz-Thr(TBDMS)-(Z)delta MeTyr(MOM)-Leu-D-Phe-OTce at a later step.

Chromatography, High Pressure Liquid↗

PubMed1997

[Prognostic factors of cervical carcinoma treated with postoperative radiotherapy].

We analyzed 119 patients with cervical carcinoma treated by postoperative radiotherapy from 1983 to 1993. Five- and 10-year survival rates of all patients were 77% and 67%, respectively. The 10-year survival rates for stage I (54 patients) and stage II (65 patients) were 76% and 58%, respectively. Ten-year survival rate for patients with both deep stromal invasion and lymph node metastasis was 37% which was much lower than in those without them (more than 90 %), indicating that they seemed to be factors related to poor prognosis. The results of multivariate analysis showed that the number of metastatic lymph nodes was the most important prognostic factor. Leg edema and intestinal and urinary insufficiency as late complications of postoperative radiotherapy occurred at low incidences and were well tolerated.

Adult↗

PubMed1996

FR901483, a novel immunosuppressant isolated from Cladobotryum sp. No. 11231. Taxonomy of the producing organism, fermentation, isolation, physico-chemical properties and biological activities.

FR901483, a novel immunosuppressant, has been isolated from the fermentation broth of Cladobotryum sp. No. 11231. The molecular formula of FR901483 has been determined as C20H31N2O6P. FR901483 exerts a potent immunosuppressive activity in vitro and significantly prolongs graft survival time in the rat skin allograft model. This compound has an intriguing tricyclic structure possessing a phosphate ester in its molecule. The ester residue may play an important role in exerting immunosuppressive activity because the desphosphoryl compound is devoid of activity. It is thought that the primary target of immunosuppression by this compound is inhibition of purine nucleotide biosynthesis.

Animals↗

PubMed1996

[Palliation of brain metastases: factors associated with survival].

One hundred ten patients with brain metastases (76 lung cancer, 11 breast cancer, 7 colorectal cancer and others) were treated with radiotherapy, and the results were retrospectively evaluated. Neurologic symptoms were improved in 66% of patients. The relationship between improvement of neurologic symptoms and several factors, including irradiation dose, primary lesion, histology, term between first treatment and recurrence, clinical symptoms and CT findings, was analyzed. There was no significant correlation between symptom relief and these factors except for the number of metastatic lesions. Four of 110 patients could survive more than 2 years after radiotherapy: one with lung cancer, one with breast cancer, one with rectum cancer and one with uterine cancer. The factors that were analyzed for symptom relief were also studied to determine their effect on the survival of lung cancer patients with brain metastases. The patients who had higher dose irradiation (50 Gy) of improvement of neurological symptoms after radiotherapy survived significantly longer than those who had 30 Gy of irradiation or no symptom relief. Multivariate analysis of these factors showed that improvement of symptoms and irradiation dose were similarly important prognostic factors, whereas the others were not correlated with survival.

Brain Neoplasms↗

PubMed1996

[Theoretical considerations of dose-rate effect for clinical application: a study of pulsed brachytherapy].

Pulsed brachytherapy is starting to be used for patients with various kinds of carcinoma, in the expectation of fair treatment results without increasing the late radiation-induced damage. We calculated the early and late biologically effective dose (BED) of this modality to predict early and late radiation damage based on the linear-quadratic model (L-Q model). The way to set the values of alpha/beta and mu (factor of repair) in the L-Q model remains controversial. The value of alpha/beta has been fixed at 3Gy for the late effect and 10Gy for the early effect in recent studies. In this study, we applied these alpha/beta values to calculate the BED, however, the effects of its alteration ranged from 0.5Gy through 15Gy on BED were also analysed. We assumed mu = 0.5 (T1/2 = 1-2 hours) in this study, but we also tried to examine the effects of its variation. Our results indicate that pulsed brachytherapy is a safe treatment procedure even if the dose-rate ranges from 1Gy/hr through 100Gy/hr, but the fractionation number should be increased when using a very high dose rate. Pulsed brachytherapy should be applied cautiously for tissues with a small mu value, such as the spinal cord, because an extremely high late BED would be expected in such cases.

Brachytherapy↗

PubMed1996

[Pitfalls of hyperfractionation: theoretical considerations of effect of repair time on late radiation damage].

The need of radiation therapy in cancer treatment is increasing, however, the dose of radiation is limited by its late effects on normal tissues. Hyperfractionated radiotherapy was applied to patients with various kinds of carcinomas in the 1980s, expecting to improve tumor control rates. Some reports showed that the total radiation dose could be increased without increasing late radiation damage by using hyperfractionated radiotherapy. We calculated the early and late biologically effective doses (BED) for conventional and hyperfractionated radiotherapy to predict the early and late radiation damage based on the linear-quadratic model (L-Q model). Setting the values of the alpha/beta ratio and t1/2 (half time repair) in the L-Q model is still controversial, and few reports deal with t1/2. Assuming t1/2 = 0.5 hour, the total radiation dose can be up to 71 Gy by hyperfractionated schedule (1.2Gy/fr. 2 times/day) without increasing the late BED of conventional schedule (2Gy/day, total 60Gy). However, the late BED should be increased in tissues with a larger t1/2 value, even if applying the same hyperfractionated schedule. Thus, it is not recommended to apply the same hyperfractionated protocol to all organs.

Humans↗

PubMed1996

[Biological effect of carbon beams on cultured human cells].

This study was performed to determine the biological effect of carbon beams on 13 human tumor cells, in comparison with 200 KVp X-rays. Carbon beams were generated by the Riken Ring Cyclotron. The RBE (relative biological effectiveness) values were distributed from 1.46 to 2.20 for LET of 20 ke V/micron, and 2.29-3.54 for 80 keV/micron. The RBEs were increased in all cell lines as the LET of carbon beams was increased from 20 to 80 keV/micron. There was no significant difference in radiosensitivity between cells from adenocarcinoma and those from squamous cell carcinoma. The relationship between the radiosensitivity of cells to X-rays and RBE was analyzed, but no significant correlation was suggested. Several survival curves of 20-40 keV/micron carbon beam irradiation showed the initial shoulders and the recovery ratios between two split doses were determined. Recovery was observed for LET of 20 keV/micron but not for that of 40 keV/micron. Furthermore, recovery ratios were 1.0-1.8, smaller than those for X-rays (1.5-2.4).

Adenocarcinoma↗

PubMed1996

[Mutation induction by heavy ion beams and X-rays: analysis with cultured cell line].

This study was performed to determine the biological effects of heavy ion beams on cultured cells. V79 cells were irradiated with carbon or neon beams or X-rays, and cell survivals was calculated by the colony assay method. The Do values for 150 kVp X-rays, 20 keV/micron and 80 keV/micron carbon beams, and 80 keV/micron neon beams were 2.2Gy, 1.8Gy, 1.0Gy and 1.4Gy, respectively. After 7-10 day expression periods, the mutation frequencies at the hypoxanthine-guanine phosphribosyl transferase (hprt) locus were analyzed from the numbers of colonies formed in media supplemented with 6-thioguanine. An extremely higher frequency of mutation was observed with heavy ion beams compared with X-rays. Both cell killing effect and the mutation induction were enhanced when the LET of carbon beams was increased from 20 to 80 keV/micron. These results mean that carbon beams had a stronger cell-killing effect than X-rays, but also carried a high risk of mutation induction. On the other hand, neon beams yielded cell survival curves similar to those of carbon beams, but they had a smaller mutation induction effect than carbon beams. The effect of fractionated irradiation (3 hr interval) on cell survival and mutation frequency were also examined. When cells were irradiated with X-rays, cell survival was increased by fractionation, but the mutation frequency was not modified. Irradiating cells with fractionated carbon beams, survival curves were not affected, but mutation frequency was reduced.

Animals↗

PubMed1996

[Prognostic factors of nasopharyngeal carcinoma treated by radiotherapy].

This study was a retrospective analysis of 82 patients with histologically confirmed nasopharyngeal carcinoma, who were treated at Keio University Hospital from March 1983 to March 1993. We studied all cases(62% of them were stage IV) with regard to their prognostic factors and chronic side effects induced by radiation. All patients were received extended-field radiotherapy from the base of the skull to the supraclavicular lymph node area. The five- and ten-year overall survival rates for the entire group were 59% and 49%, respectively. Three stage I patients were alive without recurrence or metastases for more than 5 years. The ten-year survival rate stage II-III for patients (23 patients) was 78%, and for stage IV (56 patients) 37%. The five-year survival rate of patients in T2-3 group without lymph node metastasis was 88%, while it was reduced to 55% for patients with metastasis. The prognosis of T4 patients was very poor: their five-year survival rate was 37% with lymph node metastasis, and 35%, without lymph node metastasis. Patients under 46 years old showed an increased survival rate compared with patients over 46 years old. Patients who completed less than 52 days of radiation therapy found to have a better prognosis than those receiving radiation for a longer period. Our multivariate analysis indicated that age, radiation period and N-factor were statistically significant in influencing prognosis. Hearing loss occurred relatively high with 20% of cumulative incidence in our study, probably because they received radiation therapy with the extended field including bilateral middle and inner ear. Extended-field radiotherapy for patients with nasopharyngeal carcinoma might contribute to improving their cure-rate, and precise radiation planning is warranted to avoid the late complications such as hearing loss.

Adenocarcinoma↗

PubMed1995

Cytotoxic metabolites from a fungal adherent of a marine alga.

Leptosins G, G1, G2 and H, belonging to a series of dimeric epipolythiodioxopiperazines, were isolated from the mycelium of a strain of Leptosphaeria sp. stuck on the marine alga Sargassum tortile. Their stereostructures were elucidated by chemical and spectral evidence.

Antineoplastic Agents↗

PubMed1995

Thiol and disulfide metabolites of the radiation protector and potential chemopreventive agent WR-2721 are linked to both its anti-cytotoxic and anti-mutagenic mechanisms of action.

The ability of the potential chemopreventive agent S-2-(3-aminopropylamino)ethylphosphorothioic acid (WR-2721) to protect against radiation-induced mutagenesis at the hprt locus and cell killing was studied using CHO-AA8 cells incubated for 30 min at 37 degrees C in growth medium containing its active thiol 2-[(aminopropyl)amino]ethane-thiol (WR-1065). In parallel experiments, the thiol and disulfide forms of the drug present in cells and incubation medium were determined in order to identify which, if either, of the components were associated with the observed protective effects. Treatment with 4 mM WR-1065 produced significant intracellular levels of the thiol (WRSH) and disulfide (WRSS) forms of the drug, but also caused dramatic elevation of cellular glutathione (GSH) and cysteine levels, accompanied by marked protection against 60Co gamma-photon- and neutron-induced cell killing and mutagenesis. When drug-treated cells were transferred to drug-free medium and incubated for 4 h at 37 degrees C, levels of WRSH and WRSS and protection against cell killing decreased markedly, whereas levels of GSH and cysteine and protection against mutagenesis showed little change. GSH and cysteine levels were not associated with protection against radiation-induced mutagenesis, as established by experiments performed with buthionine sulfoximine to block GSH synthesis. These data do not support the hypothesis that modulation of GSH or cysteine levels by WR-1065 is a major mechanism accounting for protection. Protection against mutagenesis was seen for cells incubated in medium with concentrations of added WR-1065 as low as 10 microM, where cellular levels of WRSH and WRSS became difficult to measure (< or = 5 microM) and no protection against cell killing was found. An unexpected observation was that cells incubated in 40 microM WR-1065 incorporated the drug much more rapidly than expected for uptake by passive diffusion and concentrated the drug to a marked degree; this indicates that a cell-mediated transport system is involved in the uptake of WR-1065 at low drug concentrations.

Amifostine↗

PubMed1995

[Palliative radiotherapy for symptomatic osseous metastases].

Bone metastases are one of the most common and serious conditions requiring radiotherapy, but there is still a considerable lack of agreement on optimal radiation schedule. We analyzed patients with symptomatic osseous metastases from lung (72 patients) and breast (63 patients) carcinoma treated by palliative radiotherapy between 1983 and 1992. In this series, the incidences of symptomatic bone metastases appearing within 2 years after the first diagnosis of the primary lesion were 96% and 36% for lung and breast carcinomas, respectively. Thirty percent of bone metastases from breast carcinoma were diagnosed more than 5 years after the first diagnosis. Thus careful follow-up must be carried out for a prolonged period. Pain relief was achieved at almost the same rate for bone metastases from lung and breast carcinomas (81% and 85%, respectively), an the rapid onset of pain relief (15 Gy or less) was obtained in about half the patients for both diseases. The rapid onset of pain relief and the lack of association between the onset of pain relief and primary tumor argued against the conventional theory that tumor shrinkage is a component of the initial response. In contrast to the fact that almost all lung carcinoma patients had very poor prognoses, one third of the breast carcinoma patients were alive more than 2 years after palliative radiotherapy. Thus, the late effects of radiation, such as radiation myelopathy, must be always considered especially in breast carcinoma patients even when it is 'just' palliative radiotherapy for bone metastases.

Adult↗

PubMed1995

Evaluation of bi-level image converting methods for nuclear medicine image database stored in the IS&C magneto-optical disk.

1. INTRODUCTION. We have developed a report and imaging management system for nuclear medicine. The report and image data are stored in the IS&C magneto-optical disk (IS&C MOD). Nuclear medicine image data are relatively small, but they are large enough to create problems when being transported over a low-speed Hospital Information System (HIS) network. Moreover, gray scale image output devices (i.e., high-resolution displays, sonoprinters) are expensive. If high quality bi-level (black and white) images were available, images could be transferred through low-speed network with inexpensive bi-level terminals or conventional page-printers. We examined images using several bi-level image conversion techniques [1, 2] in order to determine how useful the bi-level images are and to assess their suitability for use in nuclear medicine. The images were compared with original film images by ROC analysis. The modified minimized average error method was found to be superior to other methods in bone and gallium images. Its A-values are 0.83 in gallium scan and 0.85 in bone scan. 2. MATERIALS AND METHODS. Our system consists of three digital gamma cameras, a nuclear medicine data processing unit, two UNIX stations, and two personal computers. The computers and the data processing unit are connected via Ethernet and each computer has an IS&C MOD unit. Image and report data are stored in the IS&C MOD. The computers are also connected off-line through the IS&C MOD. To evaluate image quality, bone scan images and gallium scan images (1024x512, 2048x1024 pixels and 16 bits depth) were converted to bi-level images (same pixels and one bit depth) using four methods: dither method, minimized average error method (MAE)[1], modified regional adaptation method, and constrained average method. For the dither methods, three sets of dither matrix (Fatting's, Bayer's and our original matrix) were employed. The computer images were compared with original film images by ROC analysis. 3. RESULTS. The converted bi-level images were 1/16 the size of the originals. They were transferred via Ethernet, displayed on the monochrome display, and printed using a conventional page-printer (240 dpi or 300 dpi). The A-values of the original gallium and bone images on the films were 0.89 and 0.94. Modified MAE method gave better results than other methods tested, with the gallium image using this technique reading 0.83 on the A-scale; and the bone images were 0.85. 4. DISCUSSION. Generally, nuclear medicine images require lower spatial and gray level resolution than other computer imaging techniques. We compared four bi-level image conversion methods in order to know which methods are suitable to nuclear medicine images. The ROC analysis was employed to evaluate these image qualities. The MAE methods made particular texture pattern as artifacts in the intermediate gray level area. But it can express homogeneity the neutral level. The dither method images are coarse texture. It affects detectabilities of subtle abnormal findings. 5. CONCLUSIONS. The A-z values of the modified MAE images were comparable to those of original film images. Although bone images give slightly lower values, many abnormal findings can be ascertained on bi-level images. High quality bi-level image techniques are considered to be useful for nuclear medicine image database systems.

Japan↗

PubMed1995

Development of a personal computer's viewing station for the IS&C magneto-optical disk.

We developed a display station for the medical images stored in the IS&C (Image Save and Carry) magneto-optical disk. This station consisted of the personal computer and the magneto-optical disk drive. The performance of this system was evaluated and was compared with the Unix workstations. We found that the personal computer system had an adequate ability to display medical images. It provided, moreover, for portability and practical use of medical information.

Data Display↗

PubMed1994

Radiotherapy for cervical cancer with high-dose rate brachytherapy correlation between tumor size, dose and failure.

This is a retrospective analysis of 659 patients with cervical squamous cell carcinoma with a minimum follow-up of 2 years at Keio University Hospital between May 1974 and March 1990. All patients were treated with external radiation (50 Gy) and high-dose rate (HDR) intracavitary brachytherapy (20-34 Gy). The 5-year survival rates in each stage gradually decreased with the advance of the stage (I, 84%; II, 71%; III, 47%; and IVa, 12%). When 366 patients with stage III were classified into three groups according to tumor size, i.e. small (S, 102 patients), medium (M, 145) and large (L, 119), a survival gradient of small > medium > large was demonstrated and the differences between them were significant. At follow-up visits conducted between 1 and 2 months after completion of treatment, 135 patients (20%) had physical indications of residual disease. The larger the tumor size, the more likely was residual disease by 2 months. The patients with residual disease had a significantly higher rate of the pelvic failure than those without it. There was no significant correlation between radiation doses by RALS and pelvic failure rates, except in the stage III-L group. In the stage III-L group, intracavitary doses of 24 Gy or less to point A could be correlated with the higher pelvic failure rate and the lower survival rate, in contrast to doses of 27 Gy or more. The incidence of major rectosigmoid complications was 11.8% with doses of 24 Gy or less to point A, 8.1% with 27 Gy and 19.2% with doses of 30 Gy or more.(ABSTRACT TRUNCATED AT 250 WORDS)

Actuarial Analysis↗

PubMed1994

Protection against radiation-induced mutagenesis at the hprt locus by spermine and N,N"-(dithiodi-2,1-ethanediyl)bis-1,3-propanediamine (WR-33278).

The polyamine spermine and the disulfide N,N"-(dithiodi-2,1-ethanediyl)bis-1,3-propanediamine (WR-33278) are structurally similar agents capable of binding to DNA. WR-33278 is the disulfide moiety of the clinically studied radioprotective agent S-2-(3-aminopropylamino)ethylphosphorothioic acid (WR-2721). Because of their reported structural and functional similarities, it was of interest to compare their effects on cell survival and mutation induction at the hypoxanthine-guanine phosphoribosyl transferase (hprt) locus in Chinese hamster AA8 cells. WR-33278 and spermine (at concentrations of 0.01 and 0.001 mM) were electroporated into cells. Electroporation, 300 V and 125 microF, was performed either 30 min prior to or 3 h following exposure of cells to 750 cGy of ionizing radiation. Electroporation alone reduced cell survival to 75% but had no effect on hprt mutation frequency. The electroporation of either spermine or WR-33278 at concentrations greater than 0.01 mM was extremely toxic. The exposure of cells to both electroporation and irradiation gave rise to enhanced cell killing and mutation induction, with the sequence of irradiation followed 3 h later by electroporation being the more toxic protocol. Cell survival was only enhanced following electroporation of 0.01 mM of spermine and WR-33278 30 min prior to irradiation. Protection against radiation-induced hprt mutations was observed for both spermine and WR-33278 under all experimental conditions tested. Spermine at exposure concentrations of 0.01 and 0.001 mM administered 30 min before or 3 h after irradiation reduced mutation frequencies by factors of 2.2, 1.2, 1.9 and 2.2, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗