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Biomedical subjects

M Hirao

Publications and source records attributed to M Hirao.

At least 19 recordsLinked to original sources
PubMed2004

Successful vitrification of pronuclear-stage rabbit zygotes by minimum volume cooling procedure.

Rabbit zygotes at the pronuclear-stage were cryopreserved by vitrification using a gel-loading tip (GL-tip), Cryoloop or Cryotop. In GL-tip and Cryoloop methods, zygotes were first exposed to 10% ethylene glycol (EG)+10% DMSO in TCM199+20% fetal bovine serum (FBS) for 2 min, and then equilibrated for 30 s in a vitrification solution composed of 20% EG+20% DMSO+0.6 M sucrose in TCM199+20% FBS. In Cryotop method, zygotes were first exposed to 7.5% EG+7.5% DMSO+20% FBS in TCM199 for 3 min, and then equilibrated for 1 min in a vitrification solution composed of 15% EG+15% DMSO+0.5 M sucrose+20% FBS in TCM199. In vitro culture of vitrified-warmed zygotes using GL-tip and Cryoloop resulted in low cleavage rates (2 and 5%, respectively) and no development into blastocysts. In contrast, zygotes vitrified-warmed using Cryotop exhibited higher proportions of cleavage (58%) and development into blastocysts (24%). When compacted morulae or early blastocysts were vitrified by these three procedures, 80-93% of them exhibited blastocoele expansion or zona hatching during the subsequent 48 h of culture. Use of Cryotop instead of GL-tip or Cryoloop for zygote vitrification, without changing conditions of solutions and periods for exposure, equilibration and post-warm dilution, resulted in cleavage and blastocyst development rates of 88 and 45%, respectively. A longer exposure time (10 min) of zygotes to 7.5% EG+7.5% DMSO+20% FBS in TCM199 resulted in higher proportions of zygotes cleaving (94%) and developing into blastocysts (51%) after Cryotop vitrification. Proportions of post-warm zygotes (10-min exposure group) and fresh control zygotes developing into newborn offspring were 36 and 53%, respectively. Pronuclear-stage rabbit zygotes were successfully cryopreserved by vitrification using the Cryotop method.

Animals↗
PubMed2004

Determination of elastic, anelastic, and piezoelectric coefficients of piezoelectric materials from a single specimen by acoustic resonance spectroscopy.

We describe an advanced methodology to determine all the independent elastic-stiffness coefficients Cijkl, the associated internal frictions Qijkl(-1), and piezoelectric coefficients eijk of piezoelectric materials from a single monocrystal specimen using resonant-ultrasound spectroscopy with laser-Doppler interferometry. The mechanical-resonance frequencies of a piezoelectric solid depend on all of the elastic and piezoelectric coefficients, and their accurate measurement allows one to determine the elastic and piezoelectric coefficients simultaneously. Resonance-peak-width measurements yield the internal-friction tensor. Successful determination requires correct vibration-mode identification for the observed resonance frequencies. This is achieved unambiguously by measuring deformation distributions on the vibrating-specimen surface with laser-Doppler interferometry and comparing them with calculated displacement distributions. The methodology is applied to lithium niobate (LiNbO3) and langasite (La3Ga5SiO14) crystals.

Journal Article↗
PubMed2004

Phase I study of the combination of nedaplatin, adriamycin and 5-fluorouracil for treatment of advanced esophageal cancer.

This trial was conducted to determine the maximum-tolerated dose, principal toxicity, and recommended dose (RD) for the phase II study of the combination of nedaplatin (NED), adriamycin (ADM), and 5-fluorouracil (5-FU) in patients with advanced esophageal cancer. Patients with previously untreated esophageal cancer were eligible if they had performance status 0-1, were 75 years or younger and had adequate organ function. The dose of NED, the key anticancer platinum complex drug, was increased from 60 to 70, and 80 mg/m(2) on day 1. ADM and 5-FU were administered at fixed doses (30 mg/m(2) on day 1, and 700 mg/m(2) on days 1-5). The dose-limiting toxicities of NED were neutropenia and severe diarrhea, and its maximum-tolerated dose and RD were 70 mg/m(2) and 60 mg/m(2), respectively. There were four responders among the six patients administered the RD. The present study thus revealed combination chemotherapy with NED, ADM, and 5-FU to be active and well-tolerated and to warrant phase II study.

Aged↗
PubMed2001

Mammalian and viral IL-10 enhance C-C chemokine receptor 5 but down-regulate C-C chemokine receptor 7 expression by myeloid dendritic cells: impact on chemotactic responses and in vivo homing ability.

The immunosuppressive and anti-inflammatory cytokine IL-10 inhibits the phenotypic and functional maturation of dendritic cells (DC) and has been reported to confer tolerogenic properties on these important professional APC. Here, we exposed murine bone marrow-derived myeloid DC to either mouse (m) or viral (v) IL-10 early during their in vitro generation in response to GM-CSF and IL-4. Both mIL-10 and vIL-10 down-regulated the expression of CCR7 mRNA determined by RT-PCR, while mIL-10 up-regulated the expression of CCR5 transcripts. These changes in CCR7 and CCR5 expression were associated with inhibition and augmentation, respectively, of DC chemotaxis toward their respective agonists, macrophage inflammatory proteins 3beta and 1alpha, while in vivo homing of DC from peripheral s.c. sites to secondary lymphoid tissue of syngeneic or allogeneic recipients was significantly impaired. Anti-mIL-10R mAb reversed the effects of mIL-10 on CCR expression and restored DC homing ability. Retroviral transduction of mIL-10- and vIL-10-treated DC to overexpress transgenic CCR7 partially restored the cells' lymphoid tissue homing ability in allogeneic recipients. However, CCR7 gene transfer did not reinstate the capacity of IL-10-treated DC to prime host naive T cells for ex vivo proliferative responses or Th1 cytokine (IFN-gamma) production in response to rechallenge with (donor) alloantigen. These findings suggest that in addition to their capacity to subvert DC maturation/function and confer tolerogenic potential on these cells, mIL-10 and vIL-10 regulate DC migratory responses via modulation of CCR expression.

Adjuvants, Immunologic↗
PubMed2001

Effects of cryopreservation of pronuclear-stage rabbit zygotes on the morphological survival, blastocyst formation, and full-term development after DNA microinjection.

The objectives of this study were to examine the freezing sensitivity of pronuclear-stage rabbit zygotes and to produce transgenic rabbits using the cryopreserved zygotes. Zygotes were cryopreserved either by one of two vitrification protocols or by one of the two conventional freezing protocols. The morphological survival rates of zygotes subjected to two-step freezing in 1.5 M ethylene glycol and 0.1 M sucrose (74%) or to vitrification in 7.2 M ethylene glycol and 1.0 M sucrose (81%) were higher than those subjected to freezing in 1.5 M DMSO (46%) or to vitrification in a mixture of 2.0 M DMSO, 1.0 M acetamide, and 3.0 M propylene glycol (41%). But the in vitro development into blastocysts of zygotes cryopreserved by vitrification (17%) or to a lesser extent by freezing (52%) was impaired, when compared to that of fresh control zygotes (89%). Next, a fusion gene composed from bovine aS1-casein promoter and a human GH structural gene (2.8 kb) was microinjected into the pronucleus of rabbit zygotes frozen-thawed in ethylene glycol and sucrose. Then, the presence of exogenous DNA in the genome of newborn offspring was determined by PCR. The post-injection survival of frozen zygotes (97%) was the same as that of fresh control zygotes (96%). However, of 18 offspring derived from 414 frozen-thawed and DNA-injected zygotes, no transgenic rabbits were produced. Of 52 offspring derived from 403 DNA-injected fresh zygotes, 3 transgenic rabbits were found. Here we report the first rabbit offspring resulting from zygotes cryopreserved at the pronuclear-stage, although the cryopreservation procedure employed must be improved if zygotes are to be used for systematic production of transgenic rabbits.

Animals↗
PubMed2001

Relationship between mutations in the DNA gyrase and topoisomerase IV genes and nadifloxacin resistance in clinically isolated quinolone-resistant Staphylococcus aureus.

We analyzed the relationship between resistance to nadifloxacin, a fluoroquinolone antimicrobial agent, and mutations in the quinolone resistance-determining regions (QRDRs) of the A subunit of DNA gyrase and topoisomerase IV, in 24 clinical isolates of Staphylococcus aureus. Seven known mutations were found in the QRDRs. The minimum inhibitory concentration (MIC) of nadifloxacin increased in a manner that was dependent on mutations in the A subunit of DNA gyrase, and did not appear to be related to mutations in the A subunit of topoisomerase IV. The type 9 mutant, which included four mutations, was highly resistant to ofloxacin, norfloxacin and sparfloxacin, but only moderately resistant to nadifloxacin (MIC, 12.5 microg/ml). One of the norfloxacin-resistant strains that expressed high levels of norA was not resistant to nadifloxacin. To the best of our knowledge, this is the first report describing a fluoroquinolone antimicrobial agent whose primary target is suggested to be DNA gyrase in S. aureus.

Anti-Infective Agents↗
PubMed2001

Solitary neurofibroma of the nasal cavity: resection with endoscopic surgery.

We present a case of neurofibroma of the nasal cavity treated by endoscopic surgery. A 71-year-old female had complained of left-sided nasal obstruction for the past four years. Anterior rhinoscopy, computed tomography (CT) and magnetic resonance imaging (MRI) revealed a tumour involving the left nasal cavity. Histological and immunohistochemical examination showed the tumour to be a neurofibroma. The tumour was resected with endoscopic surgery. Neurofibroma arising in the area of the nose and paranasal sinuses is rare. We discuss the clinical and pathological characters of neurofibroma arising in the nasal cavity.

Aged↗
PubMed2001

Expression of bone morphogenetic protein-6 (BMP-6) in myoepithelial cells in canine mammary gland tumors.

Seventy-three mammary tumors and three mammary tissue specimens were examined to elucidate the expression of bone morphogenetic protein (BMP)-6 in the myoepithelial cells of canine mammary gland tumors. Morphologically, the myoepithelial cells were classified into four types: resting and proliferating cells inside the basement membrane, and spindle- and star-shaped cells proliferating in the outer area of the basement membrane. The characteristics of these myoepithelial cells were confirmed by immunohistochemistry using antibodies raised against keratin, cytokeratin 19, alpha-smooth muscle actin, and vimentin. In simple adenoma, a small number of resting myoepithelial cells was immunopositive for BMP-6. In complex adenomas and benign mixed tumors, all types of myoepithelial cells, depending in some cases on their specific location within the tumor, were immunopositive for BMP-6, but almost all of the tubular epithelial cells were immunonegative. Foci consisting of a proliferation of BMP-6-positive star- and spindle-shaped cells had mucinous stroma with marked hyaline and chondroid changes. In contrast, the foci with BMP-6-negative spindle- and star-shaped cells tended to have mucinous stroma without chondroid change. Several types of mesenchymal cells including chondrocytes, osteoblasts, and fibroblastlike cells in the mixed tumors, showed an intense immunopositive reaction for the BMP-6 antibody, and were located close to the ectopic cartilage and bone matrix. No significant immunoreactivity for BMP-6 was observed in most of the malignant mammary tumors; only one malignant mixed tumor was examined. All of these findings indicate that BMP-6 expression in myoepithelial cells may increase in complex adenomas and benign mixed tumors in canine mammary glands, and that BMP-6 expression is most intense in the vicinity of chondroid matrix in these tumors.

Adenocarcinoma↗
PubMed2001

A comparative study on the integration of exogenous DNA into mouse, rat, rabbit, and pig genomes.

Transgenic mammals, from small laboratory rodents to domestic animals, have been successfully produced to date, but their production efficiency within or across species has been variable. This is probably due to the differences in the type of injected DNA and/or technical procedures employed in each laboratory, as well as the reproductive characteristics of the species. Here we report the direct comparison of the efficiencies of producing transgenic mice, rats, rabbits and pigs by one technician using a fusion gene composed of the bovine alpha S1-casein promoter and human growth hormone (hGH) gene. Before the fusion gene was injected into the zygotes, high magnitude centrifugation to visualize the pronuclei was necessary for all of the pig zygotes and one-third of the rabbit zygotes, but not for mouse and rat zygotes. Post-injection survival of the mouse zygotes (67.1%) was lower than those of the rat, rabbit and pig zygotes (89.6 to 100%). The volume change of the pronucleus following DNA injection was the lowest in mice (50% increase), moderate in rabbits (148% increase), and the most prominent in rats (238% increase). The data from only 1 pig zygote indicated a 22% increase in the pronucleus volume by DNA injection. The PCR analyses of the tail DNA of new born offspring indicated that 0.8% (4/493), 4.8% (22/463), 0.8% (3/367) and 0.9% (2/221) of the injected eggs in mice, rats, rabbits and pigs, respectively, developed into transgenic offspring. Some of the founder animals in all four species expressed the transgene in the mammary gland which was confirmed in hGH mRNA by RT-PCR and/or hGH peptide in Witch's milk with ELISA. These results suggest that the maximum volume of DNA solution injectable into the pronucleus is a possible factor explaining the species differences in the production of transgenic animals.

Animals↗
PubMed2001

[A study on the process and significance of the establishment of the nurse regulations in 1915].

Believing that the origin of the complexity of the present nurse education and training courses lies in the nurse regulations enacted by the Ministry of Home Affairs in 1915, the author has studied the background and significance of the regulations through investigations and analyses of the regulations which were promulgated in prefectures prior to the 1915 nurse regulations. This study reveals that firstly, the nurse regulations made by the Ministry of Home Affairs were enacted in the intensive form of regulations locally promulgated prior to the nurse regulations, secondly that the qualification structure for nurses was the same as that for doctors, and thirdly that the two ways of nurse license acquisition; by either qualification exams or graduation from training schools, and the two level license systems; namely registered and enrolled nurses, were all decided by the nurse regulations and were the origin of the complexity of the present nurse systems, which have influenced the public health nurse midwife and nurse regulations promulgated after the second world war.

Education, Nursing↗
PubMed2000

CC chemokine receptor-7 on dendritic cells is induced after interaction with apoptotic tumor cells: critical role in migration from the tumor site to draining lymph nodes.

Dendritic cells (DCs) are very potent antigen-presenting cells and play critical roles in regulating immune responses in cancer. The migrating of DCs from the tumor site to the lymphoid organs is believed to be one of the critical events. To examine this important DC function in tumor situations, bone marrow-derived DCs, cultured for 6 days with granulocyte macrophage colony-stimulating factor and interleukin 4, were inoculated at the tumor site. We have shown (Y. Nishioka et al., Cancer Res., 59: 40354041, 1999) that DCs can migrate from tumor site to the draining lymph nodes within 24 h (approximately 0.1% of administrated DCs). The DCs then form clusters with adjacent lymphoid cells, which produce IFN-gamma (1500-3200pg/10(6) cells/48 h) in response to tumor stimulation. The number of the DCs migrating into lymph nodes were greater when they were inoculated into the tumor rather than the skin. Coculture of DCs and apoptotic tumor cells resulted in decreased expression of CC chemokine receptor (CCR) 1 and increased CCR7 expression at mRNA level without alteration in other phenotypical markers on DCs. Chemotaxis assay showed that CCR7 ligands, macrophage inflammatory protein 3beta and secondary lymphoid-tissue chemokine significantly (P < 0.05) induced the migration of DCs when cocultured with apoptotic tumor cells. To directly examine the involvement of CCR7 expression in DC migration, we investigated the functions of DCs genetically modified to express high levels of CCR7. CCR7 transduction promotes DC migration in response to relevant ligands in vitro and in vivo. These results suggest that the CCR7 expression of DCs is enhanced with direct contact with apoptotic tumor cells and may have a critical role for DC migrating to regional lymph nodes. The means to promote DC delivery to tumor and to nodal sites represent novel targets for the biological therapy of cancer.

Animals↗
PubMed2000

Production of transgenic rabbits using centrifuged pronuclear zygotes.

Superovulation of female rabbits was induced by subcutaneous injection(s) of porcine FSH. Zygotes were recovered 17 to 19 hr after hCG injection and were classified into two categories under a microscope equipped with Nomarski interference-contrast optics at x 200 magnification: (A) zygotes with clearly visible pronuclei, or (B) zygotes with visualized pronuclei after 10 min centrifugation at 12,000 x g. No significant difference between strains was found in the proportion of category-A zygotes (JW 72.6% vs NZW 79.3%). Pronuclei of category-A zygotes were located in the center of the cytoplasm, and the pronuclei of category-B zygotes were slightly moved by centrifugation toward the mass of cytoplasmic lipid droplets. Exogenous DNA solution (5 microg/ml of fusion gene composed of bovine alphaS1-casein promoter and human growth hormone structural gene) was microinjected into the pronucleus of the JW zygotes. The pronucleus of category-A zygotes with a mean volume of 7.4 pl swelled up to 16.6 pl (132% increase), while that of category-B zygotes with a mean volume of 6.1 pl swelled up to 15.9 pl (148% increase). Nevertheless, similar proportions of category-A and category-B zygotes developed into offspring after transfer to recipient females (11.1 and 11.2%, respectively). The efficiency to produce hGH-carrying transgenic rabbits was 0.9% (2/235) from category-A zygotes and 0.5% (1/215) from category-B zygotes (P>0.05). To date, transgenic rabbits have been produced without centrifugation of pronuclear zygotes. However approximately 25% of fertilized rabbit zygotes can be used for DNA microinjection after they have been centrifuged to visualize their pronuclei.

Animals↗
PubMed2000

["The Art of Nursing" by Florence Nightingale, published by Claud Morris Books Limited and printed in 1946, which is considered a draft of "Notes on Nursing"].

There are three versions of "Notes of Nursing," that is Nightingale's most famous work. Recently a book that is considered a draft of Nightingale's "Notes on Nursing," was published by Claud Morris Books Limited in 1946. This book is not contained in the "Bio-Bibliography of Florence Nightingale" by Bishop and Goldie. The publisher says about the source that this book is an entirely new and revised edition of Miss Florence Nightingale's personal notes on nursing, first written in 1859. The publisher acknowledges with gratitude the cooperation of Miss P.E. Nutt, in making available correspondence of Florence Nightingale and of Colonel C.H. Paynter in preparing a copy of the first edition of Miss Nightingale's "Notes on Nursing". This book is composed of the preface and 14 chapters. The preface's title is "Every woman is a nurse", and there is Nightingale's autograph with the date 1859 at the end of the preface. There are some differences between "The Art of Nursing" and "Notes on Nursing". In the former, there is a title to the preface and the first chapter's title is "Don't leave it to the doctor" and the last chapter's title is "Testament for a good nurse". In the latter, there is no title to the preface and the content of the former's first chapter is put only as a prologue before the first chapter. The former's last chapter's title is changed to "Conclusion". Further, each chapter has been somewhat expanded. From the above, we can see that, at what may be considered the draft stage of "Notes on Nursing", Nightingale wanted to emphasize the following things: "Every woman is a nurse", "Don't leave it to the doctor,; and "Testament for a good nurse". However, we can see that, at the stage of publishing of "Notes of Nursing", Nightingale had second thoughts. She complied with advice she received to avoid dissension among the knowlegeable and antipathy from doctors, and she settled on plain expression to make the contents clear to all.

England↗
PubMed1999

Induction of systemic and therapeutic antitumor immunity using intratumoral injection of dendritic cells genetically modified to express interleukin 12.

Bone marrow-derived dendritic cells (BM-DCs) retrovirally transduced with genes encoding murine interleukin (IL)-12 stably expressed bioactive IL-12 protein at high levels. Intratumoral injection with IL-12 gene-modified BM-DCs resulted in regression of day 7 established weakly immunogenic tumors (MCA205, B16, and D122). This antitumor effect was substantially better than that of IL-12-transduced syngeneic fibroblasts or nontransduced BM-DCs. Furthermore, intratumoral injection with IL-12-transduced dendritic cells (DCs) induced specific TH1-type responses to the tumor in regional lymph nodes and spleen at levels greater than those of IL-12-transduced fibroblasts or nontransduced BM-DCs. Trafficking studies confirmed that intratumorally injected IL-12-transduced DCs, but not fibroblasts, could migrate to the draining lymph node to the same extent as nontransduced BM-DCs. This strategy designed to deliver genetically modified DCs to tumor sites is associated with systemic and therapeutic antitumor immunity and is an alternative approach to those that use delivery of DCs loaded with tumor antigen. These results support the clinical application of IL-12 gene-modified DCs in patients with cancer.

Animals↗
PubMed1999

Metallothionein expression correlates with the pathological response of patients with esophageal cancer undergoing preoperative chemoradiation therapy.

Immunohistochemical staining for metallothionein (MT) and p53 was performed on biopsy specimens of 30 patients with esophageal squamous cell carcinoma who had received curative resection following preoperative chemoradiation. The pathologic response to chemoradiation was a partial response in 19 cases and no change was observed in 11 cases. In 16 cases with MT-positive tumor, 10 (62.5%) showed no change. In 14 cases with MT-negative tumor, 13 (92.8%) showed partial response. In 8 patients with negative staining for p53 and MT, 7 were responders, whereas in 9 patients with positive staining for p53 and MT, 6 were nonresponders. The pathologic response was significantly associated with the prognosis (p = 0. 0167). The survival rate of the responders was significantly better than that of the nonresponders. These findings suggest that MT might be a prognostic marker, and consequently we can select the patients who will benefit from preoperative chemoradiation.

Chemotherapy, Adjuvant↗
PubMed1998

Ezrin/radixin/moesin (ERM) proteins bind to a positively charged amino acid cluster in the juxta-membrane cytoplasmic domain of CD44, CD43, and ICAM-2.

CD44 has been identified as a membrane-binding partner for ezrin/radixin/moesin (ERM) proteins, plasma membrane/actin filament cross-linkers. ERM proteins, however, are not necessarily colocalized with CD44 in tissues, but with CD43 and ICAM-2 in some types of cells. We found that glutathione-S-transferase fusion proteins with the cytoplasmic domain of CD43 and ICAM-2, as well as CD44, bound to moesin in vitro. The regions responsible for the in vitro binding of CD43 and CD44 to moesin were narrowed down to their juxta-membrane 20-30-amino acid sequences in the cytoplasmic domain. These sequences and the cytoplasmic domain of ICAM-2 (28 amino acids) were all characterized by the positively charged amino acid clusters. When E-cadherin chimeric molecules bearing these positively charged amino acid clusters of CD44, CD43, or ICAM-2 were expressed in mouse L fibroblasts, they were co-concentrated with ERM proteins at microvilli, whereas those lacking these clusters were diffusely distributed on the cell surface. The specific binding of ERM proteins to the juxta-membrane positively charged amino acid clusters of CD44, CD43, and ICAM-2 was confirmed by immunoprecipitation and site-directed mutagenesis. From these findings, we conclude that ERM proteins bind to integral membrane proteins bearing a positively charged amino acid cluster in their juxta-membrane cytoplasmic domain.

Amino Acid Sequence↗
PubMed1998

Characterization of soluble E-cadherin as a disease marker in gastric cancer patients.

The soluble fragment of E-cadherin protein (S-ECD) is reported to be increased in the peripheral blood of cancer patients. In this study, we investigated the clinical significance of serum S-ECD in 81 patients with gastric cancer. The amount of serum S-ECD was significantly higher in the gastric cancer patients (4735 +/- 2310 ng ml(-1)) than in healthy volunteers (2515 +/- 744 ng ml(-1)). With the normal range cut-off at average +2 s.d., 67% patients showed abnormally high serum S-ECD levels. This frequency was significantly higher than that of other tumour markers, such as CEA (4.4%) or CA19-9 (13.3%). However, there was no significant correlation between the amount of S-ECD and clinicopathological factors. Serum S-ECD might be derived from cancer tissue, as removal of cancers by surgical treatment results in quick decline of the serum S-ECD and S-ECD can be detected by immunoblot in cancer tissues but not in normal epithelium. The serum S-ECD amount was compared with the E-cadherin expression in cancer tissues, which were classified into those showing preserved (+), partially reduced (+/-) or lost (-) expression. Interestingly, E-cadherin (+/-) tumours showed higher serum S-ECD levels than the other types, and a higher amount of S-ECD in the immunoblot analysis. Thus, the serum level of S-ECD may serve as an excellent tumour marker with high sensitivity. Furthermore, analysis of S-ECD in serum and cancer tissue can offer clues for elucidating the mechanism of reduction of E-cadherin expression in cancer cells.

Adult↗