Effect of ethylene treatment on phytochemical and ethylene-related gene expression during ripening in strawberry fruit Fragaria x ananassa cv. Camino Real.

In contrast to climacteric fruits, in which ethylene is known to be pivotal, the regulation of ripening in non-climacteric fruits is not well understood. The strawberry is a typical example of a non-climacteric fruit, which has been used as a model system of these types of fruit. In this study, the effect of exogenous ethephon on the expression of ethylene biosynthesis and signaling genes, FaERF2 and FaACO1, was analyzed in the Fragaria ananassa cultivar Camino Real by quantitative real-time polymerase chain reaction, and the physicochemical and phytochemical characteristics of fruits were evaluated in field trials and postharvest tests. Transcript accumulation was influenced by exogenous treatment with ethephon, which affected the pattern of gene expression during different stages of growth and fruit development, with the highest expression occurring during postharvest tests. In addition, ethephon significantly influenced the phytochemical profile of sugars, anthocyanins, phenolic compounds, and vitamin C contents both in the field- and postharvest-treated fruits at different stages. These results indicate that ethylene regulates the phenylpropanoid maturation pathway in strawberry fruit.

Electronic structure of charge- and spin-controlled Sr(1-(x+y))La(x+y)Ti(1-x)Cr(x)O3.

We present the electronic structure of Sr(1-(x+y))La(x+y)Ti(1-x)Cr(x)O3 investigated by high-resolution photoemission spectroscopy. In the vicinity of the Fermi level, it was found that the electronic structure was composed of a Cr 3d local state with the t(2g)3 configuration and a Ti 3d itinerant state. The energy levels of these Cr and Ti 3d states are well interpreted by the difference of the charge-transfer energy of both ions. The spectral weight of the Cr 3d state is completely proportional to the spin concentration x irrespective of the carrier concentration y, indicating that the spin density can be controlled by x as desired. In contrast, the spectral weight of the Ti 3d state is not proportional to y, depending on the amount of Cr doping.

222Rn concentrations in greenhouses in Aomori Prefecture, Japan.

Greenhouses are possible places with high 222Rn concentrations, since soil, the source of 222Rn, is directly exposed inside them. To examine this point, 222Rn concentrations in 28 greenhouses at five locations in Aomori Prefecture were measured for approximately 1 year with passive Rn detectors. For 1 week, measurements of 222Rn concentration and working level were also carried out with active detectors to get equilibrium factors and the ratio of 222Rn concentration in working time to non-working time in selected greenhouses. The geometric mean of annual 222Rn concentrations in greenhouses was 13 Bq m-3, and the same as that in dwellings and lower than that in indoor workplaces in the prefecture. However, variation of the 222Rn concentrations was far larger than in other environments, and ranged from the lowest level in outdoor workplaces to the highest level in indoor workplaces. Significant seasonal variation was also observed in 222Rn concentrations. The mean effective dose from 222Rn and its progenies was estimated to be 0.047 mSv year-1 for a farmer working in a greenhouse.

Biokinetics of radiotellurium in rats.

Radiotellurium is present in the environment primarily due to its release during nuclear reactor accidents. Little is known of tellurium metabolism in juveniles, although the element is relatively abundant and has a number of industrial uses. A biokinetic study of radiotellurium in rats was performed using gamma-ray counting. Wistar strain rats were used to determine the uptake of H2(123m)TeO3 by the whole-body retention of juvenile rats and the conceptus in relation to its gestational stages, by measurements in the placenta, fetal membranes, fetal fluid and fetus. The whole-body retention of 123mTe in juvenile rats was higher than that of adult rats. The relative concentration in the placenta and fetal membranes was higher than in the fetus. No activity was observed in the fetal fluid. These results indicate that the placenta and fetal membranes play significant roles as barriers to the transfer of 123mTe into the fetus. The ratio, relative concentration in fetus/relative concentration in mother (C(F)/C(M)), was calculated. The C(F)/C(M) ratio was dependent on the stage of gestation and ranged from 0.2 to 0.5. A little 123mTe was transferred to the suckling rats through the mother's milk when the isotope was administered intravenously to the mother.

Transfer of 14C to prenatal and neonatal rats from their mothers exposed to 14C compounds by ingestion.

The transfer of 14C through placenta or milk was investigated and the radiation dose to fetal and newborn rats was estimated. Female rats at different gestational stages or after delivery were exposed to 14C in the form of sodium bicarbonate, thymidine and lysine by a single ingestion. Radioactivity in maternal tissues and conceptuses (placenta, fetal membrane and fetus) and in the newborn was determined at various times after ingestion. After exposure to these 14C compounds, there was no significant difference between the 14C concentration in the fetus and that in the maternal tissues, suggesting that the placenta has no effect in preventing or accelerating the placental transfer of 14C. The concentration and content of 14C in the fetus and newborn were, however, dependent on the chemical form of 14C and on the prenatal or neonatal stage at the time of ingestion. The result of the dose estimation showed that 14C-lysine gave significantly higher prenatal and neonatal doses than 14C-sodium bicarbonate or 14C-thymidine.

Radon concentration in indoor occupational environments in Aomori Prefecture, Japan.

The (222)Rn concentrations in indoor workplaces were measured in Aomori Prefecture, Japan, and the results are reported here. This survey was part of a program to measure background natural radiation dose rate in the prefecture where the first Japanese nuclear fuel cycling facilities are now under construction. The survey of the (222)Rn concentrations in indoor workplaces was carried out at 107 locations from 1996 to 1998. The (222)Rn concentrations were measured for approximately one year at each site with passive Rn detectors, which used a polycarbonate film for counting alpha-rays and could separate concentrations of (222)Rn from (220)Rn. Weeklong measurements of (222)Rn concentration and working level were carried out with active detectors to get the ratio of (222)Rn concentration during working hours to non-working hours as well as equilibrium factors in selected locations. Diurnal variation of (222)Rn concentration depended on building structure, air-conditioning, time of day and day of the week (week days or weekend). The (222)Rn concentration during working hours was generally lower than that in non-working hours. Although the annual average (222)Rn concentration in indoor occupational environments was higher than that in dwellings, radiation dose for Aomori Prefecture residents from Rn in the former was 14% of the total indoor dose by Rn because of the lower concentration in working hours and lower occupancy factor.

Control of nitrogenase reactivation by the GlnZ protein in Azospirillum brasilense.

The glnZ mutant of Azospirillum brasilense (strain 7611) showed only partial recovery (20 to 40%) after 80 min of ammonia-induced nitrogenase switch-off, whereas the wild type recovered totally within 10 min. In contrast, the two strains showed identical anoxic-induced switch-on/switch-off, indicating no cross talk between the two reactivation mechanisms.

Unrelated donor marrow transplantation in children with severe aplastic anaemia using cyclophosphamide, anti-thymocyte globulin and total body irradiation.

We report a favourable outcome in 15 patients with severe aplastic anaemia (SAA) who were < 20 years of age and who underwent bone marrow transplantation (BMT) from a human leucocyte antigen (HLA)-matched unrelated donor. All patients were non-responders to intensive immunosuppressive therapy (IST) and were multiply transfused. The conditioning regimen consisted of cyclophosphamide (60 mg/kg/d, on d -4 and -3), anti-thymocyte globulin (2.5 mg/kg/d, on d -5 to -2) and total body irradiation (2.5 Gy x 2/d, on d -2 and -1). Patients received cyclosporine and methotrexate for prophylaxis of graft-versus-host disease (GVHD), except for the last four who received tacrolimus instead of cyclosporine. Donor/recipient pairs were identical for HLA class I and II antigens by serological typing, but four pairs were found to have a mismatch at the HLA-A, -B or -DRB1 locus by high-resolution typing. All patients achieved rapid engraftment and are alive at 2-86 months after transplantation (median follow-up, 51 months). Moderate to severe acute GVHD occurred in 5 out of 15 patients (33%); only one patient developed extensive chronic GVHD. Considering our encouraging results, unrelated donor transplantation for SAA is recommended as a salvage therapy in non-responders to IST.

Outcome prediction by molecular detection of minimal residual disease in bone marrow for advanced neuroblastoma.

BACKGROUND: We have determined whether sequential molecular detection of minimal residual disease (MRD) in bone marrow (BM) could predict the outcome of patients with advanced neuroblastoma (NB). PROCEDURE: Bone marrow samples from 19 patients over 12 months of age with stage 4 neuroblastoma were sequentially examined for tumor cell contamination by detecting tyrosine hydroxylase (TH) mRNA using reverse transcription-polymerase chain reaction (RT-PCR). All patients received repetitive multi-drug chemotherapy including cisplatin, cyclophosphamide or ifosphamide, adriamycin, and etoposide or vincristine. Seventeen patients received myeloablative therapy with hematopoietic stem cell transplantation after achieving complete remission. RESULTS: All but one patient were histologically positive for tumor cells in BM samples at diagnosis, and they became negative for tumor cells within 3 months histologically. By the RT-PCR analysis, all patients were positive for TH mRNA in BM samples at diagnosis, and they became negative for TH mRNA 1 to 13 months after the start of chemotherapy. Six patients whose BM samples became negative for TH mRNA within 4 months after the start of chemotherapy remained alive without evidence of disease (median 61 months, range 20-76). In contrast, 12 of 13 patients whose BM samples remained positive at that time developed relapse and 10 of them died of disease (median 24 months, range 13-43). There was a statistically significant difference in survival between the two groups (P < 0.05). No significant difference of clinical characteristics by the MRD positivity at 4 months after the start of chemotherapy. CONCLUSIONS: Persistence of MRD in BM at 4 months after the start of chemotherapy could predict poor prognosis in advanced neuroblastoma.

Rapid detection of chromosome aneuploidies by prenatal interphase FISH (fluorescence in situ hybridization) and its clinical utility in Japan.

OBJECTIVE: The purpose of this study was to assess the accuracy, informative rate, detection rate, and clinical utility of prenatal interphase fluorescence in situ hybridization (FISH) analysis of amniotic fluid samples from Japanese women. METHODS: Amniotic fluid specimens from 2,639 Japanese women were received for prenatal interphase FISH and chromosome analysis. A questionnaire was designed to evaluate FISH clinical utility by collaboration sites. RESULTS: Based on 2,319 tested samples, the accuracy (100%), informative (94%), and detection (87.6%) rates were all high. The accuracy (100%), informative (90.2%), and detection (90.0%) rates were also remarkable in third-trimester pregnancies. We perceive significant advantages from this test regarding medical management and patient satisfaction. CONCLUSIONS: This novel report shows that in Japan prenatal interphase FISH testing is highly informative and accurate, not only in second-trimester pregnancies but also in third-trimester pregnancies. This test provides advantages to both physicians and patients, provided that its capabilities and limitations are understood.

Long-term outcome of acquired aplastic anaemia in children: comparison between immunosuppressive therapy and bone marrow transplantation.

A total of 100 children under the age of 17 years with acquired aplastic anaemia (AA) were initially treated with immunosuppressive therapy (IST) (n = 63) or bone marrow transplantation (BMT) (n = 37) from an HLA-matched family donor. The projected 10-year survival rates were 55 +/- 8% and 97 +/- 3% respectively (P = 0.004). Because the IST group included 11 non-responders who were salvaged by BMT from an HLA-matched unrelated donor, we compared failure-free survival (FFS) between the groups. The probability of FFS at 10 years was 97 +/- 3% for the BMT group, compared with 40 +/- 8% for the IST group (P = 0.0001). Seven patients evolved to myelodysplastic syndrome (MDS) with monosomy 7 and the estimated cumulative incidence of MDS 10 years after diagnosis was 20 +/- 7% in the IST group. We compared the outcome of children treated with IST during the two consecutive periods of 1983-91 (group A, n = 40) and 1991-8 (group B, n = 23) to assess the impact of combined therapy with antithymocyte globulin and cyclosporin. The probability of FFS at 7 years follow-up was the same in the two groups (50 +/- 8% vs. 40 +/- 15%, P = 0.40). We recommend BMT as first-line therapy in paediatric severe AA patients with an HLA-matched family donor. Alternative donor BMT is recommended as salvage therapy in patients who relapse or do not respond to initial IST.

Real-time quantitative reverse transcription-polymerase chain reaction for the detection of AML1-MTG8 fusion transcripts in t(8;21)-positive acute myelogenous leukemia.

Quantification of AML1-MTG8 fusion transcripts was performed by using real-time reverse transcription-polymerase chain reaction (RT-PCR) and the clinical value of this method was evaluated in t(8;21)-positive acute myelogenous leukemia (AML). A t(8;21)-positive cell line, Kasumi-1, was used for constructing standard curves and the corrected AML1-MTG8 mRNA expression level relative to the expression of the GAPDH housekeeping gene was calculated. Bone marrow samples from 14 patients with t(8;21)-positive AML were sequentially examined. The corrected AML1-MTG8 expression level at diagnosis varied in the range from 0.4 to 2.7 (median, 1.5) among the patients. When samples at 1, 3 and 6 months were examined after diagnosis, the corrected AML1-MTG8 expression level was found to decrease sequentially in all but one. AML1-MTG8 fusion transcripts were also detected in four of eight samples from patients in remission for more than 1 year. In conclusion, real-time RT-PCR can provide a rapid and accurate quantification of AML1-MTG8 fusion transcripts. This system could be useful to reveal the prognostic relevance of minimal residual disease in t(8;21)-positive AML.

Prognostic value of internal tandem duplication of the FLT3 gene in childhood acute myelogenous leukemia.

BACKGROUND: Recently, an internal tandem duplication of the FLT3 gene (FLT3/ITD) was found in 20% of adult cases of acute myelogenous leukemia (AML), and this length abnormality was suggested to be associated with leukemic progression. PROCEDURE: We examined the mRNA expression of the FLT3 gene by using reverse transcription-polymerase chain reaction (RT-PCR) in 64 children with AML, and further abnormal transcripts were cloned and sequenced. RESULTS: An unexpected longer product was found in seven patients (11%) by RT-PCR of the FLT3 gene. Sequence analysis of these abnormal products revealed the presence of tandemly duplicated fragments in all seven patients. Three factors were identified to be associated with high incidence of FLT3/ITD; older patients (> or = 10 years) (P = 0.049), high WBC count (> or = 50,000/microl) at presentation (P = 0.002), and M3 in FAB subtypes (P = 0.002). Induction failure was observed in 3 (43%) of 7 patients with FLT3/ITD. Only FLT3/ITD was identified as a significant risk factor for induction failure by univariate analysis (P = 0.013), although it was not significant by multivariate analysis (P = 0.11). The Kaplan-Meier estimate of event-free survival rate at 5 years was 14% for patients with FLT3/ITD, which was significantly lower in comparison with 69% for patients without FLT3/ITD (P = 0.003). This finding was also identified by multivariate analysis (P = 0.01). CONCLUSIONS: In this study, FLT3/ITD was observed in 11% of childhood AML and identified to be associated with poor prognosis. A large prospective study with uniform treatment is necessary to confirm our results.

Retention, excretion and translocation of 239Pu in rats following inhalation of 239PuO2 calcined at 1150 and 400 degrees C.

Wistar rats inhaled 239PuO2 particles prepared by the calcination of 239Pu hydroxide at 1150 and 400 degrees C. Lung retention, fecal and urinary excretion, and translocation of 239Pu were compared between the two calcination temperatures. The clearance of 239Pu from the lungs was significantly faster in the rats exposed to 239PuO2 calcined at 400 degrees C (low-temperature group) than those exposed to 239PuO2 calcined at 1150 degrees C (high-temperature group). Both the fecal excretion of 239Pu and the ratio of fecal excretion to urinary excretion was greater in the low-temperature group than in high-temperature group. The amounts of 239Pu translocated from the lungs to the other organs were very small. Even in the liver, which accumulated the largest amount of 239Pu except for the lungs, only 0.13-0.20% of the initial lung burden was retained 1 year after inhalation. The amount of 239Pu deposited in the liver was greater in the high-temperature group than in the low-temperature group both at 1 month and 1 year after the inhalation. These findings clearly suggest that the lung retention of 239Pu in rats is significantly affected by the calcination temperature of 239PuO2.

[Clinical experiences and postoperative results with partially implantable middle ear implant].

A partially implantable middle ear implant using an ossicular vibrator of a piezoelectric ceramic bimorph has recently been developed in Japan as a new rehabilitative method for hearing. Four patients at the Miyazaki Medical College Hospital were implanted with this device between September 1994 and October 1996. Implantation of the internal component was performed under local anesthesia after confirmation of sufficient sensitivity to the ossicular vibrator in an intraoperative vibratory hearing test. All patients regained socially useful hearing with the middle ear implant without any foreign body reactions or recurrence of otitis media. None of the cases showed any elevations of bone conduction thresholds 2 to 4 years postoperatively. Natural and clear sounds without interference noise and howling due to acoustic feedback were highly satisfactory in all patients. However, the first case, who had an implant in an ear following cholesteatoma surgery using the intact canal wall technique, experienced about a 10dB deterioration of the average hearing level with the implant 2 years after implantation. Gradual deterioration appeared to be caused by the retracted tympanic membrane interfering with the mobility of the ossicular vibrator. The following three cases were implanted in ears previously operated on by radical mastoidectomy, showing stable or improving hearing results with the implant. Closure of the external canal skin at the cartilaginous portion at the time of implantation seemed appropriate to maintain a sufficient middle ear space for the ossicular vibrator to work properly. Our case study confirms that the partially implantable middle ear implant is a safe and useful tool for patients with mixed deafness which cannot be satisfactorily rehabilitated by tympanoplasty and/or a conventional hearing aid.

[Clinical trial of antithymocyte globulin for prophylaxis of acute graft-versus-host disease in pediatric recipients of bone marrow transplantation from unrelated donors].

We studied the effectiveness of antithymocyte globulin (ATG) in preventing acute graft-versus-host disease (a-GVHD) in children who received bone marrow transplants from unrelated HLA-matched donors at one institution. Of 39 patients who received transplants between 1993 and 1997, 23 were given ATG on the basis of informed consent. Either Thymoglobulin (Pasteur Merieux, 2.5 mg/kg/day) or Lymphoglobulin (15 mg/kg/day) was administered for 4 days. a-GVHD (> or = grade II) developed in 33% of the ATG group (n = 21) and in 44% of the non-ATG group (n = 16). Although a-GVHD (> or = grade II) appeared less frequent in the ATG group, the difference was not statistically significant. Among the subjects with hematological malignancies, no significant difference was observed in frequency of a-GVHD (> or = grade II) or 3-year survival rate for the ATG group (n = 10) and non-ATG group (n = 16). However, the incidence of cytomegalovirus infection was much higher (p < 0.01) in the ATG group (70%) than in the non-ATG group (19%). From this study, we were not able to confirm the benefits of ATG as described by other investigators.

Successful bone marrow transplantation in a child with X-linked hyper-IgM syndrome.

We report a case of an 11-year-old boy who underwent successful bone marrow transplantation for X-linked hyper-IgM syndrome (XHIM). The donor was an HLA-matched brother. The patient was conditioned with busulfan, cyclophosphamide and anti-thymocyte globulin. He received 4.7 x 10(8) marrow cells per kg from the donor. Prophylaxis against graft-versus-host disease consisted of cyclosporine and short-term methotrexate. The clinical course after the bone marrow transplantation was uneventful, and 12 months after transplantation the patient was doing well with no need for therapy. We examined expression of the CD40 ligand (CD40L) on the patient's activated T lymphocytes and in vitro production of immunoglobulins by his lymphocytes. Although expression of CD40L was totally absent before the bone marrow transplant, subnormal expression appeared after the transplantation. In vitro production of IgG and IgA also was improved by the transplant. Based on our experience bone marrow transplantation appears to be a reasonable therapeutic option for patients with XHIM if HLA-matched family donors are available.

Differential dose responses of pulmonary tumor types in the rat after inhalation of plutonium dioxide aerosols.

Dose responses were compared among primary lung tumors and their histological types induced by a single inhalation exposure of female Wistar strain rats to submicron-size and polydispersed aerosols of plutonium dioxide (239PuO2). While the primary lung tumors were found only in 2.3% of the unexposed control animals, the frequency of all the primary lung tumors in the exposed animals was 44% at the mean lung dose of 0.71 Gy, and increased sharply at the doses of 1.5 Gy or more, reaching the maximum of 97% at 5.4 Gy, and the dose responses around at 1.0 Gy were different between benign and malignant lung tumors. Almost all the pulmonary tumors in the exposed animals were classified into epithelial types such as adenomas, adenocarcinomas, adenosquamous carcinomas, and squamous cell carcinomas. The dose responses were different between these tumor types as shown by the peak incidence of adenomas at 0.71 Gy, adenocarcinomas at 2.9 Gy, adenosquamous and squamous cell carcinomas at 5.4-8.5 Gy, respectively. As the magnitudes of neoplastic lesions in pulmonary carcinomas were expressed by histological scores, metaplasias and adenomatous lesions most frequently appeared at doses of 1.5 Gy, while the appearance and increase of carcinomatous lesions differed in the dose ranges as shown by the peak incidence of adenocarcinomatous lesions at 2.9 Gy, and adenosquamous or squamous lesions at 5.4-6.6 Gy. These results indicate a differential dose response of pulmonary carcinogenesis in which metaplasias and benign adenomas were induced at lower doses (< 1.0 Gy), whereas malignant carcinomas were induced at relatively higher doses (> 1.5 Gy). Together with the increase of carcinomatous lesions at higher doses, the intranuclear p53 protein accumulation was detectable, but only in a few percentages of malignant carcinomas.

Allogeneic bone marrow and peripheral stem cell transplantation from a haplo-identical mother and CD34 positive selection for CML.

A 6-year-old boy with CML in blastic crisis was transplanted with BM and PBSC from his HLA-mismatched MLC-positive mother following CD34-positive selection. Preconditioning for transplant was with thiotepa, cyclophosphamide, rabbit anti-human thymocyte globulin, and TBI followed by infusion of 2.6 x 10(6)/kg of CD34-positive BM and PBSC. Engraftment was confirmed by FISH analysis, and GVHD was not observed. On day 50, he relapsed and died despite three transfusions of donor lymphocytes without GVHD prophylaxis. CD34-positive cell selection for HLA-mismatched transplantation may prevent severe acute GVHD.