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.

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.

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.

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.

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.

Conceptus uptake of the 106RuNO-nitro complex in relation to gestational stages.

106RuNO-nitro complex was administered intravenously to pregnant rats to study uptake of the radioruthenium complex by the conceptus in relation to gestational stages. Each conceptus was sampled periodically with respect to its placenta, fetal membrane, fetal fluid and fetus. Perceptible radioactivity in the fetus was detected only in the later stage of gestation and its relative concentration, defined as the ratio radioactivity per unit weight in the body tissue at sacrifice to that in the whole-body at dosing, was very low compared with other tissues. The average number of fetuses in one litter was 13 and the transfer rate of nitro complex into the fetuses 24 hr after injection to rats on the 20th day of gestation was about 1% of initial maternal dose. The relative concentration in the placenta and fetal membrane was much higher than in the fetus and decreased with time after injection. These results indicate that the placenta and fetal membrane play significant roles as barriers to the transfer of 106RuNO-nitro complex into the fetus.

Transfer of plutonium to rat embryos in vivo and in vitro.

The 239Pu distribution in the 12.5-day-old rat conceptus was compared between in vivo and in vitro experimental systems to establish a possible mechanism of cross-placental transfer of this radionuclide. In the in vivo study, plutonium citrate solution was injected intravenously to pregnant Wistar rats. In the in vitro study, either plutonium citrate or plutonium hydroxide colloid was administered, as a solution of Eagle MEM and FCS containing 239Pu at the concentration used in the maternal serum in the in vivo experiments, to rat conceptuses maintained by the whole-embryo culture method. The concentration of 239Pu in the yolk sac (239Pu activity per gram wet weight) were much higher than in the embryo in both the in vivo and in vitro experiments, suggesting that the yolk sac may be an effective barrier against the transfer of plutonium to the embryos. The ratios of the 239Pu concentration in the yolk sac to that in the embryo were relatively constant with time after administration in the in vitro system; 18-27 for plutonium citrate and 67-84 for plutonium hydroxide. In the in vivo experiment, these ratios changed with time after injection; 15 at 5 min and 62 and 60 min after injection. This suggests that in the in vivo system, the chemical form of 239Pu changed with time after injection, probably to a macromolecular form such as the hydroxide colloid or plutonium-protein complex although 239Pu was injected to the maternal blood as citrate.

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.

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.

High incidence of malignant lung carcinomas in rats after inhalation of 239PuO2 aerosol.

Female Wistar strain rats were exposed to a single inhalation of a submicron-size aerosol of high-fired 239PuO2 to investigate pulmonary carcinogenesis during lifespan periods. The absorbed lung doses of the exposed animals ranged from 0.6 to 12 Gy and were well correlated with the initial lung deposition (ILD) of 0.1 to 2.3 kBq. Survival and induction of primary lung tumors in 116 exposed rats were compared with those in 56 untreated control rats in respect to lung doses received. Mean survival time was greatly reduced, and the cumulative incidence of total lung tumors was markedly increased to 90-100% in rats that received more than 4 Gy, whereas of the controls only one animal (1.8%) died of primary lung tumors. Primary but benign adenomas were present in exposed animals given 1.0 Gy or less, and the incidence of adenomas was 22-25% at 4-5 Gy, but decreased sharply to 3-5% at 6-8 Gy. In contrast, no malignant carcinomas, including adenocarcinomas, adenosquamous carcinomas and squamous cell carcinomas, developed at a dose of less than 1.0 Gy, whereas they were present in 75% or more of the rats given 4-10 Gy, but only in 55% at 12 Gy. Although there were no clear differences in the dose and time required for induction among the carcinoma types, all tended to develop in earlier periods after inhalation than adenomas. Despite the limited number of exposed animals that received lower doses, results suggest that malignant lung carcinomas are highly and early induced and have a different dose-effect relationship than benign adenomas at doses of more than 1 Gy after inhalation exposure to 239PuO2.

Whole-body retention and tissue distribution of 60Co in rats after oral administration of freshwater fish contaminated with 60Co.

The purpose of this report is to compare the whole-body retention and tissue distribution in rats of 60Co administered by gavage as inorganic 60CoCl2 or in a form incorporated into freshwater fish. Orizias latipes were placed in vessels containing 21. of tap water with radioactive cobalt. Periodically thereafter the fish were sacrificed, homogenized, and administered to rats via a stomach tube. Control groups of rats were given the radionuclide alone or together with a homogenate of nonradioactive fish. The whole-body retention and tissue distribution of the radionuclide were determined with an Armac counter. The results revealed that rats gavaged with 60Co incorporated into the fish retained much more 60Co than control rats. This trend was notable in rats given fish kept in radioactive solution for longer periods. Marked differences in tissue distribution of 60Co were also observed between rats given 60Co incorporated into fish and control rats.

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.

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.

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.

[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.

[Serum pyridoxal concentration in children during theophylline therapy for bronchial asthma].

This study was designed to investigate the effect of sustained-release theophylline therapy on serum pyridoxal concentration in children with bronchial asthma. Forty-two children with bronchial asthma were divided into two groups according to duration of theophylline administration: The 22 children in group A had been treated with theophylline for less than 4 weeks, whereas the 20 children in group B had been treated for more than 5 weeks. The results obtained from this study were as follows: 1) The serum pyridoxal concentration in group B was significantly lower than that those in group A (p < 0.01). 2) The serum pyridoxal concentration was not significantly correlated with the serum theophylline concentration. These findings suggest that long-term theophylline therapy can depress vitamin B6 status in children with bronchial asthma. Theophylline-induced seizure may be caused by the possible decreased in gamma-aminobutyric acid concentration in the brain as a result of decrease in gamma-aminobutyric acid concentration in the brain as a result of decrease in vitamin B6 status, even if the serum theophylline concentrations are within the therapeutic range.

[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.