Prenatal diagnosis of Duchenne muscular dystrophy using single fetal cell in maternal blood

We developed a method that allows prenatal diagnosis of Duchenne muscular dystrophy using a single nucleated erythrocyte (NRBC) isolated from maternal blood. Maternal blood was obtained at 8 to 20 weeks of gestation. NRBCs were separated with Percoll using a discontinuous density gradient method and then collected by micromanipulator under microscopic observation. The entire genome of a single cell amplified by primer extension preamplification (PEP). Sex was determined from a small aliquot of the PEP reaction. After an NRBC was determined to be male and confirmed to be of fetal origin, dystrophin exons 4, 8, 12, 45, 48, 50, and 51 were determined from the same PEP reaction. This diagnostic method using maternal blood is safer than amniocentesis or cordocentesis and can be applied to other X-linked disease.     

The result of a wall failure in-pile experiment under the EAGLE project

The WF (wall failure) test of the EAGLE program, in which 2 kg of uranium dioxide fuel-pins were melted by nuclear heating, was successfully conducted in the IGR (Impulse Graphite Reactor) of NNC/Kazakhstan. In this test, a 3 mm-thick stainless steel (SS) wall structure was placed between fuel pins and a 10 mm-thick sodium-filled channel (sodium gap). During the transient, fuel pins were heated, which led to the formation of a fuel-steel mixture pool. Under the transient nuclear heating condition, the SS wall was strongly heated by the molten pool, leading to wall failure. The time needed for fuel penetration into the sodium-filled gap was very short (less than 1 s after the pool formation). The result suggests that molten core materials formed in hypothetical LMFBR core disruptive accidents have a certain potential to destroy SS-wall boundaries early in the accident phase, thereby providing fuel escape paths from the core region. The early establishment of such fuel escape paths is regarded as a favorable characteristic in eliminating the possibility of severe re-criticality events. A preliminary interpretation on the WF test results is presented in this paper.     

The integral experiment on beryllium with D-T neutrons for verification of tritium breeding

A clean benchmark experiment on beryllium was performed with D-T neutrons at the FNS facility of the Japan Atomic Energy Agency. The main objective was to verify the integral data related to the tritium production on lithium isotopes. Tritium production rates, as well as activation reaction rates were measured inside the beryllium assembly that was shaped as a pseudo-cylindrical slab with an area-equivalent diameter of 628 mm and a thickness of 355 mm. Experimental results were analyzed with a three-dimensional Monte Carlo transport code MCNP-4C and FENDL/MC-2.0, JENDL-3.2/3.3 neutron transport libraries. Evaluation of reaction rates was based on the cross section data taken from the JENDL Dosimetry File and ENDF B-VI data libraries. Analysis shows that all calculation combinations (transport and activation cross section libraries) used for evaluation of reaction rates give data that is agreeable with measured values within 10%.     

Oral absorption improvement of poorly soluble drug using solid dispersion technique

A new triazol antifungal agent, (+)-2-(2,4-difluorophenyl)-3-methyl-1-(1H-1, 2,4-triazol-1-yl)-3-[6-(1H-1,2,4-triazol-1-yl)pyridazin++ +-3-ylthio]butan-2-ol (MFB-1041), shows poor oral absorption and is practically insoluble in water (1.2 micrograms/ml). Solid dispersion systems with an enteric polymer such as hydroxypropylmethylcellulose phthalate (HP-55) and carboxymethylethylcellulose (CMEC), and a nonenteric polymer, hydroxypropylmethylcellulose (Metolose) were evaluated to improve drug absorption and solubility. The oral bioavailabilities of these solid dispersions in beagle dogs were over 6 times higher than that of a suspension system with increasing drug solubility in an alkaline medium. X-Ray powder diffraction measurement of the solid dispersion showed a complete drug phase change from a crystal to an amorphous state. Further, from the results of a stability test, the preparations were stable in a desiccated condition and the absorption profiles also showed no change. From the results, it was suggested that the oral administrative preparation of MFB-1041 having a superior absorption profile and a high stability could be obtained by a drug phase change from a crystal to an amorphous state, especially in the spray-drying method using enteric polymers.     

Sound-velocity measurements for HFC-134a and HFC-152a with a spherical resonator

A spherical acoustic resonator was developed for measuring sound velocities in the gaseous phase and ideal-gas specific heats for new refrigerants. The radius of the spherical resonator, being about 5 cm, was determined by measuring sound velocities in gaseous argon at temperatures from 273 to 348 K and pressures up to 240 kPa. The measurements of 23 sound velocities in gaseous HFC-134a (1,1,1,2-tetrafluoroethane) at temperatures of 273 and 298 K and pressures from 10 to 250 kPa agree well with the measurements of Goodwin and Moldover. In addition, 92 sound velocities in gaseous HFC-152a (1,1-difluoroethane) with an accuracy of ±0.01% were measured at temperatures from 273 to 348 K and pressures up to 250 kPa. The ideal-gas specific heats as well as the second acoustic virial coefficients have been obtained for both these important alternative refrigerants. The second virial coefficients for HFC-152a derived from the present sound velocity measurements agree extremely well with the reported second virial coefficient values obtained with a Burnett apparatus.Paper dedicated to Professor Joseph Kestin.     

Premixed flame propagating into a narrow channel at a high speed, part 2: Transient behavior of the properties of the flowing gas inside the channel

The behavior of a premixed flame propagating from a chamber into a narrow channel at a high speed was examined using high speed and instantaneous schlieren photography, ion current measurement with electrostatic probes, and temperature measurement with an optical method. From detailed measurements of flames passing through the channel, flame extinguishment in the channel, no matter whether transient or not, was found to occur due to quenching of the reacting gas by rapid turbulent mixing with cold fresh mixture at the downstream of the contraction region and by heat transfer to the cold channel walls. Extinguishment must occur when these heat losses exceed the heat released by the chemical reaction of the flowing mixture.     

Inhibition of liver metastasis of human pancreatic carcinoma by angiogenesis inhibitor TNP-470 in combination with cisplatin

The anti-tumor and anti-metastatic effects of O-(chloroacetyl-carbamoyl) fumagillol (TNP-470), an angiogenesis inhibitor, and cisplatin (CDDP), an anti-neoplastic agent, were investigated using our established liver-metastasizing pancreatic carcinoma line, HPC-3H4. HPC-3H4 was injected into the spleens of nude mice. Mice were randomly divided into 5 groups; a control group given saline solution, a group receiving 45 mg/kg TNP-470, a group receiving 90 mg/kg TNP-470, a group receiving 90 mg/kg TNP-470 in combination with 0.25 mg/kg CDDP, and a group receiving 0.25 mg/kg CDDP. In the control group, liver metastasis developed in 14 of 15 mice (93.3%). Liver metastasis developed in 9 of 11 mice (81.8%) receiving 0.25 mg/kg CDDP. It developed in 11 of 15 mice (73.3%) receiving 45 mg/kg TNP-470, in 17 of 18 mice (94.4%) receiving 90 mg/kg TNP-470, and in 4 of 10 mice (40%) receiving 90 mg/kg TNP-470 in combination with 0.25 mg/kg CDDP. TNP-470 in combination with CDDP displayed a significant inhibitory effect on liver metastasis compared to the control. Although TNP-470 alone and CDDP alone had no effect on the tumor growth in vivo, 90 mg/kg TNP-470 in combination with 0.25 mg/kg CDDP had a significant effect. In vitro examinations demonstrated that the growth of HPC-3H4 cells was only mildly inhibited by TNP-470, but the production of vascular endothelial growth factor (VEGF) by HPC-3H4 was clearly inhibited by TNP-470. The inhibitory effect on the production of VEGF was not strong with CDDP treatment. These results indicate that the angiogenesis inhibitor TNP-470 in combination with low-dose CDDP has inhibitory activity against liver metastasis of human pancreatic carcinoma.     

Adaptive functioning following traumatic brain injury and orthopedic injury: a controlled study

BACKGROUND: In an effort to intensify osteosarcoma therapy, systemic ifosfamide was added pre- and postoperatively to an already aggressive three-drug regimen. In a subgroup of patients, loco-regional treatment intensification was attempted by using the intraarterial route to give cisplatin. PATIENTS AND METHODS: Patients < or = 40 years at diagnosis of a localised, de novo high-grade central extremity osteosarcoma were eligible for inclusion into study COSS-86 if registered within three weeks from biopsy. Doxorubicin, high-dose methotrexate, and cisplatin were given to all patients. Patients who fulfilled one or more of three defined high-risk criteria received early systemic treatment intensification by adding ifosfamide as the fourth agent. Preoperatively, these high-risk patients received cisplatin either intraarterially or intravenously. RESULTS: 171 eligible patients were entered, of which 128 were stratified into the high-risk group. When all 171 were analysed by intention-to-treat, actuarial overall and event-free survival rates at ten years were 72% and 66%, respectively. No benefit of intraarterial cisplatin application was detected. Cumulative treatment toxicity was considerable. CONCLUSIONS: In a multicenter setting, intensive treatment of osteosarcoma according to protocol COSS-86 led to long-term disease-free survival for two thirds of patients. We saw no benefit of using the intraarterial route to administer cisplatin.     

Patterns of expression for the mRNA corresponding to the four isoforms of phospholipase Cbeta in mouse brain

Ligand binding to neurotransmitter and hormone receptors which couple to the Gq subclass of GTP-binding protein leads to the activation of phospholipase Cbeta (PLCbeta) which hydrolyses phosphatidyl-inositol 4,5-bisphosphate, yielding a pair of second messengers, diacylglycerol and inositol 1,4,5-trisphosphate (IP3). The expression of PLCbeta1-4 mRNAs was comparatively examined by in situ hybridization in the mouse brain. In adults, PLCbeta1 mRNA was expressed predominantly in the telencephalon, including the cerebral cortex, hippocampus, amygdala, lateral septum and olfactory bulb, with little expression in most thalamic nuclei. PLCbeta2 mRNA was distributed in the white matter, suggesting its expression in non-neuronal cells, most likely oligodendrocytes. PLCbeta3 mRNA was specifically expressed in cerebellar Purkinje cells. The highest levels of PLCbeta4 mRNA were detected in Purkinje cells. High levels of PLCbeta4 mRNA were also found in the thalamus and medial septum, whereas weak signals were detected in most telencephalic regions, thus showing an expression pattern almost reciprocal to that of PLCbeta1 mRNA. During development, such characteristic regional expression of PLCbeta1 and PLCbeta4 were observed starting in late foetal stages, while specific expression of PLCbeta2 and PLCbeta3 appeared in early postnatal stages. We conclude that despite the existence of four PLCbeta isoforms, only one or two of them is expressed in individual neurons and glial cells. The distinct expression of PLCbetas provides a molecular basis for analysing the nature of the specific signal transduction pathway leading to the production of diacylglycerol and IP3 in distinct cell types and in different regions of the brain.