Suppression of streptozotocin-induced type-1 diabetes in mice by radon inhalation.

We examined the protective effect of radon inhalation on streptozotocin (STZ)-induced type-1 diabetes in mice. Mice inhaled radon at concentrations of 1000, 2500, and 5500 Bq/m3 for 24 hours before STZ administration. STZ administration induced characteristics of type-1 diabetes such as hyperglycemia and hypoinsulinemia; however, radon inhalation at doses of 1000 and 5500 Bq/m3 significantly suppressed the elevation of blood glucose in diabetic mice. Serum insulin was significantly higher in mice pre-treated with radon at a dose of 1000 Bq/m3 than in mice treated with a sham. In addition, superoxide dismutase activities and total glutathione contents were significantly higher and lipid peroxide was significantly lower in mice pre-treated with radon at doses of 1000 and 5500 Bq/m3 than in mice treated with a sham. These results were consistent with the result that radon inhalation at 1000 and 5500 Bq/m3 suppressed hyperglycemia. These findings suggested that radon inhalation suppressed STZ-induced type-1 diabetes through the enhancement of antioxidative functions in the pancreas.

Thoron exhalation rates in areas of Japan.

Thoron exhalation rates were measured with a newly made portable instrument at 33 areas in 7 prefectures of Japan. Thoron exhalation rates ranged from 49 to 4890 mBq m(-2) s(-1). Radon exhalation rates were also measured in many of the areas at the same time and ranged from 2.1 to 11 mBq m(-2) s(-1). Thoron exhalation rates showed a rough correlation with radon exhalation rates. Both exhalation rates also showed a rough correlation with geological features.

Time-integrated monitoring of thoron progeny concentration around closed uranium mine sites in Japan.

Thoron progeny concentrations were determined using the time-integrated method around closed uranium mine sites in Japan. Because the time-integrated radon progeny monitor developed by the authors has the function to detect (212)Po, time-integrated monitoring of thoron progeny concentration is also available with the monitor. Assuming that contribution of (216)Po is negligible, equilibrium equivalent concentration of thoron (EECTn) is theoretically calculated from the etch-pit counts by (212)Po. The annual averages of EECTn observed in the investigation area were about 0.2 Bq m(-3), and they had no remarkable differences from one another.

Artifact reduction of susceptibility-weighted imaging using a short-echo phase mask.

BACKGROUND: Susceptibility-weighted imaging (SWI) is utilized in magnetic resonance (MR) venography and other applications, but can include artifacts caused by the phase-masking process. PURPOSE: To demonstrate risks of filter processes used in making phase masks for SWI, and to propose a simple method for reducing artifacts. MATERIAL AND METHODS: Phase linearity related to echo time (TE) was evaluated for the original phase and high-pass-filtered phase using a CuSO(4)-doped water phantom. Effect of filter size of the Hanning window and background homogeneity were also evaluated in a phantom study. Use of a phase mask generated by data with differing magnitudes of TE was attempted in a human study. Shorter TE was used for making the phase mask, and the number of multiplications was increased. As short and long TEs were necessary simultaneously for phase mask and T2* contrast, a dual-echo technique was used. RESULTS: Linearity of TE and phase value collapsed, and an unexpected negative phase appeared in the high-pass-filtered phase. Using a short-TE phase mask, phase-aliasing artifacts were reduced and visibility of deep veins was equivalent to that under conventional methods with an increased number of multiplications. CONCLUSION: Use of a short-echo phase mask in SWI is useful for reducing artifacts.

Blood volume of gliomas determined by double-echo dynamic perfusion-weighted MR imaging: a preliminary study.

BACKGROUND AND PURPOSE: After bolus injection, gadopentetate dimeglumine causes a T2* rate change in permeable tissue that is contaminated by the T1 shortening effect due to the leakage of contrast agent. Therefore, tumor vascularity as reported in previous single-echo perfusion-weighted MR imaging studies has been underestimated. Our aim was to quantitatively and qualitatively evaluate the degree of blood volume of glioblastoma multiformes (GBMs) underestimated by this T1 shortening effect. METHODS: We used double-echo dynamic MR imaging after a bolus injection of gadopentetate dimeglumine (double-echo perfusion-weighted MR imaging) to simultaneously determine tumor blood volume without (V(T1U)) and with (V(T1C)) T1 shortening correction. MR imaging was performed in five consecutive patients with GBMs. The ratios of V(T1U) and V(T1C) were calculated and compared by means of quantitative analysis. The degree of tumor blood volume as determined by V(T1U) and V(T1C) maps were qualitatively compared using a three-point scale. RESULTS: All GBMs showed contrast enhancement on postcontrast T1-weighted images. In all subjects, the values of V(T1U) were significantly lower than those of V(T1C) (mean +/- SD, 2.05 +/- 1.01 vs. 3.62 +/- 1.40, respectively [P <.05]), indicating that tumor blood volume obtained by double-echo perfusion-weighted MR imaging was significantly higher than that by single-echo imaging. In the qualitative analysis, tumor blood volume on the V(T1U) map was less conspicuous than that on the V(T1C) map. CONCLUSION: Careful attention should be paid to the underestimation of tumor blood volume resulting from T1 shortening effects when using single-echo perfusion-weighted MR imaging. Double-echo imaging may be more suitable for the analysis of blood volume in GBMs.

Preliminary evaluation of electrochemical PNA array for detection of single base mismatch mutations.

We report here a preliminary evaluation of a microfabricated disposable-type peptide nucleic acid (PNA) array, with a 20-channel electrode, for the detection of cancer gene c-Ki-ras. Synthetic 15-mer PNA probes complimentary to ras sequence modified with cysteine were immobilized on the gold electrodes on the array. The electrochemical PNA array was reacted with 20-mer oligonucleotide target or 128 bp PCR product for 1 h. The anodic current derived from an electrochemically active DNA binder Hoechst 33258 was measured using the PNA array in the 50 microL of reaction chamber. The anodic current from Hoechst 33258 increased with increasing the concentration of PCR product of ras gene in the range from 10(11) to 10(15) copy mL(-1). The single base mismatch mutations of c-Ki-ras/61 were also detected using the electrochemical PNA array.

Thalamic abnormalities in patients with schizophrenia revealed by proton magnetic resonance spectroscopy.

Recent investigations suggest that thalamic abnormalities may underlie symptom formation in schizophrenia. We previously demonstrated reduced concentrations of N-acetylaspartate (NAA) in tissue from the thalamus of schizophrenic patients using in vitro proton magnetic resonance spectroscopy (1H-MRS). In the present study, in vivo 1H-MR spectra of the left thalamus and frontal lobe were investigated in 20 patients with schizophrenia and 16 age-matched control subjects to replicate our previous postmortem findings and support the hypothesis of thalamic abnormality in schizophrenia. Schizophrenic patients showed significantly lower NAA/total creatine (Cr) and choline-containing compounds (Cho)/Cr ratios in the thalamus than control subjects, while no significant difference was found in the frontal lobe. There was no significant correlation in the schizophrenic patients between the NAA/Cr or Cho/Cr ratio and other clinical data including clinical symptoms or neuroleptic dosage. These findings may further support other studies suggesting decreased thalamic volume or neuronal number and/or thalamic dysfunction, and reduction in size of white matter tracts adjacent to the thalamus in schizophrenia, as well as our previous postmortem MRS study.

Vascular permeability: quantitative measurement with double-echo dynamic MR imaging--theory and clinical application.

Double-echo dynamic magnetic resonance (MR) imaging was used to evaluate both vascularity and permeability of tissues simultaneously. Vascularity was evaluated on the basis of the T2*-shortening effect due to the intravascular fraction of the contrast agent and permeability on the basis of the T1-shortening effect due to the extravascular fraction. Meningioma was characterized on the basis of higher vascularity and neurinoma on the basis of higher permeability. The proposed method enables better tissue characterization.

Research-oriented image registry for multimodal image integration.

To provide multimodal biomedical images automatically, we constructed the research-oriented image registry, Data Delivery System (DDS). DDS was constructed on the campus local area network. Machines which generate images (imagers: DSA, ultrasound, PET, MRI, SPECT and CT) were connected to the campus LAN. Once a patient is registered, all his images are automatically picked up by DDS as they are generated, transferred through the gateway server to the intermediate server, and copied into the directory of the user who registered the patient. DDS informs the user through e-mail that new data have been generated and transferred. Data format is automatically converted into one which is chosen by the user. Data inactive for a certain period in the intermediate server are automatically achieved into the final and permanent data server based on compact disk. As a soft link is automatically generated through this step, a user has access to all (old or new) image data of the patient of his interest. As DDS runs with minimal maintenance, cost and time for data transfer are significantly saved. By making the complex process of data transfer and conversion invisible, DDS has made it easy for naive-to-computer researchers to concentrate on their biomedical interest.

Sequence-specific gene detection with a gold electrode modified with DNA probes and an electrochemically active dye.

A synthesized 20-mer DNA probe complementary to a part of an oncogene v-myc region having a mercaptohexyl group at the 5'-phosphate end was immobilized on a gold electrode by chemisorption. The immobilized DNA was detected voltammetrically using Hoechst 33258 with a DNA minor groove binder and an electrochemically active dye. The modified electrode was immersed into a 100 mumol/L Hoechst 33258 solution and washed with a phosphate buffer (pH 7.0). The anodic peak current (ipa) of Hoechst 33258 on the modified electrode was higher than that on a bare gold electrode (128 and 75 nA, respectively). It was considered that Hoechst 33258 was concentrated on the electrode surface due to its association with DNA. When the modified electrode was hybridized in a solution of a model targeted gene (10(-7) g/mL), single-stranded pVM623 containing the PstI fragment of a 1.5-kilobase pair of oncogene v-myc, the ipa was 192 nA. On the other hand, the ipa was 128 nA when the modified electrode was reacted in a solution of single-stranded pUC119 without a region complementary to v-myc in pVM623. The ipa was related to the concentration of the targeted DNA in the hybridization reaction. The use of Hoechst 33258 resulted in a sequence-specific detection of the targeted DNA quantitatively ranging from 10(-7) to 10(-13) g/mL in a buffer solution.

Stable liposomes for assays of human sera.

We report a novel homogeneous immunoassay system involving protein-bearing liposome-encapsulated carboxyfluorescein as a release marker. We applied this system to determine protein antigens, e.g., ferritin, in human serum samples by a sandwich-type assay. Liposomal lysis was observed in many samples, even though no second antibody was added to the reaction mixture. We demonstrated that the functional groups used to immobilize an antibody on liposomes are related to this phenomenon. Stable liposomes in human sera were prepared by incorporating bromoacetyl groups instead of the dithiopyridyl groups used previously. A good correlation (y = 0.98x - 8.81, r = 0.98, Sx/y = 66.9, range approximately 10-2000 micrograms/L) with results by RIA was obtained in the ferritin measurement of 53 patients' sera by using these liposomes.

Ascidian entactin/nidogen. Implication of evolution by shuffling two kinds of cysteine-rich motifs.

Entactin/nidogen, a major component of the basement membrane, has a domain structure comprising three globular domains, and thread-like and rod-like domains connecting them. It contains six epidermal-growth-factor-(EGF)-like motifs and one thyroglobulin-like motif. In the present study, ascidian entactin/nidogen has been identified by a monoclonal antibody technique. We prepared anti-(ascidian entactin/nidogen)IgG, named anti-AsEnt1, then cloned the cDNA of ascidian entactin/nidogen using anti-AsEnt1 as a probe, and determined its entire sequence. Mainly because the deduced amino acid sequence exhibited high similarity to mouse entactin and human nidogen, and because the antigen localized in basement membrane of ascidian body-wall muscle, we have concluded that the antigen anti-AsEnt1 corresponds to the ascidian entactin/nidogen homologue. The deduced amino acid sequence of ascidian entactin/nidogen clearly showed that the ascidian homologue also has a domain structure. However, the ascidian homologue lacked the thread-like domain, and the rod-like domain differed from that of mouse entactin in composition, consisting of two kinds of cysteine-rich motifs, that is, the EGF-like motif and the thyroglobulin-like motif. These results suggest that entactin/nidogen have evolved by modifying the domains, especially by shuffling the two kinds of cysteine-rich motifs.

[Experimental brain tumor in adult mongrel cat].

Small animal models such as the rat have serious limitations for multiple human scale instrumentation, surgical manipulations, and computerized tomographic (CT) evaluations, so that large animal models are required for the study using them. Although brain tumors induced with Rous sarcoma virus in neonatal beagle or adult monkey had been reported, these animals are very expensive ones for tumor research. A major drawback of virally induced brain tumor model is, moreover, the need for specialized viral facilities and safety precautions for laboratory personnel. In this paper, a cat glioma model implanted with C6 glioma cells derived from rats injected with N-nitrosomethylurea is reported. For an implantation dose of 5 x 10(5) cells/50 microliters, C6 glioma cells were suspended in modified Eagle medium supplemented with 10% fetal bovine serum and 0.5% agar. Twenty adult mongrel cats were injected with 5 x 10(5) C6 glioma cells intracerebrally. Implanted cats had brain tumors of about 10 mm in diameter with a yield of 80%. The mean survival was about 3 weeks after implantation. Tumors developed as spheroidal, hemorrhagic masses with central areas of necrosis and peripheral edema. They were located within the parenchyma of the implanted region. This tumor possessed many of the histological and radiological characteristics of human glioblastoma such as the following: Areas of hemorrhage and necrosis surrounded by pseudopallisading were observed within the tumor consisting of spindle-shaped cells with pleomorphic nuclei. A mass lesion with ring or garland-like enhancement surrounded by brain edema was shown on the CT scans.(ABSTRACT TRUNCATED AT 250 WORDS)

Liposome immune lysis assay (LILA). Application of sandwich method to determine a serum protein component with antibody-bearing liposomes.

A complement-dependent liposome immune lysis assay (LILA) using carboxyfluorescein (CF)-entrapped liposomes bearing antibody was developed to measure C-reactive protein (CRP) antigen as a model of protein antigens in human sera. Goat anti-CRP antibody was covalently coupled to liposomes, and a specific lysis of the liposomes could be observed when the liposomes were incubated with both rabbit anti-CRP antibody (secondary antibody) and CRP antigen in sera in the presence of guinea pig complement. In this assay system, so-called sandwich assay, CRP (a multivalent antigen) bound to the liposomes bearing anti-CRP antibody and subsequently secondary antibody, which activated complement efficiently. The amount of CF released by a complement-dependent liposome immune lysis was proportional to CRP concentrations. This sandwich assay was simple, fast, highly sensitive, and covered the ranges 10-300 ng of CRP/ml in a homogeneous mode, that is, one where no separation step was employed. The results correlated well with those obtained by single radial immunodiffusion and enzyme immunoassay. This assay system would be applicable to the measurement of other protein antigens.

Homogeneous determination of C-reactive protein in serum using liposome immune lysis assay (LILA).

Recently, we have developed an analytical system based on a complement dependent liposome immune lysis assay (LILA) to measure antibody against protein antigens. This paper describes the application of LILA to measure protein antigens by inhibition reaction. We choose human C-reactive protein (CRP), one of acute phase reactants, for a model of antigen determination. CRP antigen is coupled to the carboxyfluorescein entrapped multilamellar liposomes by using N-hydroxysuccinimidyl 3-(2-pyridyldithio) propionate and dithiothreitol, and a specific lysis of liposomes is achieved upon the addition of anti-CRP antibody in the presence of complement. Inhibition of this assay can be observed by the addition of competing amounts of CRP in human serum. This inhibition assay is simple, fast, highly sensitive, reproducible and homogeneous. The assay covers the ranges 10-500 micrograms/l. In an experiment using 50 samples, the results by LILA correlated well with those by single radial immunodiffusion and enzyme immunoassay (correlation coefficient: 0.99 and 0.93, respectively).

Liposome immune lysis assay (LILA): a simple method to measure anti-protein antibody using protein antigen-bearing liposomes.

A new simple immunoassay technique using immune lysis of liposomes was developed to measure antibody against protein antigens. Multilamellar liposomes were composed of dipalmitoylphosphatidylcholine, cholesterol and phosphatidylethanolamine substituted with the hetero-bifunctional cross-linking reagent N-hydroxysuccinimidyl 3-(2-pyridyldithio)propionate (SPDP). The protein antigen (human IgG) was coupled to these liposomes after treatment with SPDP and mild reduction. As a release marker, carboxyfluorescein (CF) was entrapped in the liposomes. The CF release was specific to anti-human IgG antibody and depended on the presence of complement. This technique could detect 10(-15) mol of anti-human IgG antibody or human IgG. The liposomes were stable over 8 months at 4 degrees C under nitrogen gas.

Determination of microbial populations with piezoelectric membranes.

The ultrasonic determination of microbial population was carried out by using a new apparatus composed of two piezoelectric membranes. When the apparatus was immersed in a medium containing cells and an arbitrary voltage was charged on a piezoelectric membrane, an output voltage was obtained from the other membrane. The output voltage increased with increasing cell numbers. The linear relationships between the output voltage and microbial populations of Saccharomyces cerevisiae, Bacillus subtilis, and Klebsiella sp. were observed, and the output voltage was reproducible with a relative error of 6%. Furthermore, the cell population of S. cerevisiae in a fermentor could be continuously determined with this apparatus.

Successful replantation of four fingers by a single common digital artery anastomosis.

The replantation of 4 digits of 1 hand was attempted in a patient who had completely amputated his fingers at the level of the metacarpophalangeal joints. Anastomosis of the severed ends of the third common volar digital artery proved to be successful not only in re-establishing arterial circulation in the middle and ring fingers but also in the index and little fingers, as evidenced by survival of the digits and as evaluated by postoperative angiography. The reasons for this success are discussed in light of anatomic studies.