Influence of soil environmental parameters on thoron exhalation rate.

Field measurements of thoron exhalation rates have been carried out using a ZnS(Ag) scintillation detector with an accumulation chamber. The influence of soil surface temperature and moisture saturation on the thoron exhalation rate was observed. When the variation of moisture saturation was small, the soil surface temperature appeared to induce a strong effect on the thoron exhalation rate. On the other hand, when the variation of moisture saturation was large, the influence of moisture saturation appeared to be larger than the soil surface temperature. The number of data ranged over 405, and the median was estimated to be 0.79 Bq m(-2) s(-1). Dependence of geology on the thoron exhalation rate from the soil surface was obviously found, and a nationwide distribution map of the thoron exhalation rate from the soil surface was drawn by using these data. It was generally high in the southwest region than in the northeast region.

Separately measuring radon and thoron concentrations exhaled from soil using AlphaGUARD and liquid scintillation counter methods.

It was shown that radon and thoron concentrations exhaled from soil were separately measured using the AlphaGUARD and liquid scintillation counter (LSC) methods. The thoron concentrations from the RAD 7 were used to create the conversion equation to calculate thoron levels with the AlphaGUARD. However, the conversion factor was found to depend on the air flow rate. When air containing thoron of ∼60 kBq m(-3) was fed to the scintillation cocktail, thoron and thoron progeny could not be measured with the LSC method. The radon concentration of about 10 kBq m(-3) was measured with three methods, first with the LSC method and then with two AlphaGUARDs (one in the diffusion mode and the other in the flow mode (0.5 l min(-1))). There were no significant differences between these results. Finally, it was shown that the radon and thoron concentrations in air could be measured with the AlphaGUARD and LSC methods.

Expression of ATP sensitive K+ channel subunit Kir6.1 in rat kidney.

ATP-sensitive K+ (K(ATP)) channels in kidney are considered to play roles in regulating membrane potential during the change in intracellular ATP concentration. They are composed of channel subunits (Kir6.1, Kir6.2), which are members of the inwardly rectifying K+ channel family, and sulphonylurea receptors (SUR1, SUR2A and SUR2B), which belong to the ATP-binding cassette superfamily. In the present study, we have investigated the expression and localization of Kir6.1 in rat kidney with Western blot analysis, immunohistochemistry, in situ hybridization histochemistry, and immunoelectron microscopy. Western blot analysis showed that Kir6.1 was expressed in the mitochondria and microsome fractions of rat kidney and very weakly in the membrane fractions. Immunohistochemistry revealed that Kir6.1 was widely distributed in renal tubular epithelial cells, glomerular mesangial cells, and smooth muscles of blood vessels. In immunoelectron microscopy, Kir6.1 is mainly localized in the mitochondria, endoplasmic reticulum (ER), and very weakly in cell membranes. Thus, Kir6.1 is contained in the kidney and may be a candidate of mitochondrial K(ATP) channels.

Restoration by VIP of the carbachol-stimulated Cl- secretion in TTX-treated guinea pig distal colon.

To determine if vasoactive intestinal peptide (VIP) restores neural activity from tetrodotoxin (TTX) blockade, we studied the effects of VIP and related agents on carbachol (Cch)-induced Cl(-) secretion in control-isolated guinea pig distal colon and in that treated with TTX. The short circuit current (I(sc)) increased dose-dependently after serosal applications of Cch (10(-6) - 2 x 10(-5) M) and VIP (5 x 10(-9) - 10(-7) M). But no additive or synergistic increase in I(sc) was observed. Cch- and VIP-induced I(sc) was completely abolished by a serosal application of TTX (10(-6) M). However, a serosal application, not mucosal, of VIP (10(-7) M) and 8-bromo-cAMP (10(-3) M) restored the Cch-stimulated, TTX-inhibited I(sc) by 113% and 75.8%, respectively. Furthermore, mucosal and serosal applications of forskolin (aden late cyclase activator) restored the I(sc) by 43.9% and 65.3%, respectively. The restored I(sc) was completely abolished by atropine (muscarinic receptor antagonist). These results suggest that VIP may restore the cholinergic activity by increasing the level of intracellular cAMP, and that cholinergic neuron is very likely to be responsible for the regulation of Cl(-) secretion at neuroepithelial junctions. The exact mechanism of VIP's effect on the TTX-inhibited epithelial Cl(-) secretion, and its possible usefulness in the treatment of TTX-induced pathophysiological conditions, remain to be determined.

Bio-kinetics of radon ingested from drinking water.

Previous studies have identified the stomach as the most significant organ for the dose from ingested radon. An important factor in dosimetric modelling is the rate of radon loss from the stomach. In the present study, two subjects who ingested radon-rich water were measured using a NaI(Tl) detector fixed over the stomach. The counting rates for 214Pb and 214Bi peak regions were plotted as a function of time after ingestion. These data were interpreted using a compartment model that expressed biokinetics of radon and its progeny. The model was fitted to the experimental data by changing biokinetic parameters such as the rate of radon loss from the stomach. Previous models for dosimetric purposes often assumed that the half-time for radon loss from the stomach is below 20 min. The present results, however, suggest that a part of radon stayed longer in the stomach than expected in the previous models.

Combination of tumor necrosis factor alpha and interferon alpha induces apoptotic cell death through a c-myc-dependent pathway in p53 mutant H226br non-small-cell lung cancer cell line.

We investigated the role of wild-type p53 and c-myc activity in apoptosis induced by a combination of natural human tumor necrosis factor alpha (TNF-alpha) and natural human interferon alpha (IFN-alpha). Studies were performed with two human non-small-cell lung cancer cell lines, H226b, which has wild-type p53, and H226br, which has a mutant p53. The combination of IFN-alpha and TNF-alpha significantly inhibited cell growth and induced apoptotic cell death of both H226b and H226br, compared with IFN-alpha or TNF-alpha alone. Treatment with one or both cytokines did not affect the expression level of p53 in both cell lines. These results suggest that the combination of IFN-alpha/TNF-alpha induces apoptotic cell death through a p53- independent pathway. The c-myc oncogene is known to be involved in apoptosis induced by TNF. Antisense c-myc oligonucleotides have been reported to modulate cell growth or apoptosis in several cell lines. Antisense oligodeoxynucleotides were added to the culture of H226br cells before the addition of IFN-alpha/TNF-alpha. Antisense c-myc inhibited IFN-alpha/TNF-alpha cytotoxicity and apoptotic cell death. In conclusion, this study provides support for the speculation that TNF-alpha/IFN-alpha induce apoptosis through a c-myc-dependent pathway rather than a p53-dependent pathway. (c)2001 Elsevier Science.

Pentaketide melanin biosynthesis in Aspergillus fumigatus requires chain-length shortening of a heptaketide precursor.

Chain lengths and cyclization patterns of microbial polyketides are generally determined by polyketide synthases alone. Fungal polyketide melanins are often derived from a pentaketide 1,8-dihydroxynaphthalene, and pentaketide synthases are used for synthesis of the upstream pentaketide precursor, 1,3,6,8-tetrahydroxynaphthalene (1,3,6,8-THN). However, Aspergillus fumigatus, a human fungal pathogen, uses a heptaketide synthase (Alb1p) to synthesize its conidial pigment through a pentaketide pathway similar to that which produces 1,8-dihydroxynaphthalene-melanin. In this study we demonstrate that a novel protein, Ayg1p, is involved in the formation of 1,3,6,8-THN by chain-length shortening of a heptaketide precursor in A. fumigatus. Deletion of the ayg1 gene prevented the accumulation of 1,3,6,8-THN suggesting the involvement of ayg1 in 1,3,6,8-THN production. Genetic analyses of double-gene deletants suggested that Ayg1p catalyzes a novel biosynthetic step downstream of Alb1p and upstream of Arp2p (1,3,6,8-THN reductase). Further genetic and biochemical analyses of the reconstituted strains carrying alb1, ayg1, or alb1 + ayg1 indicated that Ayg1p is essential for synthesis of 1,3,6,8-THN in addition to Alb1p. Cell-free enzyme assays, using the crude Ayg1p protein extract, revealed that Ayg1p enzymatically shortened the heptaketide product of Alb1p to 1,3,6,8-THN. Thus, the protein Ayg1p facilitates the participation of a heptaketide synthase in a pentaketide pathway via a novel polyketide-shortening mechanism in A. fumigatus.

Transmural heterogeneity of myocardial integrated backscatter in diabetic patients without overt cardiac disease.

It is important to detect early changes in diabetic myocardium, because some diabetic patients suffer from diabetic cardiomyopathy, especially those with poorer glycemic control or hypertension (HT). To clarify whether ultrasonic tissue characterization can noninvasively detect ultrastructural changes in diabetic myocardium, we analyzed the transmural heterogeneity in myocardial integrated backscatter (THIB) in 20 diabetic patients and 16 normal subjects. THIB was defined as the absolute value of difference of integrated backscatter between the endocardial and epicardial half of the myocardium. THIB in diabetic patients was significantly greater than that in normal subjects. In diabetic patients, there was a significant correlation between glycosylated hemoglobin and THIB, and the greater THIB was shown in patients with HT compared with those without HT. Early changes in the myocardium, related to increased interstitial collagen deposition or other occult cardiomyopathic changes, may be detected on the basis of quantitative analysis of THIB in diabetic patients.

New approach to cell lineage analysis in mammals using the Cre-loxP system.

The Cre-loxP site-specific recombination system was used for cell lineage analysis in mammals. We constructed an expression plasmid, pCETZ-17, which consists of cytomegalovirus enhancer/chicken beta-actin promoter (CAG), a portion of the rabbit beta-globin gene, loxP-flanked DNA sequence (containing enhanced green fluorescent protein (EGFP) cDNA), and lacZ gene encoding E. coli beta-galactosidase (beta-gal). When circular pCETZ-17 plasmid DNA was microinjected into the pronuclei of fertilized eggs and these eggs were allowed to develop to two-cell stage, 62.8% (59/94) of the two-cell embryos exhibited distinct fluorescence in one or both blastomeres, but never expressed lacZ protein, as evaluated by histochemical staining with X-Gal, a substrate for beta-gal. When both circular plasmids, pCETZ-17 and pCAG/NCre (containing CAG and DNA sequences encoding nuclear location signal and Cre), were co-injected into fertilized eggs, almost all (87.0%, 47/54) embryos exhibited low or no fluorescence, but 51.9% (27/52) exhibited positive staining for beta-gal activity. This indicates that transient expression of the Cre recombinase gene removed the loxP-flanked DNA sequence in pCETZ-17 and then caused expression of the downstream lacZ sequence. We next microinjected pCETZ-17 into the pronuclei of fertilized eggs, cultured these injected eggs for 1 day, and collected only two-cell embryos expressing EGFP in both blastomeres. One blastomere of the EGFP-expressing two-cell embryos was microinjected with pCAG/NCre, and these treated embryos were cultured for 1 day up to four-cell stage. When the developing four-cell embryos were subjected to staining with X-Gal, cell lineage-related staining pattern for beta-gal activity was observed in most (77.8%, 7/9) embryos. These findings were further confirmed using two-cell embryos derived from a transgenic mouse line carrying CETZ-17 transgene. Thus, our system, which is based on transient expression of the Cre recombinase gene directly introduced into nuclei of embryonic cells by microinjection, is a powerful means for cell lineage analysis in mammals.

In vivo influence of p53 status on proliferation and chemoradiosensitivity in non-small-cell lung cancer.

Alteration of the p53 gene product is a frequent event in the progression of lung cancer. However, its importance to proliferation and response to chemoradiotherapy remains unclear. Thus, to assess its influence directly in vivo, we implanted into nude mice two kinds of human non-small-cell lung cancer (NSCLC) cells: H226br having a homozygous gene mutation in p53 (mt-p53) and H226b with intact p53 (wt-p53). We found that mt-p53 tumors grew substantially faster than wt-p53 tumors. Furthermore, treatment with cisplatin and radiation did not reduce the size of mt-p53 tumors, while wt-p53 tumors regressed by approximately 60%. Terminal-deoxytransferase-mediated dUTP-biotin nick-end labeling assay revealed apoptosis to be the mechanism responsible for the regression. Interestingly, apoptosis occurred in mt-p53 tumors although only at high doses of cisplatin and not at the magnitude detected in wt-p53 tumors. Cell labeling by staining with bromodeoxiuridine indicated that p53 is an important factor in modulating growth in NSCLC tumors. Our results are consistent with the notion that correction of a single genetic lesion enhances the therapeutic effect of chemotherapy.

[Four cases of carotid body tumor--especially the usefulness of CT and MRI in preoperative diagnosis].

Four cases of carotid body tumor are reported. Surgical removal of the tumor was performed in all cases. The internal carotid artery (ICA) was preserved in only two cases. In the other two cases ICA was unavoidably replaced by an EPTFE (expanded polytetrafluoroethylene) graft. In addition to the size and mobility in the anterior and posterior direction of the tumor, we concluded that CT and MRI were useful in decision of the surgical method. In case the ICA cannot be distinguished from the tumor at the bifurcation of the carotid artery on enhanced CT, it should be considered to be difficult to remove the tumor. MRI is suitable for discerning the inner surface of the carotid artery, but it is significant that MRI cannot necessarily reveal tumor invasion of the carotid artery wall.

Apoptosis in cultured human colon cancer cells induced by combined treatments with 5-fluorouracil, tumor necrosis factor-alpha and interferon-alpha.

Biochemical analysis using nick end-labeling was performed to investigate the effect of various combinations of 5-fluorouracil, natural human tumor necrosis factor-alpha and natural human interferon-alpha on the induction of apoptosis in RPMI 4788 human colon cancer cells. After treatment with 5-fluorouracil (1 mM) for 48 h, the number of nick end-positive cells was significantly increased in comparison to the situation without treatment. When tumor cells were treated with 1 mM 5-fluorouracil, 2.86 Japan Reference Units (JRU)/ml natural human tumor necrosis factor-alpha and 1 x 10(3) IU/ml natural human interferon-alpha in combination for 48 h, the number of nick end-positive cells was significantly higher than that after treatment with 5-fluorouracil alone. MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay revealed a significant decrease of relative viability, as compared to treatment with 5-fluorouracil (1 mM), 5-fluorouracil + natural human tumor necrosis factor-alpha, or 5-fluorouracil + natural human interferon-alpha for 48 h. Pretreatment with 5-fluorouracil (1 mM) for 24 h prior to treatment with natural human tumor necrosis factor-alpha (2.86 JRU/ml) and natural human interferon-alpha (10(3) IU/ml) for 24 h resulted in a significant increase of nick end-positive cells compared to pretreatment with natural human tumor necrosis factor-alpha and natural human interferon-alpha prior to treatment with 5-fluorouracil for 24 h (p < 0.05). These results suggest that 5-fluorouracil alone can induce apoptosis in RPMI 4788 tumor cells and that this effect can be enhanced by combination with natural human tumor necrosis factor-alpha and natural human interferon-alpha.

Anomaly in atmospheric radon concentration: a possible precursor of the 1995 Kobe, Japan, earthquake.

Clear seismic-related anomalies in the radon (222Rn) concentration of the atmosphere were observed prior to the Kobe earthquake (magnitude 7.2) on 17 January 1995. The radon anomalies were observed at a monitoring station in Kobe, which is located about 20 km away from the epicenter. The means of radon concentration in the atmosphere for each day were calculated using the data observed between April 1984 and March 1994, in order to obtain the normal radon variation. The difference between the smoothed radon concentration and the smoothed mean radon concentration is the residual value. Using the weekly averages of residual values in the historical period, the weekly residual value in the validation period were predicted. The historical period was from April 1984 to March 1994. The validation period was from April 1994 to January 1996. The seismic-related radon anomaly higher than the 99% confidence limit of the residual value of radon concentration in the atmosphere was observed beginning about 2 mo before the earthquake.

Effect of thyroid-stimulating hormone on cultured thyrocytes obtained from patients with Graves' disease and inhibitive effect by sodium iodide: a functional study.

Thyrocytes obtained from patients with Graves' disease were cultured for 3 days. This was followed by culture with 10 mU/mL thyroid stimulating hormone (TSH) (TSH group), TSH and sodium iodide (Nal group), or without (control group) for 3 additional days. On the 8th culture day, the amounts of intra- and extra-cellular cyclic adenosine monophosphate (cAMP), extracellular cAMP and thyroglobulin (TG), peroxidase (PO) activity, and cell numbers were measured. The amounts of intra- and extra-cellular cAMP correlated well. TSH increased the values of cAMP, TG and PO to levels higher than those of the control group. As the amount of Nal added to the medium increased, these values decreased. Addition of 10(-5) mol/L Nal lowered the value of cAMP only. When 10(-4) mol/L Nal was added, these three levels were lower than those of the TSH group and the value of cAMP was almost equal to that of the control group. On cell number, no difference was found between the cells cultured with TSH, TSH and Nal, and without TSH or Nal. When the thyrocytes were cultured with 1 mmol/L dibutyryl cAMP sodium salt or 8-bromoadenosine 3',5'-cyclic monophosphate instead of TSH, 10(-4) mol/L Nal did not lower the values of thyroglobulin and peroxidase activity. These results suggest that the Nal blocks the intracellular signal transduction provoked by TSH, only at the cAMP production level.

Two cases of nasopharyngeal branchial cyst.

Two cases of nasopharyngeal cyst, in a two-year-old girl and a six-year-old boy, are described. The cysts were located in the right lateral wall of the nasopharynx in both cases. Histopathological examinations revealed that the cyst walls were lined with columnar epithelium. The positions of the cysts and pathological features indicated that they were of branchial origin, and they were assumed to originate in the second branchial pouch because of their anatomic location. They differed from previously reported cases in that they extended nearly to the base of the skull, occupying the parapharyngeal space. It was considered that they might have originated from the dorsal part of the second branchial pouch or the layer of endodermal cells cut off from the lower part of the eustachian tube.

[Statistical observations of clinical cases with frontal, ethmoid and sphenoid sinus mucoceles].

Ninety six patients with paranasal mucoceles (or pyoceles) were admitted and treated at Gunma University Hospital from 1977 to 1986 (age range 15-82 years; mean 46.4 years). We statistically investigated these cases paying a special attention to 25 patients with visual disturbance. 1) In 71 cases mucocele was located in the frontal or fronto-ethmoid sinus (FE group), in 12 it was found in the ethmoid sinus (E group), and in 13 it was situated in the sphenoid or spheno-ethmoid sinus (SE group). 2) Sixty three cases had histories of previous surgery of paranasal sinuses and 29 cases were considered to have primary mucoceles. 3) The mean age of patients with primary mucocele was 56.4, while that with postoperative mucocele was 41.2. 4) In the FE group recurrence was so frequent that some device for surgical treatment should be considered (i.e., mucous membrane grafting, prolonged insertion of silicon tubes). 5) Visual disturbance was frequently observed in both the E group (7/12) and the SE group (9/13). 6) Visual disturbance may occur when mucocele compress the optic nerve and the globe at retrobulbar portion as well as at the optic canal. Twelve cases out of 25 with visual disturbance belonged in the former type. 7) Optic atrophy was most commonly seen in the SE group, but was recognized in 1 case even in the E group and the FE group respectively. 8) In patients with sudden visual disturbance, if only they were operated within a week after the onset, their visual acuity was remarkably improved except for serious cases. 9) In the cases presented with gradually attacked or varying visual disturbance, there was no correlation between the recovery of the vision and the duration of the disturbance.

[A case of bilateral nontraumatic internal carotid aneurysms presenting with recurrent massive epistaxis].

A case of bilateral nontraumatic internal carotid aneurysms presenting with recurrent massive epistaxis was reported. A 37-year-old female complaining of massive epistaxis from the left nostril was admitted to our hospital. After admission, she experienced recurrent massive epistaxis, but had no cranial nerve palsies. Carotid angiography demonstrated an aneurysm of the cavernous portion of the left internal carotid artery partially protruding into the sphenoid sinus. Neck clipping of the aneurysm was unsuccessful, therefore the left internal carotid ligation in the neck was performed with a Selverstone clamp. After the ligation, no rebleeding and neurological deficits occurred. Postoperative carotid angiography showed an aneurysm of the right internal carotid artery at the same site. The carotid angiography of 3 months later and 1 year and 3 months later revealed that the left aneurysm decreased in size and the right one remained unchanged. Twenty-one cases including ours that presented nontraumatic internal carotid aneurysm of the cavernous portion were reviewed. Twelve cases had no cranial nerve palsies, and 7 cases including ours had no other symptoms than massive epistaxis. Because massiveness of epistaxis from an internal carotid aneurysm often threatens one's life, diagnosis should be made by carotid angiography as soon as possible. There are several surgical procedures for such aneurysms. Clipping is the ideal method which can interrupt the blood flow to the aneurysm completely, but it is very difficult to be performed anatomically. Carotid ligation in the neck with little surgical invasion was an excellent method in 7 cases without rebleeding and neurological deficits. Bilateral intracavernous internal carotid aneurysms were found in our case and another case.(ABSTRACT TRUNCATED AT 250 WORDS)

Type-specific antigen of Streptococcus rattus strains (KAY1 and FA1). I. Characterization.

A serotype-specific antigen was extracted from Streptococcus rattus KAY1 strain isolated first in Japan from human dental plaque and purified on an ion exchange column to compare it chemically and immunologically with that of FA1 strain which had been examined extensively by previous workers. Antigens of both strains reacted in a double diffusion test specifically with anti-FA1 serum which had previously been demonstrated specific for the strains in the same test, agglutination reactions and/or radioimmunoassay using whole cells. After separation on a DEAE-Sephadex A-25 ion exchange column the antigen was found to be resistant to various enzymatic treatments with pronases, lipase and nucleases and produce a single precipitin band against absorbed anti-FA1 serum in immunoelectrophoresis. Chemical analysis of this antigen revealed that it composed of carbohydrate, protein and a few percentages of glycerol and phosphorus. Hapten inhibition tests between antigen and antibody showed that galactose as well as glucose were the most potent inhibitors, suggesting their involvement in the antigenic determinant. Involvement of the sugars was also supported by gas chromatographic analysis and abolishment of reactivity with antiserum after the treatment of antigens with NaIO4. Moreover, protein does not seem to be involved since after SDS-PAGE analysis an enzyme immunoassay gave a negative reaction with immunoblotted antigens.

High-speed spectroradiometer for remote sensing.

A high-speed spectroradiometer designed for spectral reflectance measurement in remote sensing is described. This instrument uses a monochromatic grating and a photomultiplier system for light detection and sweeps over the 400-850-nm wavelength spectral range with the spectral resolution of 2 nm within 1 s. The instrument has the inherent advantage of portability and speed of operation which make it particularly suitable for field work in the area of fast moving surfaces, e.g., water with wave motion. Some applications of its use in laboratory and field experiments also have been presented. The instrument would seem to be an appropriate instrument for ground data collection in remote sensing.