Radio-transparent multi-layer insulation for radiowave receivers.

In the field of radiowave detection, enlarging the receiver aperture to enhance the amount of light detected is essential for greater scientific achievements. One challenge in using radio transmittable apertures is keeping the detectors cool. This is because transparency to thermal radiation above the radio frequency range increases the thermal load. In shielding from thermal radiation, a general strategy is to install thermal filters in the light path between aperture and detectors. However, there is difficulty in fabricating metal mesh filters of large diameters. It is also difficult to maintain large diameter absorptive-type filters in cold because of their limited thermal conductance. A technology that maintains cold conditions while allowing larger apertures has been long-awaited. We propose radio-transparent multi-layer insulation (RT-MLI) composed from a set of stacked insulating layers. The insulator is transparent to radio frequencies, but not transparent to infrared radiation. The basic idea for cooling is similar to conventional multi-layer insulation. It leads to a reduction in thermal radiation while maintaining a uniform surface temperature. The advantage of this technique over other filter types is that no thermal links are required. As insulator material, we used foamed polystyrene; its low index of refraction makes an anti-reflection coating unnecessary. We measured the basic performance of RT-MLI to confirm that thermal loads are lowered with more layers. We also confirmed that our RT-MLI has high transmittance to radiowaves, but blocks infrared radiation. For example, RT-MLI with 12 layers has a transmittance greater than 95% (lower than 1%) below 200 GHz (above 4 THz). We demonstrated its effects in a system with absorptive-type filters, where aperture diameters were 200 mm. Low temperatures were successfully maintained for the filters. We conclude that this technology significantly enhances the cooling of radiowave receivers, and is particularly suitable for large-aperture systems. This technology is expected to be applicable to various fields, including radio astronomy, geo-environmental assessment, and radar systems.

Celecoxib upregulates endoplasmic reticulum chaperones that inhibit celecoxib-induced apoptosis in human gastric cells.

Nonsteroidal anti-inflammatory drugs (NSAIDs) induce apoptosis in cancer cells and this effect is involved in their antitumor activity. We recently demonstrated that NSAIDs upregulate GRP78, an endoplasmic reticulum (ER) chaperone, in gastric mucosal cells in primary culture. In the present study, induction of ER chaperones by NSAIDs and the effect of those chaperones on NSAID-induced apoptosis were examined in human gastric carcinoma cells. Celecoxib, an NSAID, upregulated ER chaperones (GRP78 and its cochaperones ERdj3 and ERdj4) but also C/EBP homologous transcription factor (CHOP), a transcription factor involved in apoptosis. Celecoxib also upregulated GRP78 in xenograft tumors, accompanying with the suppression of tumor growth in nude mice. Celecoxib caused phosphorylation of eukaryotic translation initiation factor 2 kinase (PERK) and eukaryotic initiation factor-2alpha (eIF2alpha) and production of activating transcription factor (ATF)4 mRNA. Suppression of ATF4 expression by small interfering RNA (siRNA) partially inhibited the celecoxib-dependent upregulation of GRP78. Celecoxib increased the intracellular Ca2+ concentration, while 1,2-bis(2-aminophenoxy)ethane-N,N,N'N'-tetraacetic acid, an intracellular Ca2+ chelator, inhibited the upregulation of GRP78 and ATF4. These results suggest that the Ca2+-dependent activation of the PERK-eIF2alpha-ATF4 pathway is involved in the upregulation of ER chaperones by celecoxib. Overexpression of GRP78 partially suppressed the apoptosis and induction of CHOP in the presence of celecoxib and this suppression was stimulated by coexpression of either ERdj3 or ERdj4. On the other hand, suppression of GRP78 expression by siRNA drastically stimulated cellular apoptosis and production of CHOP in the presence of celecoxib. These results show that upregulation of ER chaperones by celecoxib protects cancer cells from celecoxib-induced apoptosis, thus may decrease the potential antitumor activity of celecoxib.

Endoplasmic reticulum stress response is involved in nonsteroidal anti-inflammatory drug-induced apoptosis.

Apoptosis induced by nonsteroidal anti-inflammatory drugs (NSAIDs) is involved not only in the production of NSAID-induced gastric lesions but also in the antitumor activity of these drugs. The endoplasmic reticulum (ER) stress response is a cellular mechanism that aids in protecting the ER against ER stressors and is involved in ER stressor-induced apoptosis. Here, we examine the relationship between this response and NSAID-induced apoptosis in cultured guinea-pig gastric mucosal cells. Exposure of cells to indomethacin, a commonly used NSAID, induced GRP78 as well as CHOP, a transcription factor involved in apoptosis. Three factors that positively regulate CHOP expression (ATF6, ATF4 and XBP-1) were activated and/or induced by indomethacin. NSAIDs other than indomethacin (diclofenac, ibuprofen and celecoxib) also induced CHOP. Monitoring of the transcriptional activities of ATF6 and CHOP by luciferase assay revealed that both were stimulated in the presence of indomethacin. Furthermore, indomethacin-induced apoptosis was suppressed in cultured guinea-pig gastric mucosal cells by expression of the dominant-negative form of CHOP, or in peritoneal macrophages from CHOP-deficient mice. These results suggest that ER stress response-related proteins, particularly CHOP, are involved in NSAID-induced apoptosis.

Molecular mechanism for functional interaction between DnaA protein and acidic phospholipids: identification of important amino acids.

DnaA protein, the initiator of chromosomal DNA replication in Escherichia coli, seems to be reactivated from the ADP-bound form to its ATP-bound form through stimulation of ADP release by acidic phospholipids such as cardiolipin. We previously reported that two potential amphipathic helices (Lys-327 to Ile-344 and Asp-357 to Val-374) of DnaA protein are involved in the functional interaction between DnaA and cardiolipin. In relation to one of these helices (Asp-357 to Val-374), we demonstrated that basic amino acids in the helix, especially Lys-372, are vital for this interaction. In this study, we have identified an amino acid in the second potential amphipathic helix (Lys-327 to Ile-344), which would also appear to be involved in the interaction. We constructed three mutant dnaA genes with a single mutation (dnaAR328E, dnaAR334E, and dnaAR342E) and examined the function of the mutant proteins. DnaAR328E, but not DnaAR334E and DnaAR342E, was found to be more resistant to inhibition of its ATP binding activity by cardiolipin than the wild-type protein. The stimulation of ADP release from DnaAR328E by cardiolipin was also weaker than that observed with the other mutants and the wild-type protein. These results suggest that Arg-328 of DnaA protein is involved in the functional interaction of this protein with acidic phospholipids. We propose that acidic phospholipids bind to two basic amino acid residues (Arg-328 and Lys-372) of DnaA protein and change the higher order structure of its ATP-binding pocket, which in turn stimulates the release of ADP from the protein.

Identification of amino acids involved in the functional interaction between DnaA protein and acidic phospholipids.

DnaA protein, the initiator of chromosomal DNA replication in Escherichia coli, seems to be regulated through its binding to acidic phospholipids, such as cardiolipin. In our previous paper (Hase, M., Yoshimi, T., Ishikawa, Y., Ohba, A., Guo, L., Mima, S., Makise, M., Yamaguchi, Y., Tsuchiya, T., and Mizushima, T. (1998) J. Biol. Chem. 273, 28651-28656), we found that mutant DnaA protein (DnaA431), in which three basic amino acids (Arg(360), Arg(364), and Lys(372)) were mutated to acidic amino acids showed a decreased ability to interact with cardiolipin in vitro, suggesting that DnaA protein binds to cardiolipin through an ionic interaction. In this study, we construct three mutant dnaA genes each with a single mutation and examined the function of the mutant proteins in vitro and in vivo. All mutant proteins maintained activities for DNA replication and ATP binding. A mutant protein in which Lys(372) was mutated to Glu showed the weakest interaction with cardiolipin among these three mutant proteins. Thus, Lys(372) seems to play an important role in the interaction between DnaA protein and acidic phospholipids. Plasmid complementation analyses revealed that all these mutant proteins, including DnaA431 could function as an initiator for chromosomal DNA replication in vivo.

Expression pattern of a newly recognized protein, LECT2, in hepatocellular carcinoma and its premalignant lesion.

Leukocyte cell-derived chemotoxin 2 (LECT2) is a recently isolated protein and has been shown to be synthesized by human hepatocytes. All hepatocytes show diffuse immunostaining for LECT2 within the cytoplasm. In the present study, an attempt was made to clarify the expression pattern of LECT2 in nine cases of low-grade malignant hepatocellular carcinoma (LGM-HCC) and five cases of advanced HCC and 19 cases of premalignant lesion, termed atypical hyperplasia (AH), using the indirect immunoperoxidase technique. Variable spotty to coarsely diffuse staining in the majority of cells, a mixture of positively staining and negatively staining areas, and essentially negative staining was observed within the cellular cytoplasm of AH, LGM-HCC and advanced HCC, respectively. The expression of LECT2 became weaker with the progression of multistep hepatocarcinogenesis. The data clearly demonstrate that LECT2 becomes essentially negative in full-blown HCC cells and that the histological distinction between AH and LGM-HCC is valid. It also seems likely that LECT2 is related to hepatocyte growth.

Involvement of Arg-328, Arg-334 and Arg-342 of DnaA protein in the functional interaction with acidic phospholipids.

We reported previously that three basic amino acids (Arg-360, Arg-364 and Lys-372) of DnaA protein are essential for its functional interaction with cardiolipin. In this study, we examined the effect of mutation of some basic amino acids in a potential amphipathic helix (from Lys-327 to Ile-345) of DnaA protein on this interaction. ATP binding to the mutant DnaA protein, in which Arg-328, Arg-334 and Arg-342 were changed to acidic amino acids, was less inhibited by cardiolipin than that of the wild-type protein, as was the case for mutant DnaA protein with mutations of Arg-360, Arg-364 and Lys-372. A mutant DnaA protein with mutations of all six basic amino acids showed the most resistance to the inhibition of ATP binding by cardiolipin. These results suggest that Arg-328, Arg-334 and Arg-342, like Arg-360, Arg-364 and Lys-372, are also involved in the functional interaction between DnaA protein and acidic phospholipids.

Apoptosis of the intestinal principal cells of Xenopus larvae exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin.

In our previous work using paraffin-embedded sections, we determined that Xenopus larvae exposed to 200 ppb 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) for 5 days from shortly after fertilization to the early larval stage showed a shortening of the digestive tract and a loss of mucosal epithelium cells due to exfoliation into the gut cavity. In the current study, we ultrastructurally examined the mucosal epithelium of the gut of TCDD-exposed Xenopus larvae 12 days after fertilization. Exfoliated cell structures at the villus tip and in the lumen were equipped with a microvillus portion and occasionally had terminal web-like structures seen by ultramicroscopy. As these exfoliated structures had nuclear fragments with condensed chromatin, they were considered to be apoptotic bodies derived from the principal cells of the epithelium. In addition, many membrane-bound cell fragments-identified as apoptotic bodies derived from the principal cells based on their ultrastructural features-were observed at the basal side of the mucosal epithelium. These apoptotic bodies were phagocytized and digested chiefly by the neighboring intact principal cells. No such cells and/or cell fragments showing apoptotic features were observed in the controls. Our observations indicate that marked apoptosis occurs in the intestinal principal cells of TCDD-exposed larvae, which may result in the shortening of the gut.

Site-directed mutational analysis for the membrane binding of DnaA protein. Identification of amino acids involved in the functional interaction between DnaA protein and acidic phospholipids.

DnaA protein, the initiator of chromosomal DNA replication in Escherichia coli, interacts with acidic phospholipids, such as cardiolipin, and its activity seems to be regulated by membrane binding in cells. In this study we introduced site-directed mutations at the positions of hydrophobic or basic amino acids which are conserved among various bacteria species and which are located in the putative membrane-binding region of DnaA protein (from Asp357 to Val374). All mutant DnaA proteins showed much the same ATP and ADP binding activity as that of the wild-type protein. The release of ATP bound to the mutant DnaA protein, in which three hydrophobic amino acids were mutated to hydrophilic ones, was stimulated by cardiolipin, as in the case of the wild-type protein. On the other hand, the release of ATP bound to another mutant DnaA protein, in which three basic amino acids were mutated to acidic ones, was not stimulated by cardiolipin. These results suggest not only that the region is a membrane-binding domain of DnaA protein but also that these basic amino acids are important for the binding and the ionic interaction between the basic amino acids and acidic residues of cardiolipin and is involved in the interaction between DnaA protein and cardiolipin.

Hepatitis C virus is frequently coinfected with serum marker-negative hepatitis B virus: probable replication promotion of the former by the latter as demonstrated by in vitro cotransfection.

Patients with hepatitis C have been reported occasionally to be coinfected with serum marker-negative (silent) hepatitis B virus (HBV). The frequency and significance of such coinfection were investigated. Thirty patients with hepatitis C virus (HCV) infections (10 acute, 10 chronic, 10 cirrhotic) were selected randomly; the acute cases were without serum hepatitis B surface antigen (HBsAg) and anti-hepatitis B core IgM, and the chronic cases were without HBsAg. A nested polymerase chain reaction for the X open reading frame was used to amplify HBV DNA in serum, and immunoperoxidase staining was carried out on liver biopsy specimens. Nucleotide sequencing was carried out to characterize the amplified HBV DNAs. In order to clarify the possibility that the silent HBV mutant promotes HCV replication in the liver, the full-length HCV RNA and the cloned silent HBV DNA dimer were cotransfected into an established cell line, HuH-7, and the amount of secreted HCV RNA was quantified serially. The target HBV DNA was amplified in 26 (86.7%) of the 30 patients. Subsequent direct nucleotide sequencing in 9 selected patients revealed an 8-nucleotide deletion, characteristic of a silent HBV mutant. Immunostaining revealed hepatitis B surface antigen in 15 (50.0%). Cotransfected silent HBV DNA augmented the secretion of HCV RNA by up to 5-fold in comparison with HCV RNA transfection alone. In conclusion, HCV is coinfected frequently with the silent HBV mutant and the latter probably promotes the replication of the former in the liver.

A morphological study of liver lesions in Xenopus larvae exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) with special reference to apoptosis of hepatocytes.

Edema formation is a consistent feature of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) poisoning in experimental animals. Xenopus embryos exposed to 200 ppb of TCDD for 5 days from shortly after fertilization to early larval stage developed edema of the head, thorax, and abdomen, including ascites, from several days after the end of exposure. There has been no reports on liver lesions in TCDD-exposed anuran larvae. To elucidate the participation of liver in the edema formation, light and electron microscopic alterations in the livers from larvae that developed edema were examined. The marked change induced by TCDD was degeneration of the hepatocytes. Three types of degenerating hepatocytes were distinguished. The first type showed morphological features characteristic of apoptosis, including peripherally aggregated nuclear chromatin and overall cytoplasmic condensation; the cytoplasmic organelles retained their integrity and cytoplasmic blebbing. A considerable number of degenerating hepatocytes undergoing apoptosis and numerous apoptotic bodies derived from hepatocytes were observed by electron microscopy. The second type could not be determined as to which type of cell death it belonged to, and the third type was necrosis. Other alterations consisted of disturbances of the usual meshwork architecture of the liver, enlargements of bile canaliculi and the space of Disse, lipid accumulation, and appearance of phagocytizing macrophages. These morphological alterations in the liver generally correlated with the severity of edema with a few exception. These findings suggest that depressed hepatic function mainly due to hepatocyte death participates in the edema formation.

Pathology of livers infected with "silent" hepatitis B virus mutant.

We have discovered that non-A, non-B, non-C, non-D, non-E (so-called type F) acute and chronic hepatitis is caused by a hepatitis B virus (HBV) variant with mutations in the X open reading frame. This silent HBV mutant does not induce immunoserological markers. In the present investigation we attempted to elucidate the putative mechanism of hepatocellular necrosis and expression patterns of hepatitis B surface antigen (HBsAg) and core antigen (HBcAg) in biopsied liver tissue. The subjects consisted of 14 patients with acute hepatitis, 11 with chronic hepatitis and eight with liver cirrhosis, all of whom had been previously diagnosed as having so-called hepatitis F. Nine of the 14, 10 of the 11 and all eight, respectively, of the above patients exhibited significant positive immunostaining for HBsAg within their hepatocellular cytoplasm, diffusely or focally. HBcAg stained in a few hepatocellular nuclei in 24.2% of the patients. Histological features were characterized by necroinflammation, indicating immune-mediated hepatocellular necrosis. Despite the serological-marker negativity, the results of immunostaining for HBsAg and HBcAg support replication and expression of HBV DNA, though weak.

Mass screening for hepatocellular carcinoma: experience in Hokkaido, Japan.

Mass screening for liver cancer based mainly on abdominal ultrasound was begun in major cities of Hokkaido, Japan, in November 1981, to enable early detection and treatment of hepatocellular carcinoma (HCC). Serum alpha-fetoprotein levels were also measured to minimize false negative studies. Examinees included those who sought liver disease screening as well as high risk individuals: hepatitis B surface antigen carriers and those with a past or current liver disease, history of blood transfusion, family history of liver cancer, and more recently those with positive anti-hepatitis C antibodies. The examination was carried out on each Saturday and Sunday as one round, and by February 1992 48 rounds had been performed. A total of 8090 individuals were investigated, and HCC was detected in 91 with a detection rate of 1.12%. This rate was 1.6% among 5684 individuals who were selected for high risk. Cumulative rates of survival among these patients were 79.0% at 1 year, 43.8% at 3 years, 19.3% at 5 years and 15.4% at 7 years. These survival rates were comparable with those for the patients with HCC diagnosed during follow-up of chronic liver disease and treated at our hospital. The cost for detecting one HCC patient in this programme was 2,660,000 yen (approximately US$25,000), which was less than those for some other types of cancer in a similar setting. Considering the high detection rate in this programme, we feel that similar programmes should be encouraged and supported.

Detection of precore/core-mutant hepatitis B virus genome in patients with acute or fulminant hepatitis without serological markers for recent HBV infection.

To confirm the possibility that some hepatitis B virus (HBV) variants do not induce HB s antigen (HBsAg), anti-HB core antibody (anti-HBc) and anti-HBc IgM in a transient infection, polymerase chain reaction (PCR) was performed in 20 patients with acute hepatitis and 7 patients with fulminant hepatitis. Patients were diagnosed with non-A, non-B hepatitis by serological markers at admission. PCR successfully amplified the precore/core gene in 5 (25%) of the patients with acute hepatitis and 2 (29%) of the patients with fulminant hepatitis. Subsequent sequencing revealed frequent mutations including precore-defects in the precore/core gene.

Preliminary evaluation of sonolaparoscopy in the diagnosis of liver diseases.

A preliminary report is made on the use of sonolaparoscopy to overcome limitations of sonography and laparoscopy in the diagnosis of liver disease. The instruments used consisted of a sonolaparoscope with a mechanical radial scanner (LPSUMI/EUM1; Olympus) and the first prototype of an electronic sonolaparoscope (Olympus). Sonolaparoscopy was shown to be particularly suitable for detection and differentiation of occult liver lesions, cysts, tumors, various benign focal hyperplasias and various granulomata.

[High-resolution ultrasonography in cancer diagnosis--advances and indication of intraluminal sonography].

The present communication deals with recent developments in endoscopic sonography with special reference to the diagnosis of cancer of the upper abdominal organs. Examinations were carried out by an endoscope with a mechanical radial scanner, an Olympus GF UM1 and UM2, LPS UM1. Confirmed diagnostic applications are staging of esophageal and gastric cancer, diagnosis of submucosal tumors of the G.I. tract and differentiation between benign and malignant ulcer of the mucosa. The method also has excellent diagnostic application to malignancies involving organs adjacent to the G.I. tract, such as the mediastinum, retroperitoneum including the pancreas and adrenals, and the left lobe of the liver. Laparoscopic sonography, another endoscope-guided sonography involving the sonolaparoscope, revealed excellent ability to visualize and discriminate between benign and malignant small tumors in the liver and also the gallbladder and pancreas. The method is extremely helpful when surgical intervention is considered. Detection rate of hepatocellular cancer reached almost 90% of cases examined.

[Case of aneurysmal bone cyst of the skull].

A case of aneurysmal bone cyst of the right temporal bone was reported. The patient was a 36-year-old male who was admitted to our hospital with complaints of decreased right hearing and transient impairment of the right vision. A large tumor was palpated on the right temporal bone. Neurological examinations revealed right auditory loss, along with right facial weakness of peripheral type, and minimal pyramidal signs on the right side. The results of the laboratory examination proved to be normal. Neuroradiological examinations tended to be quite impressive. Plain x-ray films of the skull showed a blow out appearance of the right temporal bone and a bone decay in the right middle fossa, the right anterior clinoid process, and the posterior half of the right zygomatic arch. Angiograms revealed a right temporal extradural mass lesion without vascularity. CT scans showed a moderately enhanced mass lesion of soap bubble appearance in the right middle fossa. Surgical treatment was done under the diagnosis of extradural bone tumor. The operative findings disclosed many cysts containing bloody fluid and xanthochromic fluid. Numerous multinucleated giant cells, spindle cells, and foam cells were found in the microscopic examination. On the basis of macroscopic and microscopic findings, the patient was diagnosed as having an aneurysmal bone cyst. The aneurysmal bone cyst of the skull is very rare and sixteen cases have been reported in detail. This case had two interesting points, one was the transient impairment of visual acuity and the other was the CT findings.(ABSTRACT TRUNCATED AT 250 WORDS)

[Localized fibrous malignant peritoneal mesothelioma in an operated patient with massive hemorrhage to the peritoneal cavity].

We reported an operated patient with localized peritoneal mesothelioma. The patient was a 39-year-old man who had occasionally complained of left hypochondralgia for 5 years before admission. Progressive anemia due to hemorrhage into the peritoneal cavity was noted after admission. Peritoneoscopy revealed the bleeding to be located between the left lobe of the liver and the anterior wall of the stomach. On the 22nd hospital day, surgery was performed and a fist-sized tumor was resected. Microscopically, it was fibrous malignant type of the peritoneal mesothelioma. Chemotherapy was performed 3 times after the operation, and the patient is living and well 1 year and 8 months postoperatively.