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We studied the relationship between plasma alpha 2-macroglobulin (alpha 2M) and depressed cell-mediated immunity in idiopathic nephrotic syndrome (INS). Plasma alpha 2M concentrations (mumol/l) were increased during relapses of INS; however, the proteinase inhibitory activity, measured using bacterial thermolysin, was significantly decreased when calculated per 1 mol of alpha 2M, implying a reduced proteinase-inhibiting capacity of alpha 2M. The decreased proteinase-inhibiting capacity of alpha 2M was associated with the inhibitory activity of plasma on normal lymphocyte blastogenesis. Purified alpha 2M, when complexed with chymotrypsin, intensively inhibited normal lymphocyte blastogenesis induced by concanavalin A, as compared with the free form of alpha 2M. From these results it is suggested that, although the amount of alpha 2M protein has increased in the plasma of INS patients during relapse, its binding capacity to proteinases has relatively decreased. The results of this study may provide speculation for both the well-known high plasma alpha 2M concentrations and the immunodepression, both of which have been observed in INS patients in the past few decades.

A 56-year-old woman developed mixed warm and cold antibody type autoimmune hemolytic anemia (mixed AIHA) associated with systemic lupus erythematosus. The patient was admitted to our hospital for acrocyanosis and shortness of breath. High fever and jaundice were observed. Urinalysis revealed protein and hemoglobin, and the sediment contained granular and hyaline casts. Her erythrocytes agglutinated markedly at room temperature. Her hemoglobin was 5.6 g/dl and reticulocyts were 19.3%. Total bilirubin, GOT and LDH were elevated, while haptoglobin and complements were abnormally reduced. Polyclonal increase of immunoglobulin, ANA and anti-Sm antibody were detected. The direct antiglobulin test was positive; IgG1, IgG3 and C3d were detected on the red cell surface. The cold agglutinin (CA) titer was 4096, showing anti-I blood group specificity, and was still active at 30 degrees C. Upon administration of prednisolone gradual increase of hemoglobin and decrease of reticulocytes were observed, indicating the healing of hemolysis. CA disappeared but the direct antiglobulin test remained positive. Mixed AIHA has been defined as the presence of both warm and cold antibodies. In addition, the presence of symptoms of cold agglutinin disease, or low-titer and high thermal amplitude CA might be necessary for the diagnosis of mixed AIHA.

We describe a 69-year-old man who developed chylothorax after a 9-year remission of malignant lymphoma. The patient was admitted to our hospital and received exploratory laparotomy for ileus in February 1986. Bulky masses in the posterior mediastinum and the retroperitoneum, and also a jejunal tumor were observed. Fibrosis of the liver was also observed. The jejunal tumor was removed and histological findings revealed diffuse large B-cell malignant lymphoma. He was treated by combination chemotherapy and remission was achieved. He was discharged in June and remained in remission, but was readmitted for right pleural effusion in October 1994. Effusion was chylous and the chylomicron level was estimated to be 181 mg/dl. Liver cirrhosis also developed but there was no chylous ascites. Chylorrhea disappeared after continuous aspiration, but recurred in December. Continuous aspiration was ineffective, therefore 10 KE of OK-432 was administered twice into the pleural cavity, and chylorrhea again disappeared. No findings suggestive of malignant lymphoma were not detected by computerized tomography and gallium scintigram. He was discharged in March 1995 and chylothorax has not recurred since. These findings suggest that the fragility of the thoracic duct which had been infiltrated by malignant lymphoma might increase, resulting in rupture, even if in remission.

Recently, it has been reported that CDDP modifies the immune responses of tumor bearing hosts, but problem of the most appropriate treatment with CDDP to augment the host immunity remains unresolved. The purpose of this study was to determine the most advantageous administration of CDDP to increase both its cytotoxic effect and antitumor immune reactivity. Cell growth inhibition in vitro was assessed by MTT assay after treatment of U-251MG cells with various concentrations of CDDP for 24 or 120 hours. The results of this experiment showed that the growth of U-251MG cells in vitro was suppressed by CDDP in a dose-dependent as well as a time-dependent manner. Ten days after subcutaneous inoculation of human glioma cells, GL-9, in the rear flank of nude mice, they were assessed for growth suppression by CDDP. Totally, 20 ml/kg of CDDP was administered to each mouse; in one group (high-dose CDDP group), 5 ml/kg of CDDP on every fifth day, and in the other group (low-dose CDDP group), 2 ml/kg on every day. Measurement of the tumor volume in each group revealed no significant difference between the two groups in terms of the efficacy of tumor growth suppression. Twenty-one days after inoculation, we measured the splenic natural killer(NK) cell activity in each mouse. The results showed that NK cell activity was significantly increased in the low-dose cisplatin (2 ml/kg) group, and significantly decreased in the high-dose cisplatin (10 mg/ml) group, as compared to the control group. The results of our study suggest that the separate administration of low-dose CDDP is a useful treatment strategy for malignant glioma, because it increases the antitumor immune reactivity of hosts without decreasing the direct tumor cytotoxicity of CDDP.

Genomic DNAs of metallothionein I and II in Caenorhabditis elegans (CeMT-I and CeMT-II) were isolated by YAC library/polytene filter hybridization followed by subcloning of corresponding cosmid clones. Both genes are mapped at chromosome V. Although the similarities of 5'-flanking regions and coding regions have shown only 55-58%, the introns are split at the same position in both genes, indicating that these two genes are originally from the same gene. While several metal responsive elements are conserved among eukaryotes, only one metal responsive element was found in the promoter region in CeMT-II and not in CeMT-I. Indeed, neither of 5'-flanking regions of CeMT-I nor CeMT-II connected to chloramphenicol acetyltransferase reporter gene is responsive to heavy metals in mammalian culture cells by transient transfection analysis. These results would suggest that the metal regulatory factors in C. elegans might be different from those conserved in invertebrates and vertebrates, although the MTs in C. elegans revealed the similarities to mammalian MTs in several points.

In addition to biological and mechanical compatibilities for promising implant materials, a morphological compatibility is proposed by the authors. It has been reported by many investigators that implant surface with appropriate roughness and pore size exhibit better bone ingrowth activities. However, these parameters cannot characterize the complexity of surface textures. In the present study, dentulous and edentulous mandibular alveolar bones were utilized. Four segments from each mandible were subjected to the Fractal Dimension (DF) analysis. It was found that the dentulous mandible showed the DF of 1.81 +/- 0.03 while the edentulous mandible exhibited DF of 1.55 +/- 0.07, indicating that the former has more complex surface texture. It was also found that there could be a linear relationship between the surface roughness and the fractal dimension.

Complementary DNAs from HCV genome were expressed and analyzed. C8-2 protein, a part of NS5, and core protein were synthesized in E. coli. Core protein (JCC) was appeared to be very useful for diagnostic probe of HCV infection. Predicted envelope glycoproteins (E1 and E2/NS1) produced in infected insect cells were glycosylated and secreted into the culture media when the signal sequence of rabies virus G protein was introduced. An ELISA was developed using each purified recombinant protein. Anti-E1 antibody was detected in 20% of patients with non-A, non-B chronic liver diseases, whereas anti-E2/NS1 antibody was detected in more than 88%. These results indicate that the immune responses against these glycoproteins are different.

Rat lens contains two classes of glutathione S-transferase (GST) isozymes; one is class mu, Yb1-Yb1, and the other is class pi, Yp-Yp, judged from their molecular weights, immunological properties and N-terminal amino acid sequences. The expression pattern of GST isozymes in the rat lens is different from that in pig and bovine lenses which have only class pi and class mu isozymes, respectively.

Myofibroblasts (Mfs) from rat fat tissues produced a potent endothelial cell growth factor (Mf-ECGF). The growth factor activity found in the conditioned media from primary cultures of Mfs, was labile to heat (80 degrees C for 10 min) and proteinase (trypsin), and did not bind to heparin in the presence of 0.2 M NaCl. Mf-ECGF was partially purified 4760-fold with a recovery of 25% from serum-free conditioned media by sequential carboxymethyl (CM) ion-exchange column chromatography and gel filtration. This Mf-ECGF activity was recovered from the 40 kD region of a non-reducing SDS-PAGE, and from the pH region between 6.5 and 7 of isoelectric focusing, with recoveries of 20% and 65%, respectively. These results indicated that a major portion of ECGF activity in the conditioned media was clearly distinct from other well-known endothelial cell growth factors including fibroblast growth factors (FGFs).

Serotype-specific polysaccharide antigens from Actinobacillus actinomycetemcomitans ATCC 29523 (serotype a) and NCTC 9710 (serotype c) were extracted from whole cells by autoclaving and purified by ion-exchange chromatography and gel filtration. Analysis of component sugars by gas-liquid chromatography-mass spectrometry, high performance liquid chromatography, and NMR together with optical rotation data showed that the serotype a antigen was composed solely of 6-deoxy-D-talose, whereas the serotype c antigen consisted of 6-deoxy-L-talose. Structural analysis indicated that both of these antigens were composed of closely related repeating units, -3)-6-deoxy-alpha-D-Talp-(1-2)-6-deoxy-alpha-D-Talp-(1-(sero type a) and -3)-6-deoxy-alpha-L-Talp-(1-2)-6-deoxy-alpha-L-Talp-(1-(sero type c). 1H and 13C NMR analysis showed that both of these serotype antigens contained one acetyl group/2 sugar residues. These acetyl groups localized at the O-2 position of 3-linked 6-deoxy-D-talose (serotype a) or O-4 position of 3-linked 6-deoxy-L-talose residues (serotype c), respectively. These results coupled with our previous findings on the serotype b antigen (Amano, K., Nishihara, T., Shibuya, N., Noguchi, T., and Koga, T. (1989) Infect. Immun. 57, 2942-2946) showed that the serotype antigens from A. actinomycetemcomitans are a group of novel polysaccharides with structural features closely related biosynthetically.

Serotype-specific polysaccharide antigens (SPAs) were extracted from whole cells of Actinobacillus actinomycetemcomitans ATCC 29523 (serotype a), Y4 (serotype b), and NCTC 9710 (serotype c) by autoclaving and purified by chromatography on DEAE-Sephadex A-25 and Sephacryl S-300 columns. Y4 SPA induced interleukin-1 (IL-1) release by P388D1 murine macrophages. Polymyxin B had virtually no effect on the release of IL-1. Rabbit anti-murine IL-1 serum strongly suppressed the proliferation of C3H/HeJ mouse thymocytes induced with the culture supernatants of Y4 SPA-stimulated P388D1 cells and a submitogenic dose of concanavalin A. Gel filtration of the culture supernatants of Y4 SPA-stimulated macrophages on Sephacryl S-200 showed that an IL-1 peak at a point corresponding to approximately 16.5 kDa was eluted. The ability of SPAs from strains ATCC 29523 and NCTC 9710 to induce the release of IL-1 was lower than that of Y4 SPA. The IL-1-releasing ability of serotype a and c antigens was enhanced by deacetylation of both polysaccharides, suggesting that acetyl groups of these antigens might hinder the interaction between the antigens and macrophages.

We obtained highly purified fimbrae from Bordetella pertussis cells which gave a single band in SDS-polyacrylamide gel electrophoresis. Using the purified fimbriae, we prepared anti-fimbriae rabbit or goat IgGs by fimbriae-Sepharose affinity chromatography, and developed ELISA for its determination. The concentration of fimbriae in our former type of acellular vaccine (80 micrograms per ml) was determined to be 20 ng per ml, and that in our component vaccine was at an almost negligible level. To estimate the protective effects of the small levels of fimbriae, we evaluated the protective activities of three types of vaccines; the former type acellular vaccine (pertussis toxin (PT): filamentous hemagglutinin (FHA): fimbriae = 15:61:0.02 (microgram per ml)), the two-component vaccine (16:64:0.0001), and the three-component vaccine (16:63:0.8). The three types of vaccines showed no significant differences in protectivity against the experimental aerosol infection suggesting that these levels of fimbriae are not effective against the experimental aerosol infection of mice with Bordetella pertussis.

We have developed a model system for studying angiogenesis in which microvascular fragments and myofibroblasts (Mf) isolated from lipid tissues are grown in co-culture. We have found that Mf induce capillary formation by producing an endothelial cell growth factor and by secreting an extracellular matrix that causes endothelial cells to form a cordlike structure. This system appeared to be well suited for examining the effects of vasoactive substances such as the TNF and INF-gamma on capillary growth. TNF-alpha,beta, and IFN-gamma not only significantly inhibited capillary growth induced by Mf, but also blocked capillary development induced by fibroblast growth factors (FGF), well-known potent angiogenic factors. Recently, we have found that platelet-derived growth factor (PDGF) enhances in vitro capillary formation, probably at least in part by acting on Mf. Just as with FGF, capillary growth in the presence of PDGF was almost completely blocked by IFN-gamma. We examined the mode by which IFN-gamma inhibits angiogenesis and found that IFN-gamma inhibits both the proliferation of endothelial cells and collagen(s) synthesis by Mf. These actions of TNF or IFN-gamma could limit vascular formation in solid tumors.

Thiolactomycin (TLM), (4R)-(2E,5E)-2,4,6-trimethyl-3-hydroxy-2,5, 7-octatriene-4-thiolide, purified from a culture filtrate of a strain of the Nocardia species, was examined for antimicrobial activities against more than 100 strains of oral and periodontally associated bacteria. Nine other commonly used antibiotics were also included for the test. We found that TLM exhibited strong and selective antimicrobial activities against Bacteroides gingivalis and other oral black-pigmented Bacteroides species that may be etiologically associated with adult periodontitis. TLM also inhibited the growth of Actinobacillus actinomycetemcomitans, but did not affect the growth of oral streptococcal species and Eubacterium species. Strains of Eikenella corrodens were moderately susceptible to TLM, while Actinomyces viscosus strains were only slightly susceptible to it. Other antibiotics used for comparison showed a broad spectrum of antimicrobial activities in general. In conclusion, TLM exhibited highly selective antimicrobial activities to black-pigmented Bacteroides species and A. actinomycetemcomitans, both of which are implicated in the pathogenesis of human periodontal disease.

Metallothioneins in Caenorhabditis elegans (CeMT-I and II) were purified by the combination of gel filtration, ion-exchange chromatography, and high-performance liquid chromatography. Amino acid compositions and amino-terminal sequences of CeMT-I and II were slightly different from those of vertebrate MTs previously reported, although cysteine residue contents were relatively high. Enzyme immunoassay using anti-CeMT-II antibody showed the difference of antigenicity to rat MT-I and II and even to CeMT-I. Immunohistochemical staining revealed the existence of CeMT-II in the intestine and the eggs, suggesting the role of MT in detoxification and homeostasis of heavy metals.

SUN5555, (5R,6S)-6-[(R)-1-hydroxyethyl]-7-oxo-3-[(R)-2- tetrahydrofuryl]-4-thia-1-azabicyclo-[3.2.0]-hept-2-ene-carboxylic acid, is a newly developed and orally effective penem with a broad antibacterial spectrum. SUN5555 was degraded in rat tissue homogenates and serum, and its degradations in the kidney and lung homogenates were significantly inhibited by cilastatin, an inhibitor of dehydropeptidase I. The pulmonary extraction was evaluated to be 0.033 by comparing the AUCs resulting from iv and intra-arterial administration of 10 mg/kg. From the arterial-to-venous concentration differences under steady state conditions in rats, extraction ratios of SUN5555 across the kidney, hind leg, gut, and liver were determined to be 0.333, 0.104, 0.071 and 0.053, respectively, whereas no appreciable extraction across the brain was observed. Therefore, organ clearances of SUN5555 are limited by the product of plasma unbound fraction and organ intrinsic clearances in various tissues. Using a physiologically based pharmacokinetic model, unbound concentrations of SUN5555 in the tissue interstitial fluids, which are the measure of in vivo antibacterial efficacy, were estimated to be almost identical with, or higher than, those in plasma in rats. Similarly, serum concentration-time profiles of SUN5555 after iv administration in other animal species were predicted and found to be in good agreement with the observed data in dogs. A preliminary prediction of the pharmacokinetics of SUN5555 in a human subject was also attempted by animal scale-up technique.

As Zinc oxide films have many available characteristics, a large number of crystallographic studies have been reported on the films deposited by sputtering. However, most of those are about the influence of deposition conditions for (002) (c-axis) oriented films(l)–(2). The growing process of ZnOx films has not been necessarily manifested.

Single-monolayer high pentacene (Pn) dendrites grown on a hydrogen-terminated Si(111) surface [H-Si(111)] under ultrahigh vacuum were observed by low-energy electron microscopy and microbeam low-energy electron diffraction analyses. We determined the epitaxial structure (type I) inside a unique polycrystalline domain structure of such dendrites, each of which has six equivalent epitaxial orientations of Pn two-dimensional (2D) unit cells. There are three sets of these cells, which are rotated $\ifmmode\pm\else\textpm\fi{}120\ifmmode^\circ\else\textdegree\fi{}$ relative to each other. Domain boundaries inside each dendrite were successfully observed by scanning tunneling microscopy. In addition, we found another epitaxial relation (type II): the polycrystalline domain structure and lattice parameters are similar to those of the type-I dendrite; however, the 2D unit cells of the type-II dendrite are rotated approximately 90\ifmmode^\circ\else\textdegree\fi{} relative to those of the type-I dendrite. These results suggest that the crystal structure of the dendrites on H-Si(111) is determined mainly by the interaction between Pn molecules. Each dendrite is composed of domains that are exclusively of type I or II. The so-called point-on-line coincidences are found between the Pn 2D lattices of types I and II, and H-Si(111). The higher commensurability of the type-I dendrites than the type-II dendrites results in a higher probability of type-I dendrite formation. Moreover, for both the type-I and type-II dendrites, we found supercell structures. We estimated the minimum interface energy between the dendrite and H-Si(111) from an island's free energy, which is necessary to reproduce the growth of a single-monolayer high dendrite.

Based on the Japanese recommended dietary allowances for energy, the desirable physical activity level ( PAL) for Japanese is 1.67. As walking is the most common and the easiest exercise in our life, it is logical to attempt to estimate how many steps are needed daily to achieve that PAL. Data were collected from 511 healthy subjects (188 men and 323 women) who recorded their physical activities and simultaneously wore a pedometer for three days. Daily energy expenditure, daily PAL, time and PAL of different activities such as sitting and standing were estimated. Relationship between total number of steps and real walking time was studied. Men and women were different regarding daily standing and walking activities. For example, average standing time for men (96min/day) was significantly less than women (young: 144min, middle-aged: 257min). The men also walked faster (82 steps/min) than women (66 steps/min). From these results, we found that desirable number of steps for men: 30 xsleeping (min)+1700, and desirable number of steps for women: 24 xsleeping (min)+1400. Generally (standing time is less than four hours) the desirable number of steps is 13000–16000 for men, and 10000–13000 for women, corresponding to the sleeping time from 6 to 8 hours per day. Another equation was derived for men and women (especially housewives) whose standing time is more than 4 hours. Approximately 4000–9000 and 3000–7000 steps for men and women, respectively, are appropriate to achieve desirable PAL.

Maintaining the balance between energy intake (EI) and energy expenditure (EE) is essential for long term wellness of healthy adults. Until recently, measurements of EE have been difficult and inaccurate. A new accelerometer (motion sensor) which can distinguish among various postures including sitting, standing, walking and running, and can estimate daily EE has been developed. The purpose of this study was to compare estimates of total EE by this device with those obtained by EI from self reported diet records and physical activity diary (factorial method). We have developed a simple factorial method to reduce burdensome work of recording activities. Twenty six female students (out of 46) successfully completed 60 day recording of dietary intake and physical activity while wearing the accelerometer. The mean total EE obtained by the accelerometer (1696 ± 128 kcal) was approximately 94% of total EE obtained by the factorial method (1797 ± 146 kcal). Mean EI, weight gain, fat gain and physical activity level (multiples of BMR) were 1467±261 kcal, −0.2±1.1kg, −0.1 ± 1.2kg and 1.50 ± 0.06 , respectively. The mean energy balance was −330±246 kcal/day, but there was a change of body fat of −22 ± 175 kcal/day. The subjects underestimated dietary intake by about 300 kcal/day. The most serious student continued this study for 106 days and her average energy balance (+124 kcal/day) showed good agreement with her fat gain (+66 kcal/day). These results of this study demonstrated the applicability and reliability of the accelerometer in long term energy balance study.

Thermal deformation of a workpiece during surface grinding is one of the most important factors that affect a flatness of the ground surface. In this study, the thermal deformation of a workpiece attracted by a magnetic chuck is investigated by means of a finite element method. The obtained deformation pattern changes with a height to length ratio of the workpiece, and reverse shapes of these patterns coincide with the measured profiles of ground surfaces. The necessary constraining force for a magnetic chuck to prevent a vertical thermal displacement of the workpiece sole is also obtained. Consequently, the grinding conditions for avoiding the displacement by a conventional magnetic chuck is clarified.

A large scale production of human recombinant IL-5 (hIL-5) was performed by way of recombinant DNA technology. In this study, we transfected Chinese hamster ovary cells with pdKCR-hIL-5gene-dhfr plasmid and selected a cell line, with the use of methotrexate, producing large amounts of hIL-5. The recombinant hIL-5 thus obtained induced IgM production of murine B cell leukemia BCL1, and its activity was inhibited by TB13 anti-mouse IL-5 monoclonal antibody. hIL-5 could be purified from the cell-free supernatants of the transfectants with high recoveries by using anti-mouse IL-5 antibody-coupled immunoaffinity column in combination with a gel permeation column chromatography. N terminal amino acid sequence analysis of purified hIL-5 revealed that a single amino-terminal amino acid (isoleucine) is detected and hIL-5 consists of 115 amino acid residues.

We have studied the role of the membrane-associated form of IL-1 on bone resorption in vitro. Murine macrophages of the P388D1 cell line stimulated with LPS, subsequently fixed with paraformaldehyde, induced the proliferation of C3H/HeJ mouse thymocytes in the presence of a submitogenic concentration of Con A. Membrane IL-1 on P388D1 cells stimulated with LPS induced bone resorption in organ cultures of neonatal BALB/c mouse calvaria. Polyclonal antibodies directed against membrane IL-1 and soluble IL-1 from P388D1, and monospecific rabbit anti-murine rIL-1 alpha serum neutralized the membrane IL-1 activity, as measured by the thymocyte proliferation. In addition, these antibodies suppressed the bone resorption induced by membrane IL-1. The bone resorption induced by membrane IL-1 required direct contact between mouse calvaria and membrane IL-1. Salmon calcitonin strongly suppressed the calcium release from mouse calvaria in the presence of membrane IL-1. Indomethacin partially inhibited the bone resorption induced by membrane IL-1 on P388D1 cells. Moreover, membrane IL-1 on LPS-stimulated BALB/c mouse peritoneal macrophages or LPS-stimulated osteoblastic cells from BALB/c mouse calvaria induced bone resorption in vitro. These results suggest that membrane IL-1 on macrophages and osteoblastic cells may have a significant role in inflammatory bone resorption in vivo.

Serum specimens from 12 patients with type A hepatitis were analyzed for immunoglobulin M-type antibody to hepatitis A virus (IgM anti-HA). A recently developed solid-phase radioimmunoassay kit for IgM anti-HA (HAVAB-M, Abbott Laboratories) and a competitive binding radioimmunoassay kit (HAVAB, Abbott Laboratories) with or without 2-mercaptoethanol treatment, as modified by Yano et al. (Acta Hepatol. Jpn. 21, 704-712, 1980) were used to obtain an M-index. All specimens obtained within 60 days of the onset of illness and specimens from 2 of 4 patients later than 60 days after the onset were positive with the HAVAB-M test. This test gave negative results to sera which were positive for anti-HA by a standard HAVAB test in the following: 3 patients with type B hepatitis; 5 with non-A, non-B hepatitis; 11 healthy adults; and 10 sera strongly positive for rheumatoid factor. The M-index for type A hepatitis in sera within 30 days of the onset (mean value of the M-index, m, = 1.52; standard deviation, SD, = 0.25) was significantly higher than that for non-A hepatitis (m = 1.05; SD = 0.15) and for healthy adults (m = 1.02; SD = 0.10). The simplicity and usefulness of the HAVAB-M test in diagnosis of acute type A hepatitis over those measuring the M-index by HAVAB tests were shown by direct comparison of the results.

Ethidium bromide inhibits the in vitro replication of MDV-1 RNA (a small replicating RNA molecule) by reducing the rate of chain elongation. In a serial transfer experiment, in the presence of ethidium, a mutant RNA was selected that was more resistant to ethidium inhibition than is the wild-type MDV-1 RNA. The complete nucleotide sequence of the mutant RNA was determined and three nucleotides in the mutant sequence were found to be different from those in the wild type. The mixture of mutant and wild-type RNAs present in successive transfers was also sequenced. Each of the three point mutations occurred at a different time. These results show that the mutant RNA did not arise from a pre-existing strand present in the wild-type population, but rather, occurred de novo in the course of the experiment. It is probable that the chemical basis of resistance is the elimination of ethidium binding sites due to the specific alterations in the nucleotide sequence, since the mutant RNA was found to bind less ethidium than the wild-type molecules.

In 144 cases of hepatocellular carcinoma (HCC), 166 cases of cirrhosis without HCC and 142 cases of chronic hepatitis, we examined HBsAg, anti-HBs and anti-HBc in sera and compared the following factors between hepatitis B virus marker-negative and -positive patients: age, sex, alcohol consumption, family clustering of liver diseases, and histories of blood transfusion and post-transfusion hepatitis. Results of this study demonstrated several distinct differences in clinical backgrounds between non-B (negative for HBsAg, anti-HBs and anti-HBc) and B (positive for HBsAg) patients with HCC. Non-B patients were significantly older, had a lower frequency of familial tendencies for liver diseases, and more frequently had cancers other than HCC in their families. Some of these differences were also observed between non-B and B patients with cirrhosis and chronic hepatitis. Among patients with chronic hepatitis, the non-B patients had received blood transfusion or had post-transfusion hepatitis more frequently than the B patients. However, this difference was not apparent in patients with liver cirrhosis or HCC, suggesting that progression of non-A, non-B post-transfusion hepatitis to cirrhosis and HCC may not be as frequent as progression to chronic hepatitis.

A 128 K/spl times/8-b CMOS SRAM with TTL input/output levels and a typical address access time of 35 ns is described. A novel data transfer circuit with dual threshold level is utilized to obtain improved noise immunity. A divided-word-line architecture and an automatic power reduction function are utilized to achieve a low operational power of 10 mW at 1 MHz, and 100 mW at 10 MHz. A novel fabrication technology, including improved LOCOS and highly stable polysilicon loads, was introduced to achieve a compact memory cell which measures 6.4/spl times/11.5 /spl mu/m/SUP 2/. Typical standby current is 2 /spl mu/A. The RAM was fabricated with 1.0-/spl mu/m design rules, double-level polysilicon, and double-level aluminum CMOS technology. The chip size of the RAM is 8/spl times/13.65 mm/SUP 2/.

The change of motility and the presence of flagella were followed throughout growth and sporulation in a standard sporulating strain and in 19 cacogenic sporulation mutants of Bacillus subtilis. For the standard strain, the fraction of motile cells decreased during the developmental period to less than 10% at T4. Motility was lost well before the cells lose their flagella. Conditions reducing the decrease of motility also reduced sporulation: motile cells never contained spores. The decrease of motility was not coupled with a decrease in the cellular concentration of adenosine 5'-triphosphate or a decline in oxygen consumption, but an uncoupling agent immediately destroyed motility at any time. Apparently, motility decreased during development because it became increasingly uncoupled from the energy generating systems of the cell. The motility of sporulation mutants decreased after the end of growth at the same time as or earlier than the motility of the standard strain; the early decrease of motility in an aconitase mutant, but not that in an alpha-ketoglurate dehydrogenase mutant, could be avoided by addition of L-glutamate. Sporulation or related events such as extracellular antibiotic or protease production were not needed for the motility decline.

Microvariant RNA, a small self-replicating molecule (114 nucleotides long), has been isolated from Qbeta replicase reactions incubated in the absence of exogenous template. Its complete nucleotide sequence has been determined. Comparison with MDV-1 RNA, a somewhat larger endogenous Qbeta replicase product (220 nucleotides long) that had previously been characterized, revealed no significant sequence similarity. Since Qbeta replicase can mediate the synthesis of both of these disparate RNA molecules, primary sequence cannot be the sole determining factor in the processes of enzyme recognition and replication. This implies that the key is to be found in the secondary or tertiary structures. The availability of two different replicating molecules of defined sequence should aid in identifying these critical structural features.

A single-chip subscriber digital signal processor LSI (SDP) having programmable codec functions, which utilizes oversampling conversions and digital signal-processing techniques, is described. The chip is fabricated using a standard 2- mu m CMOS technology, and integrates many additional functions for PCM (pulse-code modulation) applications, such as gain control, an autobalancing network, a three-party conference, and a metering signal circuit, which are controlled by a software program. Excellent system characteristics and low power dissipation are achieved with operation from a single +5-V power supply. >

We isolated two galactosamine derivatives from Bacillus megaterium sporulating cells by lectin affinity chromatography followed by DEAE-Sephadex A-25 chromatography. From chemical analyses and measurements of these compounds, it was determined that one was uridine 5'-diphospho-N-acetylgalactosamine and that the other was uridine 5'-diphosphogalactosamine. They appeared in the middle stage of sporulation and disappeared during the period when galactosamine-6-phosphate is deposited on the forespore surface. These results suggest that uridine 5'-diphospho-N-acetylgalactosamine and uridine 5'-diphosphogalactosamine are intermediates in the biosynthesis of the galactosamine-6-phosphate polymer, a backbone structure of the exosporium.

Well orientated sputtered CoNi/Cr Films For magnetic recording disks have been prepared by a new process. A shielding mask has been used to control the incident direction of the sputtered Cr atoms. In the layered Film of the 420A thick Co 82 Ni 18 deposited over pure Cr by OC magnetron sputtering, a high coercive Force of 1430 Oe and a high squareness ratio of 0.82 in tangential direction are achieved. These magnetic properties are not isotropic. By using the shielding mask, a magnetic easy axis has been orientated in the tangential direction to the circumference of the disk. An SEM observation shows that agglomeration of the deposits into chains occurs whose long axes are parallel to the circumferential direction of the disk. The well controlled orientation is considered to be caused by the self shadowing effect.

Nephrotoxic activity of rabbit antisera to glomerular basement membrane (GBM) from three different donors (rat, gerbil, and man) was examined in mice of various strains. Animals receiving a small dose (0.1 ml) of anti-rat GBM or a large dose (0.3 ml X 2) of anti-gerbil GBM developed massive proteinuria and irreversible glomerular lesions which were identical to those seen in rats receiving anti-rat GBM, i.e. rat Masugi nephritis. Linear deposition of rabbit IgG, host's immunoglobulins and C3 was characteristically seen. Necrotizing changes and crescent formation accompanied by plasmic exudation, cellular proliferation and/or infiltration of PMNs and monocytes occurred after the injection. In animals receiving anti-human GBM, linear deposition of rabbit IgG and less marked glomerular changes were seen. The finding of no specificity in species between GBM antigens of rat and mouse indicates the usefulness of rat GBM for mouse Masugi nephritis or anti-GBM disease.

Interlayers of montmorillonite were pillared with TiO2, SiO2-TiO2and SiO2-Fe2O3sols. The pillared structures were thermally stable at least up to 500°C and retained unusually large basal spacings in the range of 24–45Å and surface areas as high as 300–500 m2/g. The TiO2pillared clay showed Type IV adsorption isotherm for nitrogen. Although SiO2-TiO2and SiO2-Fe2O3pillared clays had basal spacings much larger than that of TiO2pillared clay, these mixed oxide pillared clays had small pores and exhibited Type I isotherm. The acidic strength distributions were determined by a titration method using n-butylamine and Hammett indicators. All of the pillared clays had large acidities, but the acidic strength decreased in the following order: TiO2- SiO2» SiO2-Fe2O3pillared clays. Temperature-programmed desorption (TPD) spectra of ammonia were measured and interpreted in relation to the acidity distribution. Infrared spectra of pyridine adsorbed on TiO2pillared clay indicated that the acidity predominantly arises from Lewis acid sites.

Two patients with primary gastric lymphoma, diffuse lymphoma, a plasmacytoid variant of small cell type, with numerous intranuclear inclusions, are reported. In both cases, biopsy and surgical specimens showed diffuse proliferation of lymphoid cells having plasmacytoid features at different steps of maturation in lamina propria and submucosal layer. The cytoplasm of the lymphoma cells was stained with anti-IgM, anti-kappa chain, and anti-J chain antibodies by avidin-biotin peroxidase complex technique. A number of intranuclear inclusions in the lymphoma cells were diastase-resistant PAS positive, and also stained with anti-IgM, anti-kappa chain, and anti-J chain antibodies, suggesting a monoclonal situation. In an ultrastructural study of the first case, the inclusions were round and composed of amorphous materials surrounded by a single membrane. In many lymphoma cells, the perinuclear cisternal space was dilated and filled with materials similar to those within both inclusions and rough endoplasmic reticulum. A direct communication between the perinuclear space and the inclusion was demonstrated in some cells, suggesting the perinuclear cisternal origin of these inclusions.

Marine Pseudomonas strain 145-2 having the ability to produce extracellular protease was isolated from Uwa sea water, Ehime prefecture in Japan. This strain grows at 0 degrees - 37 degrees C, but protease production was observed only during 6-10 h cultivation at 20 degrees - 30 degrees C, when casein, as the nitrogen and carbon source in a concentration of 0.5-1.0%, was used. Maximum production of protease by the strain was also observed. Na, K, Ca and Mg ions were needed for growth and production of protease. Two extracellular proteases (proteases I and II) were purified from the culture supernatant by ammonium sulphate fractionation following Sephadex gel chromatography. Protease I had a larger molecular weight than protease II. After purification, proteases I and II were revealed as homogeneous by acrylamide gel electrophoresis and immunoelectrophoresis.

The effect of human tumor necrosis factor (TNF) on the permeability properties of liposomes containing phosphatidylserine at pH 5-6, as demonstrated by the calcein efflux. However, it did not induce any permeability change in such liposomes at neutral pH. The TNF-induced calcein efflux was also observed when an other acidic lipid was used as a component of the liposomes, i.e., phosphatidic acid or dicetyl phosphate. On the other hand, liposomes composed of neutral phospholipids such as phosphatidylcholine, phosphatidylethanolamine, and sphingomyelin showed little increases in permeability when incubated with TNF above pH 5.0. The TNF-induced permeability change was inhibited by the addition of polyaspartic acid, while it was not affected by the presence of 0.5 mM calcium ions. These data suggest that the negative charges on the liposomal surface trigger the interaction between TNF and liposomes. However, when the pH of the reaction mixture was decreased to 4.5, TNF-induced calcein efflux was observed even from neutral liposomes. When TNF was incubated with 8-anilinonaphthalene-1-sulfonic acid, the fluorescence intensity of this fluorophore increased with a decrease in the pH of the solution from 7 to 5, and a drastic increase in fluorescence was observed at pH 4.5. These data suggest that the hydrophobic region of TNF is also important for liposomal damage. Furthermore, the potencies of TNF and its derivative as to the induction of the permeability change paralleled their cytotoxic effects on mouse L929 cells, suggesting that the effect of TNF on liposomal membranes is related to its biological action.