Early vitamin K deficiency bleeding in a neonate associated with maternal Crohn's disease.

We report herein a case of early vitamin K deficiency bleeding (VKDB) in a neonate associated with maternal Crohn's disease. A female neonate was born at 37 weeks' gestation and weighed 2778 g. She developed broad purpura on her back on day 1. Laboratory data showed anemia, prolonged coagulation time and elevated protein induced by vitamin K absence or antagonist-II. Early VKDB has not been reported in a neonate born from mother with active Crohn's disease. It is essential to give vitamin K selectively as soon as possible after birth to prevent early VKDB in neonates.

Clinical and immunological profiles in 17 Japanese patients with drug-induced pemphigus studied at Kurume University.

BACKGROUND: Drug-induced pemphigus (DIP) shows clinical, histopathological and immunological features of pemphigus. However, little is known about immunological profiles in DIP. OBJECTIVES: To characterize clinical and immunological profiles in patients with DIP. METHODS: We studied 17 Japanese patients with DIP who were treated at Kurume University Hospital or who consulted from other hospitals between 1997 and 2012. Complicated diseases, clinical and histopathological manifestations, responsible drugs and findings in immunofluorescence, enzyme-linked immunosorbent assays (ELISAs), immunoblotting (IB) and prognosis were analysed. RESULTS: Eight of the 17 patients with DIP showed pemphigus foliaceus-like appearance, three showed pemphigus herpetiformis-like appearance, and six showed atypical bullous lesions. Responsible drugs were thiol-containing drugs in 16 patients (bucillamine in nine cases, d-penicillamine in four cases, and cetapril, thiopronine and captopril in one patient each), and a nonthiol drug, sulfasalazine, in one patient. By ELISAs and/or IB analyses, nine patients reacted only with desmoglein 1 (Dsg1), four reacted with Dsg1 and Dsg3, and four showed no specific reactivity. By IB of normal human epidermal extracts, in addition to positive reactivity with Dsg1, four patients with no detectable malignancy showed paraneoplastic pemphigus-like reactivity with the 210-kDa envoplakin and the 190-kDa periplakin. Four cases showed anti-Dsg3 antibodies without mucosal lesions. While 11 cases recovered after discontinuation of the causative drugs, six patients had a very protracted or intractable disease course, and might develop true pemphigus. CONCLUSIONS: The present study indicated that the majority of the patients with DIP studied showed a pemphigus foliaceus-type phenotype with anti-Dsg1 autoantibodies, caused by thiol-containing drugs.

Structural modeling of dahlia-type single-walled carbon nanohorn aggregates by molecular dynamics.

The structure of dahlia-type single-walled carbon nanohorn aggregates has been modeled by classical molecular dynamics simulations, and the validity of the model has been verified by neutron diffraction. Computer-generated models consisted of an outer part formed from single-walled carbon nanohorns with diameters of 20-50 Å and a length of 400 Å and an inner turbostratic graphite-like core with a diameter of 130 Å. The diffracted intensity and the pair correlation function computed for such a constructed model are in good agreement with the neutron diffraction experimental data. The proposed turbostratic inner core explains the occurrence of the additional (002) and (004) graphitic peaks in the diffraction pattern of the studied sample and provides information about the interior structure of the dahlia-type aggregates.

Neon and CO2 adsorption on open carbon nanohorns.

We present the results of a thermodynamics and kinetics study of the adsorption of neon and carbon dioxide on aggregates of chemically opened carbon nanohorns. Both the equilibrium adsorption characteristics, as well as the dependence of the kinetic behavior on sorbent loading, are different for these two adsorbates. For neon the adsorption isotherms display two steps before reaching the saturated vapor pressure, corresponding to adsorption on strong and on weak binding sites; the isosteric heat of adsorption is a decreasing function of sorbent loading (this quantity varies by about a factor of 2 on the range of loadings studied), and the speed of the adsorption kinetics increases with increasing loading. By contrast, for carbon dioxide there are no substeps in the adsorption isotherms; the isosteric heat is a nonmonotonic function of loading, the value of the isosteric heat never differs from the bulk heat of sublimation by more than 15%, and the kinetic behavior is opposite to that of neon, with equilibration times increasing for higher sorbent loadings. We explain the difference in the equilibrium properties observed for neon and carbon dioxide in terms of differences in the relative strengths of adsorbate-adsorbate to adsorbate-sorbent interaction for these species.

Mechanism of the dependence of hepatitis B virus genotype G on co-infection with other genotypes for viral replication.

Hepatitis B virus (HBV) is classified into several genotypes. Genotype G (HBV/G) is characterised by worldwide dispersion, low intragenotypic diversity and a peculiar sequence of the precore and core region (stop codon and 36-nucleotide insertion). As a rule, HBV/G is detected in co-infection with another genotype, most frequently HBV/A2. In a previous in vivo study, viral replication of HBV/G was significantly enhanced by co-infection with HBV/A2. However, the mechanism by which co-infection with HBV/A2 enhances HBV/G replication is not fully understood. In this study, we employed 1.24-fold HBV/A2 clones that selectively expressed each viral protein and revealed that the core protein expressing construct significantly enhanced the replication of HBV/G in Huh7 cells. The introduction of the HBV/A2 core promoter or core protein or both genomic regions into the HBV/G genome showed that both the core promoter and core protein are required for efficient HBV/G replication. The effect of genotype on the interaction between foreign core protein and HBV/G showed that HBV/A2 was the strongest enhancer of HBV/G replication. Furthermore, Western blot analysis of Dane particles isolated from cultures of Huh7 cells co-transfected by HBV/G and a cytomegalovirus (CMV) promoter-driven HBV/A2 core protein expression construct indicated that HBV/G employed HBV/A2 core protein during particle assembly. In conclusion, HBV/G could take advantage of core proteins from other genotypes during co-infection to replicate efficiently and to effectively package HBV DNA into virions.

NY-ESO-1 antibody as a novel tumour marker of gastric cancer.

BACKGROUND: NY-ESO-1 antibodies are specifically observed in patients with NY-ESO-1-expressing tumours. We analysed whether the NY-ESO-1 humoral immune response is a useful tumour marker of gastric cancer. METHODS: Sera from 363 gastric cancer patients were screened by enzyme-linked immunosorbent assay (ELISA) to detect NY-ESO-1 antibodies. Serial serum samples were obtained from 25 NY-ESO-1 antibody-positive patients, including 16 patients with curative resection and 9 patients who received chemotherapy alone. RESULTS: NY-ESO-1 antibodies were detected in 3.4% of stage I, 4.4% of stage II, 25.3% of stage III, and 20.0% of stage IV patients. The frequency of antibody positivity increased with disease progression. When the NY-ESO-1 antibody was used in combination with carcinoembryonic antigen and CA19-9 to detect gastric cancer, information gains of 11.2% in stages III and IV, and 5.8% in all patients were observed. The NY-ESO-1 immune response levels of the patients without recurrence fell below the cutoff level after surgery. Two of the patients with recurrence displayed incomplete decreases. The nine patients who received chemotherapy alone continued to display NY-ESO-1 immune responses. CONCLUSION: When combined with conventional tumour markers, the NY-ESO-1 humoral immune response could be a useful tumour marker for detecting advanced gastric cancer and inferring the post-treatment tumour load in seropositive patients.

Changes in the fluorescence spectrum of individual single-wall carbon nanotubes induced by light-assisted oxidation with hydroperoxide.

Through fluorescence-spectrum measurements, we investigated the effects of light-assisted oxidation with H2O2 (LAOx) on single-wall carbon nanotubes (SWNTs) that were individually dispersed in an aqueous solution of surfactant. The intensities of the fluorescence spectra were decreased remarkably by the LAOx when the light's wavelength was 400-500 nm and a little when 600-700 nm. The spectrum intensity did not recover even when the pH was restored to an original value of 6.5. The spectra changed little when the LAOx wavelength was 500-600 nm or the light was not irradiated. In addition, the effect of LAOx on SWNTs was related to the diameters of SWNTs. We inferred that these phenomena reflected that H2O2 was dissociated by absorbing the fluorescence light emitted from optically excited SWNTs, which, in turn, accelerated the burning out of SWNTs.

Dissociation of electrolytes in a nano-aqueous system within single-wall carbon nanotubes.

Research on material incorporation within single-wall carbon nanotubes (SWNTs) through aqueous solutions of various electrolytes is performed for the purpose of providing a foundation for future application of SWNTs to, for example, drug delivery systems. We have determined that the optical spectra of SWNTs were significantly affected when SWNTs that had opened holes or removed caps were treated through immersion in an aqueous solution of electrolytes, followed by drying at room temperature; however, the spectra of SWNTs without opened holes or removed caps were not subjected to such treatment. We infer that when the sucked solutions remained inside the tubes, even after drying (the nano-aqueous system), the electrolyte was dissociated into ions, which was likely to change the electronic states of SWNTs. On the other hand, when the SWNTs were well-dried under vacuum, no remarkable changes in their optical spectra were observed.

Isolating single-wall carbon nanohorns as small aggregates through a dispersion method.

An approach to isolating small aggregates of single-wall carbon nanohorns (SWNHs) is presented. SWNHs are ultrasonically treated in an aqueous solution of surfactant, resulting in dispersion of SWNH aggregates. Subsequent centrifuging enables the separation of small aggregates from larger aggregates or agglomerations and removal of graphitic particles (GG balls), the main impurity. The SWNHs obtained in this way were purified and formed small aggregates, thus exhibiting characteristics superior to those of SWNHs before treatment. We believe that the ability to isolate small SWNH aggregates in an aqueous solution should contribute to their application in the fields of biological sensing and drug delivery systems.

Fe-sapphire and C-Fe-sapphire interactions and their effect on the growth of single-walled carbon nanotubes by chemical vapor deposition.

We previously reported that the quantity of single-walled carbon nanotubes grown on Fe-coated sapphire by chemical vapor deposition depended on the crystallographic faces of sapphires. In this report, we show that the interaction of Fe, sapphire, and carbon depended on the sapphire faces. We deduce that the quantity of Fe available to catalyze the growth of single-walled carbon nanotubes was suppressed by the formation of Fe-Al alloys and whether the Fe-Al alloys were formed on Fe-coated sapphire or not depended on the sapphire-surface structure.

Chemically doped double-walled carbon nanotubes: cylindrical molecular capacitors.

A double-walled carbon nanotube is used to study the radial charge distribution on the positive inner electrode of a cylindrical molecular capacitor. The outer electrode is a shell of bromine anions. Resonant Raman scattering from phonons on each carbon shell reveals the radial charge distribution. A self-consistent tight-binding model confirms the observed molecular Faraday cage effect, i.e., most of the charge resides on the outer wall, even when this wall was originally semiconducting and the inner wall was metallic.

Direct imaging of Sc2@C84 molecules encapsulated inside single-wall carbon nanotubes by high resolution electron microscopy with atomic sensitivity.

Intramolecular structure of the scandium dimetallofullerene (Sc(2)@C(84)) has been clearly revealed by high resolution transmission electron microscopy with the single-atom sensitivity. Direct observation of two Sc atoms inside each fullerene molecule has led to a successful determination of the molecular symmetry among the three possible structural isomers for the Sc(2)@C(84). The present work introduces a new electron microscopic approach to investigate individual molecular structures and demonstrates the possibility for determining the molecular isomer on a single-molecular basis.

Linking chiral indices and transport properties of double-walled carbon nanotubes.

We performed in situ transport measurements in a transmission-electron microscope (TEM) on individual double-walled carbon nanotubes (DWNT). Using selected-area electron diffraction, the chiral indices of the two tubes constituting the DWNTs were determined through careful comparison with theory. We discuss the case of a DWNT whose two tubes have a gap at half filling and show a finite density of delocalized state at the Fermi level. The exact determination of chiral indices should be reachable in any transport-measurement experiment with samples that allow TEM characterization.

Distribution and partial characterisation of IgG Fc binding protein in various mucin producing cells and body fluids.

BACKGROUND AND AIMS: Mucus released from goblet cells is important in intestinal mucosal defence, and mucin glycoproteins are thought to be major components of mucus. Recently, we identified and cloned another component of human colonic mucus, IgG Fc binding protein (Fc gamma BP). Fc gamma BP is immunologically distinct from known Fc gamma receptors and its structure contains repeated cysteine rich unit sequences resembling those present in mucins. In this work, we assessed the tissue distribution of Fc gamma BP, its binding activity in various body fluids, and its ability to inhibit complement mediated haemolysis. METHODS: Immunohistochemical localisation of Fc gamma BP, using monoclonal antibodies against Fc gamma BP (K9 or K17) and labelled IgG, was conducted in various mucin producing tissues: colon, small intestine, stomach, gall bladder, cystic duct, choledochus, bronchus, submandibular gland, conjunctiva, and cervix uteri. The binding activity of Fc gamma BP in mucus extracted from colon, gastric juice, bile, nasal discharges, saliva, sputum, and tears was also examined by immunodotblot and immunoprecipitation using these monoclonal antibodies. Inhibition of complement mediated haemolysis by Fc gamma BP was investigated using sheep red blood cells (SRBC) and anti-SRBC IgG. RESULTS: The immunohistochemical study revealed that mucin secreting cells in the colon, small intestine, gall bladder, cystic duct, choledochus, bronchus, submandibular gland, and cervix uteri contained Fc gamma BP, and immunodotblot and immunoprecipitation analysis using IgG and monoclonal antibodies demonstrated that the fluids secreted by these cells were capable of binding IgG. Mucin producing cells of the conjunctiva did not express Fc gamma BP molecules or bind to IgG. The surface mucus cells in the stomach were variably positive for Fc gamma BP. Perhaps because of proteolytic degradation, Fc gamma BP in gut lavage fluid did not have IgG binding activity, although this activity was present in the mucus covering the colon. Fc gamma BP suppressed complement mediated haemolysis of SRBC. CONCLUSIONS: Fc gamma BP is widely expressed on mucosal surfaces and in external secretions. It is functionally intact in several fluids. These findings lend support to the concept that Fc gamma BP is an important component of mucosal immunological defences.

Type-specific separation of animal cells in aqueous two-phase systems using antibody conjugates with temperature-sensitive polymers.

A new type of aqueous two-phase system (ATPS) has been developed in which a temperature-sensitive polymer, poly-N-isopropylacrylamide [poly (NIPAM)] was used as a ligand carrier for the specific separation of animal cells. Monoclonal antibodies were modified with itaconic anhydride and copolymerized with N-isopropylacrylamide, and the ligand-conjugated carriers were added to the polyethylene glycol 8000-dextran T500 aqueous two-phase systems. The antibody-polymer conjugates were partitioned to the top phase in the absence or presence of 0.15 M NaCl. When ligand-conjugated carriers were used, more than 80% of the cells were specifically partitioned to the top phase in the presence of NaCl up to 0.1 M. The cells were partitioned almost completely to the bottom phase at 0.1 M NaCl or above, when no antibody-conjugate was added in the ATPS. As a model system, CD34-positive human acute myeloid leukemia cells (KG-1) were specifically separated from human T lymphoma cells (Jurkat) by applying anti-CD34 conjugated with poly-N-isopropylacrylamide in the aqueous two-phase system. By the temperature-induced precipitation of the polymer, about 90% of the antibody-polymer conjugates were recovered from the top phase, which gave approximately 75% cell separating efficiency in the next cycle of reuse.

[Structural quantification of bone from radiographs using power spectral analysis].

Bone mineral density (BMD) and bone structure are very important indices for prevention of fracture. However, it is very difficult to quantify bone structure, and only a few indices for structural quantification of bone have been reported. The purpose of this research was to investigate a new index for bone structure. The subjects were 52 women aged from 20 to 85 years. Directivity index (DI) is a new index of bone structure calculated by directivity of power spectrum from radiographs of metacarpal bone using fast Fourier transform (FFT). DI was obtained by subtracting the integral power value at 0 and 90 degree directions on the x-y plane of the two-dimensional power spectrum of bone from the integral power value at a direction of 45 degrees. A significant relationship between BMD and DI was indicated by correlation coefficient. However, no significant relationship between BMD and the first moment of the Fourier power spectrum or the fractal dimension was found. There is a possibility that DI estimates a slight deformation of bone structure. In the future, we will apply DI to the prevention of fractures and osteoarthritis.

Establishment of CHO cell lines expressing four N-methyl-D-aspartate receptor subtypes and characterization of a novel antagonist PPDC.

To develop an assay system that allows the N-methyl-D-aspartate (NMDA) receptor subtype-selective antagonistic potency of drugs, we have established Chinese hamster ovary cell lines expressing the four NMDA receptor subtypes (GluRepsilon1/zeta1-GluRepsilon4/zeta1) heat-indelibly. Using these clonal cells, we found that a novel antagonist, (1S,2R)-1-phenyl-2[(S)-1-aminopropyl]-N,N-diethylcyclopropanecarboxamide, was less selective for the GluRepsilon1/zeta1: the IC(50) values for the GluRepsilon1/zeta1-GluRepsilon4/zeta1 were 41.7, 13.3, 12.6 and 11.5 microM, respectively, while two well-known antagonists, DL-2-amino-5-phosphonovaleric acid and ifenprodil, showed the known potency and selectivity for each subtype. Thus, the established clonal cells are of use in characterizing the pharmacological properties of drugs that act on NMDA receptors.

Production of transgenic quails with high frequency of germ-line transmission using VSV-G pseudotyped retroviral vector.

We report here the production of transgenic quails using a replication-defective pantropic retroviral vector based on Moloney murine leukemia virus (MoMLV) pseudotyped with vesicular stomatitis virus G protein (VSV-G). The retroviral vector was injected into laid quail embryos at the blastodermal stage, and the embryos were incubated to hatch to produce G(0) transgenic quails. Among 134 embryos subjected to viral injection, 37 hatched. The viral vector sequence was detected in the tissues of all G(0) quails. The germ-line transmission efficiency of G(0) quails mated with nontransgenic quails was more than 80% on average. Southern blot analysis revealed that the G(1) transgenic progeny had one to three copies of the transgene. The expression of vector-encoded neomycin-resistance gene under the control of the Rous sarcoma virus (RSV) promoter was observed in several tissues including heart and muscle of both G(1) and G(2) transgenic offspring. Due to the high frequency of germ-line transmission, this method may markedly facilitate the production of transgenic avian.