Insulin like growth factor binding protein-7 reduces growth of human breast cancer cells and xenografted tumors.

Previously, we have shown that insulin-like growth factor binding protein-7 (IGFBP-7) expression is inversely correlated with disease progression in breast cancer and is associated with poor outcome. To further investigate the role of IGFBP-7 in the growth and metastatic behavior of breast cancer, primary breast tumors and metastatic tumors derived from the same patients were analyzed for IGFBP-7 expression. Immunohistochemical analysis revealed that IGFBP-7 is downregulated in half of the human metastatic breast tumors tested. IGFBP-7 has been linked to suppression of oncogenic pathways and can directly restore cellular senescence in melanomas, leading to their regression. It is possible that breast tumors with metastatic potential have escaped from IGFBP-7-induced suppression by its down-regulation. Twenty-two human primary breast tumor specimens were transplanted into human-bone NOD/SCID mice. One of the two triple negative primary breast tumors was serially xenotransplanted more than five times. Each serial transplant resulted in increased tumor take and rate of growth. Expression of IGFBP-7 was downregulated upon each serial implantation. To investigate the role of IGFBP-7 in breast tumor suppression, IGFBP-7 was overexpressed in the triple negative MDA-MB-468 human breast cancer line by stable transfection of a pSec-tag2-IGFBP-7 vector. The parental MDA-MB-468 breast cancer cells expressed extremely low levels of endogenous IGFBP-7. The production of IGFBP-7 protein by the MDA-MB-468 cells stably transfected with IGFBP-7 was confirmed by immunoblotting with anti-IGFBP-7 antibody. Ectopic overexpression of IGFBP-7 significantly reduced the growth of the IGFBP-7 transfected MDA-MB-468 cells compared to the parental MDA-MB-468 cells. We also assessed the role of IGFBP-7 on cell migration, a key determinant of malignant progression and metastasis. When parental MDA-MB-468 cells were treated with various amounts of conditioned medium derived from the IGFBP-7 overexpressing cell line, a significant difference in cell migration rate was observed between untreated and treated cells. IGFBP-7 strongly suppressed the phosphorylation of the mitogen-activated protein kinases (MAPK) ERK-1/2, suggesting that IGFBP-7 mediates its anti-proliferative effects through negative feedback signaling. Levels of phospho-ERK-1/2 were higher in the parental MDA-MB-468 than in IGFBP-7-expressing cells derived from it. When injected subcutaneously into NOD/SCID mice, the increased expression of IGFBP-7 in the MDA-MB-468 transfected cells reduced the rate of tumor growth in comparison to the parental MDA-MB-468 controls. These results suggest that the growth of breast cancer could be prevented by the forced expression of IGFBP-7 protein.

Two-dimensional pattern reverse Monte Carlo method for modelling the structures of nano-particles in uniaxial elongated rubbers.

Two-dimensional pattern reverse Monte Carlo (2D pattern RMC) analysis is performed to model the structures of nano-particles in uniaxially elongated rubbers using two-dimensional patterns of structure factor of the nano-particles obtained by time-resolved two-dimensional ultra-small angle x-ray scattering. Four spot patterns are observed for a large elongation ratio and the shapes change with increasing elongation ratio. We performed the 2D pattern RMC method for the uniaxial system in order to make a model of the structures from the two-dimensional structure factors. The preliminary results of the 2D pattern RMC analysis of the two-dimensional structure factors of silica particles in a uniaxially elongated styrene-butadiene rubber are presented.

The fish endocrine pancreas: review, new data, and future research directions in ontogeny and phylogeny.

The literature on the ontogeny and phylogeny of the endocrine pancreas of ray-finned fishes is summarized since the latest review in fish [Youson, J.H., Al-Mahrouki, A.A., 1999. Review. Ontogenetic and phylogenetic development of the endocrine pancreas (islet organ) in fishes. Gen. Comp. Endocrinol. 116, 303-335]. A basic description and a demonstration of the diversity of the fish islet organ is provided through new immunohistochemical data on islet tissue from a basal teleost, an osteoglossomorph, and a more derived teleost, a perciforme. Unlike the previous review, the present report provides a review and discussion of the utility of sequence data of insulin, somatostatin, and NPY- and glucagon-family peptides in phylogenetic analyses of jawed and jawless fishes. The present study also provides the first comparative analysis of sequences of preprohormones of endocrine peptides from closely related basal teleost species. Some nucleotide and deduced amino acid sequence data for preprosomatostatins (PPSS-I and/or -II) are compared for four species of bonytongues, Osteoglossomorpha, and with PPSSs of the white sucker, Catostomus commersoni, representing Cypriniformes, a more generalized teleost order. Phylogenetic analysis of deduced amino acid sequences of the PPSSs of these species and others from databases indicates good support for the monophyly of Osteoglossomorpha and some support for the present taxonomic grouping of the osteoglossomorphs examined, and also the white sucker. However, PPSS may have limited phylogenetic utility due to the relative short sequence, particularly in resolving relationships among lineages that diverged over a short period of time. Since in the few fish species examined we have just touched the surface in describing the diversity of structure of the islet organ, and likely the nature of the products of its cells, this report promotes the continued study of this organ.

A subthreshold MOS neuron circuit based on the Volterra system.

We present an analog neuron circuit consisting of a small number of metal-oxide semiconductor (MOS) devices operating in their subthreshold region. The dynamics of the circuit were designed to be equivalent to the well-known Volterra system to facilitate developing the circuit for a particular application. We show that a simple nonlinear transformation of system variables in the Volterra system enables designing a neuron-like oscillator, which can produce sequences in time of identically shaped pulses (spikes) by using current-mode subthreshold MOS circuits. We present experimental results of the fabricated neuron circuits as well as an application in an inhibitory neural network, where the neurons compete with each other in the frequency and time domains.

An analog CMOS central pattern generator for interlimb coordination in quadruped locomotion.

This paper proposes a neuromorphic analog CMOS controller for interlimb coordination in quadruped locomotion. Animal locomotion, such as walking, running, swimming, and flying, is based on periodic rhythmic movements. These rhythmic movements are driven by the biological neural network, called the central pattern generator (CPG). In recent years, many researchers have applied CPG to locomotion controllers in robotics. However, most of these have been developed with digital processors and, thus, have several problems, such as high power consumption. In order to overcome such problems, a CPG controller with analog CMOS circuit is proposed. Since the CMOS transistors in the circuit operate in their subthreshold region and under low supply voltage, the controller can reduce power consumption. Moreover, low-cost production and miniaturization of controllers are expected. We have shown through computer simulation, such circuit has the capability to generate several periodic rhythmic patterns and transitions between their patterns promptly.

Propofol in low doses causes redistribution of body heat in male volunteers.

BACKGROUND AND OBJECTIVE: Cardiovascular and mental stress in dental patients with phobias about dentistry may be reduced by propofol sedation. We tested the hypothesis that even low doses of propofol may have effects on body temperatures in male volunteers. METHODS: Six healthy male volunteers were given propofol over 28 min with the following infusion rates: 8 mg kg(-1) h(-1) for the first 3 min, 4 mg kg(-1) h(-1) for the next 10 min, and 2 mg kg(-1) h(-1) for the final 15 min. Body temperatures were measured at five locations: tympanic membrane, forehead, forearm, dorsum of the hand and fingertip. Thermoregulatory vasoconstriction was evaluated using the forearm minus fingertip temperature gradient. RESULTS: Tympanic membrane and forehead temperatures began to decrease at 10 and 20 min, respectively, after the start of the propofol infusion, and reached a minimum at 30 min (tympanic -0.5 +/- 0.2 degrees C) and 40 min (forehead -0.6 +/- 0.2 degrees C), respectively. Peripheral skin temperatures showed an increase between 10 and 30 min in the forearm and fingertip and between 20 and 30 min in the dorsum of the hand. After 30 min, a decrease in peripheral skin temperatures was observed. The forearm minus fingertip temperature gradient changed from negative to positive after 40 min, and increased continuously thereafter (baseline -0.3 +/- 0.4 degrees C, 90 min: 6.5 +/- 1.6 degrees C). CONCLUSIONS: A low dose of propofol impairs tonic thermoregulatory vasoconstriction and induces heat redistribution from the core to the periphery.

[A case of pleural effusion in a patient with malignant lymphoma diagnosed by thoracoscopy with local anesthesia].

A 47-year-old woman was admitted for examination of massive right pleural effusion on chest radiography, together with chief complaints of dry cough and body weight loss. Exudative effusion was aspirated by thoracentesis, revealing many lymphocytes and a high level of adenosine deaminase. Thoracoscopy with local anesthesia was performed. Elevated white lesions and miliary nodules were observed, and these lesions were examined by thoracoscopic biopsy, leading to a diagnosis of malignant lymphoma. Treatment with chemotherapy (THP-COP) resulted in partial remission. Thoracoscopy with local anesthesia in cases of pleural effusion with malignant lymphoma can be a safe and useful procedure for diagnosis.

"Devil's staircase"-type phase transition in NaV2O5 under high pressure.

The "devil's staircase"-type phase transition in the quarter-filled spin-ladder compound NaV2O5 has been discovered at low temperature and high pressure by synchrotron radiation x-ray diffraction. A large number of transitions are found to successively take place among higher-order commensurate phases with 2a x 2b x zc type superstructures. The observed temperature and pressure dependence of modulation wave number q(c), defined by 1/z, is well reproduced by the axial next nearest neighbor Ising model. The q(c) is suggested to reflect atomic displacements presumably coupled with charge ordering in this system.

X-ray magnetic circular dichroism imaging with hard X-rays.

X-ray polarization-contrast images resulting from X-ray magnetic circular dichroism (XMCD) in the hard X-ray region have been successfully recorded for the first time. The apparatus used consisted of an X-ray polarizer, double X-ray phase retarders, and a high-spatial-resolution X-ray charge-coupled-device detector. The sample used was a hexagonal-close-packed cobalt polycrystal foil having a thickness of about 4 microns. The X-ray polarization-contrast image resulting from XMCD was observed at a photon energy of 10 eV above the cobalt K-absorption edge (7709 eV). The observed contrast in the image was reversed by inversion of the magnetic field. Furthermore, the contrast was reversed again at a photon energy of 32 eV above the cobalt K-absorption edge.

X-ray double phase retarders to compensate for off-axis aberration.

An X-ray double phase retarder system composed of two transmission-type phase retarders is proposed and developed in order to compensate for off-axis aberration (phase-shift inhomogeneity due to angular divergence of incident X-rays). The scattering planes of the two phase retarders are set to be inclined by 45 degrees with respect to the plane of incident polarization, but the two phase retarders give Bragg reflections in opposite directions. By using this X-ray optical system, vertically polarized X-rays with a 0.99 degree of linear polarization were obtained from horizontally polarized synchrotron radiation with a horizontal beam divergence of 20 arcsec (0.1 mrad). This value is favorably compared with the value of 0.87 which was obtained using a conventional single phase retarder of identical total thickness, 627 microns. The comparison was made at the nickel K-absorption edge (8333 eV) with the condition that 47% of incident X-rays were transmitted through the two phase retarder crystals. The crystals were (100)-oriented diamond plates giving asymmetric 111 Laue reflections.

Fluctuations of lamellar structure prior to a lamellar-->gyroid transition in a nonionic surfactant system

Fluctuations of lamellar structure prior to a lamellar-->gyroid transition in a nonionic surfactant-water system has been investigated by means of small-angle x-ray scattering (SAXS) and differential scanning calorimeter measurements. For large DeltaT (DeltaT=T-T(LG), where T is the temperature and T(LG) the lamellar-->gyroid transition temperature) in the lamellar phase, the SAXS profiles can be described by a Caille correlation function for undulating lamellar structure. Approaching the temperature to the T(LG), an excess diffuse scattering grows at the lower Q (Q is the magnitude of scattering vector) side of the first lamellar peak. Highly oriented lamellar samples revealed that the excess diffuse scattering arises from in-plane density fluctuations. We attribute this diffuse scattering to the perforation fluctuation layer (PFL) structure and we show that the PFL is an equilibrium structure. At T(LG), the PFL transformed to the gyroid phase through a transient ordered structure having a rhombohedral symmetry.

[Clinical study of three myelodysplastic syndrome (MDS) patients with BOOP-like pulmonary disease].

Three cases of myelodysplastic syndrome (MDS) associated with BOOP-like pulmonary disease were reported. They were diagnosed from transbronchial biopsies and clinical features. In two cases, chest radiographs showed ground glass opacities or air space consolidation in both lung fields, and air space consolidation in the right lower lobe in the other case. BALF showed a marked increase of lymphocytes in one case, and organizing pneumonia and alveolitis, and alveolar spaces filled with foamy macrophages were identified histologically. Although no steroid therapy was employed, all three cases improved. However, one patient suffered a relapse 6 months later and thereafter did not respond to corticosteroid therapy.

Identification of carp proopiomelanocortin-related peptides and their effects on phagocytes.

We report the immunomodulating effects of proopiomelanocortin (POMC)-related peptides on phagocytic cells in carp. The complete amino acid sequences of two carp POMCs (I and II) were deduced from the nucleotide sequences after cDNA cloning. Both POMCs consist of 194 amino acids (91% sequence identity) including identical alpha-melanotropin (MSH) and beta-endorphin (EP). All hormonal peptides derived from two POMCs were identified by mass spectrometry after separation by high-performance liquid chromatography of an acid-acetone extract from a single pituitary. These peptides were alpha-MSH, N-Des-Ac-alpha-MSH, di-Ac-alpha-MSH, beta-MSH I, beta-MSH-II, N-Ac-beta-EP(1-29), corticotropin-like intermediate lobe peptide I and II and N-terminal peptide of POMC I and II. The immunomodulating effects of synthetic MSHs and EPs on phagocytic cells from carp head kidney were examined. Di-Ac-alpha-MSH, beta-MSH I, N-Ac-beta-EP(1-29) and beta-EP(1-29) increased the production of superoxide anion at 0.1-100 ng ml-1 for these MSHs and 1-100 ng ml-1 for EPs in RPMI 1640 medium.

Autologous peripheral blood stem cell transplantation for adults with B-lineage acute lymphoblastic leukemia: a pilot study.

We conducted a pilot study on autologous peripheral blood stem cell transplantation (PBSCT) for 11 adults with B-lineage acute lymphoblastic leukemia (ALL) in first complete remission (CR) or even in those with more advanced stages. All patients achieved CR by induction therapy, of whom 10 were treated with anthracycline, vincristine and prednisolone-based regimens. After consolidation therapy, all patients except one received high-dose cytarabine followed by granulocyte colony-stimulating factor (G-CSF) administration to collect PBSCs. Ten patients received busulfan 4 mg/kg for 4 days, etoposide 20 mg/kg for 3 days and ranimustine 200 mg/m2 for 2 days as a conditioning regimen. One received a regimen consisting of etoposide, cyclophosphamide and total body irradiation. From day 1, G-CSF was given intravenously, and no additional chemotherapy was administered. At the median follow-up time of 30.8 months, four of six patients with standard-risk B-lineage ALL survived within the range of 19.7 to 85.4 months without relapse. In contrast, only one of five with high-risk B-lineage ALL survived for 36.3 months without relapse. Autologous PBSCT as post-remission therapy may prolong CR in adults with standard-risk B-lineage ALL.

Molecular cloning of proopiomelanocortin cDNA from an elasmobranch, the stingray, Dasyatis akajei.

Recently, we have characterized a new MSH (named delta-MSH) which joins the group of MSHs (alpha, beta, gamma) in dogfish proopiomelanocortin (POMC). The present study has confirmed the presence of delta-MSH in POMC of another member of the elasmobranchian order, the stingray, Dasyatis akajei, by cDNA cloning from pituitary mRNAs. Overlapping partial cDNA clones corresponding to stingray POMC were amplified by PCR from single-strand cDNA prepared from pituitary poly (A)(+) RNA. Excluding the poly A tail, stingray POMC cDNA consists of 1077 base pairs (bp). It contains a 912-bp open reading frame encoding a signal peptide of 24 amino acids (aa) and a POMC of 280 aa. gamma-MSH, alpha-MSH, ACTH, delta-MSH, beta-MSH, and beta-endorphin are located at POMC (50-61), (115-127), (115-153), (182-193), (226-242), and (245-280), respectively. The stingray POMC is smaller than that of the dogfish POMC (294 aa) mainly due to the absence of a sequence of 11 consecutive aa between delta-MSH and beta-MSH. delta-MSH has been found only in the elasmobranchs and, therefore, delta-MSH might have evolved after the divergence of chondrichthians from the ancestral vertebrate lineage and before divergence of sharks and rays.

pH-dependent unfolding of aspergillopepsin II studied by small-angle X-ray scattering.

Aspergillopepsin II (EC 3.4.23.6) secreted from the fungus Aspergillus niger var. macrosporus is a non-pepsin-type acid proteinase. It consists of two polypeptide chains (i.e., a heavy chain and a light chain), which are bound noncovalently to each other. The pH titration analysis using small-angle X-ray scattering (SAXS) as well as circular dichroism (CD) and gel filtration indicated that the enzyme was unfolded around a neutral pH with concomitant dissociation of the two chains. Detailed analyses showed that the midpoint pH values for the unfolding are not coincident with one another (pH 6.1 in circular dichroism and gel filtration, pH 6.4 in zero-angle intensity of SAXS, pH 6.8 in radius of gyration). The difference between these values suggested the existence of an intermediate state during the unfolding. Further analyses of the SAXS data showed that the heavy chain just after the dissociation still kept molecular compactness and that it gradually increased its dimensions as the pH was further raised. Noncoincidence of the two phenomena (i.e., chain dissociation and swelling) led to elucidation of a novel intermediate state during unfolding, which was confirmed by the subsequent singular value decomposition (SVD) analysis.

Isolation, cDNA cloning, and growth promoting activity of rabbitfish (Siganus guttatus) growth hormone.

We report the isolation, cDNA cloning, and growth promoting activity of rabbitfish (Siganus guttatus; Teleostei; Perciformes; Siganidae) growth hormone (GH). Rabbitfish GH was extracted from pituitary glands under alkaline conditions, fractionated by gel filtration chromatography on Sephadex G-100, and purified by high-performance liquid chromatography. The fractions containing GH were identified by immunoblotting with bonito GH antiserum. Under nonreducing conditions, the molecular weight of rabbitfish GH is about 19 kDa as estimated by SDS-PAGE. The purified hormone was potent in promoting growth in rabbitfish fry. Weekly intraperitoneal injections of the hormone significantly accelerated growth. This was evident 3 weeks after the start of the treatment, and its effect was still significant 2 weeks after the treatment was terminated. Rabbitfish GH cDNA was cloned to determine its nucleotide sequence. Excluding the poly (A) tail, rabbitfish GH cDNA is 860 base pairs (bp) long. It contained untranslated regions of 94 and 175 bp in the 5' and 3' ends, respectively. It has an open reading frame of 588 bp coding for a signal peptide of 18 amino acids and a mature protein of 178 amino acid residues. Rabbitfish GH has 4 cysteine residues. On the amino acid level, rabbitfish GH shows high identity (71-74%) with GHs of other perciforms, such as tuna, sea bass, yellow tail, bonito, and tilapia, and less (47-49%) identity with salmonid and carp GHs.

Demonstration of X-ray linear dichroism imaging with hard X-rays.

X-ray polarization-contrast images resulting from X-ray linear dichroism (XLD) in the hard X-ray region have been successfully recorded for the first time. The apparatus used consisted of an X-ray polarizer, double X-ray phase retarders and a high-spatial-resolution X-ray charge-coupled device (CCD) detector. The sample used was a hexagonal close packed (h.c.p.) cobalt single-crystal foil of thickness about 12 microm. The experiment was performed at X-ray energies of 23 and 29 eV above the cobalt K edge (7709 eV), at which the maximum linear dichroisms (approximately 3%) were observed, with their signs reversed, in the XLD spectrum measured with quadruple X-ray phase retarders. The contrasts in the images at the two X-ray energies were reversed as a result of the XLD in the sample. Furthermore, the values of the contrast in the images arising from the linear dichroism (approximately 3%) were in good agreement with those yielded by the XLD spectrum.