The production of coagulation factor VII by adipocytes is enhanced by tumor necrosis factor-α or isoproterenol.

BACKGROUND: A relationship has been reported between blood concentrations of coagulation factor VII (FVII) and obesity. In addition to its role in coagulation, FVII has been shown to inhibit insulin signals in adipocytes. However, the production of FVII by adipocytes remains unclear. OBJECTIVE: We herein investigated the production and secretion of FVII by adipocytes, especially in relation to obesity-related conditions including adipose inflammation and sympathetic nerve activation. METHODS: C57Bl/6J mice were fed a low- or high-fat diet and the expression of FVII messenger RNA (mRNA) was then examined in adipose tissue. 3T3-L1 cells were used as an adipocyte model for in vitro experiments in which these cells were treated with tumor necrosis factor-α (TNF-α) or isoproterenol. The expression and secretion of FVII were assessed by quantitative real-time PCR, Western blotting and enzyme-linked immunosorbent assays. RESULTS: The expression of FVII mRNA in the adipose tissue of mice fed with high-fat diet was significantly higher than that in mice fed with low-fat diet. Expression of the FVII gene and protein was induced during adipogenesis and maintained in mature adipocytes. The expression and secretion of FVII mRNA were increased in the culture medium of 3T3-L1 adipocytes treated with TNF-α, and these effects were blocked when these cells were exposed to inhibitors of mitogen-activated kinases or NF-κB activation. The ß-adrenoceptor agonist isoproterenol stimulated the secretion of FVII from mature adipocytes via the cyclic AMP/protein kinase A pathway. Blockade of secreted FVII with the anti-FVII antibody did not affect the phosphorylation of Akt in the isoproterenol-stimulated adipocytes. CONCLUSION: Obese adipose tissue produced FVII. The production and secretion of FVII by adipocytes was enhanced by TNF-α or isoproterenol via different mechanisms. These results indicate that FVII is an adipokine that plays an important role in the pathogenesis of obesity.

Cloning and sequence analysis of an esterase gene from Pseudomonas sp. KWI-56.

The gene encoding an esterase from Pseudomonas sp. KWI-56 was cloned and sequenced. The nucleotide sequence contained an open reading frame encoding a polypeptide comprising 262 amino acids, whose molecular weight agreed well with the value obtained by SDS-PAGE. Comparison of the amino acid sequence with those of the other homologous enzymes suggested that Ser-92 and His-24l might be included in the catalytic triad.

Use of technetium-99m as a radioactive label to study migratory patterns of leukocytes.

Technetium-99m had been used as a radioactive label to study the migratory patterns of neutrophils. In a previous in vitro study, neutrophils were labeled with Tc-99m and infused into patients with and without various hematological disorders. Increased pulmonary localization was detected by scintillation camera within 10 min; this decreased gradually within 3 hr. Accumulation was seen in the liver and spleen at 3 hr. The same results were noted by using neutrophils labeled with Tc-99m sulfur colloid. In a patient with severe ulceration in the oral cavity, due to acute leukemia, Tc-99m-labeled transfused neutrophils that were collected by filtration leukopheresis were concentrated in the infected lesions.

Prolongation of platelet survival in idiopathic thrombocytopenic purpura by high-dose intravenous gamma globulin.

The mechanism of the increase of platelet counts after the intravenous injection of high dose gammaglobulin was evaluated by measuring platelet survival before and after the injection. The platelet survival after treatment prolonged more significantly than that before treatment. There was no evidence of platelet release from bone marrow to peripheral blood. These findings might suggest the inhibition of platelet destruction by transient blockade of reticuloendothelial system.

Chain conformational analysis of beijeran by n-h map calculations.

Possible regular helix models of beijeran, a new acidic heteropolysaccharide consisting of a trisaccharide as a repeating unit, were investigated. Conformational analyses of the three component disaccharides were first carried out by calculating their relaxed-residue energy maps with respect to the glycosidic bond rotations, phi and psi. A search for possible beijeran chain conformations was carried out by calculating the two-dimensional map of the helix parameters; n (the number of asymmetric units per a fiber repeat) and h (the axial rise per the unit), on the basis of the phi-psi conformations taken from the low energy regions of each of the three energy maps. The n-h values of the helix models with low steric energies were mostly found to be around the experimental values (n = 2 and h = 1.20-1.25 nm), which may support the present methodology. It was also suggested that the internal flexibility of beijeran chain allowed it to conform in diverse helical structures, each of which were reasonably in accord with the observed n-h values. The three representative helix models were finally proposed for the beijeran chain conformation in the crystal structure.

Molecular and crystal structure of the regenerated form of (1----3)-alpha-D-mannan.

The crystal structure of the hydrated form of (1----3)-alpha-D-mannan, obtained by solid-state deacetylation of the partially O-acetylated mannan, was analyzed by combined X-ray diffraction and stereochemical-model refinement techniques. The structure crystallizes in a four-chain, monoclinic unit cell with parameters a = 11.33 A, b = 18.36 A, c (fiber repeat) = 8.25 A, and gamma = 101.75 degrees, and the most probable space group is P2(1). In the most probable structure the chain-backbone conformation is a two-fold helix, but with all four O-6 rotational positions nonequivalent. The chains pack with antiparallel polarity and are connected by pairs of intermolecular hydrogen bonds that form an infinite, zig-zag sheet. There are 16 water molecules in the unit cell, generally embedded between the sheets in crystallographic positions, providing additional hydrogen bonding and establishing a three-dimensional hydrogen-bond network in the crystal structure. The reliability of the structure analysis is indicated by the X-ray residual R" = 0.281, based on 98 hkl reflection intensities.

Molecular and crystal structure of konjac glucomannan in the mannan II polymorphic form.

A probable crystal structure of konjac glucomannan (mannose:glucose ratio = 1.6) is proposed based on X-ray data and constrained linked-atom least-squares model refinement. The structure crystallizes in the mannan II polymorphic form, in an orthorhombic unit-cell with a = 9.01 A, b = 16.73 A, c (fiber axis) = 10.40 A, and a probable space group I222. The backbone conformation of the chain is a two-fold helix stabilized by intramolecular O-3-O-5' hydrogen bonds, with the O-6 rotational position gt. The unit cell contains four chains with antiparallel packing polarity and eight water molecules which reside in crystallographic positions. Intermolecular hydrogen bonds occur exclusively between chains and water molecules, establishing a three-dimensional hydrogen-bond network in the crystal structure. The glucose residues replace mannoses in the structure in isomorphous fashion, although some disorder appears possible. A structure having alternating gg-gt O-6 rotational positions and conforming to space group P222 appears to describe the disorder regions of the crystal. The reliability of the structure analysis is indicated by the X-ray residuals R = 0.276 and R" = 0.223.

X-ray study of beijeran sodium salts, a new galacturonic acid-containing exo-polysaccharide.

X-Ray fiber diffraction patterns were obtained from oriented films of sodium salts of a new uronic acid-containing polysaccharide (beijeran) both in its native, poly [-->3)-alpha-D-GalA-(1-->3)-beta-L-Rha-(1-->3)-alpha-D-Glc-O6Ac-(1 -->], and deacetylated forms. Initially the stretched films of both polysaccharides were amorphous, but the crystallinity was much improved by annealing at high temperature. The deacetylated specimen had higher crystallinity than the native. Both films showed similar X-ray fiber patterns indicating that these polysaccharides had similar unit cell dimensions and that the O-acetyl groups in the native beijeran chain did not disturb the regular array in the crystal having space group P21. All the visible reflections could be indexed in terms of a monoclinic unit cell with dimensions a = 1.277, b = 1.611, c (fiber axis) = 2.437 nm, and gamma = 96.79 degrees. The fiber axis length and the presence of (002) and (006) reflections indicated that the conformation was made up of two trisaccharide residues, in an extended two-fold helix.

Molecular and crystal structure of (1----3)-alpha-D-glucan triacetate.

A crystal and molecular structure for GTA I, the low temperature polymorph of (1----3)-alpha-D-glucan triacetate, is proposed on the basis of X-ray diffraction analysis of well-oriented films, combined with stereochemical model refinement. The unit cell is monoclinic with parameters a = 30.17 A, b = 17.42 A, c (fibre axis) = 12.11 A, and beta = 90 degrees C. The probable space group is P2(1) with b axis unique. Six molecular chains pass through the unit cell with alternating polarity and with three independent chains comprising the asymmetric unit. The chain axes are located in a hexagonal packing arrangement. The chain backbone conformation is a left-handed, three-fold helix, but all nine O(6) acetyl groups of the asymmetric unit are in non-equivalent rotational positions. The most probable structure is indicated by X-ray residuals R = 0.261 and R" = 0.283, based on 62 reflection intensities (41 observed and 21 unobserved).

An evaluation of crystal structure of mannan I by X-ray powder diffraction and molecular mechanics studies.

The present study reports the re-examination of the crystal structure of mannan I by the X-ray powder diffraction study combined with the miniature crystal model simulation. A primary objective of this study was to investigate an adequate chain staggering position along the fiber axis. Among the several crystal structure models proposed for mannan I, that derived from the electron diffraction study of the single crystals was adopted as a starting model. The X-ray crystallographic residuals were calculated for the single crystal structure at different chain staggering positions, while the ab projection was kept invariant. In a similar fashion, the miniature crystal models consisting of seven mannotetraoses were constructed, each having different chain staggering values and the three-dimensional structures of all the models were subsequently optimized by using the MM3(92) program without introducing any constraint. Both the X-ray residual and lattice energy plots with respect to the chain staggering position showed the common minima around the -0.25 x c chain staggering. The minimum models were subjected to a search for preferred orientations of O2 and O6 hydroxyl groups. It was found that the hydroxyl groups present inside the minicrystal tended to rotate into particular orientations during the structure optimizations to form the O5-H-O2 and O3-H-O6 intermolecular hydrogen bonds between the adjacent oligomers in an alternative direction.

[Clinical evaluation of cefotiam in internal medicine, with special reference to infectious disease associated with hematological disorders (author's transl)].

Clinical evaluation of cefotiam (panspolin), a new cephem antibiotics, was performed in the infectious disease associated with hematological disorders and in the respiratory system. In hematological dis orders, 40% of good and 25% of fair results were obtained in clinical effects. In respiratory infections, however, 92% of good results were obtained. Opportunistic infection due to Gram-negative bacilli are often experienced in patients with leukemia. Since cefotiam has sufficient bacteriocidal effects in broad spectrum, it would be a good therapeutic agent against infectious diseases associated with hematological disorders.

Immune thrombocytopenia in an elderly patient treated successfully by pulse therapy with cyclophosphamide.

Thrombocytopenia due to immune mechanisms is rare and difficult to manage in elderly patients. We describe a case of an 89-year-old female with severe immune thrombocytopenia (ITP) who rapidly improved by pulse therapy with cyclophosphamide. She was admitted to our hospital because she had arthralgia in both sides of her femoral region since January 1999, aphthous stomatitis and ecchymosis of the leg since April 1999, and bloody phlegm in July 1999. On admission, her peripheral blood count revealed severe thrombocytopenia (0.1 x 10(4)/microl). Her megakaryocyte count from bone marrow was increased to 512/microl without abnormal cells. Systemic lupus erythematosus was suspected because of strong positive protein in the urine in addition to the clinical and hematological findings described above, but she was negative for all the autoantibodies examined. Finally, she was diagnosed as having ITP on the basis of high platelet associated immunoglobulin G in addition to hematological and physical findings and she was treated with prednisolone. It was difficult to maintain her platelet count with only prednisolone, but 600 mg of cyclophosphamide rapidly increased her platelet count in spite of tapering the prednisolone. In September 2000, her platelet count was kept within normal limits by administration of 15 mg/day of prednisolon. It is suggested that immunosuppressive therapy for ITP using high-dose cyclophosphamide is useful in elderly patients as well as in juvenile adult patients.

[Acute promyelocytic leukemia with a history of RAEB in transformation and the 15/17 translocation].

A 34-year old female was admitted to our clinic because of fever and general fatigue on March 26, 1987. On admission, peripheral blood (PB) revealed pancytopenia. Bone marrow smears revealed 9. 0% of promyelocytic cells with or without Auer rods. Diagnosis of RAEB in transformation was made. Chromosome study of the bone marrow cells showed t(15; 17) in 3 out of 20 cells analysed. After 3 months, the leukemic cells were observed in PB and increased in number. Then the patient showed bleeding tendency and fibrin degradation products (FDP) increased up to 40 micrograms/ml. And the leukemic cells were over 30% in PB at the end of July, 1987. The diagnosis of APL with DIC was made. To our knowledge, this is the first case of APL with a history of MDS with t(15; 17).

[Bone marrow aplasia without blast crisis in a case of CML of 10-year survival].

A case of chronic myelogenous leukemia (CML) of 10-year survival in described. A 44-year old male was admitted to our hospital because of general malaise, abdominal fullness and fever in February, 1977. On physical examination, giant splenomegaly and hepatomegaly were detected. Peripheral blood examination revealed leukocytosis without hiatus leukemia , normochromic macrocytic anemia and thrombocytosis. NAP rate and score were 16% and 22. Cytogenetic analysis of PB without stimulator revealed 46, XY, Ph1. Then he was diagnosed as having a typical type of Ph1-positive CML. He had been successfully treated over 9 years by intermittent administration of busulfan. However, anemia suddenly progressed in February, 1986 followed by leukopenia and thrombocytopenia. Hemorrhage was not detected by the examination. Though he had been received blood transfusion, the anemia progressed rapidly. He was died of cachexia on 4th of August, 1987. The postmortem examination revealed bone marrow aplasia with no signs of blast crisis nor myelofibrosis. Secondary hemochromatosis was seen in the liver, spleen, pancreas and some other organs.