Autonomous fluorescence of ascidian blood cells with special reference to identification of vanadocytes

Summary A tunichrome that has been suggested to be involved in the accumulation of vanadium ions in ascidian blood cells produces an autonomous fluorescence upon excitation with blue-violet light. However, we have found that signet ring cells, which contain large amounts of vanadium, do not fluoresce upon such excitation. The strongest fluorescence due to the tunichrome was observed in morula cells, which do not contain vanadium.     

Peripheral erythrocyte levels, hemolysis and three vanadium compounds

Three vanadium compounds of different valence states were administered to adult mice. Two, four, and eight days following treatment of vanadium, cardiac blood was collected. The blood sample was used to ascertain the peripheral erythrocyte count (cell/mm3) and to determine the in vitro hemolytic index of erythrocytes obtained from mice treated in vivo with either the tri-, tetra-, or pentavalent vanadium compound. Data indicate that the tetravalent form was the most effective test substance in 1) promoting rupture of isolated erythrocytes compared to red cells retrieved from control mice and 2) depressing the erythrocyte count obtained from heart blood; maximum effects were manifest four days post-treatment. For all treatments there appeared to be a good correlation between the degree of vanadium-induced hemolysis and the peripheral erythrocyte count reduction following exposure to the vanadium.     

Peripheral erythrocyte levels,hemolysis and three vanadium compounds

Summary Three vanadium compounds of different valence states were administered to adult mice. Two, four, and eight days following treatment of vanadium, cardiac blood was collected. The blood sample was used to ascertain the peripheral erythrocyte count (cell/mm³) and to determine the in vitro hemolytic index of erythrocytes obtained from mice treated in vivo with either the tri-, tetra- or pentavalent vanadium compound. Data indicate that the tetravalent form was the most effective test substance in 1) promoting rupture of isolated erythrocytes compared to red cells retrieved from control mice and 2) depressing the erythrocyte count obtained from heart blood; maximum effects were manifest four days post-treatment. For all treatments there appeared to be a good correlation between the degree of vanadium-induced hemolysis and the peripheral erythrocyte count reduction following exposure to the vanadium.     

Vanadium biochemistry: The unknown role of vanadium-containing cells in ascidians (sea squirts)

Summary This article reviews several new developments in vanadium biochemistry, as elucidated from studies of ascidians. A hypothesis correlating ascidian blood cell function to anaerobiosis, via two prominent redox constituents, namely vanadium(III) and the tunichromes, a family of metal ion complexing/reducing hydroquinonoid peptides, is presented.     

Toxicokinetic study of vanadium after intramuscular and intratracheal administration to rats]

The toxicokinetic study, made in the rat with 48V under VOCl2, shows that vanadium does not accumulate in the system. 66% of the radioactivity is found in urines 24 hrs. after the inoculation by intramuscular method. After intratracheal deposit, a part of the vanadium is eliminated very rapidly. The other which is lower is fixed on the pulmonary tissues.     

Ultrastructural localization of vanadium in the blood cells of Ascidiacea.

X-ray histospectrographic analysis at the scanning and transmission electron microscope (STEM) are made on the blood cells of Phallusia mamillata Cuvier and Ciona intestinalis, to study the 'direct' intracellular sites of accumulation of vanadium. The results show a clear accumulation of vanadium on the membrane and in the granules of vacuoles of amebocytes, signet ring cell, compartment cell and traces of metal in the 'vanadophores' of vanadocytes.     

Induction of vanadium accumulation and nuclear sequestration causing cell suicide in human Chang liver cells

Very little is known about the modulation of vanadium accumulation in cells, although this ultratrace element has long been seen as an essential nutrient in lower life forms, but not necessarily in humans where factors modulating cellular uptake of vanadium seem unclear. Using nuclear microscopy, which is capable of the direct evaluation of free and bound (total) elemental concentrations of single cells we show here that an NH₄Cl acidification prepulse causes distinctive accumulation of vanadium (free and bound) in human Chang liver cells, concentrating particularly in the nucleus. Vanadium loaded with acidification but leaked away with realkalinization, suggests proton-dependent loading. Vanadyl(4), the oxidative state of intracellular vanadium ions, is known to be a potent source of hydroxyl free radicals (OH^.). The high oxidative state of nuclei after induction of vanadyl(4) loading was shown by the redox indicator methylene blue, suggesting direct oxidative damage to nuclear DNA. Flow cytometric evaluation of cell cycle phase-specific DNA composition showed degradation of both 2N and 4N DNA phases in G₁, S and G₂/M cell cycle profiles to a solitary 1N DNA peak, in a dose-dependent manner, effective from micromolar vanadyl(4) levels. This trend was reproduced with microccocal nuclease digestion in a time response, supporting the notion of DNA fragmentation effects. Several other approaches confirmed fragmentation occurring in virtually all cells after 4 mM V(4) loading. Ultrastructural profiles showed various stages of autophagic autodigestion and well defined plasma membrane outlines, consistent with programmed cell death but not with necrotic cell death. Direct intranuclear oxidative damage seemed associated with the induction of mass suicide in these human Chang liver cells following vanadium loading and nuclear sequestration.     

Ultrastructural localization of vanadium in the blood cells of Ascidiacea

Summary X-ray histospectrographic analysis at the scanning and transmission electron microscope (STEM) are made on the blood cells ofPhallusia mamillata Cuvier andCiona intestinalis, to study the direct intracellular sites of accumulation of vanadium. The results show a clear accumulation of vanadium on the membrane and in the granules of vacuoles of amebocytes, signet ring cell, compartment cell and traces of metal in the vanadophores of vanadocytes.We thank Dr C. Weichan and Mr G. Hubert for kind hospitality and for technical assistance in the Department of Electron Microscopy of Siemens Laboratory in Karlsruhe (BRD). We thank also Dr V. Andria of Siemens Elettra in Rome (Italy).     

建立炼钢快速分析系统

结合提钒炼钢工艺的特点,在确保分析质量的前提下,以缩短分析反馈时间为目标,合理配置资源,优化布局,改进操作,提高检测一次成功率,建立符合攀钢实际的快速分析系统,为炼钢缩短冶炼周期,提高产品质量,增加产量提供了有力的技术支持。系统投用后效果明显,平均缩短冶炼周期2 min以上。     

Distribution of tunichrome and vanadium in sea squirt blood cells sorted by flow cytometry

Specialized blood cells of many tunicates accumulate high concentrations of vanadium and phenolic peptide pigments called tunichromes (TC). In order to determine whether V and TC reside in the same cells, Ascidia nigra and Ascidia ceratodes blood cell subpopulations were isolated by fluorescence-activated cell sorting (flow cytometry) and chemically analyzed. V was found in the spherical, green/grey signet ring cells, and to a lesser degree in the mulberry-shaped, yellow/green morula cells (MRs), whereas free TC was detected mainly in MRs.     

Sur la réaction du chlorure de nitrosyle avec le tétrachlorure de vanadium (IV)

Summary The reaction of excess nitrosyl chloride with vanadium tetrachloride under specific conditions furnishes the previously unreported compound VCl₄, 2NOCl. The formation of a 12 adduct shows that vanadium (IV) can behave as a typical tetravalent metal in the M^IVCl₄-NOCl system. Spectroscopic measurements reveal both nitrosonium and hexachlorometallate ions in the addition compound and indicate structural analogy with (NO)₂-TiCl₆. The complex may thus be designated as dinitrosonium hexachlorovanadate, (NO)₂VCl₆. However, this compound is probably not purely ionic, since the relatively low frequency observed for the IR-absorption of the NO+ ion (2146 cm^–1 compared with the normal range 2150 to 2400 cm^–1) and the appreciable volatility of the compound suggest a significant degree of covalent bonding.

---

---

L'auteur remercie M. le Professeur R.Rohmer pour l'interêt qu'il a porté à ce travail.     

Vanadium induced hemolysis of vitamin E deficient erythrocytes in Hepes buffer

Several vanadium compounds were tested for their ability to induce in vitro hemolysis of vitamin E-deficient hamster erythrocytes. Free vanadyl caused hemolysis in Hepes buffer but not in Tris or phosphate buffer, while hemolysis was inhibited by catalase, chelators such as deferoxamine mesylate and EDTA, and hydroxyl radical scavengers such as ethanol andd-mannitol. Although metavanadate itself could not induce hemolysis, metavanadate with NAD(P)H caused hemolysis in Hepes buffer only, and superoxide dismutase prevented it. Hydrogen peroxide, hydroxyl radical and Hepes radical were involved in vanadyl-induced hemolysis; superoxide anion was further involved in metavanadate plus NAD(P)H-induced hemolysis. Vitamin E prevented hemolysis under both conditions.     

Vanadium induced hemolysis of vitamin E deficient erythrocytes in Hepes buffer

Several vanadium compounds were tested for their ability to induce in vitro hemolysis of vitamin E-deficient hamster erythrocytes. Free vanadyl caused hemolysis in Hepes buffer but not in Tris or phosphate buffer, while hemolysis was inhibited by catalase, chelators such as deferoxamine mesylate and EDTA, and hydroxyl radical scavengers such as ethanol andd-mannitol. Although metavanadate itself could not induce hemolysis, metavanadate with NAD(P)H caused hemolysis in Hepes buffer only, and superoxide dismutase prevented it. Hydrogen peroxide, hydroxyl radical and Hepes radical were involved in vanadyl-induced hemolysis; superoxide anion was further involved in metavanadate plus NAD(P)H-induced hemolysis. Vitamin E prevented hemolysis under both conditions.     

Biologische und chemische Wege zur Anreicherung von Spurenelementen

Summary Using as examples hemovanadin, the vanadium-containing blood pigment of the ascidianPhallusia mamillata, and ferritin, the protein for iron storage in mammals, pathways for the concentration of vanadium and of iron are described: These have been realizedin vitro and may occurin vivo. The uptake and concentration of heavy metals in nature seems to occur by means of complexing agents.High molecular weight, insoluble chelating substances which are able to bind metal ions, especially uranium and copper selectively, have been synthesized. Recovery of the metals is possible without destruction of the chelating groups, and, as in the case of ion-exchange resins, repeated cycles can be carried out without diminishing the metal-binding capacity.     

Über die Molekulargrösse des Hämovanadins

Summary The red-brown haemovanadin which is contained in the haemolysate (Henze Solution) of blood cells of the ascidiaPhallusia mamillata Cuvier is a chromoproteid with trivalent vanadium as the central atom and sulphuric acid bound coordinatively. It is found to have a molecular weight of 24,400±1,900 according to the estimation of the diffusion coefficient (D_5°=6.87±0.2 · 10^–7 cm² · s^–1 at pH 2.5–2.8).

---

---

Dem Entdecker des Hämovanadins zum 80. Geburtstag gewidmet.     

Helminthosporoside,a host-specific toxin fromHelminthosporium sacchari: A structure revision and a new partial structure

Summary Reevaluation of the previously proposed structure of helminthosporoside, a host-specific toxin fromHelminthosporium sacchari, reveals a sesquiterpenoid bis-digalactoside. The carbohydrate portion of the toxin was characterized by¹³C-NMR spectroscopy, methylation analyses, FD and FAB mass spectroscopy. The ring size and anomeric configuration of the galactose moieties were determined by utilizing a¹³C-NMR structural analysis method. A new partial structure is proposed.Acknowledgment. We extend special thanks to C.E. Costello, Department of Chemistry, M.I.T., Cambridge, MA, for FD mass spectral analyses, and to C.E. Ballou, Department of Biochemistry, University of California, Berkeley, CA, for FAB mass spectral analyses. We submit sincere appreciation to both V.N. Reihold, Harvard Medical School, Boston, MA, and P. Albersheim, Department of Chemistry, University of Colorado, Boulder, CO, for methylation analyses. We thank J.S. Frye at the Colorado State University Regional NMR-Center, funded by National Science Foundation grant No. DHE 78-18581, Department of Chemistry, Colorado State University, Fort Collins, Co, for high resolution CMR and NMR, and we are grateful to P.W. Jennings, Department of Chemistry, Montana State University, Bozeman, Mt, for total carbon analyses. This work was supported in part by a Herman Frasch Foundation grant to G.A. Strobel, by NSF grant PCM-78-22517, and funds from the Montana Agricultural Experiment Station. B.P. Mundy acknowledges the partial support of NSF (ISP-801149).     

基因工程改造噬菌体应用于细菌感染的研究进展

噬菌体作为细菌病毒,在细菌性感染尤其是多重耐药菌感染的治疗方面具有抗生素无法比拟的优势。目前人工改造噬菌体的理论和技术已趋成熟,研究者通过基因工程技术解决了噬菌体特异性高、半衰期短、释放内毒素等问题,使基因工程改造噬菌体具有了较强的临床应用潜力。本文主要就基因工程改造噬菌体在扩大宿主范围、增强抗生素疗效、延缓免疫清除、避免内毒素释放等方面所具有的优势,及其在剂量确定、细菌耐受、宿主安全性等方面可能会出现的问题进行了阐述。     

Melatonin,clock genes and mitochondria in sepsis

After the characterization of the central pacemaker in the suprachiasmatic nucleus, the expression of clock genes was identified in several peripheral tissues including the immune system. The hierarchical control from the central clock to peripheral clocks extends to other functions including endocrine, metabolic, immune, and mitochondrial responses. Increasing evidence links the disruption of the clock genes expression with multiple diseases and aging. Chronodisruption is associated with alterations of the immune system, immunosenescence, impairment of energy metabolism, and reduction of pineal and extrapineal melatonin production. Regarding sepsis, a condition coursing with an exaggerated response of innate immunity, experimental and clinical data showed an alteration of circadian rhythms that reflects the loss of the normal oscillation of the clock. Moreover, recent data point to that some mediators of the immune system affects the normal function of the clock. Under specific conditions, this control disappears reactivating the immune response. So, it seems that clock gene disruption favors the innate immune response, which in turn induces the expression of proinflammatory mediators, causing a further alteration of the clock. Here, the clock control of the mitochondrial function turns off, leading to a bioenergetic decay and formation of reactive oxygen species that, in turn, activate the inflammasome. This arm of the innate immunity is responsible for the huge increase of interleukin-1β and entrance into a vicious cycle that could lead to the death of the patient. The broken clock is recovered by melatonin administration, that is accompanied by the normalization of the innate immunity and mitochondrial homeostasis. Thus, this review emphasizes the connection between clock genes, innate immunity and mitochondria in health and sepsis, and the role of melatonin to maintain clock homeostasis.     

Examination of the potential mutagenicity of hair dye constituents using the micronucleus test

Summary 12 compounds which are constitutents of hair dyes or chemically related aromatic amines, aminophenols, their nitroderivatives and aromatic hydroxyderivatives were examined for evidence of mutagenic potential by means of the micronucleus test. None of the compounds tested caused an increase in the incidence of micronucleated erythrocytes after oral dosing.Acknowledgment. These investigations were conducted on behalf and with the assistance of Elida-Gibbs GmbH, Hamburg, Goldwell GmbH, Darmstadt, Thera-Chemie GmbH, Dusseldorf, L'Oreal-Haarkosm, und Parf. GmbH, Karlsruhe, Kadus-Werk K.G., Lenzkirch, Hans Schwarzkopf GmbH, Hamburg, Wella AG, Darmstadt.