James H. Swinehart

THE VANADIUM AND SELECTED METAL CONTENTS OF SOME ASCIDIANS

The vanadium and other selected metal contents of primarily California ascidians have been determined. The species Ascidia ceratodes and Perophora annectens have large vanadium contents as has been predicted for members of the families Ascidiidae and Perophoridae from the order Phlebobranchia. Several species in the order Aplousobranchia have large vanadium contents: the vanadium being present as vanadium (IV) whereas it is vanadium (III) that is found in the order Phlebobranchia. Molgula manhattensis, a species from the order Stolidobranchia, shows a large iron content: the metal being localized in the fluid from the heart.Three dominant fractions were chromatographed from the cells contained in the fluid of Ascidia ceratodes. The roles of the compounds present in these fractions are discussed. The spectra of these fractions are correlated with the spectrum of the cells.

Vanadium content and oxidation state related to ascidian phylogeny

Abstract 1. 1. New atomic absorption and electron spin resonance (e.s.r.) analyses of 50 ascidian species together with published information on other species demonstrates that when vanadium is found in the ascidian suborder Aplousobranchia, it is present as vanadium(IV); in the Phlebobranchia, vanadium is present as vanadium(III); and no significant amounts of vanadium are present in the Stolidobranchia. 2. 2. Vanadium e.s.r. analysis confirms Cionidae as a family of the Aplousobranchia, and an independent origin of the Didemnidae from an early ancestral stock is implied by the high level of vanadium(IV) in the genus Leptoclinides ; loss of vanadium is associated with evolutionary trends. 3. 3. A reconstruction of ascidian phylogeny is based on morphology and vanadium chemistry.

The kinetics of the hexacyanoferrate (III)-sulphite reaction

Abstract The kinetics of the hexacyanoferrate (III)-sulphite reaction have been examined in order to elucidate the detailed mechanism of the reaction. A radical mechanism best satisfies the kinetic data: Fe(CN) 6 3− +SO 3 2− →Fe(CN) 6 4− +SO 3 − slow SO 3 2− +Fe(CN) 6 3− →SO 3 (SO 4 2− )+Fe(CN) 6 4− 2SO 3 − →S 3 O 6 2− The effect of the “inert” electrolyte on the rate of the reaction is discussed. The ratio of bimolecular rate constants in H2O and D2O is 1·4 ± 0·1. The enthalpies of activation when K+ and Na+ are the “inert” cationic species are 12·6 ± 1·0 and 11·7 ± 1·0 kcal/mole respectively.

The distribution of vanadium and sulfur in the blood cells, and the nature of vanadium in the blood cells and plasma of the ascidian, Ascidia ceratodes

Abstract Blood cells:| 1. 1. At least three blood cell types contain vanadium and sulfur: morula, compartment, and pigment cells. 2. 2. The S contents of the pigment and morula cells are nearly the same, but less than that of the compartment cells; while the V contents of the pigment and compartment cells are nearly the same, but greater than that of the morula cells. 3. 3. S/V ratios suggest an ontological relation between compartment and morula cells, if the change in cell type only involves a dilution or concentration of the S- and V-containing constituents. 4. 4. Vanadium (III) in the blood cells is partly present as the dimer VOV4+, but the dimer is not found in the V(IV)- containing aplousobranch ascidian Ciona intestinalis or the low V-containing stolidobranch ascidian Pyura haustor. Plasma: 5. 1. Vanadium(V) is present in the blood plasma at 0.01 – 0.06 mM. 6. 2. The form of V(V) in the plasma is different than in seawater at the same concentration. 7. 3. Vanadium(V) levels in the plasma appear to be regulated.

IRON AND MANGANESE DEPOSITION IN THE PERIOSTRACA OF SEVERAL BIVALVE MOLLUSCS

The presence of high levels of iron (Fe) and manganese (Mn) in the shells of freshwater and marine molluscs is a consistent observation in the literature. It has been reported that the shell of a freshwater lamellibranch in the genus Anodonta has Fe and Mn levels of 2600 and 560 ppm (0.26 and 0.056%, respectively) . Seven marine lanaellibranchs and four marine gastropods had Fe and Mn levels ranging between 4.5 and 1600 ppm and 1.1 and 18 ppm, respectively (Segar, Collins and Riley, 1971 ) . It was not determined whether the Mn or Fe was concentrated in a particular region of the shell. In some cases Mn is found concentrated on the outer surface of shells. Allen ( 1960) examined the Mn present in the surface deposits on a variety of marine molluscs. For Nucula sulcata, Mn levels in surface deposits were reported to be 8.5 and 17.9%. In regions where N. sulcata was found, the bottom sediments contained Mn at 0.1%. Zavarzin ( 1964) reported that Mn is deposited on the shell of the marine mollusc Maconia baltica L. Both Allen and Zavarzin suggested that the process by which Mn is concentrated is biological and bacterial. Zavarzin concluded that Mn2@ exchanges into the CaCO3 structure of the shell and microorganisms cause the oxidation of Mn2@on the shell surface to manganese oxides. The bacterial oxida tion of Mn2@ to oxides of higher oxidation states of Mn is well documented (Ehrlich, 1968 ; Branter, 1970 ; Buchanan and Gibbons, 1974). The specific experiments described here are focused on the incorporation mechanism and on the chemical characteristics of the metals found in the shells of molluscs. The Mn and Fe levels in the shells of several freshwater and marine molluscs were examined by the electron microprobe technique and a variety of physical and chemical methods. It was found in the cases of the freshwater molluscs, Anodonta californiensis and Unio novahollandae, that Mn and Fe were concentrated locally at high levels (up to 13 and 40%, respectively) in their periostraca, while in the case of the marine mollusc, Mytilus californianus, Mn and Fe were not @detectable (<0.05%) by electron niicroprobe methods in the periostracum or shell.

Comparative study of the blood plasma of the ascidians Pyura stolonifera and Ascidia ceratodes

The components of the blood plasma of the ascidians, Pyura stolonifera and Ascidia ceratodes, have been separated by Sephadex G75 chromatography and gel electrophoresis. The absorption spectra of the plasma and the fractions for both species and the corresponding circular dichroisni spectra for Pyura stolonifera have been recorded. The components for each species consisted of a number of proteins and two N-acetylaminosugar compounds, probably aminopolysaccharides. Although there were similarities in the properties of these components between the two species, a number of important differences were observed, especially with regard to the uptake of metals as studied by 59Fe, “?�Mn, 48V, and 51Cr radiotracers. Electron spin resonance measurements detected manganese ( I I ) in a N-acetylamino sugar component of the plasma of Pyura stolonif era.

Intracellular acidity in the ascidian

1. 1. Morula cells of three Ascidiidae and two Perophoridae from the suborder Phlebobranchia, and one Holozoidae and one Clavelinidae from the suborder Aplousobranchia were acidic to neutral red; where methyl red was used, the stain corresponded to pH values between 4.6 and 6.0 depending on species. Five Didemnids (Aplousobranchia), and two Pyuridae, two Styelidae, and one Molgulidae (Stolidobranchia) had neutral morula cells. All compartment cells were neutral. 2. 2. Phosphorus-31 NMR spectrum of blood cells of Pyura stolonifera (Stolidobranchia) had Pi signals corresponding to pH 6.4 and 6.1, probably from phosphate within the globules and cytoplasm of the morula cells. The 31P NMR spectra of whole colonies of Diplosoma similis and Diplosoma virens had signals corresponding to Pi at pH values of 5.58 and 5.65. 3. 3. In Phlebobranchia, acid released by vanadocytes upon lysis could not be accounted for solely by the hydrolysis of V(III) and Fe(III), but within the vanadocytes the pH is too high for the metals to be in hydrated forms. 4. 4. Tunic acid was examined. The tunic cells of many species of Aplousobranchia and Phlebobranchia released acid on lysis but the intact cells were neutral.

The effect of potential aqueous pollutants on the solubility of Pb+2 in cerussite—Calcite phase

The results of this work show:1.Pb+2 is rapidly immobilized from aqueous solutions using calcite or aragonite, CaCO3, and the Pb+2 is precipitated as PbCO3 (cerussite) on the surface of the CaCO3 (the Pb+2-CaCO3 phases),2.using CaCO3, the concentration of Pb+2 in certain solutions can be reduced below environmentally tolerated concentrations, and3.organics present in solutions and natural waters can mobilize Pb+2 from Pb+2-calcite phases; with effluent waters from a paper factory, and 1:1 Ca+2-EDTA being more effective than lignin sulfonates, 1:1 Ca+2-tartrate, soap, detergent, water from a sewage treatment plant, and ordinary river water.ZusammenfassungDie Ergebnisse dieser Arbeit zeigen, daß:1.Pb+2 aus wäßrigen Lösungen mit Calcit oder Aragonit, CaCO3, rasch entfernt werden kann, wobei Pb+2 also PbCO3 (Cerussit) an der Oberfläche von CaCO3 (den Pb+2-CaCO3-Phasen) niedergeschlagen wird,2.sich die Pb+2-Konzentration in bestimmten Lösungen mit CaCO3 unter die nach Ö-Norm erlaubte Grenze senken läßt,3.organische Substanzen in wäßrigen Lösungen und natürlichen Wässern zunächst in Pb+2-Calcit-Phasen fixiertes Pb+2 wieder mobilisieren können, wobei die Abwässer einer Papierfabrik und 1:1 Ca+2-EDTA stärker wirken als Ligninsulfonate, 1:1 Ca+2-Tartrat, Seife, Waschmittel, Wasser einer Kläranlage und gewöhnliches Flußwasser.

2,4-Dichlorophenoxyacetic acid (2,4-D) and paranitrophenol (PNP) interactions with gills of Anodonta californiensis and Mytilus californianus: Uptake and effects on membrane fluxes

Abstract The effects of pesticides and their breakdown products on membrane fluxes is a subject of interest in assessing the potential ecological impact of these substances. As part of a continuing program we have examined the effects of 2,4-dichlorophenoxyacetic acid (2,4-D) and paranitrophenol (PNP) on divalent cation and primary amine losses from and glycine uptake by gills of the bivalve molluscs Anodonta californiensis (fresh water) and Mytilus californianus (marine). Both 2,4-D and PNP reduce glycine influx into gills of M. californianus . This observation is consistent with the view that glycine is bound to the gill surface as a Mg 2+ complex prior to active transport. For gills of A. californiensis , Ca 2+ and Mg 2+ losses are increased by 10 −3 M 2,4-D relative to that into distilled water. Primary amine losses are increased for both A. californiensis and M. californianus gills at low 2,4-D concentrations. For A. californiensis and M. californianus gills the uptake of both 2,4-D and PNP are reduced by increasing concentrations of Ca 2+ and Mg 2+ . In the case of A. californiensis , Ca 2+ has a larger effect than Mg 2+ , which is consistent with the demonstrated stabilizing effect of Ca 2+ on biological membranes. The uptake of PNP is larger than that of 2,4-D and glycine for gills of A. californiensis . The uptake of 2,4-D is reduced by the presence of an excess concentration of glycine but this is probably a physical effect. The uptake of 2,4-D, PNP, and glycine are all passive processes for A. californiensis gills. Glycine does not reduce 2,4-D uptake into M. californianus gills. For M. californianus the transport mechanism for glycine is not the same as that for 2,4-D; the former being active transport and the latter passive transport. However, 2,4-D does interfer with the active transport of glycine.

The vanadium(III) chloride-dioxygen reaction in anhydrous pyridine: Evidence for a vanadium(III)-dioxygen adduct

Abstract The kinetics of the reaction between vanadium(III) chloride and dioxygen in pyridine has been examined. In both excess O2 and V(III) the forms of the rate law are constent with the mechanism V(III)+O 2 ⇌ K V(III)O 2 , V(III) → k products Due to equilibria involving VCl3 in pyridine, the values of K and k depend on the concentration of VCl3 used. At large concentrations of V(III) when (py)3VCl3 is the dominant species: [V(III)] in excess, k = (9.2 ± 6.3) × 10−2 sec−1 and K = 40 ±24 M−1 at 25°C. At small concentrations of V(III) when an appreciable amount of (py)4VCl2+ is present: [O2] in excess, k = (7.1 ± 0.5) × 10−2 sec−1 and K = 265 ± 18 M−1 at 25°C. Once the V(III)−O 2 reaction is initiated, “reacted” V(III) can be regenerated, supporting the view that a V(III)-O2 adduct is formed.