Hypoxia tolerance of the mummichog: the role of access to the water surface

Low dissolved oxygen (DO) had a significant effect on specific growth rate (G_S), length increment (I_L) and haematocrit (Hct) of the mummichog Fundulus heteroclitus. Regardless of access to the water surface, F. heteroclitus maintained high growth rates (G_S and I_L) at DO concentrations as low as 3 mg O₂ l^?1. With access to the water surface, both G_S and I_L of F. heteroclitus decreased by c. 60% at 1·0 mg O₂ l^?1 compared to all higher DO treatments. When denied access to the water surface, a further decrease in G_S (c. 90%) and I_L (c. 75%) was observed at 1 mg O₂ l^?1. There was no effect of diel‐cycling DO (1–11 mg O₂ l^?1) with or without surface access on G_S, I_L or Hct of F. heteroclitus. Similar trends between G_S and faecal production across DO treatments suggest that decreased feeding contributed significantly to the observed decrease in growth rate. Haematocrit was significantly elevated at 1 mg O₂ l^?1 for fish with and without access to the water surface. Increased Hct, however, was not sufficient to maintain high G_S or I_L at severely low DO. When permitted to respire in the surface layer, however, F. heteroclitus was capable of maintaining moderate growth rates at DO concentrations of 1 mg O₂ l^?1 (c. 15% saturation). Although aquatic surface respiration (ASR) was not quantified in this study, F. heteroclitus routinely swam in contact with the water surface and performed ASR at DO concentrations ≤3 mg O₂ l^?1. No hypoxia‐related mortality was observed in any DO or surface access treatment for as long as 9 days. This study demonstrates that surface access, and thus potential for ASR, plays an important role in providing F. heteroclitus substantial independence of growth rate over a wide range of low DO conditions commonly encountered in shallow estuarine environments.     

Colonization and Degradation of Thermally Oxidized High-Density Polyethylene by Aspergillus niger (ITCC No. 6052) Isolated from Plastic Waste Dumpsite

Plastic materials, particularly polyethylene, are the potential source of environmental pollution. In the present study, a fungal strain was isolated from plastic waste dumpsites capable of adhering to high-density polyethylene (HDPE) surface. The fungal strain was identified as Aspergillus niger (ITCC no. 6052). A visible increase in the growth of the fungi was observed on the surface of the polyethylene when cultured in minimal medium at 30°C and 120 rpm, for 1 month. Approximately 3.44% reduction (gravimetrically) in mass and 61% reduction in tensile strength of polyethylene was observed after 1 month of incubation with fungal isolate. Scanning electron microscope analysis showed hyphael penetration and cracks on the surface of polyethylene. A thick network of fungal hyphae forming a biofilm was also observed on the surface of the plastic pieces. The efficient biofilm formation on polyethylene surface by Aspergillus niger (ITCC no. 6052) is attributed to its high cell surface hydrophobicity. This study indicated that Aspergillus niger (ITCC no. 6052) has ability to degrade thermally oxidized polyethylene.     

Lactones Produced through Thermal Oxidation of Higher Fatty Acids

Inhibitors of plant virus infection with systemic effects were found in the culture filtrates of Basidiomycetes such as Fomes fomentarius and Schizophyllum commune. These inhibitors were widely distributed in Agaricales and Polyporales. The inhibitors designated as BAS (Basidiomycete Antiviral Substance) were highly active against the mechanical transmission of tobacco mosaic virus (TMV). No toxic effect was observed on the host plants. BAS-F, a polysaccharide produced by F. fomentarius, almost completely inhibited infection, when BAS-F at 2 μg/ml was applied to the same surface of leaves of Xanthi-nc tobacco 24 h before TMV inoculation to the upper surface of the leaves, and 500/0 inhibition was shown when BAS-F at 10 μg/ml was applied to the under surface of leaves. BAS-F also induced systemic resistance to the non-treated leaves when it was applied to only one leaf of the plant. BAS-F also had similar effects against the infection of TMV on bell pepper and tomato plants.     

Localization of adenylate cyclase activity in Populus: its relation to pollen-pistil recognition and incompatibility

Summary Adenylate cyclase has been localized cytochemically in female and male parents as well as during the pollen-stigma interaction with an original technique employing strontium as the capture ion and adenyl imidodiphosphate as the specific substrate. The specificity of the reaction was checked by using several controls. No final specific reaction product was detected in unpollinated P. deltoides stigmas or in the P. deltoides or P. alba pollen grains used for compatible and incompatible pollinations. In the compatible cross between P. deltoides × P. deltoides, fine dense precipitates were observed in the dictyosomes and the plasma membrane and exterior to the exine of hydrated pollen grains adhering to the stigma surface. Labeling of the stigmatic pellicle was also observed after pollen adhesion and hydration. This was accompanied by a strong reactivity of the cell wall and plasmalemma of the stigma papillae at the sites of pollen tube germination on the stigma surface and at the sites of penetration of pollen tubes between adjacent papillae. In the incompatible cross between P. deltoides x P. alba, adenylate cyclase activity was still present but reduced at the stigma surface following adhesion, hydration, and germination of P. alba pollen. This activity was completely abolished after the penetration of pollen tubes between stigma papillae. These findings suggest that in Populus, adenylate cyclase activity is correlated to pollen adhesion, hydration, and germination at the stigma surface, and that the abolition of this enzyme activity could be one of the cellular events governing the gametophytic phenotype of incompatibility in the cross between P. deltoides and P. alba.     

Exploring the Cadmium Tolerance and Removal Capability of a Filamentous Fungus Fusarium solani

Abstract

Cadmium is one of the most toxic contaminant causing many problems to human health and the environment. These days the world is moving toward ecofriendly and efficient techniques to remove the pollutant from the wastewater. The present study aims to investigate the tolerance of Fusarium solani toward cadmium (Cd), nickel (Ni), and lead (Pb). Maximum tolerance was observed with Cd. Cadmium removal ability of F. solani was examined from contaminated PDB medium. pH, initial concentration and time optimization for maximum removal of Cd by F. solani was also studied. The maximum removal (92.4%) was recorded at initial concentration of 50?mg/L after 144?h of incubation. Cadmium exposure increased the level of glutathione (GSH) and oxidized glutathione (GSSG) contents and the activity of catalase (CAT) in F. solani. Fourier-transform infrared spectroscopy (FTIR) analysis indicated the involvement of the different surface functional group in biosorption of Cd while Scanning Electron Microscopy coupled with Energy Dispersive Spectroscopy (SEM-EDS) analysis revealed the presence of Cd on the surface of fungal cell. The changes observed in compositions of S, P, and Cd using EDS analysis on biomass surface indicated the precipitation of Cd as CdS and Cd₃(PO₄)₂. The XRD analysis revealed the presence of Cd₃(PO₄)₂ on mycelia surface of F. solani.     

Trichoderma species form endophytic associations within Theobroma cacao trichomes

Trichoderma species are usually considered soil organisms that colonize plant roots, sometimes forming a symbiotic relationship. Recent studies demonstrate that Trichoderma species are also capable of colonizing the above ground tissues of Theobroma cacao (cacao) in what has been characterized as an endophytic relationship. Trichoderma species can be re-isolated from surface sterilized cacao stem tissue, including the bark and xylem, the apical meristem, and to a lesser degree from leaves. SEM analysis of cacao stems colonized by strains of four Trichoderma species (Trichoderma ovalisporum-DIS 70a, Trichoderma hamatum-DIS 219b, Trichoderma koningiopsis-DIS 172ai, or Trichoderma harzianum-DIS 219f) showed a preference for surface colonization of glandular trichomes versus non-glandular trichomes. The Trichoderma strains colonized the glandular trichome tips and formed swellings resembling appresoria. Hyphae were observed emerging from the glandular trichomes on surface sterilized stems from cacao seedlings that had been inoculated with each of the four Trichoderma strains. Fungal hyphae were observed under the microscope emerging from the trichomes as soon as 6 h after their isolation from surface sterilized cacao seedling stems. Hyphae were also observed, in some cases, emerging from stalk cells opposite the trichome head. Repeated single trichome/hyphae isolations verified that the emerging hyphae were the Trichoderma strains with which the cacao seedlings had been inoculated. Strains of four Trichoderma species were able to enter glandular trichomes during the colonization of cacao stems where they survived surface sterilization and could be re-isolated. The penetration of cacao trichomes may provide the entry point for Trichoderma species into the cacao stem allowing systemic colonization of this tissue.     

Donor‐Acceptor Interfacial Interactions Dominate Device Performance in Hybrid P3HT‐ZnO Nanowire‐Array Solar Cells

The adsorption of self‐assembled monolayers (SAMs) on metal oxide surfaces is a promising route to control electronic characteristics and surface wettability. Here, arylphosphonic acid derivatives are used to modulate the surface properties of vertically oriented ZnO nanowire arrays. Arylphosphonate‐functionalized ZnO nanowires are incorporated into hybrid organic‐inorganic solar cells in which infiltrated poly(3‐hexylthiophene) (P3HT) serves as the polymer donor. Strong correlations between device short‐circuit current density (J _sc) and power conversion efficiencies (PCEs) with ZnO surface functionalization species are observed and a weak correlation in the open‐circuit voltage (V _oc) is observed. Inverted solar cells fabricated with these treated interfaces exhibit PCEs as high as 2.1%, primarily due to improvements in J _sc. Analogous devices using untreated ZnO arrays having efficiencies of 1.6%. The enhancement in J _sc is attributed to surface passivation of ZnO by SAMs and enhanced wettability from P3HT, which improve charge transfer and reduce carrier recombination at the organic‐inorganic interface in the solar cells.     

Inter‐ and intraspecific variation in the surface pattern of the dermal bones of two sturgeon species

Archaeological bone remains of sturgeon (Acipenser sturio/Acipenser oxyrinchus) from northwestern Europe are often identified to species on the basis of their surface morphology and then used to reconstruct the spatial distribution of the two species through time. The dermal bones of A. sturio are said to have an exterior surface pattern consisting of tubercles, while those of A. oxyrinchus are said to display alveoli. In the present paper, the validity of the surface pattern as a species‐specific characteristic is critically assessed. To this purpose, dermal plates from modern, genetically identified museum specimens were studied and the surface morphology observed in a series of archaeological remains was compared with the genetic identifications obtained on these same remains. The analyses show that the surface pattern of dermal bones is related to the size of the individual, with the pattern of small A. oxyrinchus being similar to that of A. sturio. In addition, variations in the surface pattern among the bones of a single individual and within the same bone have been observed. These findings explain previous conflicting results between morphological and genetic identifications and allow the formulation of some recommendations for more accurate morphological identification of isolated archaeological sturgeon dermal bones.     

Coccolithophores in the equatorial Atlantic Ocean: response to seasonal and Late Quaternary surface water variability

The present study was initiated to ascertain the significance of coccolithophores as a proxy for paleoceanographic and paleoproductivity studies in the equatorial Atlantic. Data from a range of different samples, from the plankton, surface sediments as well as sediment cores are shown and compared with each other.In general, the living coccolithophores in the surface and subsurface waters show considerable variation in cell numbers and distribution patterns. Cell densities reached a maximum of up to 300×10³ coccospheres/l in the upwelling area of the equatorial Atlantic. Here, Emiliania huxleyi is the dominant species with relatively high cell numbers, whereas Umbellosphaera irregularis and Umbellosphaera tenuis are characteristic for oligotrophic surface waters. Although they are observed in high relative abundances, these species only occur in low absolute numbers. The lower photic zone is dominated by high abundances and considerable cell numbers of Florisphaera profunda.The geographical distribution pattern of coccoliths in surface sediments reflects the conditions of the overlying surface water masses. However, abundances of the oligotrophic species Umbellosphaera irregularis and Umbellosphaera tenuis are strongly diminished, causing an increase in relative abundance of the lower photic zone taxa Florisphaera profunda and Gladiolithus flabellatus.During the past 140,000 years the surface water circulation of the equatorial Atlantic has changed drastically, as can be seen from changes in the coccolithophore species composition, absolute coccolith numbers, as well as coccolith accumulation rates. Significant increases in coccolith numbers and accumulation rates is observed in the southern equatorial Atlantic during the last glacial interval (oxygen isotope stages 2–4), which we attribute to enhanced upwelling intensities and advection of cool nutrient rich waters at this site. In the western equatorial Atlantic we observe an opposite trend with decreasing numbers of coccoliths during glacial periods, which probably is caused by a deepening of the thermocline.     

Studies on conjugation of <Emphasis Type="Italic">Spirogyra</Emphasis> using monoclonal culture

We succeeded in inducing conjugation of Spirogyra castanacea by incubating algal filaments on agar plate. Conjugation could be induced using clone culture. The scalariform conjugation was generally observed, while lateral conjugation was rarely. When two filaments formed scalariform conjugation, all cells of one filament behaved as male and those of other filament did as female. Very rarely, however, zygospores were formed in both of pair filaments. The surface of conjugation tube was stained with fluorescently labeled-lectins, such as Bandeiraea (Griffonia) simplicifolia lectin (BSL-I) and jacalin. BSL-I strongly stained the conjugation tubes, while weakly did the cell surface of female gamete first and then that of male gamete. Jacalin stained mainly the conjugation tubes. Addition of jacalin inhibited the formation of papilla, suggesting some important role of jacalin-binding material at the initial step of formation of the conjugation tubes.     

Construction of an <Emphasis Type="Italic">Aspergillus oryzae</Emphasis> cell-surface display system using a putative GPI-anchored protein

A novel cell-surface display system was constructed in Aspergillus oryzae. Each of the five genes encoding the putative cell-wall-localized protein from the A. oryzae genome was cloned and these cell-surface anchor functions were examined by fusion to the C-terminal of the green fluorescent protein (GFP). Using the MP1 and CWP proteins as anchor proteins, GFP signals were strongly observed on the cell surface of recombinant A. oryzae. When these proteins were used as anchor proteins for cell-surface display of β-glucosidase from A. oryzae, enzyme activity was detected on the cell surface. In particular, β-glucosidase activity of recombinant A. oryzae using MP1, a putative glycosylphosphatidylinositol (GPI) anchor protein was higher than CWP. Based on these results, it was concluded that the MP1 protein can act as a GPI-anchor protein in A. oryzae, and the proposed cell-surface display system using MP1 allows for the display of heterogeneous and endogenous proteins.     

Spatial distribution of aquatic birds in Anavilundawa Ramsar wetland sanctuary in Sri Lanka

Biotic and abiotic factors that influence the avian distribution in a dry zone wetland was investigated by studying the distribution of Asian Openbill (Anastomus oscitans), Cotton Pygmy-goose (Nettapus coromandelianus) and Pheasant-tailed Jacana (Hydrophasianus chirurgus) in Anavilundawa Ramsar sanctuary in Sri Lanka in 2006. Their distribution was recorded in Anavilundawa, Suruwila and Maiyawa reservoirs, their catchments and respective paddy fields, through line transects. The floral cover of surface water was recorded by floating quadrates. Water lily (Nymphea spp.) was the dominant flora in Anavilundawa reservoir, invasive water hyacinth (Eichhornia crassipes) in Suruwila reservoir and a native lotus (Nelumbo nucifera) in Maiyawa reservoir, respectively. Anavilundawa area had a higher distribution of birds than in the other two areas. Among the three species observed, Asian Openbill distribution was mainly restricted to Anavilundawa whereas Pheasant-tailed Jacana was present in all three areas. Nesting of Asian Openbill occurred only in Anavilundawa while nesting of Pheasant-tailed Jacana was only observed in Maiyawa. Among the three species recorded, Cotton Pygmy-goose numbers were the least. Asian Openbill preferred dead trees and trees with no leaves for nesting and perching over live trees. It is concluded that the spread of invasives such as Water hyacinth and Salvinia (Salvinia molesta) forming mats over surface water have reduced the abundance of habitat specialists like Cotton Pygmy-goose.     

Observation by SEM of the attachment ofAgrobacterium tumefaciens to the surface of vinca,asparagus and rice cells

Transformation of vinca cells was performed by the co-cultivation of cell-wall regenerated vinca protoplasts withAgrobacterium tumefaciens. Using thisin vitro and single cell system, attachment of the bacteria to the surface of vinca cells was observed by scanning electron microscopy (SEM). Figures of the bacteria polarly binding to the plant cell wall were often observed. AsEscherichia coli does not attach to the plant cells at all, the observed attachment ofA. tumefaciens is suggested as a characteristic feature in crown gall induction. Even though no evidence of transformation was obtained by the co-cultivation methods, a similar attachment was observed in the cell-wall regenerated protoplasts of rice. The bacteria also attached to the surface of isolated mesophyll cells of asparagus and root hairs of rice. From these observation, we concluded that the attachment is not the limiting step of crown gall induction byA. tumefaciens in monocotyledonous plants. Extracellular fibrils like pili were observed with a few strains of A.tumefaciens for the first time. These fibrils were observed regardless of their ability of attachment and infectivity.     

Cell surface hydrophobicity ofBifidobacterium bifidum subsp.pennsylvanicum

The possible role of lipoteichoic acid with respect to cell surface properties ofBifidobacterium bifidum subsp.pennsylvanicum was studied. Standard suspensions of bacteria were mixed with octane or xylene.B. bifidum subsp.pennsylvanicum was shown to possess a strongly hydrophobic cell surface. Hydrophobicity of the bacteria could be reduced by treatment with trypsin, pepsin (at pH 4.5), HCl and penicillin. The latter treatment resulted in an increased excretion of lipoteichoic acid. Albumin was capable of inhibiting the adherence to octane when it was present in the assay buffer. The data suggest that both protein and lipoteichoic acid may be involved in cell surface hydrophobicity. A great divergence in cell surface properties was observed within the genusBifidobacterium.     

Inactivation of bacterial opportunistic skin pathogens by nonthermal DC-operated afterglow atmospheric plasma

Aims: Multidrug‐resistant opportunistic pathogens are clinically significant and require the development of new antimicrobial methods. In this study, Acinetobacter baumannii, Pseudomonas aeruginosa and Staphylococcus aureus cells were exposed to atmospheric plasma on agar plates and in vitro on porcine skin for the purpose of testing bacterial inactivation. Methods and Results: Microbial inactivation at varying exposure durations was tested using a nonthermal plasma jet generated with a DC voltage from ambient air. The observed reduction in colony forming units was quantified as log₁₀ reductions. Conclusions: Direct plasma exposure significantly inactivated seeded bacterial cells by approx. 6 log₁₀ on agar plates and 2–3 log₁₀ on porcine skin. On agar plates, an indirect ‘bystander’ inactivation outside the plasma delivery area was also observed. The reduced inactivation observed on the skin surface was most likely due to cell protection by the variable surface architecture. Significance and Impact of Study: Atmospheric plasma has potential for clinical application as a disinfectant of patient skin and medically relevant surfaces.     

Expression of Fimbriae and Host Response in Branhamella catarrhalis Respiratory Infections

Sputum during the acute exacerbation of chronic respiratory diseases were observed under the electron microscope, to determine the in vivo expression of surface structures of Branhamella catarrhalis (B. catarrhalis), the polymorphonuclear neutrophil (PMN) response to B. catarrhalis infections, and the composition of sputum. It was found that during infection fimbriae are expressed in B. catarrhalis. However, there were sparsely to densely fimbriated bacteria in each sputum sample. The length of the fimbriae were from 50 to 76 nm. In the sparsely fimbriated B. catarrhalis, external to the cell wall, a thin, granular, electron-dense layer was observed. Due to the presence of fimbriae, this layer was not seen in densely fimbriated B. catarrhalis. Blebs were also found in B. catarrhalis. PMNs were found to phagocytose both B. catarrhalis and debris. Evidence was found that debris were formed mainly by the destruction of PMNs. Bacteria as well as debris were phagocytosed by PMNs.     

Bacterial quorum sensing and cell surface electrokinetic properties

The hypothesis tested in this paper is that quorum sensing influences the microbial surface electrokinetic properties. Escherichia coli MG1655 and MG1655 LuxS- mutant (lacking quorum-sensing gene for Autoinducer synthase AI-2) were used for this study. AI-2 production (or lack of) in both strains was analyzed using the Vibrio harveyi bioassay. The levels of extracellular AI-2 with and without glucose in the growth medium were consistent with previously published work. The surface electrokinetic properties were determined for each strain of E. coli MG1655 by measuring the electrophoretic mobility using a phase amplitude light-scattering (PALS) Zeta potential analyser. The findings show that the surface charge of the cells is dependent upon the stage in the growth phase as well as the ability to participate in quorum sensing. In addition, significant differences in the electrophoretic mobility were observed between both strains of E. coli. These findings suggest that quorum sensing plays a significant role in the surface chemistry of bacteria during their growth.     

Synthesis,characterization and in vitro activity of a surface-attached antimicrobial cationic peptide

Infection associated with implanted biomaterials is common and costly and such infections are extremely resistant to antibiotics and host defenses. Consequently, there is a need to develop surfaces which resist bacterial adhesion and colonization. The broad spectrum synthetic cationic peptide melimine has been covalently linked to a surface via two azide linkers, 4-azidobenzoic acid (ABA) or 4-fluoro-3-nitrophenyl azide (FNA), and the resulting surfaces characterized by X-ray photoelectron spectroscopy and contact angle measurements. The quantity of bound peptide was estimated by a modified Bradford assay. The antimicrobial efficacy of the two melimine-modified surfaces against Pseudomonas aeruginosa and Staphylococcus aureus was compared by scanning electron microscopy (SEM) and fluorescence microscopy. Attachment of melimine via ABA gave an approximately 4-fold greater quantity of melimine bound to the surface than attachment via FNA. Surfaces melimine-modified by either attachment strategy showed significantly reduced bacterial adhesion for both strains of bacteria. P. aeruginosa exposed to ABA–melimine and FNA–melimine surfaces showed marked changes in cell morphology when observed by SEM and a reduction of approximately 15-fold (p < 0.001) in the numbers of adherent bacteria compared to controls. For the ABA–melimine surface there was a 33% increase in cells showing damaged membranes (p = 0.0016) while for FNA–melimine there was no significant difference. For S. aureus there were reductions in bacterial adhesion of approximately 40-fold (p < 0.0001) and 5-fold (p = 0.008) for surfaces modified with melimine via ABA or FNA, respectively. There was an increase in cells showing damaged membranes on ABA–melimine surfaces of approximately 87% (p = 0.001) compared to controls, while for FNA–melimine there was no significant difference observed. The data presented in this study show that melimine has excellent potential for development as a broad spectrum antimicrobial coating for biomaterial surfaces. Further, it was observed that the efficacy of antimicrobial activity is related to the method of attachment.     

The surface microstructure of marine and terrestrial isopoda (Crustacea,Peracarida)

Summary The microstructure of the surface of thirteen marine littoral and two terrestrial isopods was investigated by scanning electron microscopy. A great diversity of surface ornamentation is present, including non-sensory microscales, pits, tubercles, and ridges, and sensory tricorns, pit organs, pores, papillae and setae. Microscales are common features of the integument surface; their shape and size are highly variable. Tricorns were not observed on the marine littoral isopods. Several hitherto undescribed structures were observed including spade-like projections from the tergite surface of Oniscus asellus, hair-like filaments associated with the microscales of Jaera and ridged conical protuberances on Edotea triloba and E. montosa. The possible function of certain surface microstructures is discussed.     

Potassium and phosphorus uptake by corn genotypes grown in the field as influenced by root characteristics

Summary Root parameters of three corn (Zea mays L.) genotypes influencing P and K uptake were investigated in solution culture and field experiments. The data for these parameters were used to simulate P and K uptake by plants grown in the field using the Claassen-Barber model⁵. Root characteristics for ion influx, maximum rate of influx,Imax; Michaelis-Menten constant,Km; and minimum concentration of solution below which no further net influx occurs,Cmin were determined in solution culture. These kinetic parameters varied 2 to 3 fold among genotypes. Variations among genotypes were different for K than for P.Three corn genotypes were grown in the field and harvested 47, 54 and 68 days after emergence. Yield and root surface per plant increased about 3 fold during this time. At 47 days, 2/3 of the total root surface was in the top soil whereas 3 weeks later, it was less than 50%. Genotypes differed in distribution of roots between the topsoil and subsoil as well as in root surface per unit of shoot.K uptake predicted by the Claassen-Barber model was 2 to 3 times the observed. The overprediction could be related to high root density (length of root per unit soil volume) which indicated that competition between roots occurred that was not considered in the simulation model. The predicted P uptake (y) was correlated (r=0.91) to observed uptake (x) byy=0.98+0.67x, indicating underprediction of P uptake. The presence of root hairs may have been the cause of the underprediction. The calculated contribution of the subsoil to the observed uptake was 10% for K and 1% in the case of P. It was concluded that the plant parameters used to simulate nutrient uptake were rated accurately when allowance was made for root competition and presence of root hairs.Journal Paper No. 7608. Purdue University, Agric. Exp. Station, West Lafayette, IN 47907. Contribution from the Department of Agronomy. This research was supported in part by the Tennessee Valley Authority and the Deutsche Forschungsgemeinschaft.