Chaos Theology: A New Creation Theology and Its Applications

The problems inherent in creatio ex nihilo have led the author to the development of a new creation theology: chaos theology. Its main points are creation from an unexplained initial chaos, a remaining chaos element that is the source of physical and moral evil, and continuing creation toward fulfilment on the Last Day. Chaos theology can be reconciled with the scientific account of cosmic and biological evolution. Combining chaos theology with the physical theory of chaos helps in the understanding of God\'s action in the world. Jesus Christ is shown to be the cosmic Christ, who reconciles the entire cosmos, not only humanity. The problem of evil is readily solved in chaos theology as the effect of the remaining chaos element. From chaos theology and scientific insight in cancer, a theology of illness can be derived

Effects of Altering Freshwater Chemistry on Physiological Responses of Rainbow Trout to Silver Exposure

The influence of different water Cl- (50–600 μM), Ca2+ (50–1,500 μM), Na+ (50–1,500 μM), or dissolved organic carbon (DOC, 0.31–5 mg/L) levels on silver-induced physiological and biochemical perturbations of rainbow trout were investigated. Fish were acclimated to soft water (50 μM; Cl-, Ca2+, and Na+), then exposed to 3.7 μg/L Ag (as AgNO3) for 6 h, which resulted in a reduction in Na+ influx from the water, an inhibition of gill sodium- and potassium-activated adenosine triphosphatase (Na+/K+-ATPase) activity, and an accumulation of silver on the gills. Increasing the water Cl- or DOC levels ameliorated the silver toxicity. However, increasing water Ca2+ or Na+ concentration did not reduce the silver-induced physiological and biochemical perturbations. The free silver ion (Ag+) concentrations (calculated from MINEQL+, a geochemical speciation computer program) showed a negative correlation with the Na+ influx rates and gill Na+/K+-ATPase activity. However, gill silver levels did not correlate to Ag+ concentrations and no correlation was found between gill silver levels and either Na+ influx rates or gill Na+/K+-ATPase activity. These results support the notion that the [Ag+] concentration is of major importance when assessing silver toxicity in fish, and that this should be taken into account in regulatory strategies for silver in the natural environment

Polyphosphate metabolism in Acinetobacter johnsonii 210A

Since the seventies, there is a growing interest in the process of biological phosphate removal in which microorganisms able to accumulate large amounts of phosphate play a central role. Over the years many bacteria have been isolated from sludge systems showing enhanced biological phosphate removal, either to investigate the bacterial population or to obtain pure cultures which were used as model organisms in studies on enhanced biological phosphate removal. Often, bacteria of the genus Acinetobacter were found. Therefore, it was suggested that these organisms probably play an important role in biological phosphate removal. In 1980, Acinetobacter strain 210A was isolated from sludge of a wastewater treatment plant in Renkum. This organism was able to accumulate large amounts of phosphorus in the form of polyphosphate (Van Groenestijn, 1988). The aim of this study was to examine various aspects of the metabolism of polyphosphate in this organism. This thesis describes next to the identification of Acinetobacter strain 210A, properties of bacterial polyphosphate, degradation and in vivo synthesis of polyphosphate and the regulation of polyphosphate metabolism by external phosphate.Acinetobacter strain 210A was identified as Acinetobacter johnsonii by using a combination of biochemical and genetic properties. The organism was able to synthesize two polymers, polyphosphate and poly-β-hydroxybutyric acid. Polyphosphate was formed during growth at excess phosphate whereas poly-β-hydroxybutyric acid was synthesized when phosphate was the limiting nutrient. Intact cells were able to oxidize a variety of monosaccharides in the presence of PQQ (pyrollo-quinoline quinone). Ibis ability was also found in other Acinetobacter strains and was used in the examination of the bacterial population of two activated sludge types showing enhanced biological phosphate removal. It was found that the bacterial community structure of these sludges differed strongly. Therefore, it was suggested that the presence of Acinetobacter sp. in activated sludge systems showing enhanced biological phosphate removal depends on process design and influent composition of the treatment system (Chapter 2).Polyphosphate synthesized by Acinetobacter johnsonii 210A was localized in the cytoplasm mostly complexed in one or two large and several small granules. Electron microscopy and energy dispersive X-ray micro-analysis were used to examine the elemental composition of the large polyphosphate granules in unfixed and unstained intact cells of A. johnsonii 210A. When the organism was grown in standard medium, the granules were composed of phosphorus, magnesium and potassium. By modifying the amount of Ca 2+and Me 2+in the medium, the intracellular concentration of Ca 2+and Mg 2+as well as the elemental composition of the polyphosphate granules could be changed. A high Mg/Ca-ratio in the medium resulted in a high Mg/Ca-ratio in the cytoplasm and in the presence of Mg as counterion in the polyphosphate granule. Ca became the major cation in the polyphosphate bodies during growth in a medium with a low Mg/Ca-ratio. These results were at variance with previous studies dealing with the elemental composition of large polyphosphate granules which revealed Ca as the dominant counterion in the large polyphosphate bodies (Buchan 1981; Buchan 1983). This discrepancy could be ascribed to fixation and embedding procedures. Fixation of cells in glutaraldehyde and embedding in EPON have a profound effect on the elemental composition of polyphosphate granules (Chapter 3). Polyphosphate accumulated by A.johnsonii 210A consisted of about 700 Presidues as determined by gelelectrophoresis. In vivo31P-NMR was used to follow polyphosphate formation in intact cells. The amount of polyphosphate synthesized, correlated positively with the intracellular ATP concentration, suggesting an involvement of ATP in polyphosphate formation. In contrast to polyphosphate synthesis, polyphosphate degradation is a slow process (Chapter 4). Two polyphosphate degrading enzymes, polyphosphate:AMP phosphotransferase and polyphosphatase, have been found in A. johnsonii 210APolyphosphate: AMP phosphotransferase phosphorylates AMP to ADP with polyphosphate: PP n (polyphosphate) + AMP _>PP n-1 + ADP. The enzyme was purified more than 1,500-fold from A.johnsonii 210A by streptomycin sulfate precipitation, and by Mono- Q Phenyl Superose, and Superose column chromatography. Kinetic studies showed apparent K m values of 0.26 mM for AMP and 0. 8 μM for polyphosphate with an average chain length of 35 P-groups. The highest activities were found with polyphosphate molecules of 18 to 44 phosphate residues. The polyphosphate chain was degraded completely to ADP via a processive mechanism. No activity was obtained with ortho-, pyro-, tri-, and tetraphosphate. The enzyme was inhibited by pyro-, tri-, and tetraphosphate (Chapter 5).Chapter 6 describes the purification and characterization of polyphosphatase of A. johnsonii 210A This enzyme hydrolyzes polyphosphate to P i : PP n + H 2 O _>PP n-1 + P i . It was purified 77-fold by Q-Sepharose, hydroxylapatite and mono-Q column chromatography. The enzyme was specific for high polymeric polyphosphates and showed no activity towards pyrophosphate and organic phosphate esters. Analysis of kinetic properties revealed an apparent K m -value for polyphosphate with an average chain length of 64 residues of 30 ing polyphosphate per liter and for tetraphosphate of 1.2 mM. Polyphosphate chains were degraded to short chain polymers via a processive mechanism. The enzyme was inhibited by iodoacetamide and, in the presence of high Mg 2+-concentrations, by pyro-and triphosphate. The activating effect of Mg 2+on polyphosphatase was enhanced by K +and NH4+.Pyro-and triphosphate were hydrolyzed by pyrophosphatase, an enzyme which is in contrast to the polyphosphate degrading enzymes, widely distributed in nature. It was purified from A.johnsonii 210A and showed except against pyroand triphosphate no activity towards polyphosphates and a wide variety of organic phosphate esters. The enzyme is composed of 6 identical subunits of 23 kDa, giving a molecular mass of 141 kDa for the native enzyme. Mg 2+was required for activity. The enzyme was heat-stable and inhibited by fluoride and iodoacetamide. The apparent K m value for pyrophosphate was estimated to be 0.26 mM (Chapter 7).The effect of varying phosphate concentrations on biomass, cellular composition, phosphate uptake rate and activities of enzymes involved in (poly)phosphate metabolism of A.johnsonii 210A was investigated in P-or C-limited chemostat cultures. The organism accumulated poly-β-hydroxybutyric acid under P-deprivation, at phosphate concentrations ranging from 0.1 to 0.7 mM. The amount of biomass was proportional to the phosphate concentration in the medium and no polyphosphate was formed.When shifting a culture from P-to C-limitation, phosphate was accumulated as polyphosphate. No poly-β-hydroxybutyric acid could be detected in these cells. As soon as polyphosphate synthesis was possible, the specific activities of polyphosphate:AMP phosphotransferase and polyphosphatase increased about four fold. The specific activities of alkaline phosphatase and the P-uptake system were induced at residual phosphate concentrations below the detection limit. The effect of phosphate on the cellular polyphosphate content and on the P-uptake rate showed a hysteresis behaviour. When chemostat cultures were shifted from low to high phosphate concentrations, polyphosphate reached a maximum of about 60 ing P per gram of dry weight at a phosphate influent concentration of 2.5 mM. In the reverse case (high to low), polyphosphate did never exceed 45 mg P per gram of dry weight (Chapter 8).To optimize the process of biological phosphate removal, a thorough understanding of all processes involved in polyphosphate metabolism of bacteria present inactivated sludge is needed. These processes comprise: polyphosphate synthesis, polyphosphate degradation, and phosphate transport. In this thesis, properties of polyphosphate and polyphosphate degradation in A. johnsonii 210A were examined extensively. However, polyphosphate synthesis and phosphate transport were not clarified. From the positive correlation between intracellular ATP levels and polyphosphate synthesis, it was suggested that polyphosphate kinase may be involved in the process of polyphosphate formation. However, in cell extracts of A. johnsonii 210A, no polyphosphate kinase activity could be detected. This discrepancy can be interpreted in two ways. Either the polyphosphate kinase enzyme looses its activity during extract preparation or other polyphosphate synthesizing enzymes are involved in the formation of polyphosphate in A.johnsonii 210A. Two enzyme systems different from the polyphosphate kinase can be hypothesized: (1) a system in which a phosphorylated compound other than ATP is the P-donor or (2) a membrane bound proton translocating enzyme system. However, no positive indications for either of these two systems were found. Recently, the transport of phosphate in A. johnsonii 210A was characterized. Two transport systems could be demonstrated, (i) an inducible, ATP-driven, binding protein-dependent system, and (ii) a constitutive, low-affinity uptake system. It was suggested that the low- affinity system may be involved in the anaerobic metabolism of A.johnsonii 210A. Metabolic energy could be conserved by the generation of an electrochemical gradient across the cytoplasmic membrane, when phosphate is excreted together with ions (Van Veen et al. 1993). A more detailed knowledge of polyphosphate synthesis and phosphate transport would not only enlarge the insight in polyphosphate metabolism of phosphate accumulating bacteria, but would be of major importance for a better understanding of the process of biological phosphate removal. A better understanding would allow the optimization of its application based on information on physiological and biochemical mechanisms

Re‐evaluation of phosphoric acid–phosphates – di‐, tri‐ and polyphosphates (E 338–341, E 343, E 450–452) as food additives and the safety of proposed extension of use

The Panel on Food Additives and Flavourings added to Food (FAF) provided a scientific opinion re‐evaluating the safety of phosphates (E 338–341, E 343, E 450–452) as food additives. The Panel considered that adequate exposure and toxicity data were available. Phosphates are authorised food additives in the EU in accordance with Annex II and III to Regulation (EC) No 1333/2008. Exposure to phosphates from the whole diet was estimated using mainly analytical data. The values ranged from 251 mg P/person per day in infants to 1,625 mg P/person per day for adults, and the high exposure (95th percentile) from 331 mg P/person per day in infants to 2,728 mg P/person per day for adults. Phosphate is essential for all living organisms, is absorbed at 80–90% as free orthophosphate excreted via the kidney. The Panel considered phosphates to be of low acute oral toxicity and there is no concern with respect to genotoxicity and carcinogenicity. No effects were reported in developmental toxicity studies. The Panel derived a group acceptable daily intake (ADI) for phosphates expressed as phosphorus of 40 mg/kg body weight (bw) per day and concluded that this ADI is protective for the human population. The Panel noted that in the estimated exposure scenario based on analytical data exposure estimates exceeded the proposed ADI for infants, toddlers and other children at the mean level, and for infants, toddlers, children and adolescents at the 95th percentile. The Panel also noted that phosphates exposure by food supplements exceeds the proposed ADI. The Panel concluded that the available data did not give rise to safety concerns in infants below 16 weeks of age consuming formula and food for medical purposes

Modification of Experimental Protocols for a Space Shuttle Flight and Applications for the Analysis of Cytoskeletal Structures During Fertilization, Cell Division , and Development in Sea Urchin Embryos

To explore the role of microgravity on cytoskeletal organization and skeletal calcium deposition during fertilization, cell division, and early development, the sea urchin was chosen as a model developmental system. Methods were developed to employ light, immunofluorescence, and electron microscopy on cultures being prepared for flight on the Space Shuttle. For analysis of microfilaments, microtubules, centrosomes, and calcium-requiring events, our standard laboratory protocols had to be modified substantially for experimentation on the Space Shuttle. All manipulations were carried out in a closed culture chamber containing 35 ml artificial sea water as a culture fluid. Unfertilized eggs stored for 24 hours in these chambers were fertilized with sperm diluted in sea water and fixed with concentrated fixatives for final fixation in formaldehyde, taxol, EGTA, and MgCl2(exp -6)H2O for 1 cell to 16 cell stages to preserve cytoskeletal structures for simultaneous analysis with light, immunofluorescence, and electron microscopy, and 1.5 percent glutaraldehyde and 0.4 percent formaldehyde for blastula and plueus stages. The fixed samples wre maintained in chambers without degradation for up to two weeks after which the specimens were processed and analyzed with routine methods. Since complex manipulations are not possible in the closed chambers, the fertilization coat was removed from fixation using 0.5 percent freshly prepared sodium thioglycolate solution at pH 10.0 which provided reliable immunofluorescence staining for microtubules. Sperm/egg fusion, mitosis, cytokinesis, and calcium deposition during spicule formatin in early embryogenesis were found to be without artificial alterations when compared to cells fixed fresh and processed with conventional methods

Effects of some analgesics and antidepressants on the (Na+ + K + )-adenosine triphosphatase from cortices of brain and kidney

The effects of benzomorphans, tricyclic antidepressants, monoamine oxidase (MAO) inhibitors and chlorpromazine on the microsomal (Na+ + K+)-adenosine triphosphatase (ATPase) from beef cerebral cortex were studied. As a comparison, the interaction of these drugs with the corresponding enzyme from kidney cortex was also investigated. In addition to chlorpromazine, the benzomorphans and the tricyclic antidepressants inhibited the brain enzyme considerably, whereas the MAO inhibitors had little effect. The (Na+ + K+)-ATPase from kidney was not affected by the benzomorphans or MAO inhibitors, but its activity diminished in the presence of the tricyclic antidepressants and chlorpromazine.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/34064/1/0000342.pd

Polarity of the blood-brain barrier: Distribution of enzymes between the luminal and antiluminal membranes of brain capillary endothelial cells

The subcellular distribution in brain capillaries of alkaline phosphatase and Na+, K+-ATPase was investigated by two methods. Cytochemical studies using whole brain perfusion and electron microscopic examination indicated that alkaline phosphatase activity was located in both the luminal and antiluminal cytoplasmic membranes of the brain capillary endothelial cells. By contrast, the K+-dependent phosphatase activity associated with Na+, K+-ATPase was located in only the antiluminal membrane. Biochemical studies using membranes prepared by homogenization of isolated brain capillaries and density gradient centrifugation resulted in identification of two plasma membrane fractions. The light fraction contained alkaline phosphatase but very little Na+, K+-ATPase while the heavier fraction contained both enzyme activities. In addition, [gamma]-glutamyl transpeptidase showed a distribution similar to alkaline phosphatase while 5'-nucleotidase activity was distributed with the Na+, K+-ATPase activity. We conclude that the luminal and antiluminal membranes of brain capillaries are biochemically and functionally different. This polarity should permit active solute transport across brain capillary endothelial cells which are the cells responsible for the blood-brain barrier.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/23218/1/0000147.pd

"Self-screening" of rhodopsin in rod outer segments

Microspectrophotometry (MSP) shows rhodopsin highly concentrated (about 3.0mmol/l) in rod outer segments (ROS). Calculation of the in vivo absorption spectrum of human rhodopsin from such data reveals a striking failure to agree with the action spectrum of human rod vision. Agreement is good between the spectral distribution of absorption coefficients and the action spectrum, but the "concentration-broadening" (or "self-screening") introduced by the high end on absorbance at this concentration results in a misfit among the largest in the 93 years comparisons of this kind have been made! To deal with this anomaly, it has been suggested that "concentration-broadening" is inappropriate for rhodopsin in rod vision. This proposal was tested by comparing rod action spectra of 15-day-old and adult rats, since the lengths of ROS increase by a factor of about two in maturation. Three lines of evidence are inconsistent with it. Although the conundrum remains unexplained, it cannot be dismissed by supposing "self-screening" inappropriate for night vision.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/27007/1/0000574.pd