Agonist-specific desensitization of PGI2-stimulated cyclic AMP accumulation by PGE1 in human foreskin fibroblasts

Agonist-specific desensitization of prostaglandin I₂-stimulated (PGI₂)¹ adenosine 3′:5′-monophosphate (cyclic AMP) accumulation can be demonstrated in intact human foreskin fibroblasts (HFF) following a single exposure to PGE₁ or a stable PGI₂ analog (nitrilo-PGI₂). A single PGI₂-stimulation of HFF cells does not result in desensitization. Continuous re-addition of PGI₂ over a 4 hr period does induce desensitization to subsequent PGI₂-stimulation. HFF cells that are desensitized to PGI₂ are also desensitized to PGE₁ or nitrilo-PGI₂ stimulation indicating that these agonists share a common adenylate cyclase complex. Desensitization to PGI₂ can be measured after a 60 min, but not after a 30 min, exposure to PGE₁ or nitrilo-PGI₂. Once HFF cells are desensitized, a 12–24 hr period is required for the recovery of PGI₂ sensitivity.The adenylate cyclase in membranes prepared from intact cells that were preincubated with PGE₁ is also desensitized to subsequent PGI₂-stimulation. Preincubation of cells with PGI₂ does not induce desensitization of PGI₂-stimulated adenylate cyclase. These data suggest that HFF cells must be constantly exposed to a biologically active prostaglandin for desensitization to occur. The intrinsic chemical lability of PGI₂ may be a biochemical protection mechanism against desensitization in cells that normally respond to PGI₂.     

Photorespiration in Air and High CO(2)-Grown Chlorella pyrenoidosa

Oxygen inhibition of photosynthesis and CO₂ evolution during photorespiration were compared in high CO₂-grown and air-grown Chlorella pyrenoidosa, using the artificial leaf technique at pH 5.0. High CO₂ cells, in contrast to air-grown cells, exhibited a marked inhibition of photosynthesis by O₂, which appeared to be competitive and similar in magnitude to that in higher C₃ plants. With increasing time after transfer to air, the photosynthetic rate in high CO₂ cells increased while the O₂ effect declined. Photorespiration, measured as the difference between ¹⁴CO₂ and ¹²CO₂ uptake, was much greater and sensitive to O₂ in high CO₂ cells. Some CO₂ evolution was also present in air-grown algae; however, it did not appear to be sensitive to O₂. True photosynthesis was not affected by O₂ in either case. The data indicate that the difference between high CO₂ and air-grown algae could be attributed to the magnitude of CO₂ evolution. This conclusion is discussed with reference to the oxygenase reaction and the control of photorespiration in algae.     

High-spin binuclear cyclopentadienyliron chlorides: a density functional theory study

Theoretical studies on the cyclopentadienyliron chlorides Cp₂Fe₂Cl _n (n?=?6???1) with iron in the formal oxidation states from +1 to +4 indicate that all the high-spin species are predicted to be the lowest energy structures and they are paramagnetic complexes with magnetic moments between 2.8μ _B and 5.9μ _B. The mixed oxidation state derivatives with odd number of chloride atoms have larger magnetic moments than other species. In addition to Cp₂Fe₂Cl, which has the largest magnetic moment, these high-spin species have terminal Cp rings and bridging Cl atoms up to a maximum of two bridges. The Cp₂Fe₂Cl₄, Cp₂Fe₂Cl₃ and Cp₂Fe₂Cl₂ derivatives are predicted to be thermodynamically stable molecules with respect to exothermic reactions for the loss of one Cl atom from Cp₂Fe₂Cl _n . Moreover, the lowest energy Cp₂Fe₂Cl _n (n?=?3, 4) derivatives can be derived by the oxidative addition reactions of Cp₂Fe₂Cl _n?2 + Cl₂ → Cp₂Fe₂Cl _n .

Theoretical study on the functionalization of BC2N nanotube with amino groups

Using density functional theory calculations, we investigated properties of a functionalized BC₂N nanotube with NH₃ and five other NH₂-X molecules in which one of the hydrogen atoms of NH₃ is substituted by X = ?CH₃, ?CH₂CH₃, ?COOH, ?CH₂COOH and ?CH₂CN functional groups. It was found that NH₃ can be preferentially adsorbed on top of the boron atom, with adsorption energy of ?12.0 kcal mol^?1. The trend of adsorption-energy change can be correlated with the trend of relative electron-withdrawing or -donating capability of the functional groups. The adsorption energies are calculated to be in the range of ?1.8 to ?14.2 kcal mol^?1, and their relative magnitude order is found as follows: H₂N(CH₂CH₃) > H₂N(CH₃) > NH₃ > H₂N(CH₂COOH) > H₂N(CH₂CN) > H₂N(COOH). Overall, the functionalization of BC₂N nanotube with the amino groups results in little change in its electronic properties. The preservation of electronic properties of BC₂N coupled with the enhancement of solubility renders their chemical modification with either NH₃ or amino functional groups to be a way for the purification of BC₂N nanotubes.     

Induced Protein Synthesis during the Adaptation to H2 Production in Chlamydomonas moewusii

Gas production by Chlamydomonas moewusii in the light has been followed by manometric techniques during the adaptation to anaerobiosis. The only detectable gases produced are CO₂ and H₂ CO₂ is produced at a rather constant rate whereas H₂ evolution increase with time. This increase of H₂ evolution during the adaptation period can be inhibited by cycloheximide and by chloral hydrate, two inhibitors of protein synthesis. If the inhibitors are added to already adapted cells there is no effect on H₂ evolution. Adapted cell suspensions are sensitive to oxygen. Incubation under O₂ for 10 min inhibits the H₂ evolution to 100%. After removal of oxygen the capability to evolve H₂ can be restored only by a new adaptation period. This second adaptation to H₂ evolution can also be inhibited by cycloheximide.     

Generation of hydrogen peroxide on oxidation of NADH by hepatic plasma membranes

The oxidation of NADH by mouse liver plasma membranes was shown to be accompanied by the formation of H₂O₂. The rate of H₂O₂ formation was less than one-tenth the rate of oxygen uptake and much slower than the rate of reduction of artificial electron acceptors. The optimum pH for this reaction was 7.0 and theK _m value for NADH was found to be 3×10^–6 M. The H₂O₂-generating system of plasma membranes was inhibited by quinacrine and azide, thus distinguishing it from similar activities in endoplasmic reticulum and mitochondria. Both NADH and NADPH served as substrates for plasma membrane H₂O₂ generation. Superoxide dismutase and adriamycin inhibited the reaction. Vanadate, known to stimulate the oxidation of NADH by plasma membranes, did not increase the formation of H₂O₂. In view of the growing evidence that H₂O₂ can be involved in metabolic control, the formation of H₂O₂ by a plasma membrane NAD(P)H oxidase system may be pertinent to control sites at the plasma membrane.     

Organic derivatives of aluminium. Part I. Reactions of alkanolamines with aluminium isopropoxide

Reactions of alkanolamines [R₁R₂NXOH; R₁ = H, CH₃, C₂H₅; R₂ = H, CH₃, C₂H₅ and X = -CH₂CH₂-, -CH₂CH₂CH₂-, -CH₂CHCH₃, -C₆H₄CH₂CH₂-] with aluminium isopropoxide in different molar ratios (1 to 3) yield compounds of the type Al(OPrⁱ)_3?n(OXNR₁R₂)_n, where ‘n’ can be 1, 2 and 3. Most of the derivatives are distillable liquids, soluble in common organic solvents and susceptible to hydrolysis even by atmospheric moisture. The new derivatives are characterized by elemental analysis, IR and ¹H NMR spectra. Molecular weight measurements of Al(OPrⁱ)_3?n(OXNR₁R₂)_n reveal them to be tetrameric in nature.     

The effects of nitroxyl (HNO) on H2O2 metabolism and possible mechanisms of HNO signaling

Nitroxyl (HNO) possesses unique and potentially important biological/physiological activity that is currently mechanistically ill-defined. Previous work has shown that the likely biological targets for HNO are thiol proteins, oxidized metalloproteins (i.e. ferric heme proteins) and, most likely, selenoproteins. Interestingly, these are the same classes of proteins that interact with H₂O₂. In fact, these classes of proteins not only react with H₂O₂, and thus potentially responsible for the signaling actions of H₂O₂, but are also responsible for the degradation of H₂O₂. Therefore, it is not unreasonable to speculate that HNO can affect H₂O₂ degradation by interacting with H₂O₂-degrading proteins possibly leading to an increase in H₂O₂-mediated signaling. Moreover, considering the commonality between HNO and H₂O₂ biological targets, it also seems likely that HNO-mediated signaling can also be due to reactivity at otherwise H₂O₂-reactive sites. Herein, it is found that HNO does indeed inhibit H₂O₂ degradation via inhibition of H₂O₂-metaboilizing proteins. Also, it is found that in a system known to be regulated by H₂O₂ (T cell activation), HNO behaves similarly to H₂O₂, indicating that HNO- and H₂O₂-signaling may be similar and/or intimately related.     

Diffusion effects in the metabolism of hydrogen peroxide by rat liver peroxisomes.

Rat liver peroxisomes are membrane-bounded organelles containing catalase and oxidases producing H₂O₂. Diffusion effects in the metabolism of H₂O₂ and the physiological significance of the structure of peroxisomes are explored on the basis of two models. Model I considers the liver cell as consisting of two rapidly mixed compartments, the peroxisomal contents and the rest of the cell, separated by a membrane. On the basis of model I, it is concluded that in order to maintain a minimal H₂O₂ concentration in the cytoplasm, there must be an H₂O₂ destroying system in the cytoplasm, but the capacity of this system need be only a small fraction of that of the catalase in the peroxisomes. Model II takes account of the detailed morphology of peroxisomes and includes the effect of peroxisomal membrane permeability to H₂O₂ and H₂O₂ diffusion inside and outside the peroxisomes. On the basis of previously published experimental data and model II, it is concluded that the latency of catalase activity in intact peroxisomes is due chiefly to a permeability barrier to H₂O₂ at the peroxisomal membrane rather than to a restriction of H₂O₂ diffusion within the peroxisomes. Peroxisomes are calculated to be very efficient at destroying the H₂O₂ produced within them, whether the H₂O₂ is produced in the catalase-free core or in the catalase-containing matrix. Less than 2% of the H₂O₂ produced in peroxisomes leaves the particles. The efficiency of H₂O₂ trapping is the consequence of the membrane permeability barrier. A similar H₂O₂ trapping efficiency could be achieved by particles without a membrane barrier only if H₂O₂ diffusion within such particles were reduced by many orders of magnitude.     

Lethality of hydrogen peroxide in wild type and superoxide dismutase mutants of Escherichia coli. (A hypothesis on the mechanism of H2O2-induced inactivation of Escherichia coli)

The toxicity of H₂O₂ in Escherichia coli wild type and superoxide dismutase mutants was investigated under different experimental conditions. Cells were either grown aerobically, and then treated in M9 salts or K medium, or grown anoxically, and then treated in K medium. Results have demonstrated that the wild type and superoxide dismutase mutants display a markedly different sensitivity to both modes of lethality produced by H₂O₂ (i.e. mode one killing, which is produced by concentrations of H₂O₂ lower than 5 mM, and mode two killing which results from the insult generated by concentrations of H₂O₂ higher than 10 mM). Although the data obtained do not clarify the molecular basis of H₂O₂ toxicity and/or do not explain the specific function of superoxide ions in H₂O₂-induced bacterial inactivation, they certainly demonstrate that the latter species plays a key role in both modes of H₂O₂ lethality. A mechanism of H₂O₂ toxicity in E. coli is proposed, involving the action of a hypothetical enzyme which should work as an O₂^-• generating system. This enzyme should be active at low concentrations of H₂O₂ (<5 mM) and high concentrations of the oxidant (>5 mM) should inactivate the same enzyme. Superoxide ions would then be produced and result in mode one lethality. The resistance at intermediate H₂O₂ concentrations may be dependent on the inactivation of such enzyme with no superoxide ions being produced at levels of H₂O₂ in the range 5–10 mM. Mode two killing could be produced by the hydroxyl radical in concert with superoxide ions, chemically produced via the reaction of high concentrations of H₂O₂ (>10 mM) with hydroxyl radicals. The rate of hydroxyl radical production may be increased by the higher availability of Fe²⁺ since superoxide ions may also reduce trivalent iron to the divalent form.     

RuBPCO kinetics and the mechanism of CO2 entry in C3 plants

Abstract. The CO₂ partial pressure in the chloroplasts of intact photosynthetic C₃ leaves is thought to be less than the intercellular CO₂ partial pressure. The intercellular CO₂ partial pressure can be calculated from CO₂ and H₂O gas exchange measurements, whereas the CO₂ partial pressure in the chloroplasts is unknown. The conductance of CO₂ from the intercellular space to the chloroplast stroma and the CO₂ partial pressure in the chloroplast stroma can be calculated if the properties of photosynthetic gas exchange are compared with the kinetics of the enzyme ribulose 1,5-bisphosphate carboxylase/oxygenase (RuBPCO). A discrepancy between gas exchange and RuBPCO kinetics can be attributed to a deviation of CO₂ partial pressure in the chloroplast stroma from that calculated in the intercellular space. This paper is concerned with the following: estimation of the kinetic constants of RuBPCO and their comparison with the CO₂ compensation concentration; their comparison with differential uptake of ¹⁴CO₂ and ¹²CO₂; and their comparison with O₂ dependence of net CO₂ uptake of photosynthetic leaves. Discrepancy between RuBPCO kinetics and gas exchange was found at a temperature of 12.5 °C, a photosynthetic photon flux density (PPFD) of 550 μmol quanta m^?2 s^?1, and an ambient CO₂ partial pressure of 40 Pa. Consistency between RuBPCO kinetics and gas exchange was found if CO₂ partial pressure was decreased, temperature incresed and PPFD decreased. The results suggest that a discrepancy between RuBPCO kinetics and gas exchange is due to a diffusion resistance for CO₂ across the chloroplast envelope which decreases with increasing temperature. At low CO₂ partial pressure, the diffusion resistance appears to be counterbalanced by active CO₂ (or HCO₃) transport with high affinity and low maximum velocity. At low PPFD, CO₂ partial pressure in the chloroplast stroma appears to be in equilibrium with that in the intercellular space due to low CO₂ flux.     

Identification of the major urinary metabolite of thromboxane B2 in the monkey

Thromboxane B₂ (TxB₂) was biosynthesized from prostaglandin endoperoxides (PGG₂, PGH₂) using guinea pig lung microsomes and infused into an unanesthetized monkey. Urine was collected and TxB₂ metabolites were isolated by reversed phase partition chromatography and high performance liquid chromatography. A major metabolite (TxB₂-M) was found to be excreted in greater than two-fold abundance relative to other metabolites. Its structure was determined by gas chromatography-mass spectrometry to be dinorthromboxane B₂. $\text{In vitro}$ incubation of TxB₂ with rat liver mitochondria yielded a C₁₈ derivative with a mass spectrum identical to that of TxB₂-M, substantiating that the major urinary metabolite of TxB₂ in the monkey is a product of a single step of β-oxidation.     

Number of floral organs in Circaeaster agrestis (Circaeasteraceae) and possible homeosis among floral organs

98.9% of 5092 flowers from 1041 individuals of Circaeaster agrestis have five floral organs, the formula is P₃A₁G₁ (73.13%), P₂A₂G₁ (25.59%), and P₂A₁G₂ (0.22%). Only 0.4% of the flowers have six floral organs and the formula is P₃A₁G₂ (20 flowers) or P₃A₂G₁ (one flower). All these flowers have one vascular bundle in the pedicel and were considered to be normal ones. There are 33 flowers (0.65%) with six or more floral organs and two vascular bundles in the pedicel and we found traces of fusion of different degree of two flowers into one. These flowers were considered as abnormal. Therefore the normal number of floral organs of C. agrestis is five and occasionally six, and the floral formulas are P₃A₁G₁ or P₂A₂G₁, sometimes P₂A₁G₂, and occasionally P₃A₁G₂ or P₃A₂G₁. A tepal in P₃A₁G₁ may be replaced by a stamen in P₂A₂G₁ or by a carpel in P₂A₁G₂ or in reverse. A carpel in P₃A₁G₂ may be replaced by a stamen in P₃A₂G₁ or in reverse. We hypothesize that there are two possibilities for the number of the floral organs to be five (six), the result of reduction from P₃A₂G₂, or there exists homeosis among floral organs.     

Titanium complexes with chiral amino alcohol ligands: synthesis and structure of complexes related to hydroamination catalysts

Titanium complexes with chiral amino alcohol ligands are useful precatalysts for the intramolecular hydroamination of aminoallenes. They can be synthesized via protonolysis of titanium dimethylamide starting materials with the free ligand. In most cases, the resulting materials are not isolable due to their oily nature. However, several complexes were prepared in pure form and isolated as solid materials. [Ti(Cl)(NMe₂)(-OCH₂CH(Ph)N(CHMe₂)-]₂ was prepared at room temperature from TiCl(NMe₂)₃ and the corresponding N-substituted d-amino alcohol; the dimeric nature of the complex was established by X-ray crystallography. [Ti(NMe₂)₂(-OCH₂CH(Ph)N(2-Ad)-)]₂ (2-Ad = 2-Adamantyl) was prepared from Ti(NMe₂)₄ and the corresponding N-substituted l-amino alcohol after prolonged heating. An intermediate complex that could not be purified or isolated is believed to be Ti(NMe₂)₃(-OCH₂CH(Ph)NH(2-Ad)). Two complexes with the composition TiCl₂(-OCH₂CH(R*)N(CHMe₂)-)(HNMe₂) (where R* = CH₂Ph or CHMe₂) were prepared at room temperature by protonolysis of TiCl₂(NMe₂)₂ with the corresponding N-substituted l-amino alcohols. These two complexes exhibit dynamic behavior on the NMR timescale that is believed to be a dimer-monomer equilibrium, but they decompose at elevated temperatures.     

The limitations of heart rate as a predictor of metabolic rate in fish

Although telemetered heart rate (f_H) has been used as a physiological correlate to predict the metabolic rate (as oxygen consumption, V?O₂) of fish in the field, it is our contention that the method has not been validated adequately for fish. If f_H in fish is to be used to estimate V?O₂, a single linear (or log-linear) relationship must be established for each species between the two variables which allows V?O₂ to be predicted accurately under all environmentally relevant conditions. Our analyses of existing data indicate that while a good linear (or log-linear) relationship can be established between f_H and V?O₂, the conditions under which the relationship applies may be quite restricted. Physiological states and environmental factors affect the relationship between f_H and V?O₂ significantly such that several curves can exist for a single species. In addition, there are situations in which f_H and V?O₂ do not covary in a significant manner. In some situations f_H can vary over much of its physiological range while V?O₂ remains constant; in others V?O₂ may vary while f_H is invariate. The theoretical basis for this variability is examined to explain why the use of telemetered f_H in predicting V?O₂ of fish may be limited to certain specified applications.     

In situ formation of H2O2 for P450 peroxygenases

An in situ H₂O₂ generation approach to promote P450 peroxygenases catalysis was developed through the use of the nicotinamide cofactor analogue 1-benzyl-1,4-dihydronicotinamide (BNAH) and flavin mononucleotide (FMN). Final productivity could be enhanced due to higher enzyme stability at low H₂O₂ concentrations. The H₂O₂ generation represented the rate-limiting step, however it could be easily controlled by varying both FMN and BNAH concentrations. Further characterization can result in an optimized ratio of FMN/BNAH/O₂/biocatalyst enabling high reaction rates while minimizing H₂O₂-related inactivation of the enzyme.     

Mechanism leading to N<Subscript>2</Subscript>O production in wastewater treating biofilm systems

Wastewater treatment plants are known to be important point sources for nitrous oxide (N₂O) in the anthropogenic N cycle. Biofilm based treatment systems have gained increasing popularity in the treatment of wastewater, but the mechanisms and controls of N₂O formation are not fully understood. Here, we review functional groups of microorganism involved in nitrogen (N) transformations during wastewater treatment, with emphasis on potential mechanism of N₂O production in biofilms. Biofilms used in wastewater treatment typically harbour aerobic and anaerobic zones, mediating close interactions between different groups of N transforming organisms. Current models of mass transfer and biomass interactions in biofilms are discussed to illustrate the complex regulation of N₂O production. Ammonia oxidizing bacteria (AOB) are the prime source for N₂O in aerobic zones, while heterotrophic denitrifiers dominate N₂O production in anoxic zones. Nitrosative stress ensuing from accumulation of NO₂ ^? during partial nitrification or denitrification seems to be one of the most critical factors for enhanced N₂O formation. In AOB, N₂O production is coupled to nitrifier denitrification triggered by nitrosative stress, low O₂ tension or low pH. Chemical N₂O production from AOB intermediates (NH₂OH, HNO, NO) released during high NH₃ turnover seems to be limited to surface-near AOB clusters, since diffusive mass transport resistance for O₂ slows down NH₃ oxidation rates in deeper biofilm layers. The proportion of N₂O among gaseous intermediates (NO, N₂O, N₂) in heterotrophic denitrification increases when NO or nitrous acid (HNO₂) accumulates because of increasing NO₂ ^?, or when transient oxygen intrusion impairs complete denitrification. Limited electron donor availability due to mass transport limitation of organic substrates into anoxic biofilm zones is another important factor supporting high N₂O/N₂ ratios in heterotrophic denitrifiers. Biofilms accommodating Anammox bacteria release less N₂O, because Anammox bacteria have no known N₂O producing metabolism and reduce NO₂ ^? to N₂, thereby lowering nitrosative stress to AOB and heterotrophs.     

Dimeric HER2-specific affibody molecules inhibit proliferation of the SKBR-3 breast cancer cell line

HER2-specific affibody molecules in different formats have previously been shown to be useful tumor targeting agents for radionuclide-based imaging and therapy applications, but their biological effect on tumor cells is not well known. In this study, two dimeric ((Z_HER2:4)₂ and (Z_HER2:342)₂) and one monomeric (Z_HER2:342) HER2-specific affibody molecules are investigated with respect to biological activity. Both (Z_HER2:4)₂ and (Z_HER2:342)₂ were found to decrease the growth rate of SKBR-3 cells to the same extent as the antibody trastuzumab. When the substances were removed, the cells treated with the dimeric affibody molecules continued to be growth suppressed while the cells treated with trastuzumab immediately resumed normal proliferation. The effects of Z_HER2:342 were minor on both proliferation and cell signaling. The dimeric (Z_HER2:4)₂ and (Z_HER2:342)₂ both reduced growth of SKBR-3 cells and may prove therapeutically useful either by themselves or as carriers of radionuclides or other cytotoxic agents.     

The structures and stabilities of small diarsenic-doped silicon clusters

The structures and stabilities of As₂-doped Si_n (n = 1-7) clusters have been investigated at the B3LYP level of theory, incorporating the 6-311+G^∗ basis set. An isosceles triangle is predicted to be the lowest-energy structure of the As₂Si cluster, whereas the global minimum of As₂Si₂ possesses an As-As-butterfly structure. The ground state structures for As₂Si₃, As₂Si₄ and As₂Si₅ are all bipyramids: trigonal, tetragonal and pentagonal, respectively, which could have important applications as building blocks to synthesize silicon nanowires. The most stable isomer of As₂Si₆ possesses a tricapped trigonal bipyramid structure. The lowest energy structure of As₂Si₇ can be viewed as a substitutional structure of the tricapped trigonal prism Si₉ isomer. In the majority of the lowest energy isomers, the two As atoms tend to be separated from each other, in order to maximize the number of Si-As bonds, and therefore locate at the axial vertex or face-capping atomic positions, especially for As₂Si₄-As₂Si₇. According to results of the incremental binding energies, the HOMO-LUMO gaps and the vertical ionization potentials, the As₂Si₃ and As₂Si₆ clusters are relatively stable compared to their neighbors. Natural bond orbital analyses suggest that delocalized electrons and multi-centered bonds play an important role in stabilizing the low-energy As₂Si_n structures.