The Ene Reaction of Singlet Oxygen with Olefins

The reactions of singlet oxygen (¹O₂) with cis and trans butenes-1,1,1-d₃, at—80°C in Freon-11, show a product isotope effect (k_H/k_D) of 1.38 and 1.25 respectively. Isomerization of the starting materials or formation of dioxetanes were not observed during the course of the photooxygenation. Together with the isotope effects on the reactions of tetramethylethylene-d₆ isomers with singlet oxygen, these results require the reversible formation of a perepoxide or charge transfer intermediate.     

Control of acid-base status in active and dormant land snails,Otala lactea (Pulmonata,Helicidae)

Summary Control of extracellular acid-base status was examined during activity and dormancy inOtala lactea (Pulmonata, Helicidae). Active snails showed little variation in hemolymph pH and at constant temperature. With increase of temperature, hemolymph increased from about 6 Torr at 5°C to 13 Torr at 24°C and pH decreased by about 0.017 pH units/°C, a pattern consistent with alphastat regulation of pH via ventilatory control of .During dormancy, mean hemolymph increased to about 50 Torr. Venous pH declined by about 0.4 units due to hypercapnia and fluctuated more widely than in active snails due to variability of . Hemolymph pH declined further in prolonged dormancy due to progressive metabolic acidosis; after one year of dormancy the mean hemolymph pH was about 0.8 units lower than that of active snails at similar temperature.Active snails exposed experimentally to high showed a large increase in hemolymph [HCO ₃ ^– ]. However, [HCO ₃ ^– ] declined by up to 50% during dormancy, despite the naturally occurring hypercapnia. Hemolymph osmolality and the concentrations of solutes other than [HCO ₃ ^– ] increased with increasing duration of dormancy. Concentrations of magnesium and calcium increased about 2.5 times more rapidly than those of sodium and chloride, indicating that acidosis is partially offset by the dissolution of carbonates from the shell or tissues.     

Intracellular reaction rates,enzyme activities and biomass yields in Methylomonas L3: growth rate and substrate composition effects

Summary The effects of specific growth rate and medium feed composition on the metabolic reactions of methanol incorporation and oxidation have been studied in carbon-limited, chemostatic cultures of Methylomonas L3. An in situ radioisotopic tracer technique was employed. The in vivo rates of substrate-carbon flow and the corresponding steady-state levels of several key RuMP-type methylotrophic enzymes were determined over a range of dilution rates from 0.19 to 0.41 h^-1 on methanol and methanol/formaldehyde substrates. It was determined that an absolute correlation does not exist between the in vivo specific carbon flux and the in vitro specific activity of any of the key enzymes studied. Oxidation of substrate-carbon via 6-phosphogluconate dehydrogenase is not stringently regulated in this methylotroph and the extent of its operation may be dependent on kinetic factors which make immediate cellular detoxification of formaldehyde imperative. As such, the cyclic oxidation mechanism in this methylotroph does not appear to be coupled to efficient energy utilization, since it was observed that high levels of the cyclic oxidation flux are commensurate with depressed biomass yields.     

The disappearance of isocitrate lyase from the green alga Chlorella fusca studied by immunoprecipitation

When acetate-adapted cultures of Chlorella fusca were transferred to nitrogen-free medium containing glucose, isocitrate lyase activity was lost over a period of about 25 h. Using a combination of in vivo isotope labelling and immunoprecipitation with anti-isocitrate lyase IgG it was shown that: 1. The onset of loss of enzyme activity preceeded the complete cessation of enzyme synthesis. 2. Disappearance of isocitrate lyase activity was accompanied by loss of enzyme protein, without accumulation of antigenic protein distinguishable from the normal subunit polypeptide of the enzyme, as judged by SDS gel electrophoresis of immunoprecipitated samples from supernatant cell-free extracts. 3. SDS gel electrophoresis of immunoprecipitated isocitrate lyase revealed the presence of antigenic protein bands of M_r about twice that of the normal subunit polypeptide, but the appearance of these apparent dimer forms did not obviously correlate with enzyme degradation. 4. Isoelectric focusing of immunoprecipitated isocitrate lyase showed that the enzyme became progressively more oxidised during the period of its degradation in vivo. 5. By titrating crude broken cell suspensions with anti-isocitrate lyase antibody, preliminary evidence was obtained for transfer of the enzyme from the soluble fraction to an insoluble form as part of the process of disappearance.     

Acceptors for cyclic AMP-dependent and calcium ion-dependent protein kinases in rat brain cytosol fractions: A comparison of occluded (synaptosomal) cytosol with non-occluded cytosol

Endogenous protein phosphorylation patterns were compared in occluded and non-occluded cytosol fractions prepared from rat forebrain. The occluded fraction was taken as representative of synaptosomal cytosol. One- and two-dimensional autoradiographs revealed the presence in non-occluded cytosol of a substrate for cAMP- and Ca²⁺/calmodulin-dependent protein kinase activities of M_r 300kD, corresponding to phosphorylated microtubule-associated protein-2 (MAP-2); this protein was absent in occluded cytosol. In contrast, a major substrate for protein kinase C was observed exclusively in occluded cytosol after phosphorylation under basal conditions. However, after phosphorylation in the presence of exogenous lipids, approximately equal amounts of the 82kD substrate were detected in both fractions, suggesting that protein kinase C in the occluded fraction was present in a partially activated state. Other minor differences in phosphorylation patterns between the two fractions were observed.Special Issue dedicated to Prof. Eduardo De Robertis.     

Kinetic studies on the chymotrypsin Aα-catalyzed hydrolysis of a series of N-acetyl-l-phenylalanyl peptides

A series of N-acetyl-l-phenylalanyl peptides of general formula Ac-Phe-(Gly)_n-NH₂ (n = 0–2) has been synthesized to study the effect of leaving group chain length on the efficiency of chymotrypsin A_α amidase and peptidase activities. The effect upon catalysis of hydrophobic side chains on the leaving group was investigated using similar substrates with one of the glycine residues selectively substituted by an alanine residue as in AcPheAlaNH₂, AcPheAlaGlyNH₂, and AcPheGlyAlaNH₂. Values of k_cat and K_m have been obtained from kinetic measurements at pH 8.00 and 25 °C. The results are shown to be consistent with binding schemes postulated from published model building studies. The catalytic reactions were studied over a range of temperature (15–35 °C) and in each case the Arrhenius law was obeyed. It was thus possible to obtain meaningful values for the thermodynamic functions of activation for the acylation step of the catalytic reaction. The results are shown to confirm the findings of postulated binding schemes but indicate that conclusions drawn from kinetic measurements at a single temperature may sometimes be misleading.     

Population Dynamics of Meloidogyne incognita and Rotylenchulus reniformis Alone and in Combination,and Their Effects on Sweet Potato

Meloidogyne incognita (Mi) and Rotylenchulus reniformis (Rr) interactions on sweet potato were studied in naturally and artificially infested field plots for 3 years. In a naturally infested field, early season counts of Mi or Rr were positively correlated with later counts of the same nematode, but negative correlations were found between early Mi and subsequent Rr, and early Rr and subsequent Mi counts. In field plots fumigated with methyl bromide and then infested with low levels of Rr, Mi, and Rr + Mi, final population densities of Mi juveniles were reduced by Rr, but Rr was not affected by Mi. In field plots with a high natural population density of Rr, artificial infestation with high levels of Mi in both fumigated and nonfumigated treatments inhibited Rr, while the final Mi juvenile population density was not affected. Results indicate that a competitive interaction exists with each species capable of inhibiting the other and becoming the dominant population. The nematodes had no apparent effect on yield at the inoculum densities used, either alone or mixed. Both nematodes increased cracking of sweet potatoes, but mixed populations did not differ in incidence of cracking from either Rr or Mi alone.     

Spatial relationships between fibroblasts during the growth of rat-tail tendon

Sections of tendons from the base of the tail of rats were taken at eight time intervals from 18 days in utero until 244 days after birth and were examined in the electron microscope. For each time period, measurements were made of the relative area of fibroblasts, collagen and interstitial material, of the number of fibroblasts per unit area of tendon and of the average area of individual fibroblasts. The spatial arrangement of fibroblasts in the tendon sections was described quantitatively using the "nearest neighbor" method. Initially there was a rapid increase in the area of collagen accompanied by a decrease in the area occupied by fibroblasts but after 104 days of age these values changed very little. The numbers of fibroblasts per unit area decreased steadily from the embryo until 104 days whereas the average size of each cell increased to reach a maximum area at 40 days of age and then declined. At all time intervals cells were arranged in a regular, dispersed pattern across the tendon fascicles. Growth in width of the rat tail appears to involve the secretion of collagen and other intercellular material symmetrically around each fibroblast, so as to gradually separate the cells until a stage is reached at which cells are sufficiently far apart that there is little contact between adjacent cell processes. This may interfere with the integration of metabolic activity in the tissue. As a consequence, there is shrinkage of the cell bodies and a reduction in secretory activity so that, between 55 and 104 days of age, the tendon enters a period of terminal senescence.