Cultural characteristics and fatty acid composition of propionibacteria

The cultural characteristics and cellular fatty acid composition of 40 strains representing 7 species of Propionibacterium and of 9 cultures of anaerobic corynebacteria were studied. The cultures were characterized by means of 23 separate cultural and biochemical tests. Cultures of the two genera differed consistently in only two reactions; the propionibacteria did not produce indole or liquefy gelatin, whereas the anaerobic corynebacteria were consistently positive with these tests. The fatty acids were extracted from whole cells and examined as methyl esters by gas-liquid chromatography. The most abundant acid in the seven Propionibacterium species was a C(15)-saturated branched-chain acid which was present in both the iso-and anteiso-form. Based on a comparison of the relative abundance of these isomers (i-C(15) and a-C(15)), the species were separated into two groups. P. freudenreichii and P. shermanii (group one) were similar and contained the a-C(15) isomer as the predominant acid. The i-C(15) isomer was the most abundant acid in the second group (P. arabinosum, P. jensenii, P. pentosaceum, P. thoenii, and P. zeae). The fatty acid profiles of the anaerobic corynebacteria were somewhat similar to those of the second group of propionibacteria, but were distinct from the profiles of P. freudenreichii and P. shermanii. The addition of branched-chain amino acids (l-leucine and l-isoleucine) to the growth medium increased the synthesis of the specific fatty acid(s) structurally related to the added amino acid.     

Distribution of Neuraminidase among Food-poisoning Strains of Clostridium perfringens

A survey was made to determine the distribution of the enzyme neuraminidase among 76 strains of Clostridium perfringens. Representative strains from each toxigenic type (A to F) and atypical C. perfringens type A food-poisoning strains of both American and English (Hobbs types) origin were tested. Both the American food-poisoning and nonfood-poisoning associated cultures consisted of both neuraminidase-positive and -negative strains. Furthermore, American strains which could not be differentiated from the original Hobbs cultures consisted of both neuraminidase-positive and -negative representatives. In contrast, the English (Hobbs) strains uniformly failed to produce an active intracellular or extracellular neuraminidase. No enzyme activity was detected in these strains when cultures were grown in different growth media, when grown in the presence of substrate (neuraminlactose), or upon extended incubation of enzyme preparations with substrate. With the exception of a type F strain, representative strains of the other toxigenic types (A to F) produced neuraminidase; 85% of the typical type A strains contained the enzyme.     

Fat metabolism and temperature acclimatization in the fly Phormia terraenovae R.-D.

When larvae of the fly Phormia terraenovae were fed on diets containing fats with different melting points and degrees of saturation, the fat laid down in the depots were effected, though the range of the depot fats was much narrower than that of the fat in the diet. Larvae reared at high temperatures also laid down fat which had a higher melting point and a lower iodine number than did larvae reared at low temperatures.No relation between the properties of the fat and the thermal death point was discovered, though the temperature of rearing had an effect.

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Zusammenfassung Larven der Fliege Phormia terraenovae R.-D. wurden im Insektarium bei annähernd 18° C gezogen und mit folgenden Nährstoffen gefüttert: Hefe/Milch, Schweine-, Hammel-, Rindfleisch, Hering.Wenn die Larven völlig erwachsen waren, wurden sie getötet und analysiert. Die Larven wiesen nach allen Ernährungsformen ähnliche Zusammensetzung auf, mit Ausnahme der Jodzahl des Fettkörpers. Diese variierte folgendermaßen: Milch-Hefe-Diät=62, Schwein =70, Hammel=71, Rind=69, Hering=90. Die Unterschiede zwischen mit Schwein, Hammel und Rind ernährten Larven waren nicht signifikant, die anderen Differenzen jedoch stark signifikant.Die Unterschiede zwischen den Jodzahlen der Fette in den verschiedenen Nährstoffen waren größer als diejenigen, die in den mit ihnen gefütterten Larven gefunden wurden (Milch-Hefe=30, Schwein=60, Hering=130).Mit Hammelfleisch bei 35° C ernährte Larven enthielten Fett mit einer Jodzahl von 64 (gegenüber 71 bei den unter 18° C gehaltenen).Der thermale Todespunkt war für alle bei 18° C gezüchteten Larven unabhängig von ihrer Ernährung ungefähr der gleiche. Die bei 35° C gehaltenen Larven wiesen einen annähernd um einen Grad höheren Todespunkt auf.Es scheint also wenig oder gar keine Beziehung zwischen der Zusammensetzung des Fettkörpers der Phormia-Larven und ihrer Resistenz gegen höhere Temperaturen zu bestehen.

     

Effect of membrane potential on band 3 conformation in the human erythrocyte membrane detected by triplet state quenching experiments.

The triplet lifetime and absorption anisotropy decay of eosin-labeled band 3 was measured in resealed erythrocyte ghosts. Membrane potentials were generated by the addition of valinomycin in the presence of a K+ gradient. Neither negative nor positive membrane potentials had any detectable effect on the rotational diffusion of band 3 nor on the eosin triplet lifetime. The membrane potential did, however, affect quenching of the eosin triplet state by I- and TEMPO (2,2,6,6-tetramethylpiperidine-N-oxyl). Quenching was enhanced by a negative membrane potential (negative inside) and reduced by a positive membrane potential. In addition, it was found that a negative membrane potential enhanced the efficiency of eosin labeling of band 3 in intact erythrocytes. A positive membrane potential had the opposite effect. These results indicate that the eosin binding site on band 3 becomes more accessible to the extracellular aqueous phase in the presence of a negative membrane potential and less accessible in the presence of a positive membrane potential. Quenching by I- and TEMPO of the triplet state of eosin-labeled band 3 was further investigated as a function of pH. Quenching by TEMPO and its dependence on membrane potential were relatively insensitive to pH. In contrast, the rate of quenching by I- showed a marked decrease over the range pH 5.5-9.5. Moreover, the effect of a negative membrane potential on I- quenching also varied with pH. These results are discussed on the supposition that the eosin probe is located in the anion access channel of band 3.(ABSTRACT TRUNCATED AT 250 WORDS)     

Cytogenetic characterization of 20 lymphoblastoid lines derived from human individuals differing in bleomycin sensitivity

Summary Forty lymphoblastoid (lymphoid) lines were established from 42 volunteer blood donors, including healthy individuals and patients with head and neck carcinomas. Each peripheral blood sample was split into two portions, one for the establishment of a lymphoid line and the other for short-term culture, which was used to estimate bleomycin sensitivity by cytogenetic procedures. Twenty lymphoid lines were selected at random to compare bleomycin sensitivity with data obtained from short-term lymphocyte cultures. In each set, bleomycin sensitivity of lymphoid cells was similar to that of the lymphocytes. The lymphoid lines, which can be propagated for an unlimited supply of relatively homogeneous cellular material, will be useful for a variety of future investigations. This investigation was supported by grants from the John S. Dunn Foundation, Houston, TX, the Esther Knispel Fund administered by The University of Texas M. D. Anderson Cancer Center, Houston, TX, and Department of Health and Human Services PHS grant DE 07007.     

Carboxy-terminal deletion analysis of oat phytochrome A reveals the presence of separate domains required for structure and biological activity.

A series of seven carboxy-terminal deletion mutants of oat phytochrome A were stably expressed in transgenic tobacco to localize phytochrome domains involved in chromophore attachment, spectral integrity, photoreversibility between the red light (Pr)- and far-red light (Pfr)-absorbing forms, dimerization, and biological activity. Amino acids necessary for chromophore attachment in vivo were localized to the amino-terminal 398 residues because mutant proteins this small had covalently bound chromophore. Deletion mutants from the carboxy terminus to residue 653 were spectrally indistinguishable from the full-length chromoprotein. In contrast, further truncation to residue 399 resulted in a chromoprotein with a bleached Pfr absorbance spectrum, Pr and Pfr absorbance maxima shifted toward shorter wavelengths, and reduced Pfr to Pr phototransformation efficiency. Thus, residues between 399 ad 652 are required for spectral integrity but are not essential for chromophore attachment. The sequence(s) between residues 919 and 1093 appears to be necessary for dimerization. Carboxy-terminal mutants containing this region behaved as dimers under nondenaturing conditions in vitro, whereas truncations without this region behaved as monomers. None of the plants expressing high levels of deletion mutants lacking the 35 carboxy-terminal amino acids displayed the light-exaggerated phenotype characteristic of plants expressing biologically active phytochrome A, even when the truncated phytochromes were expressed at levels 6- to 15-fold greater than that effective for the full-length chromoprotein. Collectively, these data show that the phytochrome protein contains several separable carboxy-terminal domains required for structure/function and identify a domain within 35 residues of the carboxy terminus that is critical for the biological activity of the photoreceptor in vivo.     

Toxicity versus avoidance response of golden shiner, Notemigonus crysoleucas, to five metals

Avoidance thresholds and 96-h LC50 values were determined for golden shiners, Noiemigonus crysoleucas , for five individual elements: chromium, Cr; copper, Cu; cadmium, Cd; arsenic, As; selenium, Se. The avoidance concentrations were 73, 26 and 28 μgl^-1 for Cr-VI, Cu and As-III, respectively. Cadmium and Se were not avoided at experimental concentrations up to 68 and 3489 μg1^-1, respectively. Acute flow-through 96-h LC50 values were Cr-VI 55·0, Cu 84·6, and As-III 12·5 mg 1^-1, which were more than two orders of magnitude above avoidance concentrations. The acute flow-through 96-h LC50 values for Cd and Se were 2·8 and 11·2 mg 1^-1, respectively. These concentrations are 31 and 2·2 times the highest concentration employed in the avoidance tests, neither of which were avoided by the test organisms. Thus, simple toxicity tests do not identify the environmental hazard of some elements, and the most toxic elements may not elicit a behavioural response. When used in concert with tests of organism function, more realistic indicators of environmental hazard or safety may be determined.     

The Actions of Calmodulin Antagonists W-7 and TFP and of Calcium on the Gating Kinetics of the Calcium-activated Large Conductance Potassium Channel of the Chara Protoplasmic Drop: A Substate-sensitive Analysis

The effects of the calmodulin antagonists W-7 and trifluoperazine have been measured on the Ca²⁺-activated potassium channel in the membrane surrounding protoplasmic drops expressed from internodal cells of charophyte plants. The large-conductance (170 pS), voltage- and Ca²⁺-dependent gating, and prominent conductance substrate of this channel shows a strong kinetic resemblance to those of the Maxi-K channel from animal cells. This is the first study of the action of calmodulin antagonists which measures their effects on the most populated substates as well as the closed and main open states of Maxi-K channels. The substate analysis provides new evidence for different modes of action of- and different bindings sites for these calmodulin antagonists. Neither antagonist produces the simple closure of the channel reported previously as its effect on the Maxi-K channel, though both do induce flicker-block, reducing the mean current to near zero at high concentrations following an inverted Michaelis-Menten curve. W-7 reduces residence time in the fully open state, thus raising, in the same proportions, the probabilities of finding the channel in the closed state or a pre-existing substate. Its binding to the channel is voltage- and calcium-dependent. In contrast, trifluoperazine reduces residence in the open state and promotes an apparently new substate which overlaps the closed state at −50 mV but is distinguishable from it at voltages more negative than −100 mV. This substate may represent times that trifluoperazine is bound to the channel. Both antagonists have effects clearly distinguishable from that of withdrawing calcium from the channel, which does not affect open state residence time but increases closed state residence time. Thus neither antagonist reverses the activating effect of Ca²⁻. This is good kinetic evidence against the view that the channel is activated by Ca²⁺-calmodulin and that the effect of a calmodulin antagonist is to reverse this process by making Ca²⁻-calmodulin less available. Received: 26 August 1996/Revised: 7 October 1996