The Escherichia coli cytochrome b556 gene, cybA, is assignable as sdhC in the succinate dehydrogenase gene cluster

Abstract The cytochrome b556 -deficient mutant Escherichia coli K12 strain TK3D11 [7] could not grow with succinate as the sole carbon source, but could grow well on dl -lactate. This finding suggested that cytochrome b556 is primarily responsible for oxidative metabolism and utilization of succinate. 24 Amino acid residues at the amino-terminal of purified cytochrome b556 were determined. This sequence coincided completely with amino acid residues 4 to 27, predicted from the DNA sequence of the sdhC gene, one of the unassigned open reading frames of the sdh gene cluster recently reported by Wood et al. [16]. Based on these and other results, we concluded that cybA , the gene for cytochrome b556 , is assignable as sdhC .     

Epitope expression on primate lymphocyte surface antigens

The cross-reactivity of peripheral blood mononuclear cells from 28 nonhuman primates was investigated with ten kinds of Leu series of monoclonal antibodies specific to human T-, natural killer/killer-, and B-cells. The chimpanzees possessed all ten epitopes examined but the orangutan lacked Leu4 and Leu7 epitopes and the gibbons lacked Leu4, Leu7, and Leu12 epitopes. In addition to the above epitopes, the Old World monkeys lacked Leu1 and Leu10 epitopes. The Leu3a/Leu2a cell ratios varied from 0 to 1.56 among the 12 macaque species and this enabled classification of these species into three groups. In the New World monkeys, Leu2a epitope was absent, whereas Leu11a epitope was detected in several species and Leu3a epitope was found only in the owl monkeys. The prosimians expressed only HLA-DR epitope.     

The effects of short-chain fatty acids on the neuronal membrane functions ofHelix pomatia. I. electrical properties

The effects of short-chain fatty acids on the membrane excitability, current-voltage (I-V) characteristics, and cell volume of Helix pomatia neurons were studied. 2-Decenoic acid (DA), having 10 carbon atoms in the hydrocarbon chain, suppressed the excitability of bursting neurons RPa1 (Sakharov and Salanki, 1969) for 30-60 min, while valeric acid (VA), having 5 carbon atoms, had no significant effect on excitability. DA had three different effects on the excitability of beating neurons: in some neurons DA suppressed excitability as in bursting neurons; in a second type of neuron DA had a negligible effect on excitability; and in the neuron located near RPa1 DA had a pentylentetrazol (PTZ)-like effect, i.e., it converted the discharge of the neuron from beating to bursting. DA decreased the peak value of the current, inducing a negative-resistance region in the I-V curve of the bursting neuron without any change in the level of the voltage at which the current reaches its maximal value. DA inhibited the hyperpolarization induced by activation of the Na+ pump, tested after preliminary enrichment of neurons with Na+ ions by incubation in a potassium-free solution for 20 min. DA caused a swelling of the neuron by about 10% which was independent of the Na+ pump. In all the above-mentioned cases VA had no significant effect.     

Ferredoxins in Developing Spinach Cotyledons: The Presence of Two Molecular Species

An antibody for ferredoxin was used to investigate the developmentof ferredoxin during the greening of spinach cotyledons. Ferredoxinwas present in 8-day-old etiolated cotyledons and increasedwith illumination, which means that the synthesis of ferredoxinwas both light dependent and independent. The ferredoxin purified from etiolated cotyledons, greeningcotyledons, and mature leaves was a mixture of two chemicallydistinct molecular species; ferredoxin I and II. The relativecontents of these two species varied with the stage of developmentand the conditions used. Ferredoxin I was identical with that isolated previously asvalidated by its amino acid sequence [Matsubara and Sasaki (1968)J. Biol. Chem. 243: 1732]. The complete amino acid sequenceof the second component, ferredoxin II, was determined as well.It was composed of 97 amino acid residues and differed fromferredoxin I by 25 residues. (Received October 16, 1982; Accepted December 14, 1982)     

Electrocardiographic studies in the Japanese monkey (Macaca fuscata) with special reference to the effect of anesthesia with barbiturates

The electrocardiograms of 157 healthy Japanese monkeys (Macaca fuscata), covering a wide range of ages in both sexes, were recorded under light pentobarbital (Nembutal) anesthesia. Although results were generally similar to those reported for other macaque species, some quantitative differences were observed.The heart rate was about 160 per minute in all monkeys examined; the P-Q interval was 0.11±0.06 sec.; the duration of QRS was 0.04±0.01 sec.; the Q-T interval was 0.24±0.06 sec. The mean axis of QRS was +59° and the pattern of the QRS complex was qR type in most cases.The comparison with the human electrocardiogram shows that the heart rate ofM. fuscata is about twice that of man, while the P-Q, QRS, and Q-T intervals were about one-half of those found in human subjects. In the monkey, however, the P wave was sharp and the T wave flat.In order to estimate the effect of anesthesia on the electrocardiogram, the records of several monkeys before, during, and after intravenous administration of barbiturates were compared. Although some animals showed extrasystoles after barbiturate was administered, generally no essential changes were noted in the records, except for the retardation of the rate and proportional prolongation of intervals.This work was presented at the 10th Annual Meeting of the Primate Research Association held in Inuyama, March 13, 1966.     

Stain Technology: A Simplified Aluminum Stain in Paraffin Sections of Bone from Hemodialysis Patients

A new simplified method has been devised for staining aluminum and has been tested in paraffin sections of bone from 60 patients who have undergone hemodialysis. Iliac crest bone fragments were fixed in 20% phosphate-buffered formalin for less than a day and demineralized at room temperature in 10% phosphate-buffered formalin containing 5% formic acid for only 2 to 3 hr. Four-micron paraffin sections, accompanied by positive controls, were stained with Maloney's aluminum stain, the Berlin blue reaction for iron, dylon or Congo red for amyloid and von Kossa's reaction for calcium. Aluminum and iron were demonstrated particularly at the mineralizing front of bony tissues; aluminum in 52 cases, iron in 45. Dylon staining also gave positive results in 52 cases. It is important in determining whether aluminum deposition is present that the von Kossa reaction remains positive even after demineralization. This method may be more useful for demonstrating aluminum in bony tissues than the complicated and time-consuming resin-embedding method currently used.     

In situ DNA-RNA hybridization using in vivo bromodeoxyuridine-labeled DNA probe

Summary An in vivo 5-bromodeoxyuridine (BrdUrd) labeled DNA probe was used for in situ DNA-RNA hybridization. BrdUrd was incorporated into plasmid DNA by inoculating E. coli with Luria-Bertani (LB) culture medium containing 500 mg/L of BrdUrd. After purification of the plasmid DNA, specific probes of the defined DNA fragments, which contained the cloned insert and short stretches of the vector DNA, were generated by restriction endonuclease. The enzymatic digestion pattern of the BrdUrd-labeled plasmid DNA was the same as that of the non-labeled one. BrdUrd was incorporated in 15%–20% of the total DNA, that is, about 80% of the thymidine was replaced by BrdUrd. Picogram amounts of the BrdUrd-labeled DNA probe itself and the target DNA were detectable on nitrocellulose filters in dot-blot spot and hybridization experiments using a peroxidase/diaminobenzidine combination. The BrdUrd-labeled DNA probe was efficiently hybridized with both single stranded DNA on nitrocellulose filters and cellular mRNA in in situ hybridization experiments. Through the reaction with BrdUrd in single stranded tails, hybridized probes were clearly detectable with fluorescent microscopy using a FITC-conjugated monoclonal anti-BrdUrd antibody. The in vivo labeling method did not require nick translation steps or in vitro DNA polymerase reactions. Sensitive, stable and efficient DNA probes were easily obtainable with this method.