Active K+ transport in Mycoplasma mycoides var. Capri. Net and unidirectional K+ movements.

Analysis of the cation composition of growing Mycoplasma mycoides var. Capri indicates that these organisms have a high intracellular K+ concentration (Ki: 200--300 mM) which greatly exceeds that of the growth medium, and a low Na+ concentration (Na+i: 20 mM). Unlike Na+i,K+i varies with cell aging. The K+ transport properties studied in washed organisms resuspended in buffered saline solution show that cells maintain a steady and large K+ concentration gradient across their membrane at the expense of metabolic energy mainly derived from glycolysis. In starved cells, K+i decreases and is partially compensated by a gain in Na+. This substitution completely reverses when metabolic substrate is added (K+ reaccumulation process). Kinetic analysis of K+ movement in cells with steady K+ level shows that most of K+ influx is mediated by an autologous K+-K+ exchange mechanism. On the other hand, during K+ reaccumulation by K+-depleted cells, a different mechanism (a K+ uptake mechanism) with higher transport capacity and affinity drives the net K+ influx. Both mechanisms are energy-dependent. Ouabain and anoxia have no effect on K+ transport mechanisms; in contrast, both processes are completely blocked by dicyclohexylcarbodiimide, an inhibitor of the Mg2+ -dependent ATPase activity.     

INTERFERENCE OF SEX-RELATED FACTORS IN THE RESPONSE OF LIVER CELLS TO EXPERIMENTAL MITOTIC STIMULI

Stimulation of liver cell multiplication was obtained under two different experimental conditions.

• 1 A single injection of casein solution resulted in (a) an identical synchronized mitotic wave response in 10-day old male and female rats and (b) a significantly lower response in adult male rats compared to females, a difference which was reduced by castration of males at birth but essentially maintained if animals were operated when 10 days old.
• 2 Partial hepatectomy shortly after puberty resulted in active hepatocyte multiplication occurring 3 hr earlier in females than in males. This difference was suppressed when females were ovariectomized at birth and significantly reduced when they were spayed at a later age. Hepatocytes of castrated females entered actively into S phase 2 hr later than the sham-operated controls. Unilateral ovariectomy on the other hand indicated that during compensatory and/or hyper-compensatory activity of the single ovary there was a maximum difference between the male and female rate of [³H]thymidine uptake in liver nuclei 20 hr after hepatectomy. A further kinetic study (t= 25, 30, 40, 65, 90 hr) indicated no significant sex-related difference in the number of S phases per 10,000 cells.

The DNA content of regenerating versus control livers was comparable in both sexes at t= 22 and 90 hr but higher in females at t= 40 and 65 hr. A possible early postnatal interference of certain hormonal mechanisms in the receptivity to mitotic stimuli is postulated and discussed.     

Displacements of Simulium damnosum and strategy of control against onchocerciasis.

Regular aerial treatment of 14 000 km of watercourses has achieved and maintained, over an area of 700 000 km2 of West African savannah, a very high degree of control of the larvae of Simulium damnosum sensu stricto and S. sirbanum, the vectors of onchocerciasis in this area. However, particular and relatively restricted parts of this area, mainly in northern Ivory Coast and neighbouring parts of Upper Volta, experience regular and prolonged reinvasions by parous female vectors, which have already taken bloodmeals (and many of them carrying the parasites) and arrive from unknown sources probably hundreds of kilometres away, from directions probably between southwest and north. This reinvasion, now experienced in three successive years, represents the outstanding scientific, epidemiological and logistic problem still facing the WHO Onchocerciasis Control Programme. An outline is presented of the multidisciplinary investigations being undertaken to find a solution.     

DNA bifunctional intercalators. I. Synthesis and conformational properties of an ethidium homodimer and of an acridine ethidium heterodimer.

An ethidium homodimer and an acridine ethidium heterodimer have been synthesized. The ethidium and the acridine chromophore were introduced in such bifunctional intercalators in order to allow the fluorometric study of the interaction of such molecules with DNA, which is reported in the companion paper (Gaugain, B., Barbet, J., Capelle, N., Roques, B.P., & Le Pecq, J.B.(1978) Biochemistry 17 (following paper in this issue)). In the preparation of the acridine-ethidium dimer, we report the use of acetyl groups as new protecting agents in the phenanthridine series. Conformational studies of these molecules by visible absorption and NMR spectroscopy indicate that these dimers exist in equilibrium between folded and unfolded conformations and that this equilibrium is pH and temperature dependent. Models for the geometry of the folded forms are proposed.     

Measurement of the expected DNA lengthening caused by mono-and bisintercalating drugs using electron microscopy

PM₂ DNA molecules were treated with intercalating reagents (ethidium bromide, ethidium dimer, acridine dimer) and observed by electron microscopy. The adaptation of different electron microscopy techniques has enabled the determination of DNA lengthening upon drug intercalation. A 50% length increase was generally obtained for DNA saturated with the drugs. This result is in agreement with the intercalation model proposed by Lerman. In some cases (ethidium dimer), an increase of length larger than 50% can be obtained. Experimental conditions of DNA spreading strongly interfere with the DNA–drug interaction. In some cases it was possible to estimate the apparent binding constants and also to distinguish the mono- from the bisintercalating derivatives in their reaction with DNA.     

A novel type of endotoxin structure present in Bordetella pertussis. Isolation of two different polysaccharides bound to lipid A.

The endotoxin of Bordetella pertussis was cleaved by mild acidic hydrolysis to yield a polysaccharide (polysaccharide I, 15%), a glycolipid (63%) and lipid X (2%). Further treatment of the glycolipid with stronger acid released a second polysaccharide (polysaccharide II, 9%) and material similar to lipid A present in enterobacterial endotoxins. Both polysaccharides possess a single molecule of 3-deoxy-2-octulosonic acid as the reducing, terminal sugar. In polysaccharide II the octulosonic acid is phosphorylated in position 5 and presumably substituted in position 4; in polysaccharide I the octulosonic acid is not phosphorylated, but is substituted in position 5. Following treatment of the endotoxin with strong base, a fragment was isolated that contained bound, non-phosphorylated 3-deoxy-2-octulosonic acid, glucosamine phosphate and fatty acids. This indicated that polysaccharide I, like polysaccharide II, was bound to the lipid region of the endotoxin. The endotoxin structure thus defined is different from that proposed for the lipopolysaccharides of enterobacteria.     

Transmission experiments with pike fry (Esox Indus L.) rhabdovirus

Experimental infection of fertilized pike eggs with 'red-disease' virus produced 100% mortality in the fry. This mortality was associated with a disease that had previously been described as hydrocephalus internus, indicating that 'red-disease' and hydrocephalus are different manifestations of the same disease. The name pike fry rhabdovirus disease (PFRD) is suggested for the disease complex, and the name pike fry rhabdovirus (PFR) for the causative agent. Exposure of PFR to a Wescodyne * solution containing 25 ppm of iodine resulted in an inactivation of at least 99–99% of viral activity within 30 sec. Experimental egg transmission of PFR could be interrupted by disinfecting the eggs in a Wescodyne solution, suggesting that the virus was located on the egg surface. Conclusive evidence of a naturally occurring vertical transmission in pike culture is still lacking because, using FHM cells as a detection system, PFR could not be found in spawners and their sexual products. The susceptibility of pike fry to PFR rapidly decreases at increasing age.     

Effect of 50 S subunit proteins L5, L18 and L25 on the fluorescence of 5 S RNA-bound ethidium bromide

Among the three Escherichia coli 50 S subunit proteins L5, L18 and L25, which have an affinity for 5 S RNA, only protein L18 exerts a strong effect on the fluorescence of 5 S RNA-ethidium bromide complexes, without changing the quantum yield of the fluorescence. Proteins L5 and L25, although they have little effect on the fluorescence, have a strong stabilizing influence on the 5 S RNA-L18 complex. The results are discussed in terms of the secondary and tertiary structures of 5 S RNA in relation to ribosomal protein binding.