Loss of the PGE1 requirement for MDCK cell growth associated with a defect in cyclic AMP phosphodiesterase

Prostaglandin E₁(PGE₁), one of the components in the hormone-supplemented, serum-free medium for Madin Darby Canine Kidney (MDCK) cells (Medium K-1), is required for both long-term growth and for dome formation. Variant cells have been isolated from MDCK populations, which lack the PGE₁, requirement for long-term growth in Medium K-1. These variants will be useful in identifying the molecular events initiated by PGE₁ which are necessary for the growth response to be observed. The growth and functional properties of five independently isolated PGE₁ independent clones have been examined. Normal MDCK cells grew at an equivalent rate in Medium K-1 and in serum-supplemented medium; the growth rate was lower in Medium K-1 lacking PGE₁. In contrast, PGE₁ independent clone 1 grew at an equivalent rate in Medium K-1 minus PGE₁, and in serum-supplemented medium. When PGE₁ was added to K-1 minus PGE₁, less growth of PGE₁ independent clone 1 was observed. A similar observation was made with one other PGE₁ independent clone which was studied. A hormone deletion study indicated that PGE₁ independent clone 1 still retained growth responses to the other four supplements in Medium K-1 (insulin, transferrin, T₃, and hydrocortisone). The molecular alterations associated with loss of the PGE₁ requirement for long-term growth were examined. At confluency, all of the PGE₁ independent clones studied had higher intracellular cyclic AMP levels following PGE₁ treatment, as compared with normal MDCK cells. The increased cyclic AMP levels in the variant cells could result from a number of different types of defects, including reduced cyclic adenylic acid (cyclic AMP) efflux, an increased affinity of PGE₂ for the PGE₁ receptor, or a defect in cyclic AMP metabolism. However, in all of the variant clones studied a decreased rate of cyclic AMP degradation by cyclic AMP phosphodiesterase was observed. Thus, the increased cyclic AMP levels in the PGE₁ independent variants may result from alterations which affect cyclic AMP metabolism. The effect of PGE₁ on dome formation by the variant cells was also examined. The frequency of dome formation by PGE₁ independent clone 1 was enhanced in a dosage-dependent manner, like normal MDCK cells. This observation suggests that PGE₁ affects MDCK cell growth and dome formation by different mechanisms.     

Study of Chromosomes in the Newborn after Ultrasonic Fetal Heart Monitoring in Labour

The effect of continuous-wave ultrasound on the chromosomes of newborn infants has been investigated. Twenty-four women were studied during labour. The fetal heart was monitored by a Sonicaid FM2 monitor applied to the abdomen, and continuous monitoring undertaken for intervals varying from 1 hour 5 minutes to 9 hours 25 minutes. There was no increase in the number of chromosome aberrations in cultures of blood taken from the insonated babies when compared with controls.     

Acetamide Agar Medium Selective for Pseudomonas aeruginosa

A synthetic base of acetamide and salts in agar permitted isolation of small numbers of Pseudomonas aeruginosa from swarming Proteus and other gram-negative species.     

Adhesive apparatus in certain Indian hill stream fishes

The results of a detailed study on the adhesive apparatus of Indian hill stream cyprinid and sisorid fishes are presented. The structure of the mental suctorial disc in the genus Garra is described. Histological study of the mental region of Crossocheilus and Psilorhynchus has also been made and compared with the mental disc of Garra . The muscles of the disc which control its movements in bringing a partial vacuum during adhesion have been investigated. The complexity of the geniohyoideus muscles in Garra is interesting. Variations in the degree of development and histological modifications of the mental disc are exhibited by the fishes of the genus Garra .
A thoracic adhesive apparatus in the form of longitudinal or transverse ridges and grooves is present in the sisorids, Laguvia, Glyptothorax and Pseudecheneis except Gagata . A close study has revealed that the adhesive apparatus of the catfishes works mainly on the principle of friction. The variations in shape, size and location of the adhesive apparatus in various species are correlated to their respective habitats.
Structural variation in the adhesive apparatus of Pseudecheneis is maximum in comparison to that in Laguvia and Glyptothorax . It is discovered that the modifications are associated with the presence of multi-spinous layers, formation of caps by the basal or holdfast cells and with the transformation of loose areolar tissue into thick collagenous dermis. The presence of a pad of adipose tissue in the region of the adhesive apparatus has been reported and its physiological significance as a source of stored food suggested.
The anterior part of the protractor ischii muscle is modified to control the action of the adhesive apparatus. In Pseudecheneis , besides the m. protractor ischii, the ventral part of m. mesioventral is also associated with the adhesive apparatus.     

Comparison of Direct Plating Media for the Isolation and Enumeration of Enterococci in Certain Frozen Foods

A total of 15 agar media were examined for their yield, selectivity, readability, and simplicity of preparation and use. A thallium medium of Barnes was selected as the better of the high yield-fair selectivity type of medium and an azide-citrate medium of Reinbold appeared to be the better of the low yield-high selectivity type of medium. Sodium carbonate (optimal concentration, 0.20%) was found to increase recovery substantially when added to certain media, especially in the presence of 0.05% Tween 80. When these two ingredients were incorporated into a medium modified after Slanetz and Bartley, the resultant medium was superior to other media for the isolation and enumeration of enterococci in certain frozen foods, such as peas and hamburger, by the direct plating method.     

Freezing-Out Technique Applied to the Concentration of Biologically Active Materials

When applied to a dilute solution of folic acid and glucose, a freezing-out (with agitation) technique was shown to be an effective method of achieving a 20-fold reduction in volume with a loss of 10% of the active material being concentrated. Concentration of a stimulatory factor for Lactobacillus casei produced by Candida albicans in a complex medium was limited by the total solute concentration. Salts in the medium were concentrated to levels inhibitory for L. casei. The process is not selective and all solutes are concentrated.