Myosin light-chain expression during avian muscle development

Monoclonal antibodies to adult chicken myosin light chains were generated and used to quantitate the types of myosin light-chain (MLC) isoforms expressed during development of the pectoralis major (PM), anterior latissimus dorsi (ALD), and medial adductor (MA) muscles of the chicken. These are muscles which, in the adult, are composed predominantly of fast, slow, and a mixture of fiber types, respectively. Three distinct phases of MLC expression characterized the development of the PM and MA muscles. The first identifiable pase occurred during the period of 5-7 d of incubation in ovo. Extracts of muscles from the pectoral region (which included the presumptive PM muscle) contained only fast MLC isoforms. This period of exclusive fast light-chain synthesis was followed by a phase (8- 12 d of incubation in ovo) in which coexpression of both fast and slow MLC isoforms was apparent in both PM and MA muscles. During the period, the composition of both fast and slow MLC isoforms in the PM and MA muscles was identical. Beginning at day 12 in ovo, the ALD was also subjected to immunochemical analyses. The proportion of fast and slow MLCs in this muscle at day 12 was similar to that present in the other muscles studied. The third development phase of MLC expression began at approximately 12 d of incubation in ovo and encompassed the transition in MLC composition to the isoform patterns incubation in ovo and encompassed the transition in MLC composition to the isoform patterns typical of adult muscle. During this period, the relative proportion of slow MLC rose in both the MA and ALD and fell in the PM. By day 16, the third fast light chain, LC(3f), was apparent in extracts of both the PM and MA. These results show that there is a developmental progression in the expression of MLC in the two avian muscles studied from day 5 in ovo; first, only fast MLCs are accumulated, then both fast and slow MLC isoforms are expressed. Only during the latter third of development in ovo is the final MLC isoform pattern characteristic of a particular muscle type expressed.     

Mechanism for leukotriene C4 stimulation of chloride transport in cornea

Summary The leukotriene, LTC₄, exerts a stimulatory effect on chloride transport in the frog cornea. In the work described here, the mechanism of action of LTC₄ to stimulate chloride transport was studied.In corneas pretreated with indomethacin, the effect of LTC₄ was abolished, suggesting the involvement of cyclo-oxygenase products in the response. Incubation of corneas with LTC₄ resulted in a significant stimulation in PGE₂ synthesis, as determined by TLC-autoradiography and radioimmunoassay. In addition, LTC₄ was found to stimulate cAMP synthesis in the cornea, and this stimulation was blocked with indomethacin. PGE₂ was previously shown by us to be the dominant cyclo-oxygenase product formed in the frog cornea, and is capable of stimulating cAMP and chloride transport. We suggest that LTC₄ stimulation of chloride transport is mediated via activation of the cyclooxygenase pathway, resulting in enhanced PGE₂ synthesis. Elevated PGE₂ levels induce cAMP synthesis, and ultimately, the stimulation of chloride transport. Further, the activation of cyclo-oxygenase was found to be dependent on phospholipase A₂ activity. This was shown by the inhibition of the LTC₄ effect in the presence of quinacrine. Similarly, inhibition of the LTC₄ effect in the presence of trifluoperazine suggests that cyclo-oxygenase activation by LTC₄ may be mediated via calmodulin. We have previously demonstrated that the frog cornea has the biosynthetic capacity to produce LTC₄. Therefore LTC₄ may function as an endogenous regulator of chloride transport in this tissue.     

Bacterial effect of hydrogen peroxide on urinary tract pathogens.

Bacterial contamination of urinary drainage bags is a frequent source of bladder bacteriuria in patients with indwelling catheters. Previous work demonstrated that the addition of 30 ml of 3% H2O2 prevented bacterial contamination of urinary drainage bags for up to 8 h in patients with urinary infections (greater than 10(5) colony-forming units per ml). Survival curves of a variety of organisms in filter-sterilized urine with various concentrations of H2O2 (0.6 to 0.01%) were constructed. Organisms with high cellular catalase activity (Staphylococcus aureus, Serratia marcescens, and Proteus mirabilis) required 30 to 60 min of exposure to 0.6% H2O2 for a reduction of 10(8) to less than 1 colony-forming unit per ml, whereas Escherichia coli, Streptococcus sp., and Pseudomonas sp. required only 15 min of exposure. The efficacy of H2O2 in urine was maintained despite exposure to room temperature for 5 days and reinoculation with bacterial suspensions. H2O2 is inexpensive and relatively nontoxic, and these data suggest that periodic instillation of H2O2 into urinary drainage bags may eliminate a source of bladder bacteriuria and environmental contamination.     

GROWTH FACTOR INTERACTIONS IN THE TISSUE CULTURE OF TUMOROUS AND NONTUMOROUS NICOTIANA GLAUCA-LANGSDORFFII

Schaeffer , Gideon W., Harold H. Smith and Marion P. Perkus . (Brookhaven Natl. Lab., Upton, N. Y.) Growth factor interactions in the tissue culture of tumorous and nontumorous Nicotiana glauca-langsdorffii. Amer. Jour. Bot. 50(8): 766–771. Illus. 1963.—Tissues representing tumorous and nontumorous Nicotiana glauca-langsdorffii were cultured on high (5 ×) and low (1 ×) concentrations of a modified White's basal medium containing 2.9 × 10^–6m indoleacetic acid. The growth responses of tissues of both the tumorous and nontumorous genotypes to supplements of kinetin, glutamine, inositol and nucleic acid constituents added singly and in all combinations were noted on high-salt media. The nucleic acid components inhibited growth and were omitted from low-salt media. The best growth response was observed with glutamine and inositol for tissues from the tumorous hybrid and with glutamine, inositol and kinetin in the nontumorous type. Kinetin was a distinct and consistent requirement for rapid growth of nontumorous tissues, but no appreciable kinetin effect could be observed with tissues from the tumorous genotype.     

Scopoletin and Polyphenol-Induced Lag in Peroxidase Catalyzed Oxidation of Indole-3-acetic Acid

The rate of initial indole-3-acetic acid (IAA) oxidation with horseradish peroxidase is modified with scopoletin, scopolin and other phenolic derivatives. In the presence of phenolics there is an initial lag phase in the oxidation. The early lag is dissipated enzymatically after which the rale of IAA oxidation again returns to normal. Chlorogenic and sinapic acids produce the longest lag periods of the compounds reported here, whereas the glucoside, scopolin, produced the least inhibition. Scopoletin is more than 10× as inhibitory as scopolin.     

Interaction of staphylococcal enterotoxin B with cell cultures of human embryonic intestine

Schaeffer, W. I. (Massachusetts Institute of Technology, Cambridge), J. Gabliks, and R. Calitis. Interaction of staphylococcal enterotoxin B with cell cultures of human embryonic intestine. J. Bacteriol. 91:21-26. 1966.-The cytotoxic effect of staphylococcal enterotoxin B upon human embryonic intestine cell cultures is characterized by retraction of cells from the monolayer. This is followed by clumping of the retracted cells to form clear areas in the monolayer and finally by sloughing of the clumps from the glass surface. The 50% effective dose of the toxin, determined by protein analysis of the cultures used in titration studies, was found to be between 40 and 60 mug/ml. The cytotoxic property of the enterotoxin was completely neutralized by 3.9 x 10(-5) ml of specific antitoxin per mug of toxin. The cytotoxicity was found to be slightly enhanced by 2.2 g of bicarbonate per liter of Eagle's basal medium (Earle's salt solution level), the absence of serum, the absence of penicillin and streptomycin, and the presence of 2.8 mmoles of calcium in the medium. The cytotoxicity was profoundly influenced by the age of the culture. No cytotoxicity was evident until after 2 days of growth had taken place, when the cell number was approximately 4.0 x 10(5) cells per culture.