Keratinocyte growth-promoting activity from human placenta

Extracts of term human placenta were tested for enhancement of proliferative growth of primary cultures of human keratinocytes. Saline extracts or supernatants from homogenates were dialyzed extensively, lyophilized, and tested in subcultures of keratinocytes in MCDB 153 medium with 0.1 mM Ca++ containing only defined supplements (insulin, hydrocortisone, transferrin, ethanolamine, phosphoethanolamine). Cells plated in the absence of EGF at moderately high densities (1000-3000 cells per cm2) formed colonies and grew in the presence of placental extract at 25-500 micrograms/ml. Extracts of cord serum or maternal serum were inactive, suggesting that the activity is derived from placental tissue. The activity is not EGF, since the activity in the placental extract, unlike EGF, did not promote growth at low cell density, was synergistic with EGF under some conditions, and did not produce changes in colonial morphology which occurred in the presence of EGF. Unlike keratinocyte growth-promoting activity in bovine hypothalamic extract, the activity is non-dialyzable and is destroyed at 100 degrees C. Placental extract could not replace any of the defined components of the medium and is therefore distinct from them. The presence of activity in the placenta with distinctive properties suggests that this is a previously undescribed material with growth-promoting properties for epithelium.     

Histological demonstration of capillaries, interstitial space and muscle fibers in heart and skeletal muscle with fluorescent dyes

A method is described which allows a clear demonstration of capillaries and muscle fibers in the heart and skeletal muscle of experimental animals. The fluorescent dyes fluorescein isothiocyanate (FITC) and lissamine rhodamine B 200 (RB 200) were conjugated with a protein of high (gamma-globulin) and low (myoglobin) molecular weight, respectively, and were intravitally injected into the vascular system of rats. FITC globulin distributes itself in the intravasal space and RB 200 myoglobin in the extracellular. In histological sections the capillary lumina and the borderlines of the muscular fibers can be clearly identified and quantitatively evaluated because of the selective fluorescence in the respective structures.     

Structure of a hemoglobin gene cluster and nucleotide sequence of three hemoglobin genes from the midge Chironomus thummi piger (Diptera, Insecta)

The aquatic larvae of the genus Chironomus (Diptera, Insecta) contain at least 12 different hemoglobin (Hb) variants in their hemolymph. In the present study we have analysed the structure and part of the nucleotide sequence of a Hb gene cluster cloned from the genomic DNA of Chironomus thummi piger. The cluster contains probably 6 different genes, separated by intergenic regions of various lengths. The nucleotide sequence of three putative Hb genes including the intergenic regions is presented. The inferred amino-acid sequences show clearly that two of these putative genes code for subvariants of the Hb variant VIIB. The third gene codes for a so far unknown Hb protein. As known already for other chironomid Hb genes, there are no intron sequences present in the coding regions.