Fibroblast heterogeneity in glucocorticoid regulation of collagen metabolism: Genetic or epigenetic?

Summary Cultured fibroblasts derived from normal human dermis show a consistent 62% inhibition of collagen synthesis by hydrocortisone, whereas cultures derived from keloids average only 30% inhibition and show a much larger strain to strain variation ranging from 75% inhibition to 49% stimulation. Examination of fibroblast clones indicates that this high variation among keloid strains is not due to differences in the proportion of normal and keloid cells in the mass culture populations. Small but significant differences in the effect of hydrocortisone on collagen deposition are also seen among these clonal populations, but are not related to the type of tissue from which cultures were derived. Two to three-fold differences among clones derived from a single individual were observed, possibly suggesting functional heterogeneity of dermal fibroblasts with regard to collagen metabolism under control conditions and in response to hydrocortisone. However, this variation among clones may simply reflect differences in clonal growth, inasmuch as both collagen synthesis and deposition, and the effect of hydrocortisone on these processes, are strongly affected by population density. This work was supported in part by PHS grants, CA-17229 from the National Cancer Institute and AG-02046 from the National Institute on Aging, DHHS; and by Grant RIM 78-17313 from the National Science Foundation.     

Calorimetric studies of the structural transitions of the human erythrocyte membrane. Studies of the B and C transitions

Differential scanning calorimetry has been used to study several structural transitions of the human erythrocyte membrane. Earlier studies have shown that one of these transitions (the A transition) is due to the thermal unfolding of spectrin on the membrane. In this paper, it is shown that two of the other transitions (B and C) exhibit a high sensitivity to a local anesthetic, benzyl alcohol. Increasing the ionic strength of the suspending medium results in a splitting of the B transition into two independent transitions (B₁ and B₂). It is found that one of these (B₂) is associated with titrating groups, since the midpoint for the transitions shifts by about 20°C, with an apparent $\text{p}\text{K}$ near 7.5. Extensive bilateral proteolysis by papain causes a drastic decrease in the size of all transitions except the C transition, which remains unaltered. On the other hand, treatment with phospholipase A₂ largely affects the C transition, causing its disappearance. Because of the lack of sensitivity to proteolysis and the high sensitivity to phospholipase, it appears that the C transition has a large extent of ‘lipid involvement’. It might result from the melting of a small fraction of phospholipid which exists in a crystalline state under physiological conditions. Alternatively, the C transition could arise from changes in protein-lipid interactions or from lipid-dependent changes in protein-protein interactions, providing one assumes that only protease-resistant portions of membrane proteins are participating.     

Interaction of lipopolysaccharide with detergents and its possible role in the detergent resistance of the outer membrane of gram-negative bacterua

In the presence of MgCl₂, amounts of detergents which disrupted phospholipid vesicles caused lipopolysaccharide I from Proteus mirabilis to aggregate and form vesicular, membrane-like structures. Vesicle formation with P. mirabilis lipopolysaccharide II containing longer O-polysaccharide chains was extremely poor. Lipopolysaccharides of Salmonella minnesota R mutants (chemotypes Ra, Rc and Re) displayed a growing tendency for vesicle formation with increasing deficiency of the R core polysaccharide. Lipopolysaccharides of chemotypes Rc and Re produced vesicles even in the absence of MgCl₂ and detergent. Spherical aggregates consisting of P. mirabilis lipopolysaccharide I, MgCl₂ and detergent were unable to either entrap or retain [¹⁴C]-sucrose, [³H]inulin or [³H]dextran. On the other hand, S. minnesota R mutant lipopolysaccharides of chemotypes Rc and Re could entrap all three saccharides and retain them for at least short periods of time. Leakage of [³H]-inulin out of Re-lipopolysaccharide vesicles was greatly retarded by addition of MgCl₂ to the vesicle system. Incorporation of P. mirabilis lipopolysaccharide I or S. minnesota Rc lipopolysaccharide into phospholipid vesicles protected these model membranes from disruption by detergent. This suggested a similar protective function of lipopolysaccharide in the outer membrane of enteric bacteria against the action of surfactants occuring in their normal intestinal habitat.     

Thymidylate synthetase: Studies on the peptide containing covalently bound 5-fluoro-2′-deoxyuridylate and 5,10-methylenetetrahydrofolate

Studies are reported on the FdUMP-CH₂-H₄ folate-peptide obtained upon proteolysis of the complex formed from thymidylate synthetase, FdUMP and 5,10-CH₂-H₄folate. Contrary to a previous report from this laboratory, the peptide does contain a cysteine residue. The sequence of the largest peptide obtained is Ala-Leu-Pro-Pro-(His,Cys)-Thr. Quantitative modification of the histidine residue with the Pauly reagent indicates that imidazole is not directly linked to the nucleotide. The stability of the peptide indicates the covalent bond to the cofactor involves its 5-nitrogen; from this, it may be concluded that the reactive form of the cofactor is the 5-iminium ion.     

Intersexual variation in the foraging ecology of sexually monochromatic Western Wood‐Pewees

ABSTRACT Investigators generally pool observations of males and females in studies of the foraging behavior of sexually monochromatic songbirds. However, such pooling can obscure possible intersexual differences. We compared the foraging behavior of male and female Western Wood‐Pewees (Contopus sordidulus), a sexually monochromatic species, in the Sierra Nevada Mountains of California during the breeding seasons of 2007 and 2008. We recorded 143 foraging observations (male N= 74, female N= 69). Overall, mean foraging rates of females (2.8 attacks/min) were higher (P < 0.001) than those of males (1.1 attacks/min). In addition, female foraging rates were significantly higher during incubation than during the nest building, nestling, and fledgling periods. When foraging, males perched higher above ground than females (means = 17.1 and 6.7 m, respectively). Differences between male and female Western Wood‐Pewees in foraging rates and perch heights suggest that males may spend more time on vigilance while females focus on foraging quickly during incubation and when feeding nestlings. Because metrics such as foraging attack rates are sometimes used as indicators of habitat quality and we found that rates can differ between the sexes and among nesting stages, investigators should consider the possibility of such differences when assessing habitat quality, especially for sexually monochromatic species of birds.