The stabilities of mammalian apomyoglobins vary over a 600-fold range and can be enhanced by comparative mutagenesis.

Apomyoglobins from 13 different mammals were examined for resistance to denaturation by guanidinium chloride. Unfolding was followed by circular dichroism and tryptophan fluorescence and analyzed globally using the two-step, three-state mechanism first described by Barrick and Baldwin (Barrick, D., and Baldwin, R. L. (1993) Biochemistry 32, 3790-3796). With one exception, the rise and fall of Trp fluorescence intensity correlates quantitatively with the native to intermediate to unfolded steps seen in the CD curves. Although the O(2) binding properties of the holoproteins are nearly identical, the unfolding transitions of the apomyoglobins show 600-fold differences in resistance to guanidinium chloride denaturation. Apomyoglobins from diving mammals, particularly from sperm whales, are the most stable, whereas the apoproteins from pig, horse, and sheep are the least stable, indicating selective pressure for resistance to denaturation in the whale proteins. Sequence comparisons suggest that the key stabilizing residues in whale globins are Ala(5), His(12), Ile(28), Thr(51), Ala(53), Ala(74), Lys(87), Lys(140), and Ile(142). Combinations of these residues were substituted into pig myoglobin. The resultant multiple mutants showed stabilities approaching that of recombinant sperm whale apomyoglobin. Thus, comparative mutagenesis can be used to increase heme protein stability and improve expression yields in bacteria without compromising function.

Effects of prenatal stress on development in mice: maturation and learning.

Female CD-1 mice were stressed during the final week of gestation. Beginning 3 days after birth, until weaning, their pups were examined for eye opening, startle response, tooth eruption, surface righting, ability to cling to and climb an incline, tail pull reflex, rotation, linear movement and exploration. At 3 months of age, they were tested in a Morris Water Maze. Stressed animals were significantly lighter and shorter than non-stressed animals the first week after birth. By 3 days after birth, significantly fewer stressed animals could rotate or right themselves. By 6 days after birth, significantly fewer stressed animals could cling to or climb an inclined screen, or show the tail pull reflex. By 9 days of age, significantly fewer stressed animals had teeth. In contrast, by day 12 of age, significantly more stressed animals demonstrated exploratory behavior than did non-stressed animals. There were no sex differences in the ability of animals to perform these tasks at the same age. Stressed animals were significantly slower than non-stressed animals to reach the hidden platform in the water maze on all trials, and this difference was due to stressed females being slower to find the platform than non-stressed females, with no main effect of stress on males. This study supports and expands previous findings in rodents that prenatal stress can cause deficiencies in some early indices of physical maturation and also that these deficiencies can be continued into adulthood.

[Effect of an extract containing esterol on plasma cholesterol and oxidants].

16 patients with hypercholesterolemia were treated with an extract of alpha-alpha leaves (esterol) while on a low-fat, low-cholesterol diet. Esterol is believed to inhibit the absorption of cholesterol and bile acids and may interfere with the absorption of essential nutrients. As oxidative modification of lipoproteins is required for the process of atherosclerosis, plasma antioxidant vitamins were followed. After 4 months of treatment, plasma cholesterol decreased by 10% from 282 to 250 mg/dl (p < 0.001) and LDL cholesterol by 13%, from 203 to 177 mg/dl (p < 0.001). Plasma antioxidant vitamins E, A and beta-carotene were unchanged. Thus, esterol has a cholesterol-lowering effect but apparently does not lower fat-soluble, plasma antioxidant vitamins. Both cholesterol-lowering and plasma antioxidant vitamins are important for the primary prevention of coronary artery disease in hypercholesterolemia.

[The effect of alpha/beta and gamma interferon preparations on the functional activity of macrophages in experimental staphylococcal infections]. / Vlianie preparatov interferon alpha/beta i gamma-tipov na funktsional'nuiu aktivnost' makrofagov pri éksperimental'noi stafilokokkovoi infektsii.

The preparations of alpha/beta- and gamma-interferons have been shown to stimulate the functional activity characteristics of mouse macrophages (phagocytosis, spreading, contacts with lymphocytes, bactericidal properties) obtained from the peritoneal exudate of intact animals and those infected with staphylococci. The immunomodulating action of gamma-interferon is more pronounced than that of the preparation of alpha/beta-interferon.