Noninvolvement of Acyl Carrier Protein with Citrate Synthase and Malate Synthase

Acyl carrier protein (ACP coli) was isolated from commercially grown Escherichia coli B and was acetylated by chemical methods. Biological activity of the synthesized acetyl-ACP coli was checked in an in vitro fatty acid-synthesizing system isolated from E. coli B. Since acetyl-ACP is preferred over acetyl-coenzyme A (CoA) as a substrate in these reactions, the possibility that it may substitute for acetyl-CoA in biosynthetically and oxidatively important cellular pathways (glyoxylate and Krebs cycles, respectively) was examined. Acetyl-ACP was tested for substrate activity with the enzyme of each cycle which has been found to utilize acetyl-CoA. Crystalline citrate synthase (EC 4.1.3.7) of porcine origin (Calbiochem) was found to be inactive with acetyl-ACP coli, which acted neither as a substrate nor as an inhibitor in the presence of acetyl-CoA. Malate synthase (EC 4.1.3.2) of the acetate type was isolated from acetate-grown cells of a mutant of E. coli K-12 (VGD(3)H(5)) and was also found to be inactive with acetyl-ACP coli. The significance of these results and of the recent discovery of another phospho-pantetheine-containing protein are discussed.     

DYNAMICS OF ACRIDINE ORANGE-CELL INTERACTION : I. Interrelationships of Acridine Orange Particles and Cytoplasmic Reddening

The in vitro localization of acridine orange (AO) in living cells was monitored by means of fluorescence microscopy, quantitative cell viability studies, and photofluorimetric measurements following dye-cell interaction. The parameters, pH, time, dye concentration, and the metabolic state of the cell were found to exert a profound influence on the time course and distribution of staining. The parameters studied are mutually interdependent, and intracellular dye localization may be predictably altered by their appropriate manipulation. Conditions are defined whereby two morphologically distinct but physiologically interrelated reactions, namely, acridine orange particle (AOP) formation and cytoplasmic reddening (CR) may be caused, prevented, reversed, or modified. These results are explained in terms of the facilitation or inhibition of an intracytoplasmic dye-segregating mechanism, in turn affected by the rate of dye ingress and the physiological state of the cell. Whereas the accumulation of AO in AOP is compatible with cell viability, the appearance of CR is correlated with cell death. It is pointed out that meaningful interpretation of vital staining requires precise regulation of many parameters in the extracellular milieu. A scheme of cell compartmentalization with respect to AO is proposed to satisfactorily account for the effects of environmental variations on the distribution and ultimate fate of intracellular dye. The AOP are viewed as normally present acid phosphatase-positive multivesicular bodies.     

Studies on Bacterial Pyrogenicity: III. Specificity of the United States Pharmacopeia Rabbit Pyrogen Test

The specificity of the first or “presumptive” portion of the USP rabbit pyrogen test was investigated by use of a new absolute standard of reference. The reference standard was a 0.9% sodium chloride solution prepared to be pyrogen-free. Details of the preparation were described. The hypothesis was explored that the temperature response of rabbits after intravenous injection of the standard solution was independent of exogenous pyrogen. Reactions observed among the rabbits in our colony allowed a classification of these animals ranging from “consistently reliable” to “consistently unreliable.” Details of the experimental results and implications for pyrogen testing are discussed. The recommendation was made that all rabbit test animals be “screened” in sham and actual tests before being used for pyrogen testing.     

Repetitive proline-rich proteins in the extracellular matrix of the plant cell

Several classes of hydroxyproline-rich proteins have been found in the cell walls of plants. Monomeric forms of the proteins can be solubilized from the walls, but the majority of the proteins are insolubilized by as yet unidentified crosslinks. The proteins have repeated sequences and are often rich in basic amino acids. The repeat proline-rich region may be serving to generate an elongated rod-like structure while the regularly spaced lysines probably interact with the acidic pectins. Several of the genes coding for these proteins have been isolated, and their expression has been found in some cases to be tissue specific and in others inducible by hormones and various types of stress.     

Understanding osteoporosis.

Considerable progress has been achieved recently in our understanding of the normal process by which bone mass is regulated. Age-related trabecular bone loss is characterized not simply by a global loss of bone but also by cortical porosity and loss of trabecular connections. Because bone strength depends on architectural as well as material properties, bone quantity alone cannot define fracture risk with precision. Traditional therapies for osteoporosis increase bone mass, and estrogen therapy, in particular, profoundly decreases fracture risk. The pharmacologic restoration of bone quantity and quality, however, remains elusive. Modern biotechnology offers the hope that progress may come about through the development of growth factors and other osteotropic compounds for clinical use.     

Comparative localization of three classes of cell wall proteins.

The localization of the cell wall proline-rich proteins (PRPs), and the gene expression of the cell wall glycine-rich proteins (GRPs) and the hydroxyproline-rich glycoproteins (HRGPs) were examined in several dicot species. The PRPs are accumulated in the corner walls of the cortex where several cells are joined together and in the protoxylem cell walls of 3-day-old soybean root. In 1-month-old soybean plants, the PRPs are specifically deposited in xylem vessel elements of the young stem, and they are accumulated in both phloem fibers and xylem vessel elements and fibers of the older stem. Likewise, the PRPs are localized in xylem vessel elements and fibers in tomato, petunia, potato and tobacco stems. They are also found in outer and inner phloem fiber cell walls of tomato stem and in outer phloem fiber cell walls of petunia stem. The gene expression of the HRGPs and the GRPs is developmentally regulated in tomato, petunia and tobacco stems. HRGP mRNAs are abundant in outer and inner phloem regions, while GRP mRNAs are present mostly in primary xylem and in the cambium region. Immunocytochemical localization showed that the GRPs have a localization pattern similar to that of the PRPs in tomato, petunia and tobacco stems.     

Muscle hypertrophy response to resistance training in older women.

We conducted a 12-wk resistance training program in elderly women [mean age 69 +/- 1.0 (SE) yr] to determine whether increases in muscle strength are associated with changes in cross-sectional fiber area of the vastus lateralis muscle. Twenty-seven healthy women were randomly assigned to either a control or exercise group. The program was satisfactorily completed and adequate biopsy material obtained from 6 controls and 13 exercisers. After initial testing of baseline maximal strength, exercisers began a training regimen consisting of seven exercises that stressed primary muscle groups of the lower extremities. No active intervention was prescribed for the controls. Increases in muscle strength of the exercising subjects were significant compared with baseline values (28-115%) in all muscle groups. No significant strength changes were observed in the controls. Cross-sectional area of type II muscle fibers significantly increased in the exercisers (20.1 +/- 6.8%, P = 0.02) compared with baseline. In contrast, no significant change in type II fiber area was observed in the controls. No significant changes in type I fiber area were found in either group. We conclude that a program of resistance exercise can be safely carried out by elderly women, such a program significantly increases muscle strength, and such gains are due, at least in part, to muscle hypertrophy.     

Some aspects of the reproductive biology of the clupeid Ilisha africana (Bloch) off the Lagos Coast, Nigeria

The maturation development, spawning period, oogenesis, fecundity and sex ratio of Ilisha africana off the Lagos Coast, Nigeria were investigated. Six macroscopic and seven microscopic stages of gonad maturity were identified. Maturity was attained at 12.0 cm total length. Breeding occurred throughout the year with the peak spawning between May and December. Oogenesis consisted of seven stages and breeding marks were observed in the species. Fecundity ranged between 2098 and 11 687 eggs. There was a slightly higher correlation between fecundity and weight than between fecundity and length. The egg diameter varied from 0.77 to 1.35 mm. The overall sex ratio was 1 male to 0.97 female. The females however predominated over the males during most months in the peak breeding period.