DNA damage and repair in brain: relationship to aging.

The usefulness of conducting DNA damage and repair studies in a postmitotic tissue like brain is emphasized. We review studies that use brain as a tissue to test the validity of the DNA damage and repair hypothesis of aging. As far as the accumulation of age dependent DNA damage is concerned, the data appear to overwhelmingly support the hypothesis. However, attempts to demonstrate a decline in DNA repair capacity as a function of age are conflicting and equally divided. Possible reasons for this discrepancy are discussed. It is suggested that assessment of the repair capacity of neurons with respect to a specific type of damage in a specific gene might yield more definitive answers regarding the role of DNA repair potential in the aging process and as a longevity assurance system.     

Interactions of surfactin,a biosurfactant fromBacillus subtilis,with inorganic cations

Summary The properties of surfactin, a biosurfactant lipopeptide, were highly modified in the presence of inorganic cations. The micellization of surfactin was favoured by monovalent and, especially by divalent cations with a modification of the molecular area at the air-water interface. Haemolysis of erythrocytes by surfactin was enhanced by low concentrations of divalent cations with an increase of the binding of the lipopeptide to membrane. Inorganic ions induced conformational rearrangements probably due to ion-surfactin associations which modify the surface-active properties.     

Regulation of glucose transport in Clone 9 cells by thyroid hormone.

Triiodothyronine (T3) is found to stimulate cytochalasin B-inhibitable glucose transport in Clone 9 cells, a 'non-transformed' rat liver cell line. After an initial lag period of more than 3 h, glucose transport rate is significantly increased at 6 h and reaches more than 3-times the control rate at 24 h. The enhancement of glucose transport by T3 is due to an increase in transport Vmax and occurs in the absence of a change in either the Km for glucose transport (approximately 3 mM) or the Ki for inhibition of transport by cytochalasin B ((1-2).10(-7) M). Consistent with the observed Ki for cytochalasin B, Northern blot analysis of RNA from control and T3-treated cells employing cDNA probes encoding GTs of the human erythrocyte/rat brain/HepG2 cell transporter (GLUT-1), rat muscle/fat cell transporter (GLUT-4), and rat liver transporter (GLUT-2) types indicates expression of only the GLUT-1 mRNA isoform in these cells. The abundance of GLUT-1 mRNA increases approx. 1.9-fold after 24 h of T3 treatment and is accompanied by an approx. 1.3-fold increase in the abundance of GLUT-1 in whole-cell extracts as demonstrated by Western blot analysis employing a polyclonal antibody directed against the 13 amino acid C-terminal peptide of GLUT-1. The more than 3-fold stimulation of glucose transport at 24 h substantially exceeds the fractional increment in transporter abundance suggesting that, in addition to increasing total GLUT-1 abundance, exposure to T3 may result in a translocation of transporters to the plasma membrane or an activation of pre-existing membrane transporter sites.     

Isolation and characterization of two individual glycoprotein components from human milk-fat-globule membranes.

Two individual glycoprotein components from human milk-fat-globule membranes (MFGM) has been purified by selectively extracting the membrane glycoproteins followed by lectin affinity chromatography and gel filtration on Sephadex G-200 in the presence of protein-disaggregating agents. The purified glycoprotein components, termed 'epithelial-membrane glycoprotein' (EMGP-155 and EMGP-39) have estimated molecular weights of 155 000 and 39 000 respectively, and yield a single band under reducing conditions on sodium dodecyl sulphate/polyacrylamide gel. EMGP-155 and EMGP-39 contain 21.0% and 7.0% carbohydrate by weight, with fucose (13.5%, 12.4%), mannose (3.7%, 6.2%), galactose (28.5%, 22.6%), N-acetylglucosamine (17.8%, 7.4%) and sialic acid (36.4%, 51.4%) of the carbohydrate moiety respectively. For both the glycoprotein components, aspartic and glutamic acid and serine are the major amino acid residues.     

Luteinizing hormone stimulates the production of prostacyclin by isolated ovarian cell invitro

Granulosa cells isolated from mature Graafian follicles of swine produced significant quantities of immunoreactive 6-keto-PGF₁α under chemically defined conditions in vitro. Luteinizing hormone elicited a dose-dependent stimulation of 6-keto-PGF₁α accumulation, but follicle stimulating hormone, prolactin, L-epinephrine, estradiol-17B, or PGE₂ were devoid of effect. The time-dependent in vitro production of 6-keto-PGF₁α by ovarian cells was susceptible to inhibition by indomethacin, U-51506, cycloheximide, and actinomycin D. These observations implicate granulosa cells in the specific and hormonally regulated production of prostacyclin.     

Variation and taxonomy of Aesculus pavia L. (Hippocastanaceae) in Texas

The status of Aesculus pavia var. flavescens, a set of yellow-flowered populations endemic to the Edwards Plateau of central Texas, was evaluated on the basis of comparative morphology, geography, flowering phenology, and pollination ecology. Plants of typical A. pavia have red, tubular flowers with exserted stamens and are effectively pollinated by ruby-throated hummingbirds. Plants of var. flavescens have yellow, campanulate flowers with included stamens that can be pollinated effectively by large bees. These ethological reproductive isolating mechanisms are further augmented by a difference in flowering time between the two sets of populations and by geographical restriction of the yellow-flowered plants to localities on the Edwards Plateau. The yellow and red-flowered plants constitute two morphologically distinct groups, despite evidence of limited introgression in counties along the Balcones Escarpment. A yellow-flowered plant discovered in coastal east Texas may indicate the mechanism by which var. flavescens originated, but this plant is much more closely comparable to typical red-flowered var. pavia than to the yellow-flowered plants on the Edwards Plateau. An evolutionary history of section Pavia of Aesculus is presented, and it is concluded that the yellow-flowered plants of central Texas are best regarded as a taxonomic variety.     

The galactan sulfate from the edible,red alga Porphyra columbina

A galactan sulfate has been isolated from the seaweed Porphyra columbina, and its structure established by a combination of methylation, methanolysis, treatment with alkali followed by methylation, and ¹³C-n.m.r. spectroscopy. The polysaccharide belongs to the porphyran class, and consists of 3-linked β-d-galactosyl residues and 4-linked α-l-galactosyl residues. 3,6-Anhydro-l-galactose and l-galactose 6-sulfate residues total approximately half of the sugar units, the other half being made up of d-galactose and 6-O-methyl-d-galactose residues. Some evidence is presented that suggests that the galactan sulfate does not have a completely alternating structure.     

Cell proliferation kinetics of epidermis and sebaceous glands in relation to chalone action.

Median S-phase lengths of pinna epidermis and sebaceous glands, and of epithelia from the oesophagus and under surface of the tongue of Albino Swiss S mice were estimated by the percentage labelled mitoses method (PLM). The 18.4 and 18,8 hr for the median length of S-phase for pinna epidermis and sebaceous glands respectively made it possible for these two tissues to be used experimentally for testing tissue specificity in chalone assay experiments. The 10.0 and 11.5 hr for oesophagus ang tongue epithelium respectively made experimental design for chalone assay difficult when pinna epidermis was the target tissue. The results of the Labelling Index measured each hour throughout a 24-hr period showed no distinct single peaked diurnal rhythm for pinna epidermis and sebaceous glands. Instead a circadian rhythm with several small peaks occurred which would be expected if an S-phase of approximately 18 hr was imposed on the diurnal rhythm. This indicates that there may be very little change in the rate of DNA synthesis. The results are given for the assay in vivo of purified epidermal G1 and G2 chalones, and the 72--81% ethanol precipitate of pig skin from which they could be isolated. These experiments were performed over a time period which took into account the diurnal rhythm of activity of the mice as well as the S-phase lengths. Extrapolating the results with time of action of the chalone shows that the G1 chalone acts at the point of entry into DNA synthesis and that the S-phase length was approximately 17 hr for both the pinna epidermis and sebaceous glands. This may be a more correct value since the PLM method overestimates the median S-phase length as it is known that in pinna skin the [3H]TdR is available to the tissues for 2 hr and true flash labelling does not take place. The previous reports that epidermal G1 chalone acts some hours prior to entry into S-phase resulted from experiments on back skin where the S-phase is shorter and there is a pronounced diurnal rhythm which could mask the chalone effect. The epidermal G2 chalone had no effect on DNA synthesis even at different times in the circadian rhythm. Thus the circadian rhythms and S-phase lengths of the test tissues need to be considered when experiments are performed with chalones. Ideally, the target tissues selected for cell line specificity tests should have the same cell kinetics for the easier and more accurate assessment and interpretation of results. When the tissues have markedly different cell kinetics, experimental procedures and results need to be evaluated accordingly. The point of action of G1 chalone can only be assessed if the effect is measured over the peak of incorporation of [3H]TdR into DNA. The results of the effects of skin extracts are analysed in relation to changes in the availability of [3H]TdR for the incorporation into DNA and to the possibility of there being two distinct populations of proliferating cells.