Dolichol and Dolichyl Phosphate Levels in Brain Tissue from English Setters with Ceroid Lipofuscinosis

Human ceroid lipofuscinosis (CL) is an inherited disease marked by cerebromacular degeneration and early death. We have utilized the canine model to investigate the possible role of dolichol and dolichyl phosphate in the developmental pathology of CL. We found that while brain levels of dolichol increase with age in both affected and unaffected dogs, the amount in the diseased animal was similar to that in controls. Brain levels of dolichyl phosphate ranged from 20 to 35 micrograms/g in control dogs at all ages examined, but increased substantially during development in the affected dogs, a value of 113 +/- 24 micrograms/g (mean +/- SD) being obtained in the end-stage animals. In addition to the results obtained in the canine model, dolichyl phosphate levels in human brain tissues from a 5-year-old with late infantile CL and from a 19-year-old with juvenile CL were found to be 153 and 382 micrograms/g, respectively, compared with a control that assayed 26 micrograms/g. The preliminary findings with human tissues provide further evidence for an association of elevated brain dolichyl phosphate levels with CL. Whether the increase is primary or secondary remains to be determined.     

Theoretical effects of transmembrane electroneutral exchange on membrane potential

Transmembrane electroneutral transport mechanisms [e.g., Na/H exchange, Cl/HCO3 exchange, (K + Cl) cotransport] have recently been identified in a wide variety of cell types. If these exchanges sum to give a net electroneutral Na/K exchange, they may hyperpolarize the membrane potential beyond the value calculated from the Mullins-Noda equation, provided the cell maintains steady state intracellular ionic concentrations. In extreme circumstances, the membrane potential could hyperpolarize beyond the potassium reversal potential. This effect is mediated by the electrogenic Na/K pump. If either Na or K exchanges electroneutrally against a third ion (e.g., Na/Ca exchange), then the exchange may depolarize the membrane potential.     

Sucrose density gradient centrifugation of human myometrial and myomal progestin receptors

Progestin receptors present in cytosols of myomal and myometrial origin were analyzed on sucrose density gradients of low ionic strength using a vertical tube rotor. Short term incubations (90 min) contained mainly 8S receptors. Most myometrial but few myomal preparations contained an additional 4S component, the presence of which was hormone dependent. None could be detected if the serum concentration of estradiol was high. Myometrial receptors incubated over night exhibited only a 4S peak, whereas many myomal ones still sedimented as 8S peaks. The 8 to 4S shift represented a transformation of the receptors and not a sequential interaction of the hormone with first an 8S and then a 4S entity. The transforming activity contained in the myometrial cytosol could also attack the myomal receptor as shown by mixing experiments. Only the 8S peak was observed, whenever the cytosol was first fractionated and then incubated with the hormone. Thus the transformation occurred only in presence of [3H]promegestone. It could be suppressed with protease inhibitors. It is concluded that these tissues contain a receptor processing protease which (1) is much less prominent in myomal than myometrial cytosol, which (2) attacks only the occupied receptors, and (3) the activity of which is hormone dependent.     

Sequence specific antibodies to a deoxyribodinucleotide

Antibodies were elicited in rabbits to the dinucleotide, dpTpA, conjugated to a carrier protein. Among a few deoxyribomono- and oligonucleotides tested for binding to the antibodies by nitrocellulose tilter binding assay, only dpTpA and dpTpA containing oligonucleotides showed binding. The inhibition of the binding of 3H-dpTpA to the antibodies by various nonradioactive mono- and oligonucleotides showed that the antibodies were sequence specific and recognized the whole molecule of dpTpA. dpTpA specific antibodies were purified by a two step affinity chromatography. By competition studies, it was found that the purified antibodies bound denatured DNA at dpTpA sequences. The antibodies did not bind RNA.     

Heliobacterium chlorum: cell organization and structure

The basic cellular organization of Heliobacterium chlorum is described using the freeze-etching technique. Internal cell membranes have not been observed in most cells, leading to the conclusion that the photosynthetic apparatus of these organisms must be localized in the cell membrane of the bacterium. The two fracture faces of the cell membrane are markedly different. The cytoplasmic (PF) face is covered with densely packed particles averaging 8 nm in diameter, while the exoplasmic (EF) face contains far fewer particles, averaging approximately 10 nm in diameter. Although a few differentiated regions were noted within these fracture faces, the overall appearance of the cell membrane was remarkably uniform. The Heliobacterium chlorum cell wall is a strikingly regular structure, composed of repeating subunits arranged in a rectangular pattern at a spacing of 11 nm in either direction. We have isolated cell wall fragments by brief sonication in distilled water, and visualized the cell wall structure by negative staining as well as deep-etching.Abbreviations PF protoplasmic fracture face - EF exoplasmic fracture face     

Structural forms, stabilities and transitions in double-helical poly(dG-dC) as a function of hydration and NaCl content. An infrared spectroscopic study.

The poly(dG-dC) helical duplex forms a modified, B-family structure (B*) at very high hydration and a normal B structure at slightly lower hydration. The B* structure is slightly different in sugar-phosphate and base-stacking conformations than the B structure. Increasing the hydration or decreasing the NaCl content stabilizes B* with respect to B. Poly(dG-dC) forms the Z structure at low NaCl contents when the hydration is sufficiently reduced. At moderate NaCl content, the B to Z transition is sharp and cooperative for hydration with D2O. Hydration with H2O broadens the transition which occurs at lower hydration. This suggests that hydrogen bonding is stronger in the Z structure and helps stabilize Z over B. IR spectra may be used to quantitatively estimate the fractions of B and Z structures present in a sample. Some new indicator bands are described.