The Medusa of Velella velella (Linnaeus, 1758) (Hydrozoa, Chondrophorae)

A mature medusa of Velella velella (Linnaeus, 1758) is reportedfor the first time from the North Atlantic; previously adultmedusae were known only from the Mediterranean. This specimen,collected by SCUBA divers, is the largest specimen recordedto date. Distinctive features are: two opposite adaxial-axialpairs of perradial capitate tentacles; two marginal bulbs lackingtentacles; conical manubrium with 4 perradial longitudinal gonads;perradial exumbrellar cnidae tracts; cnidome of stenoteles andmacrobasic euryteles; and zooxanthellae within the subumbrella.Since this specimen was collected near the surface and has zooxanthellae,it is likely that V. velella medusae are epipelagic.     

Purification of the protein employed in an immunodiffusion test to diagnose infectious bovine rhinotracheitis.

The antigen used in an immunodiffusion test to diagnose infectious bovine rhinotracheitis has been purified by affinity chromatography. The homogeneity of the antigen was indicated by sedimentation rate and sedimentation equilibrium experiments. A So20,w of 0.749 was determined and a molecular weight of 8900 was calculated from sedimentation equilibrium analysis. The purified antigen formed precipitin lines of identity with crude diagnostic antigen. Purified antigen remained serologically active in the immunodiffusion test after lyophilization and subsequent reconstitution.     

Alcohol-inducible cytochrome P-450IIE1 lacking the hydrophobic NH2-terminal segment retains catalytic activity and is membrane-bound when expressed in Escherichia coli

We have expressed in Escherichia coli a cDNA encoding rabbit liver cytochrome P-450IIE1, the ethanol-inducible P-450. The expressed P-450 is located primarily in the bacterial inner cell membrane and comprises 3% of the E. coli total membrane protein. The partially purified cytochrome exhibits a reduced CO difference spectrum with a maximum at 452 nm, characteristic of P-450IIE1, and solubilized membranes or partially purified P-450 preparations reconstituted with NADPH-cytochrome P-450 reductase and phosphatidylcholine catalyze the deethylation of N-nitrosodiethylamine with a turnover number equal to that of purified liver P-450IIE1 (approximately 4.5 nmol/min/nmol of P-450). A modified IIE1 cDNA that encodes a protein lacking amino acids 3-29, a proposed membrane anchor for cytochrome P-450, was also expressed in E. coli and, unexpectedly, the shortened protein was also found to be predominantly located in the bacterial inner membrane rather than the cytosol. Like the full-length protein, this truncated cytochrome has a reduced CO difference spectrum characteristic of P-450IIE1 and is fully active in the deethylation of N-nitrosodiethylamine. These results demonstrate that the NH2-terminal hydrophobic segment is not solely responsible for attachment to the membrane and evidently is not required for proper protein folding or catalytic activity.     

Isolation of selective 3H-chlornaltrexamine-bound complexes,possible opioid receptor components in brains of mice

The nonequilibrium narcotic antagonist, chlornaltrexamine (CNA) was used to bind selectively and covalently pioid specific sites on brain membrane preparations. Selective binding of [³H]CNA occured with a saturation maximum of 185 fmol/mg protein. Bound [³H]CNA was extracted with Triton X-100, dialyzed against Brij 36T, precipitated with trichloroacetic acid and chromatographed on an ultrogel AcA 22 column. The elution profile suggests that this extract contains a minimum of four selective [³H]CNA complexes. At least two of these complexes migrate in a single large peak. Column calibration showed that this peak eluted at 590,000 daltons. One of these specific [³H]CNA complexes elutes at the elution volume of the column and is dialyzable. Finally, putative aggregate of these complexes elutes with the void volume.     

DEVELOPMENT AND ORGANIZATION OF THE SECONDARY VESSEL SYSTEM IN POPULUS GRANDIDENTATA

The apical 22 cm of a dormant, first-year sprout of Populus grandidentata was sectioned serially, and the primary and secondary xylem systems were studied microscopically and graphically reconstructed. A total of 15 nodes was present on the mature stem and 14 foliar primordia in the dormant bud. The vascular traces in the lower portion of the mature stem conformed to a 2/5 phyllotaxy while those of the upper portion and within the dormant bud conformed to a 3/8 phyllotaxy. The 2/5 to 3/8 phyllotactic transition occurred in an extremely precise and systematic two-step pattern: (1) The lateral traces shifted to a new point of origin on the parent central trace, and (2) three new central traces were initiated in sequence by divergences from left-traces. Metaxylem, when followed downward, conformed to the arrangement of the procambial trace system only within one orthostichy. Below this point, the metaxylem components of lateral traces physically separated from those of the protoxylem and continued downward on a new course. Metaxylem vessels produced by the trace cambium originated from a postulated vessel-generating center at the stem-petiole junction. Each metaxylem vessel developing basipetally through the primary body was continuous with a secondary vessel developing basipetally in the secondary body. Because secondary development closed the vascular cylinder, vessels originating from developing leaves or primordia situated at higher levels in the shoot were displaced radially outward when they entered the secondary xyelm. The distribution of vessels in the secondary xylem can therefore be accounted for by a knowledge of the production and distribution of metaxylem vessels in the primary body.     

An NO Donor Approach to Neuroprotective and Procognitive Estrogen Therapy Overcomes Loss of NO Synthase Function and Potentially Thrombotic Risk

Selective estrogen receptor modulators (SERMs) are effective therapeutics that preserve favorable actions of estrogens on bone and act as antiestrogens in breast tissue, decreasing the risk of vertebral fractures and breast cancer, but their potential in neuroprotective and procognitive therapy is limited by: 1) an increased lifetime risk of thrombotic events; and 2) an attenuated response to estrogens with age, sometimes linked to endothelial nitric oxide synthase (eNOS) dysfunction. Herein, three 3^rd generation SERMs with similar high affinity for estrogen receptors (ERα, ERβ) were studied: desmethylarzoxifene (DMA), FDMA, and a novel NO-donating SERM (NO-DMA). Neuroprotection was studied in primary rat neurons exposed to oxygen glucose deprivation; reversal of cholinergic cognitive deficit was studied in mice in a behavioral model of memory; long term potentiation (LTP), underlying cognition, was measured in hippocampal slices from older 3×Tg Alzheimer''s transgenic mice; vasodilation was measured in rat aortic strips; and anticoagulant activity was compared. Pharmacologic blockade of GPR30 and NOS; denudation of endothelium; measurement of NO; and genetic knockout of eNOS were used to probe mechanism. Comparison of the three chemical probes indicates key roles for GPR30 and eNOS in mediating therapeutic activity. Procognitive, vasodilator and anticoagulant activities of DMA were found to be eNOS dependent, while neuroprotection and restoration of LTP were both shown to be dependent upon GPR30, a G-protein coupled receptor mediating estrogenic function. Finally, the observation that an NO-SERM shows enhanced vasodilation and anticoagulant activity, while retaining the positive attributes of SERMs even in the presence of NOS dysfunction, indicates a potential therapeutic approach without the increased risk of thrombotic events.     

Proteins searching for their target on DNA by one-dimensional diffusion: overcoming the “speed-stability” paradox

The sequence dependence of DNA-protein interactions that allows proteins to find the correct reaction site also slows down the 1D diffusion of the protein along the DNA molecule, leading to the so-called “speed-stability paradox,” wherein fast diffusion along the DNA molecule is seemingly incompatible with stable targeting of the reaction site. Here, we develop diffusion-reaction models that use discrete and continuous Gaussian random 1D diffusion landscapes with or without a high-energy cut-off, and two-state models with a transition to and from a “searching” mode in which the protein diffuses rapidly without recognizing the target. We show the conditions under which such considerations lead to a predicted speed-up of the targeting process, and under which the presence of a “searching” mode in a two-state model is nearly equivalent to the existence of a high-energy cut-off in a one-state model. We also determine the conditions under which the search is either diffusion-limited or reaction-limited, and develop quantitative expressions for the rate of successful targeting as a function of the site-specific reaction rate, the roughness of the DNA-protein interaction potential, and the presence of a “searching” mode. In general, we find that a rough landscape is compatible with a fast search if the highest energy barriers can be avoided by “hopping” or by the protein transitioning to a lower-energy “searching” mode. We validate these predictions with the results of Brownian dynamics, kinetic Metropolis, and kinetic Monte Carlo simulations of the diffusion and targeting process, and apply these concepts to the case of T7 RNA polymerase searching for its target site on T7 DNA.