Expression of the plasma membrane proteolipid in mouse neuroblastoma cells: transient increase in synthesis during differentiation with N6,O2-dibutyryl adenosine 3',5'-cyclic monophosphate

We have examined the regulation of plasma membrane proteolipid (PM-PLP) synthesis and steady-state levels in mouse NB2a/d1 neuroblastoma cells during differentiation with dibutyryl cyclic AMP (dbcAMP) and retinoic acid (RA), agents which have been previously shown to induce the elaboration of exclusively axonal or dendritic neurites, respectively. We report that a PM-PLP-immunoreactive species is expressed by this neuroblastoma cell line, and that its expression is regulated by specific states of differentiation. Differentiation of cells with dbcAMP was accompanied by an initial 2-fold increase in this PM-PLP immunoreactive species at 24 h after treatment, which returned to control levels by 96 h after treatment. By contrast, no significant increase in synthesis was detected when cells were treated with RA. Protein blot analysis of PM-PLP in dbcAMP-treated cells indicated that there was little change in its steady-state level until 96 h following treatment, at which time a reduction of 40% was observed. Throughout induced differentiation with dbcAMP, NB2a/d1 cells continued to express a PM-PLP-immunoreactive species which comigrated on immunoblot analysis with PM-PLP form characteristic of embryonic brain (14-16 kDa), and apparently did not express the PM-PLP form characteristic of adult brain (18 kDa).     

Red yeast rice fermentation with Bacillus subtilis B2 under blue light-emitting diodes increases antioxidant secondary products (Manuscript ID: BPBSE-18-0387)

Bioprocess and Biosystems Engineering - Light and bacteria can be used in combination to enhance secondary metabolite production during fermentation. Red yeast rice powder (RYRP) was inoculated...     

Suppression of interferon synthesis by the pesticide carbaryl as a mechanism for enhancement of goldfish virus-2 replication

Interferon production was demonstrated by the goldfish-derived CAR cell line in response to infection by goldfish virus-2. Supernatants of infected cultures provided antiviral protection to CAR cells and another cell line derived from goldfish, ABIII. The protective factor retained activity after ultracentrifugation, dialysis, freezing and thawing, acid treatment (pH 2), or heating to 56 degrees C but was sensitive to trypsin. Supernatants of infected cultures did not affect adsorption of virus. Previous studies have shown that replication of goldfish virus type 2 is enhanced by pretreatment of cultures with subcytotoxic concentrations of carbaryl. In the present study, pesticide-treated cultures were found to synthesize reduced levels of interferon.     

Effects of germanium dioxide,an inhibitor of diatom growth,on the microscopic laboratory cultivation stage of the kelp, <Emphasis Type="Italic">Laminaria saccharina</Emphasis>

The kelp, Laminaria saccharina, is an economically and biologically valuable seaweed used in integrated multi-trophic aquaculture. The development of the initial microscopic phase of the life cycle of this species is carried out in the laboratory. A treatment of germanium dioxide (GeO₂) can be applied to algal laboratory cultures to inhibit the growth of contaminating diatoms. Previous studies reported inhibitory effects also affecting the macroscopic stage of the life cycle of L. saccharina, the sporophyte, but the effects on the microscopic gametophytic life stage are unknown. To determine the effects, cultures of L. saccharina were treated with varying concentrations of GeO₂ and the resulting lengths and surface area of the juvenile sporophytes produced were measured. It was determined that GeO₂ follows a typical dose-effect pattern, increasing the growth rate of juvenile sporophytes until a critical point at which an inhibitory effect was observed. To obtain juvenile sporophytes ready for deployment to aquaculture sites in the shortest time and to successfully reduce diatom contamination, it was determined that a concentration between 0.10 and 0.50mL of GeO₂ per litre of seawater, applied at day 8, was the most efficient.     

Mechanistic Positron Emission Tomography Studies of 6-[18F]Fluorodopamine in Living Baboon Heart: Selective Imaging and Control of Radiotracer Metabolism Using the Deuterium Isotope Effect

Abstract: Mechanistic positron emission tomography (PET) studies using the deuterium isotope effect and specific pharmacological intervention were undertaken to examine the behavior of 6-[¹⁸F]fluorodopamine (6-[¹⁸F]FDA; 1 ) and (?)-6-[¹⁸F]fluoronorepinephrine {(?)-6-[¹⁸F]FNE; 2 } in the baboon heart. Two regiospecifically deuterated derivatives of 6-[¹⁸F]FDA [α,α-D₂(3 ) and β,β-D₂ (4 )] were used to assess the contributions of monoamine oxidase (MAO) and dopamine β-hydroxylase, respectively, to the clearance kinetics of 6-[¹⁸F]FDA. Compound 3 showed a reduced rate of clearance, consistent with MAO-catalyzed cleavage of the α C-D bond, whereas compound 4 showed no change, indicating that cleavage of the β C-D bond is not a rate-limiting step. Pretreatment with pargyline, an MAO inhibitor, also decreased the rate of clearance. Desipramine and tomoxetine [norepinephrine (NE) uptake inhibitors], but not GBR-12909 (a dopamine uptake inhibitor), blocked the uptake of both (?)-6-[¹⁸F]FNE and 6-[¹⁸F]FDA, with (?)-6-[¹⁸F]FNE showing a higher degree of blockade. Chiral HPLC demonstrated that 6-[¹⁸F]FDA is stereoselectively converted to (?)-6-[¹⁸F]FNE in vivo in the rat heart. These studies demonstrate that (a) the more rapid clearance of 6-[¹⁸F]FDA relative to (?)-6-[¹⁸F]FNE can be largely accounted for by metabolism by MAO; (b) selective deuterium substitution can be used to protect a radiotracer from metabolism in vivo and to favor a particular pathway; (c) 6-[¹⁸F]FDA and (?)-6-[¹⁸F]FNE share the NE transporter; (d) 6-[¹⁸F]FDA is stereoselectively converted to (?)-6-[¹⁸F]FNE in vivo; and (e) the profile of radioactivity in the heart for 6-[¹⁸F]FDA is complex, probably including labeled metabolites as well as neuronal and nonneuronal uptake.     

Freezing of bovine embryos: Effects of embryo quality,time from thawing to transfer and number frozen per vial

Over a ten-month period 736 embryos were collected from 103 cows induced to superovulate. Embryos were frozen in 10% glycerol and, after thawing, 655 appeared viable and were transferred nonsurgically to synchronous recipients. When embryos did not appear degenerative before freezing and transfers were performed within 3 hr of thawing, 40% of the embryos resulted in pregnancies. If embryos showed signs of degeneration before freezing, only 27% resulted in pregnancies after thawing and transfer. Similarly, when non-degenerative embryos were kept in Casou straws for more than 3 hr after thawing, fewer produced pregnancies (25%) following transfer. The number of embryos frozen in each vial ranged from one to 18 and this had no significant effect on success. By selecting embryos to be frozen and transferring them soon after thawing, the proportion of embryos surviving the freeze-thaw process can be very high.     

Effects of Deer Browsing, Fabric Mats, and Tree Shelters on Quercus rubra Seedlings

We examined the effects of deer browsing, fabric mats, and tree shelters on the growth and survival of Quercus rubra (northern red oak) seedlings planted as part of a reforestation project in southcentral Minnesota. Browsing by white-tailed deer occurred on 68.6% of the seedlings growing without tree shelters, whereas only 3.6% of the trees were browsed by mice or voles and 2.5% were browsed by rabbits. Fabric mats, used to control competition from herbaceous plants for roughly half of the seedlings included in the study, had a detrimental effect overall. Seedlings grown with mats had a greater frequency of deer browsing and a greater chance of dying than seedlings grown without mats. Stem height for seedlings browsed by deer was less overall than for nonbrowsed seedlings, although this pattern varied with use of fabric mats and plot location. The use of plastic tree shelters effectively prevented deer browsing and reduced the mortality rate from 34.6% to 3.2%. Our results indicate that fabric mats should not be used in restoration projects with large deer populations. They also suggest that planting seedlings away from existing forest edges and using seedling protection measures such as tree shelters will increase seedling survival and growth in future restoration projects.