II. Selective accumulation of MPP+ in the substantia nigra: A key to neurotoxicity?

The neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) is rapidly metabolized to a 1-methyl-4-phenylpyridinium species (MPP+) in the squirrel monkey. After administration of toxic doses of MPTP, the concentration of MPP+ in the substantia nigra appears to increase during the first 72 hours, reaching the highest concentration of any central nervous system (CNS) tissue studied. In contrast, the concentration of this compound in other brain areas suggested time dependent elimination during the same period. Pretreatment of animals with the monoamine oxidase (MAO) inhibitor pargyline blocks both the neurotoxic action and the biotransformation of MPTP. In animals given pargyline and MPTP, initial MPTP levels are much higher in all brain regions than in those not receiving pargyline, but by 12 hours, MPTP levels had fallen rapidly in all regions except the substantia nigra and the eye. It may be that the selective toxicity of MPTP is related in some way to the accumulation of its oxidized metabolite in the substantia nigra.     

Studies on Galalpha3-binding proteins: comparison of the glycosphingolipid binding specificities of Marasmius oreades lectin and Euonymus europaeus lectin

The carbohydrate binding preferences of the Galalpha3Galbeta4 GlcNAc-binding lectins from Marasmius oreades and Euonymus europaeus were examined by binding to glycosphingolipids on thin-layer chromatograms and in microtiter wells. The M. oreades lectin bound to Galalpha3-terminated glycosphingolipids with a preference for type 2 chains. The B6 type 2 glycosphingolipid (Galalpha3[Fucalpha2]Galbeta4GlcNAcbeta3Galbeta4Glcbeta1Cer) was preferred over the B5 glycosphingolipid (Galalpha3Galbeta4GlcNAcbeta3Galbeta4Glcbeta1Cer), suggesting that the alpha2-linked Fuc is accommodated in the carbohydrate binding site, providing additional interactions. The lectin from E. europaeus had broader binding specificity. The B6 type 2 glycosphingolipid was the best ligand also for this lectin, but binding to the B6 type 1 glycosphingolipid (Galalpha3[Fucalpha2]Galbeta3GlcNAcbeta3Galbeta4Glcbeta1Cer) was also obtained. Furthermore, the H5 type 2 glycosphingolipid (Fucalpha2Galbeta4GlcNAcbeta3Galbeta4Glcbeta1Cer), devoid of a terminal alpha3-linked Gal, was preferred over the the B5 glycosphingolipid, demonstrating a significant contribution to the binding affinity by the alpha2-linked Fuc. The more tolerant nature of the lectin from E. europaeus was also demonstrated by the binding of this lectin, but not the M. oreades lectin, to the x2 glycosphingolipid (GalNAcbeta3Galbeta4GlcNAcbeta3Galbeta4Glcbeta1Cer) and GlcNAcbeta3Galbeta4GlcNAcbeta3Galbeta4Glcbeta1Cer. The A6 type 2 glycosphingolipid (GalNAcalpha3[Fucalpha2]Galbeta4GlcNAcbeta3Galbeta4Glcbeta1Cer) and GalNAcalpha3Galbeta4GlcNAcbeta3Galbeta4Glcbeta1-Cer were not recognized by the lectins despite the interaction with B6 type 2 glycosphingolipid and the B5 glycosphingolipid. These observations are explained by the absolute requirement of a free hydroxyl in the 2-position of Galalpha3 and that the E. europaea lectin can accommodate a GlcNAc acetamido moiety close to this position by reorienting the terminal sugar, whereas the M. oreades lectin cannot.     

Messenger ribonucleoprotein complexes isolated with oligo(dT)-cellulose chromatography from kidney polysomes

As an initial step towards understanding the role of mRNP complexes in translational regulation during compensatory renal hypertrophy, characteristics of polysome-associated mRNP isolated by affinity chromatography were studied. Renal mRNP contained 15–30% of the counts after a 1 hr pulse with ³H-orotic acid; it sedimented mainly between 10S and 100S and had a buoyant density of 1.42–1.44 g/cm³. RNA derived from the mRNP sedimented between 5S and 40S on sucrose density gradients, with the greatest radioactivity in the region of 15S. After labeling with ³H-adenine for 1 hr, up to 17% of the radioactivity present in the mRNP-associated RNA was resistant to digestion by pancreatic and T1 ribonucleases. The mRNP protein moiety contained six polypeptides with molecular weights 69,000, 75,000, 80,000, 100,000, 109,000, and 118,000 daltons, which were undetected in the material not binding to oligo(dT)-cellulose.     

Increased diversity and concordant shifts in community structure of coral‐associated Symbiodiniaceae and bacteria subjected to chronic human disturbance

Both coral‐associated bacteria and endosymbiotic algae (Symbiodiniaceae spp.) are vitally important for the biological function of corals. Yet little is known about their co‐occurrence within corals, how their diversity varies across coral species, or how they are impacted by anthropogenic disturbances. Here, we sampled coral colonies (n = 472) from seven species, encompassing a range of life history traits, across a gradient of chronic human disturbance (n = 11 sites on Kiritimati [Christmas] atoll) in the central equatorial Pacific, and quantified the sequence assemblages and community structure of their associated Symbiodiniaceae and bacterial communities. Although Symbiodiniaceae alpha diversity did not vary with chronic human disturbance, disturbance was consistently associated with higher bacterial Shannon diversity and richness, with bacterial richness by sample almost doubling from sites with low to very high disturbance. Chronic disturbance was also associated with altered microbial beta diversity for Symbiodiniaceae and bacteria, including changes in community structure for both and increased variation (dispersion) of the Symbiodiniaceae communities. We also found concordance between Symbiodiniaceae and bacterial community structure, when all corals were considered together, and individually for two massive species, Hydnophora microconos and Porites lobata, implying that symbionts and bacteria respond similarly to human disturbance in these species. Finally, we found that the dominant Symbiodiniaceae ancestral lineage in a coral colony was associated with differential abundances of several distinct bacterial taxa. These results suggest that increased beta diversity of Symbiodiniaceae and bacterial communities may be a reliable indicator of stress in the coral microbiome, and that there may be concordant responses to chronic disturbance between these communities at the whole‐ecosystem scale.     

D-periodic distribution of collagen type IX along cartilage fibrils

It has recently become apparent that collagen fibrils may be composed of more than one kind of macromolecule. To explore this possibility, we developed a procedure to purify fibril fragments from 17-d embryonic chicken sternal cartilage. The fibril population obtained shows, after negative staining, a uniformity in the banding pattern and diameter similar to the fibrils in situ. Pepsin digestion of this fibril preparation releases collagen types II, IX, and XI in the proportion of 8:1:1. Rotary shadowing of the fibrils reveals a d-periodic distribution of 35-40-nm long projections, each capped with a globular domain, which resemble in form and dimensions the aminoterminal globular and collagenous domains, NC4 and COL3, of type IX collagen. The monoclonal antibody (4D6) specific for an epitope close to the amino terminal of the COL3 domain of type IX collagen bound to these projections, thus confirming their identity. Type IX collagen is therefore distributed in a regular d-periodic arrangement along cartilage fibrils, with the chondroitin sulfate chain of type IX collagen in intimate contact with the fibril.     

Effects of ultraviolet-B radiation on plants during mild water stress. IV. The insensitivity of soybean internal water relations to ultraviolet-B radiation

The combined effects of ultraviolet-B (UV-B, 280–320 nm) radiation and water stress were investigated on the water relations of greenhouse grown soybean [ Glycine max (L.) Merr. cv. Essex]. On a weighted (Caldwell 1971), total daily dose basis, plants received either 0 or 3 000 effective J m² UV-B_BE supplied by filtered FS-40 sunlamps. The latter dose simulated the solar UV-B radiation anticipated at College Park, Maryland, U.S.A. (39°N latitude) in the event that the global stratospheric ozone column is reduced by 25%. Plants were either well-watered or preconditioned by drought stress cycles. Diurnal measurements of water potential and stomatal conductance were made on the youngest fully expanded leaf. Various internal water relations parameters were determined for detached leaves. Plants were monitored before, during and after water stress. There were no significant differences in leaf water potential or stomatal conductance between treatments before plants were preconditioned to water stress. However, drought stress resulted in significantly lower midday and afternoon leaf water potentials and lower leaf conductances as compared to well-watered plants. UV-B radiation had no additional effect on leaf water potential; however, UV did result in lower leaf conductances in plants preconditioned to water stress. Turgid weight:dry weight ratio, elastic modulus, bound water and relative water content were unaffected by UV-B radiation. Osmotic potentials at full and zero turgor were significantly lower in the drought stressed treatments as compared to well-watered plants.