In vitro regeneration of apomictic bluestem grasses

Explants from immature inflorescences of four genotypes of Old World bluestem grasses, (Bothriochloa spp.), produced callus tissue on Linsmaier and Skoog (RM) and 1/2 Murashige and Skoog (1/2 MS) media containing high levels of growth regulators. Callus masses were composed of two distinct tissue types, one a compact, white, embryogenic portion (E calli), the other soft, translucent, gelatinous and nonembryogenic (NE calli). When transferred to medium with a reduced level of 2,4-D, and/or supplemented with zeatin, E callus underwent further organization culminating in shoot production. Light and scanning electron microscopy confirmed the embryogenic pathway of differentiation. Genotype significantly affected callus induction frequency and the number of plants regenerated. The RM medium induced more explants to initiate callus compared to the 1/2 MS medium. Age of the inflorescence explant, as indicated by size, was critical for callus induction. Inflorescences with racemes 8 mm in length were superior to older ones. Five-hundred-twenty-two plantlets were regenerated and grown to maturity.     

Photolabile and paramagnetic reagents for the investigation of transmembrane signaling events

To investigate the dynamics of membrane processes that may be integral components of specific transmembrane signaling events we have synthesized several novel paramagnetic probes and their photoreactive counterparts. The structure of these probes was designed to (1) restrict “flipping” across the membrane bilayer; (2) contain paramagnetic or photoreactive moieties that could be placed at specific depths within the bilayer; (3) provide information about membrane structure as well as dynamics of protein movement; and (4) in the case of the photoreactive probes, be of high specific radioactivity. The molecules described in this paper consist of amino acid, dipeptide, or carbohydrate groups attached to arylazide- or nitroxide-bearing fatty acids. The synthesis and initial characterization of these membrane probes is described.     

Excitotoxic Brain Injury Suppresses Striatal High-Affinity Glutamate Uptake in Perinatal Rats

In immature rodent brain, the glutamate receptor agonist N-methyl-D-aspartate (NMDA) is a potent neurotoxin. In postnatal day (PND)-7 rats, intrastriatal injection of 25 nmol of NMDA results in extensive ipsilateral forebrain injury. In this study, we examined alterations in high-affinity [3H]glutamate uptake (HAGU) in NMDA-lesioned striatum. HAGU was assayed in synaptosomes, prepared from lesioned striatum, the corresponding contralateral striatum, or unlesioned controls. Twenty-four hours after NMDA injection (25 nmol), HAGU declined 44 +/- 8% in lesioned tissue, compared with the contralateral striatum (mean +/- SEM, n = 6 assays, p less than 0.006, paired t test). Doses of 5-25 nmol of NMDA resulted in increasing suppression of HAGU (5 nmol, n = 3; 12.5 nmol, n = 3; and 25 nmol, n = 5 assays; p less than 0.01, regression analysis). The temporal evolution of HAGU suppression was biphasic. There was an early transient suppression of HAGU (-28 +/- 4% at 1 h; p less than 0.03, analysis of variance, comparing changes at 0.5, 1, 2, and 3 h after lesioning); 1 or 5 days postinjury there was sustained loss of HAGU (at 5 days, -56 +/- 11%, n = 3, p less than 0.03, paired t test, lesioned versus contralateral striata).(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects on phytoplankton of nutrients added in conjunction with acidification

1. The effects of nitric acidification on phytoplankton were studied in a small, eperimentally manipulated, oligotrophic lake (L302N) in the Eperimental Lakes Area of Canada. The focus was altered after 9 years of acidification to investigate the possibility of using nutrient additions to stimulate recovery, followed by a controlled incremental recovery, in which the pH was increased to a predetermined target level. 2. Five years of additions of HNO₃ to L302N reduced its pH from 6.5 to 6.1. Nitrate concentration increased because the algal community was severely P deficient. The phytoplankton community structure and productivity were not significantly affected by these additions. 3. The phytoplankton community was significantly affected when pH was subsequently decreased over three successive years from 6.1 to 5.1 by the addition of HCl. Dominance shifted from chrysophytes to a co-dominance of chlorophytes and dinoflagellates, which altered the size structure of the community. Species diversity significantly decreased, although phytoplankton productivity remained unchanged. 4. At pH 5.1 nitrate and sulphate additions were made, creating conditions like those in lakes in eastern North America, which receive high loadings of nitrogen from the atmosphere. The phytoplankton assemblage shifted to dominance by small coccoidal chlorophytes. However, biomass and productivity were unaffected. 5. Finally, phosphate, as phosphoric acid, was added, along with nitrate and sulphate, to the epilimnion, which stimulated internal alkalinity generation and productivity. It is concluded that CO₂ concentrations and the form of N (nitrate vs. ammonia) affect algal composition but that P determines algal biomass and productivity. Chlorophytes were found to be good competitors for P when N and CO₂ were high; it is epected that cyanobacteria would be more competitive for P in low CO₂ systems. Conversely, dinoflagellates are most competitive in systems with low pH and high P, such as that which occurred in L302N. Although the P additions reduced N concentrations and created alkalinity, this is not a recommended remedial procedure in acidified lakes because it enhanced dinoflagellate abundance, which has been associated with fish kills. 6. When all additions ceased, the pH of L302N recovered from 5.1 to 5.8, chrysophytes and chlorophytes became more abundant and dinoflagellates decreased in abundance. Phytoplankton biomass decreased and species diversity increased. Phytoplankton productivity remained unchanged     

Hb Mobile [α2β2 73(E17)Asp → Val]: A new variant

A new hemoglobin variant found in a mother and her child was characterized by column chromatography of the tryptic hydrolysate of the aminoethylated, glycinamidated -chain, followed by chymotryptic digestion of the abnormal T-9 peptide and amino acid analyses. It was shown to be _{2 2} 73(E17) Asp Val and named Hb Mobile.This work was supported in part by Research Grants AM0780 and AM13173 from the National Institute for Arthritis and Metabolic Disease.     

Structures of peptide agonists for human protease activated receptor 2

Protease activated receptor 2 (PAR2) is an unusual G-protein coupled receptor in being self-activated, after pruning of the N-terminus by serine proteases like trypsin and tryptase. Short synthetic peptides corresponding to the newly exposed N-terminal hexapeptide sequence also activate PAR2 on immunoinflammatory, cancer and many normal cell types. (1)H nuclear magnetic resonance (NMR) and circular dichroism (CD) spectroscopy were used here to search for structural clues to activating mechanisms of the hexapeptide agonists SLIGRL (rat), SLIGKV (human) and the peptidomimetic analogue, 2-furoyl-LIGRLO. Either with a free or acetyl capped N-terminus, these agonist peptides display significant propensity in aprotic (DMSO) or lipidic (water-SDS) solvents for turn-like conformations, which are predicted to be receptor-binding conformations in the transmembrane or loops region of PAR2. These motifs may be valuable for the design of small molecule PAR2 agonists and antagonists as prospective new drugs for regulating inflammatory and proliferative diseases.     

Preliminary survey of carnivore hemoglobin compositions

Summary The amino acid differences among 12 chains and 10 chains of carnivore hemoglobins are given. These hemoglobins conform to the substitution rate found for hemoglobins in general. A table compares the differences among known-chain sequences when deduced from tryptic peptide compositions and when taken from actual sequences. Among the carnivore hemoglobins studies, tryptic peptide compositions are about 14 % low in giving the number of sequence differences.