Acid-catalyzed hydration of reduced nicotinamide adenine dinucleotide and its analogues.

The rate of the primary acid modification reaction of 1,4-dihydronicotinamide adenine dinucleotide (NADH) and 1,4-dihydro-3-acetylpyridine adenine dinucleotide (APADH) and their analogues has been studied over a wide pH range (pH 1-7) with a variety of general acid catalysts. The rate depends on [H+] at moderate pH and becomes independent of [H+] at low pH. This behavior is attributed to substrate protonation at the carbonyl group (pK of NADH = 0.6). The reaction is general acid catalyzed; large solvent deuterium isotope effects are observed for the general acid and lyonium ion terms. Most buffers cause a linear rate increase with increasing buffer concentration, but certain buffers cause a hyperbolic rate increase. The nonlinear buffer effects are due to complexation of the buffer with the substrate, rather than to a change in rate-limiting step. The rate-limiting step is a proton transfer from the general acid species to the C5 position of the substrate. Anomerization is not a necessary first step in the case of the primary acid modification reaction of beta-NADH, in which beta to alpha anomerization takes place.     

Wiring diagrams of MAPK regulation by MEKK1, 2, and 3.

Mitogen-activated protein kinase (MAPK) pathways are activated by a plethora of stimuli. The literature is filled with papers describing the activation of different MAPKs by almost any stimulus or insult imaginable to cells. In this review, we use signal transduction wiring diagrams to illustrate putative upstream regulators for the MAPK kinase kinases, MEKK1, 2, and 3. Targeted gene disruption of MEKK1, 2, or 3 defined phenotypes for each MEKK associated with loss of specific MAPK regulation. Genetic analysis of MEKK function clearly defines specific components of the wiring diagram that require MEKK1, 2, or 3 for physiological responses. We propose that signal transduction network wiring diagrams are valuable tools for hypothesis building and filtering physiologically relevant phenotypic responses from less connected protein relations in the regulation of MAPK pathways.     

ULTRASTRUCTURE AND ECOLOGY OF AUREOCOCCUS ANOPHAGEFERENS GEN. ET SP. NOV. (CHRYSOPHYCEAE): THE DOMINANT PICOPLANKTER DURING A BLOOM IN NARRAGANSETT BAY,RHODE ISLAND,SUMMER 19851

Observations of a marked cessation of feeding in filter feeding animals maintained in flowing Narragansett Bay seawater in June 1985 drew our attention to a bloom of a golden alga 2 μm in diameter at unprecedented populations of 10⁹ cells. L^?1. This picoplankter lacked morphological features useful in discriminating it from other similar sized forms with either phase contrast or epifluorescence light microscopy. Natural populations of picoplankton, obtained from the height of the bloom until its decline, were examined in thin section with transmission electron microscopy. A cell with a single chloroplast, nucleus, and mitochondrion and an unusual exocellular polysaccharide-like layer was apparently the bloom alga. The ultrastructure of this alga is consistent with that of the Chrysophyceae, and a new genus and species, Aureococcus anophagefferens is described. Attempts to grow this previously unrecognized picoplanktonic alga as an obligate phototroph failed and only yielded cultures of other previously described picoalgae. Facultative and obligate phagotrophic protists with ingested cells of Aureococcus were only observed as the bloom waned and minute diatoms became common. Cells of A. anophagefferens with virus particles typical for picoalgae occurred throughout the bloom. Populations of the usually dominant photosynthetic picoplankter, the cyanobacterium Synechococcus Nägeli, were depressed during the bloom. This could be due in part to selective grazing on Synechococcus rather than Aureococcus by elevated populations of Calycomonas ovalis Wulff which accompanied the algal bloom.     

Effect of elevating serum lipids on luteinizing hormone response to gonadotrophin releasing hormone challenge in energy-deficient anestrous heifers

A study was conducted to evaluate the effect of feeding a bypass fat on luteinizing hormone (LH) response to gonadotrophin releasing hormone (GnRH) in noncyclic Holstein heifers. Twelve cyclic Holstein heifers were fed a complete diet at 40% net energy for maintenance (NE(m)) until cessation of ovarian activity. Based on weights and condition scores, heifers were assigned to either a control or treatment diet containing 0.45 kg bypass fat and fed at an energy level of 85% NE(m). Diet adjustments were made following weekly weighings. GnRH challenges were conducted at four periods: prior to initial energy deprivation, at termination of 40% NE(m) feeding, and twice more at 21-d intervals after 85% NE(m) feeding began. Blood was sampled via a jugular catheter every 15 min for 5 h, and GnRH was injected after the fourth sample. None of the heifers exhibited estrous activity after the initial energy deprivation. Heifers on the bypass fat diet continued to lose weight during the treatment period, while the control heifers gained a slight amount of weight. Baseline and peak concentrations of LH were not significantly affected by time or diet. Time to GnRH-induced LH peak was longer (53 vs 130 min, P < 0.01) after 40% NE(m) and remained greater at all times thereafter. Serum lipid levels increased 82.5% among heifers being fed the bypass fat. Energy restriction had no effect on the magnitude of LH response to GnRH but did delay response time.     

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.     

Ethylene production by loblolly pine seedlings associated with water stress

Effects of water stress on production of ethylene and its precursor, 1-aminocyclopropane-1-carboxylic acid (ACC), by loblolly pine ( Pinus taeda L.) seedlings from a Texas drought-hardy and a Virginia Coastal Plain source were investigated. Ethylene production rates in needles from the Virgnia source increased slightly with initial stress (-1.3 MPa), declined until water potential reached -1.6 MPa and then increased sharply at -2.5 MPa. The ethylene production rates in needles from the Texas also increased slightly with initial stress, then decreased with decreasing water potential. Ethylene production by root tissue was two to three times higher than needle tissue and decreased with decreasing water potential. ACC concentrations in needles of both seed sources decreased as water potential began decreasing. Below -1.4 MPa, ACC levels started increasing (Texas source) or remained constant until -2.8 MPa (Virginia source) at which time its level increased three-fold. Mean ACC levels in root tissue [122 nmol (g dry weight)⁻¹] were slightly higher than the mean levels in the needle tissue [92 nmol (g dry weight) ⁻¹]; roots apparently were more efficient in converting it to ethylene since ethylene production was two to three times higher than needle tissue. The modulation of ethylene synthesis by ACC synthase and ethyleneforming enzyme appeared to be influenced by stress level, organ and seed source.