Primary Structure and Phylogenetic Relationships of Glyceraldehyde-3-Phosphate Dehydrogenase Genes of Free-Living and Parasitic Diplomonad Flagellates

ABSTRACT. Complete nucleotide sequences have been established for two genes (gap1 and gap2) coding for glyceraldehyde-3-phosphate dehydrogenase (GAPDH, EC 1.2.1.12) homologs in the diplomonad Giardia lamblia. In addition, almost complete sequences of the GAPDH open reading frames were obtained from PCR products for two free-living diplomonad species, Trepomonas agilis and Hexamita inflata, and a parasite of Atlantic salmon, an as yet unnamed species with morphological affinities to Spironucleus. Giardia lamblia gap 1 and the genes from the three other diplomonad species show high similarity to each other and to other glycolytic GAPDH genes. All amino-acyl residues known to be highly conserved in this enzyme are also conserved in these sequences. Giardia lamblia gap2 gene is more divergent and its putative translation reveals the presence of a cysteine and serine-rich insertion resembling a metal binding finger. This motif has not yet been noted in other GAPDH molecules. All sequences contain an S-loop signature with characteristics close to those of eukaryotes. In phylogenetic reconstructions based on the derived amino acid sequences with neighborjoining, parsimony and maximum-likelihood methods the four typical GAPDH sequences of diplomonads cluster into a single clade. Within this clade, G. lamblia gap1 shares a common ancestor with the rest of the genes. The latter are more closely related to each other, indicating an early separation of the lineage leading to the genus Giardia from the lineage encompassing the morphologically less differentiated genera, Trepomonas, Hexamita and that of the unnamed species. This result is discordant with the orthogonal evolution of diplomonads suggested on the basis of comparative morphology. In neighbor-joining reconstructions G. lamblia gap2 occupies a variable position, due to its great divergence. In parsimony and maximum likelihood analysis however, it shares a most recent common ancestor with the typical G. lamblia gap1 gene, suggesting that it diverged after the separation of the Giardia lineage. The position of the diplomonad clade in broader phylogenetic reconstructions is firmly within the typical cytosolic glycolytic representatives of GAPDH of eukaryotes.     

IMPRINTING EFFECTS OF THREE AMINO ACIDS (ALANINE,LYSINE AND GLYCINE) AND THEIR OLIGOPEPTIDES INTETRAHYMENA PYRIFORMIS. DATA FROM THE HORMONE AND HORMONE RECEPTOR EVOLUTION

Hormonal imprinting takes place at the first encounter of the hormone and receptor, and results in a changed binding capacity and reaction of the cell and its progeny generations. The imprinting effect of three amino acids and their oligopeptides is studied using fluorescent-labelled peptides. Glycine and lysine could provoke positive imprinting (increased binding in the progeny generations) for their own peptides, but alanine could not. Mostly positive imprinting was provoked by glycine and lysine peptides for their own peptides of different chain length. The optimal chain length provoking self-imprinting was four for glycine, two for lysine and three for alanine. Except in this case, alanine was neutral or provoked mostly negative imprinting. After reaching the optimal chain length, there is a decline in binding. Evolutionary conclusions are discussed.     

Mycobacterium kansasii infection in squirrel monkeys (Saimiri sciureus sciureus)

Abstract: This report documents asymptomatic infections of Mycobacterium kansasii in four of five tuberculin positive squirrel monkeys (Saimiri sciureus sciureus). The mycobacterial DNA amplified by polymerase chain reaction (PCR) from a bronchial lymph node had no affinity for the species specific probes of M. tuberculosis, M. avium, and M. intracellular, thus allowing the presumptive diagnosis of an atypical mycobacterial infection. Infection by Mycobacterium kansasii was confirmed by culture of bronchial lymph nodes from three monkeys. The source of the infection was never identified.     

Photomorphogenesis and canopy dynamics. Phytochrome-mediated proximity perception accounts for the growth dynamics of canopies of Populus trichocarpa × deltoides'Beaupré'

The phytochrome family of signal-transducing photoreceptors provides plants with the capacity to perceive variations in the relative fluxes of red (R) and far-red (FR) radiation. This capacity has been proposed to be of ecological value in the perception of the proximity of neighbouring plants and the consequent induction of shade avoidance responses. The work reported here has evaluated this potential by determining quantitatively the effect of neighbour proximity on the growth of canopies of Populus trichocarpa×deltoides‘Beaupré’ trees, and relating the measured variables to the long-term vectoral radiation quality inside each canopy. The spectral distribution of radiation inside four canopies of Populus trichocarpa×deltoides‘Beaupre’ of different densities was monitored throughout the growing season. Spectral distributions inside the canopies were measured in 10° wedges at different heights and angles. The results are presented as PFD over 400–700 nm (PFD400–700) and PFD over 400–800 nm (PFD400–700). Results are also presented for the calculated phytochrome photoequilibrium (Pfr/P) and red:far-red ratio (R:FR). Data are presented as in-canopy angular and height profiles, and as diurnal and seasonal variations. PFD_400–700 and Pfr/P were found to be reduced inside each canopy, the reduction being greatest in the most dense canopy, and least in the most open canopy. At any height within each canopy, calculated Pfr/P decreased linearly with time throughout the growing season, until leaf senescence began. The reduction was greater in the denser canopies and was found to be similar for three consecutive field seasons. Linear relationships were found between plant stem growth rate, plant spacing and Pfr/P calculated from radiation propagated approximately horizontally within the canopies. The findings support the role of phytochrome in proximity perception in the natural environment and provide a quantitative basis for investigating the competitive interactions between plants growing in dense stands. The hypothesis is proposed that the dynamics of developing or regenerating canopies can be accounted for on the basis of phytochrome-mediated perception of the proximity of neighbouring plants.     

Effect of open-air fumigation with sulphur dioxide and ozone on phyllosphere and endophytic fungi of conifer needles

The phyllosphere microbial populations inhabiting the needles of three conifer species, Scots pine (Pinus sylvestris L.), Sitka spruce (Picea sitchensis L.) and Norway spruce (Picea abies (L.) Karst.), exposed to SO₂ and O₃, in an open-air fumigation experiment were analysed over a 3 year period using serial dilution after washing, direct plating and a fluorescein diacetate (FDA) enzyme assay. Total fungal populations ranged from 10² to 10⁵ colonyforming units (CPU) g^?1 fresh weight of needles. The dominant fungi isolated from needles varied with tree species and isolation technique; Aureobasidium pullulans (de Bary) Arnaud was most common on Scots pine and Norway spruce and white yeasts on Sitka spruce using the dilution plating method. However, direct plating of needle segments onto culture media indicated that Sclerophoma pythiophila (Corda) Hohnel was dominant on Scots pine and A. pullulans on Sitka and Norway spruce. Green needles of Sitka spruce were found to be endophytically colonized by Rhizosphaera kalkhoffii Bubak, but seldom by Lophodermium piceae (Fuckel) Hohn during extensive sampling in 1990. Statistical analyses revealed significant differences (P<0.05) between plots in the 3 year mean of the total fungal populations or the fungal biomass (FDA assay) on all three tree species. Differences between plots were also observed for a number of dominant component species. Data were also analysed for treatment effects. A significant effect of SO₂ treatment was observed on the total fungal populations on Sitka spruce (P<0.05) which were reduced markedly by the low-SO₂ treatment, while the O₃ treatment caused a significant increase in total fungal numbers on Scots pine (P<0.05). The FDA activity on needles of both Scots pine and Sitka spruce was noticeably higher in the 0₃-only treatment plot, but the overall O₃ effect was not significant. Treatment effects were also detected on the occurrence of component species. The serial dilution method revealed an SO₂ effect (P<0.05) of a reduction in the occurrence of pink yeasts on Sitka spruce and an O₃ effect (P<0.05) of an increase in the occurrence of S. pythiophila on Sitka spruce (P<0.01) but a decrease of Epicoccum nigrum Link and Cladosporium spp. on Scots pine. The direct-plating method revealed an SO₂ effect of an increase in S. pythiophila on Norway spruce (P<0.05). Ozone treatment caused a significant increase in the isolation of a black strain of A. pullulans on Norway spruce (P<0.05). Endophytic colonization of Sitka spruce needles by R. kalkhoffii was found to be increased on two occasions by O₃ exposure.     

Occurrence of unstable PEP carboxylase in C4 grasses in the genus Spartina Schreb.

Abstract The aims of this work were to discover the distribution within the C₄ grass Spartina anglica of a PEP carboxylase which is very unstable during and after extraction, and to determine whether this unstable form occurs in other members of the genus. In S. anglica, only the leaf contains an unstable PEP carboxylase. Within the leaf only the major one of two isoenzymes is unstable, and this is located in the mesophyll cells. The unstable isoenzyme is inactivated during extraction and storage unless protected by bovine serum albumin or Triton X-100, and is inactivated in assay mixtures at optimum pH in the absence of PEP. Evidence is presented that inactivation is not due to degradation or inhibition during extraction and storage. The enzyme from leaves of Spartina species taxonomically closely related to S. anglica is also very unstable during and after extraction, but that from less closely related species is much more stable.     

Archaeological Inference and Inductive Confirmation

The limitations of the hypothetico-deductive (H-D) method of inductive confirmation are described, and an alternate method, the hypothetico-analog (H-A) method is described in detail. The H-A method can be characterized as a modified and supplemented form of the simple H-D method, and is proposed as being more appropriate for archaeological inference. Aspects of the H-A method that are given particular attention include the establishment of boundary conditions for reference classes, plausibility considerations, multiple working hypotheses, bridging arguments, and criteria for selecting alternative hypotheses . [scientific method, archaeological inference, hypothetico-analog method, inductive confirmation]     

Effect of Temperature and External pH on the Cytoplasmic pH of Chara corallina

Cytoplasmic pH (pH_c) in Chara corallina was measured (from [¹⁴C]stribution)as a function of external pH (pH₀)and temperature. With pH₀near 7, pH_c at 25?C is 7.80; pH_cincreases by 0.005 pH units?C^–1 temperature decrease, i.e. pH_c at 5 ?C is 7.90. WithpH? near 5.5, the increase in pH_c with decreasing temperatureis 0.015 units ?C^–1 between 25 and 15?C, but 0.005 units?C^–1 between 15 and 5?C. This implies a more precise regulationof pH_c with variations in pH_o at 5 or 15 ?C compared with 25?C. The observed dp H_c/dT is generally smaller than the –0.017units ?C^–1 needed to maintain a constant H⁺/OH^–1,or a constant fractional ionization of histidine in protein,with variation in temperature. It is closer to that needed tomaintain the fractional ionization of phosphorylated compoundsor of CO₂–HCO₃^– The value of dpH_c/dT has importantimplications for several regulatory aspects of cell metabolism.These include (all as a function of temperature) the rates ofenzyme reactions, the _H+ at the plasmalemma(and hence the energy available for cotransport processes),and the mechanism for pH_c regulation by the control of bidirectionalH⁺ fluxes at the plasmalemma.