Effects of exercise on young and adult cattle at low and high altitude

Nine calves and nine oxen walked on a treadmill in a climatized low pressure chamber for one hour each day, 2 weeks at 400 m and 4 weeks at 3,500 m. The overall effects of walking were: increases in heart rate, pulmonary arterial pressure, rectal temperature, respiratory rate, blood-pH and lactate/pyruvate ratio. Haemoglobin, haematocrit, blood specific gravity and blood viscosity increased in the oxen but decreased in the calves. Blood lactate and blood pyruvate declined in both age groups, plasma viscosity only in the calves. The exercise effects were more pronounced at 3,500 m than at 400 m as exemplified by the following percentile differences (3,500-400 m): in heart rate 26%, mean pulmonary arterial pressure: 22%, respiratory rate: 11%, blood pH: 0.3%, blood lactate: 39%, blood pyruvate: 56%, haemoglobin: 4%, blood viscosity: 5%. Compared with the calves, the oxen experienced larger increases in heart rate and respiratory rate in response to exercise, suggesting a greater rise in metabolic rate: they also showed a more pronounced respiratory alkalosis. Thus, exercise seems to have strained the oxen more than the calves. In the oxen, there was a training effect as judged by reductions in exercising heart rate, respiratory rate and rectal temperature.     

Differential seed predation on two species of Arctostaphylos (Ericaceae)

Summary The fire-prone California chaparral contains two sympatric species of shrubs: Arctostaphylos glauca and A. glandulosa. A previous study showed that in a stand where both species had similar amounts of coverage, A. glauca had fewer seeds in the soil. We attempt to answer the questions: 1) Could ground-foraging seed predators produce the lower population of A. glauca seeds in the soil? 2) Do predators select fruits randomly with respect to fruit size? 3) Do the fruits of the two species differ in the proportions of fruit components (i.e. seeds, endocarp, mesocarp, and exocarp) in ways that could be important to seed predators? Predation was measured on artificial caches of fruits, for 17 weeks. Selection by predators was examined by comparing weights of fruits recovered from soil samples with newlymatured fruits on the shrubs. Fruits components were characterized by dividing fruits into 3 fractions and weighing. More fruits of A. glauca were removed from the caches. Fruits of both species recovered from the soil were lighter than those on the shrubs. The weights of seeds, stony and fleshy fruit layers were all larger in A. glauca. Within fruits of A. glandulosa, the weights of the three components, various combinations, and ratios were all significantly correlated, while in A. glauca no other component, combination of components, or ratio examined was significantly correlated with the weights of the seeds.     

Retention of antidiuretic hormone-induced particle aggregates by luminal membranes separated from toad bladder epithelial cells

Aggregates of intramembrane particles appear in the luminal membranes of renal collecting duct and amphibian bladder cells after stimulation by antidiuretic hormone (ADH). We undertook this freeze-fracture study to determine whether particle aggregates, once in place, remain in the luminal membrane of the amphibian bladder after the membrane is physically separated from the rest of the cell. We found that the aggregates do remain in high yield in isolated membranes stabilized with a bifunctional imidoester (DTBP) followed by fixation with glutaraldehyde, or unfixed but stabilized with DTBP. These findings support the view that the particles are intrinsic membrane components and that their organization in the form of aggregates does not depend on the presence of the intact cell. In addition, the availability of isolated membranes containing particle aggregates provides a starting point for the isolation of the water-conducting proteins.     

Giemsa-11 technique. Applications in the chromosomal characterization of hematologic specimens

Summary Use of the Giemsa-11 procedure for the localization of heterochromatic regions of human chromosomes and for differentiation of primate and rodent chromosomes has been somewhat limited since its discovery in 1972. An adaptation of this technique to the cytogenetic characterization of hematologic specimens has aided in the interpretation of translocations, deletions, and inversions involving human chromosome 9. The chromosomal analyses of 10% of over 100 patients, principally leukemic, were aided through the use of this auxiliary procedure. The diseases of these patients are given and portions of karyotypes are presented to show clarification of abnormalities made possible through the use of the Giemsa-11 technique.     

Role of D1-His190 in the proton-coupled oxidation of tyrosine YZ in manganese-depleted photosystem II.

To further characterize the role of D1-His190 in the oxidation of tyrosine Y(Z) in photosystem II, the pH dependence of P(680)(*)()(+) reduction was measured in H190A and Mn-depleted wild-type PSII particles isolated from the cyanobacterium, Synechocystis sp. PCC 6803. Measurements were conducted in the presence and absence of imidazole and other small organic bases. In H190A PSII particles, rapid reduction of P(680)(*)()(+) attributed to electron transfer from Y(Z) increased dramatically above pH 9, with an apparent pK(A) of approximately 10.3. In the presence of ethanolamine and imidazole, this dramatic increase occurred at lower pH values, with the efficiency of Y(Z) oxidation correlating with the solution pK(A) value of the added base. We conclude that the pK(A) of Y(Z) is approximately 10.3 in D1-H190A PSII particles. In Mn-depleted wild-type PSII particles, P(680)(*)()(+) reduction was accelerated by all exogenous bases examined (substituted imidazoles, histidine, Tris, and 1,4-diazabicyclo[2.2.2]octane). We conclude that Y(Z) is solvent accessible in Mn-depleted wild-type PSII particles and that its pK(A) is near that of tyrosine in solution. In Mn-depleted wild-type PSII particles, over 80% of the kinetics of P(680)(*)()(+) reduction after a flash could be described by three kinetic components. The individual rate constants of these components varied slightly with pH, but their relative proportions varied dramatically with pH, showing apparent pK(A) values of 7.5 and 6.25 (6.9 and 5.8 in the presence of Ca(2+) and Mg(2+) ions). An additional pK(A) value (pK(A) < 4.5) may also be present. To describe these data, we propose (1) the pK(A) of His190 is 6.9-7.5, depending on buffer ions, (2) the deprotonation of Y(Z) is facilitated by the transient formation of a either a hydrogen bond or a hydrogen-bonded water bridge between Y(Z) and D1-His190, and (3) when protonated, D1-His190 interacts with nearby residues having pK(A) values near 6 and 4. Because Y(Z) and D1-His190 are located near the Mn cluster, these residues may interact with the Mn cluster in the intact system.     

Binding of MutS mismatch repair protein to DNA containing UV photoproducts, "mismatched" opposite Watson--Crick and novel nucleotides, in different DNA sequence contexts

Mismatch-repair (MMR) systems suppress mutation via correction of DNA replication errors (base-mispairs) and responses to mutagenic DNA lesions. Selective binding of mismatched or damaged DNA by MutS-homolog proteins-bacterial MutS, eukaryotic MSH2.MSH6 (MutSalpha) and MSH2.MSH3-initiates mismatch-correction pathways and responses to lesions, and may cumulatively increase discrimination at downstream steps. MutS-homolog binding selectivity and the well-known but poorly understood effects of DNA-sequence contexts on recognition may thus be primary determinants of MMR specificity and efficiency. MMR processes that modulate UV mutagenesis might begin with selective binding by MutS homologs of "mismatched" T[CPD]T/AG and T[6--4]T/AG photoproducts, reported previously for hMutSalpha and described here for E. coli MutS protein. If MMR suppresses UV mutagenesis by acting directly on pre-mutagenic products of replicative bypass, mismatched photoproducts should be recognized in most DNA-sequence contexts. In three of four contexts tested here (three substantially different), T[CPD]T/AG was bound only slightly better by MutS than was T[CPD]T/AA or homoduplex DNA; only one of two contexts tested promoted selective binding of T[6--4]T/AG. Although the T:G pairs in T[CPD]T/AG and T/G both adopt wobble conformations, MutS bound T/G well in all contexts (K(1/2) 2.1--2.9 nM). Thus, MutS appears to select the two mismatches by different mechanisms. NMR analyses elsewhere suggest that in the (highly distorted) T[6--4]T/AG a forked H-bond between O2 of the 3' thymine and the ring 1-imino and exocyclic 2-amino guanine protons stabilizes a novel planar structure not possible in T[6--4]T/AA. Replacement of G by purines lacking one (inosine, 2-aminopurine) or both (nebularine) protons markedly reduced or eliminated selective MutS binding, as predicted. Previous studies and the work here, taken together, suggest that in only about half of DNA sequence contexts could MutS (and presumably MutSalpha) selectively bind mismatched UV photoproducts and directly suppress UV mutagenesis.     

First records of dive durations for a hibernating sea turtle

The first published record, from the early 1970s, of hibernation in sea turtles is based on the reports of the indigenous Indians and fishermen from Mexico, who hunted dormant green turtles (Chelonia mydas) in the Gulf of California. However, there were no successful attempts to investigate the biology of this particular behaviour further. Hence, data such as the exact duration and energetic requirements of dormant winter submergences are lacking. We used new satellite relay data loggers to obtain the first records of up to 7h long dives of a loggerhead turtle (Caretta caretta) overwintering in Greek waters. These represent the longest dives ever reported for a diving marine vertebrate. There is strong evidence that the dives were aerobic, because the turtle surfaced only for short intervals and before the calculated oxygen stores were depleted. This evidence suggests that the common belief that sea turtles hibernate underwater, as some freshwater turtles do, is incorrect.     

Arabidopsis thaliana AtPOLK encodes a DinB-like DNA polymerase that extends mispaired primer termini and is highly expressed in a variety of tissues

Cell survival after DNA damage depends on specialized DNA polymerases able to perform DNA synthesis on imperfect templates. Most of these enzymes belong to the recently discovered Y-family of DNA polymerases, none of which has been previously described in plants. We report here the isolation, functional characterization and expression analysis of a plant representative of the Y-family. This polymerase, which we have termed AtPolkappa, is a homolog of Escherichia coli pol IV and human pol kappa, and thus belongs to the DinB subfamily. We purified AtPolkappa and found a template-directed DNA polymerase, endowed with limited processivity that is able to extend primer-terminal mispairs. The activity and processivity of AtPolkappa are enhanced markedly upon deletion of 193 amino acids (aa) from its carboxy (C)-terminal domain. Loss of this region also affects the nucleotide selectivity of the enzyme, leading to the incorporation of both dCTP and dTTP opposite A in the template. We detected three cDNA forms, which result from the alternative splicing of AtPOLK mRNA and have distinct patterns of expression in different plant organs. Histochemical localization of beta-glucuronidase (GUS) activity in transgenic plants revealed that the AtPOLK promoter is active in endoreduplicating cells, suggesting a possible role during consecutive DNA replication cycles in the absence of mitosis.     

The design and synthesis of sulfonamides as caspase-1 inhibitors

A series of sulfonamides (1) has been prepared as inhibitors of interleukin-1beta converting enzyme (ICE), also known as caspase 1. These compounds were designed to improve potency by rigidifying the enzyme bound molecule through an intramolecular hydrogen bond. An X-ray crystal structure of a representative member of this series bound to the active site of ICE, confirms the presence of the hydrogen bonding interaction.