Diel Surface Temperature Range Scales with Lake Size

Ecological and biogeochemical processes in lakes are strongly dependent upon water temperature. Long-term surface warming of many lakes is unequivocal, but little is known about the comparative magnitude of temperature variation at diel timescales, due to a lack of appropriately resolved data. Here we quantify the pattern and magnitude of diel temperature variability of surface waters using high-frequency data from 100 lakes. We show that the near-surface diel temperature range can be substantial in summer relative to long-term change and, for lakes smaller than 3 km², increases sharply and predictably with decreasing lake area. Most small lakes included in this study experience average summer diel ranges in their near-surface temperatures of between 4 and 7°C. Large diel temperature fluctuations in the majority of lakes undoubtedly influence their structure, function and role in biogeochemical cycles, but the full implications remain largely unexplored.     

Single crystals of bacteriophage T7 RNA polymerase

Single crystals of T7 RNA polymerase have been grown to a maximum size of 1.8 x 0.3 x 0.3 mm. The crystals are composed of fully intact T7 RNA polymerase which is enzymatically active upon dissolution. These crystals belong to the monoclinic space group P2(1) and have unit cell parameters a = 114.5 A, b = 139.6 A, c = 125.7 A, and beta = 98.1 degrees. Self-rotation function studies indicate that there are three molecules per asymmetric unit. The crystals diffract to at least 3.0 A resolution. These are the first crystals of a DNA-dependent RNA polymerase suitable for high-resolution X-ray structure determination.     

Fc receptor isoforms exhibit distinct abilities for coated pit localization as a result of cytoplasmic domain heterogeneity

Mouse macrophages and lymphocytes express two distinct isoforms of a single class of Fc receptor for IgG. The macrophage isoform (FcRII-B2) is identical to the lymphocyte isoform (FcRII-B1) except for an inframe insertion in the cytoplasmic tail of FcRII-B1 that increases its length from 47 to 94 amino acids. To determine the functional significance of this cytoplasmic domain variation, presumably the result of alternative mRNA splicing, we expressed both isoforms in receptor-negative fibroblasts. While FcRII-B2 mediated the efficient ligand internalization and delivery to lysosomes, endocytosis via FcRII-B1--and via a tailminus mutant--was relatively inefficient. This difference reflected the inability of FcRII-B1 (and the tailminus mutant) to accumulate in clathrin-coated pits. Thus, the FcRII-B2 cytoplasmic tail contains a domain needed for accumulation in coated pits, and this domain is disrupted by the 47 amino acid insertion in FcRII-B1.     

Preliminary crystallographic analysis of class 3 rat liver aldehyde dehydrogenase

NAD-linked aldehyde dehydrogenases (A1DH) (EC 1.2.1.3) catalyze the irreversible oxidation of a wide variety of aldehydes to their respective carboxylic acids. Crystals of a class 3 AIDH (from an Escherichia coli expression system) suitable for X-ray analysis have been obtained. These crystals, which can be grown to a size of 0.8 x 0.3 x 0.2 mm, diffract to 2.5 A resolution. Analysis of the diffraction pattern indicates that the crystals belong to the monoclinic space group P21, with cell parameters a = 65.11 A, b = 170.67 A, c = 47.15 A, and beta = 110.5 degrees. Assuming one dimer per asymmetric unit, the value Vm is calculated to be 2.45 and the solvent content of the crystal is estimated to be 50%. A self-rotation function study produced significant rotation peaks (58% of the origin) on the kappa = 180 section at psi = 90 degrees and phi = 71 degrees and 341 degrees, indicating that the pseudo-dimer axis is (or is very nearly) perpendicular to the b-axis.     

PH dependence of the alpha-glucose 1,6-diphosphate inhibition of hexokinase II.

α-Glucose 1,6-diphosphate is a much better inhibitor of hexokinase II than 1,5-anhydroglucitol 6-phosphate or glucose 6-phosphate (Glc-6-P) at pH 6–7 and poorer at higher pH. Because the K_i of Glc-6-P is pH independent, the observed pH effects are attributed to the phosphate group at C-1 which is bound as a monoanion to a specific site but which is excluded as a dianion. None of the following kinetic properties of the hexokinase II reaction varies greatly with pH: V, K_m of glucose and K_m of ATP.     

Concentration and partitioning of intermediates in the fructose bisphosphate aldolase reaction. Comparison of the muscle and liver enzymes

Chemical analysis of enzyme reaction intermediates has been used to compare the liver and muscle isozymes of rabbit aldolase at equilibrium and in their steady states to determine if they have properties that favor the direction of flow of glycolytic intermediates in their tissues of origin. For both enzymes at saturating concentrations of fructose 1,6-P2, the sum of intermediates in the steady state agreed with the total active enzyme calculated to be present. The two half-reactions, characterized by fructose 1,6-bisphosphate(Fru-P2):aldehyde exchange and DHAP:proton exchange were found to be of different importance in determining the rate of reaction with Fru-P2 with the liver enzyme being much more limited in the processing of DHAP. The chemical interconversions within each half-reaction are generally rapid compared with the release of products. The greater sensitivity of liver aldolase to inhibition by aldehydes in Fru-P2 cleavage seems to be a normal consequence of the higher level of the eneamine of DHAP in the forward steady state with the liver enzyme and probably should not be ascribed to a greater intrinsic affinity. An earlier report (Grazi, E., and Trombetta, G. (1979) Eur. J. Biochem. 100, 197-202) purporting to show a special interaction of glyceraldehyde-3-P with liver enzyme prior to proton abstraction from DHAP could not be reproduced. Examples are presented from the data that validate the use of the analytical methods used for analysis of intermediates in the case of the Schiff's base aldolases.     

Anesthetic and supportive management during experimental pulsatile flow perfusion studies in calves

The purpose of this study was to determine the factors influencing successful experimental cardiopulmonary bypass studies using pulsatile flow perfusion and the medications and methodology necessary to produce successful bypass in calves. In six calves showing no cardiopulmonary pathology prior to bypass procedures, successful anesthesia and surgical intervention was accomplished. Animals were maintained on 5 hours of pulsatile flow bypass perfusion. Successful recovery from the procedures was accomplished. In two calves with pre-existing pulmonary pathology, anesthetic and surgical intervention was accomplished with the utilization of extensive anesthetic management and cardiac supportive medications until the animals could be initiated into 5 hours of pulsatile flow bypass perfusion, in spite of major pulmonary dysfunction. In these two animals, attempts to resuscitate upon termination of pulsatile flow perfusion were unsuccessful due to pre-existing excessive lesions in the lungs. This study shows a contrast between complete success of a pulsatile flow system in normal subjects versus the ultimate failure in experimental animals with pre-existing pulmonary pathology. The inability of experimental calves with a diseased lung to resume spontaneous cardiopulmonary function after the challenges of thoracic intervention indicates the unsuitability of animals with marked pre-existing pulmonary disease status for use in cardiopulmonary bypass studies.     

Biochemical genetics of resistance to MGBG in tobacco: Mutants that alter SAM decarboxylase or polyamine ratios, and floral morphology

Summary Previously we reported the isolation of Nicotiana tabacum cell lines resistant to methylglyoxal-bis(guanylhydrazone) (MGBG) an inhibitor of the polyamine synthetic enzyme s-adenosylmethionine decarboxylase (SAMde). Here we report that these mutants fall into several distinct classes on the basis of their biochemical characteristics. At least two lines, Mgr12 and Mgr16, have a SAMdc that displays increased resistance to MGBG in in vitro enzyme assays, suggesting that these two alleles have altered the SAMdc enzyme itself, and thus they may serve to identify the SAMdc structural gene. Other cell lines have elevated levels of some of the polyamines or polyamine-conjugates: Mgr11 and Mgr14 have high levels of putrescine and high levels of activity of putrescine synthesizing enzymes; Mgr23 and Mgr3 have elevated spermidine and spermidine-conjugate levels, with Mgr23 also having elevated putrescine levels. Mgr12 and Mgr3 have been analyzed genetically through F₁ crosses with wild-type tobacco, and subsequently by a backcross of an F₁ plant to wild type; however, the total number of seeds obtained in each cross was very small. The results of the genetic analysis are consistent with Mgr12 and Mgr3 being nuclear dominant traits. The floral abnormalities previously reported as associated with these mutations display linkage with the MGBG resistances. At least for Mgr12 we have thus obtained evidence as to the precise nature of the mutation, an altered SAMdc, and demonstrated that this is likely to be genetic cause of the altered flower phenotype.