Accumulation of scandium in the shoots of aquatic bryophytes in acid water

The contents of scandium (Sc) in the shoots of aquatic bryophytes from rivers, streams, lakes and springs were determined using inductively coupled plasma. The shoots of aquatic bryophytes were found to accumulate scandium in acid water, attaining a level as 33 g g^–1 in Scapania undulata (L.) Dum. in an acid stream with a pH of about 4.2. The content of scandium was highly correlated with that of aluminum in the shoots of S. undulata (r = 0.986, 0.967; p < 0.01).     

Alteration of 1,2-diacylglycerol content in ischemic and reperfused heart

Human term placenta contains an ATP diphosphohydrolase activity which hydrolyses ATP to ADP and inorganic phosphate and ADP to AMP and a second mole of inorganic phosphate. The activity has a pH optimum between 8.0 and 8.5. Magnesium or calcium ions are required for maximum activity. Other nucleoside phosphates, p-nitrophenyl phosphate or sodium pyrophosphate, are not hydrolysed. The activity is not due to ATPases, or to myokinase, as determined by the use of inhibitors. NaF and NaN₃ were found to inhibit strongly the activity thus identifying it as an ATP diphosphohydrolase.A sensitive enzymatic assay for measurement of AMP, one of the products of the reaction, was established, based on the strong inhibition of muscle fructose 1,6-biphosphatase by AMP. The range of the assay was 0.05–0.8 µM AMP. ATP diphosphohydrolase was found to have a rate of AMP production from ADP twice the rate from ATP. Under the same conditions, the assay for Pi release, on the other hand, gave velocities similar to each other for the two substrates.The activity appears to be identical to the ADP-hydrolysing activity in placenta reported by others.Abbreviations Ap₅A P¹ - P⁵-di(adenosine-5) Pentaphosphate - ATP-DPH ATP Diphosphohydrolase - DCCD N,N Dicyclohexycarbodiimide - Fru-P₂ase Fructose 1,6-biphosphatase - SDS Sodium Dodecyl Sulfate - TLC Thin Layer Chromatography     

Kinetic study on oxygenation of Lingula unguis hemerythrin using the stopped-flow 02-jump method

O₂-jump experiments with an improved stopped-flow apparatus have been used to study oxygenation and deoxygenation processes in Lingula unguis hemerythrin. With an O₂ electrode set in the observation cell, O₂ concentration conld be obtained directly. The reliability of this method has been compared with other conventional methods.O₂-jump (up and down) experiments were carried out with L. unguis hemerythrin at pH 6.8 (non-cooperative pH) and at pH 7.6 (cooperative pH). At pH 6.8, both O₂-jump (up) and O₂-jump (down) experiments showed single exponential processes which were consistent with the following scheme: . The value of k _on was estimated to be (4.4 ± 0.5) × 10⁵ M^–1 s ^–1, and k _off was (15 ± 5) s^–1. These values are consistent with those obtained by the temperature-jump method (Zimmer et al. 1986). At pH 7.6, O₂-jump (up) experiments showed two relaxation processes, whereas O₂-jump (down) experiments showed a single exponential process. The faster process in the O₂-jump (up) experiments could be attributed to the same process as that seen in the temperature-jump experiments (Zimmer et al. 1986). The slower process in the O₂-jump (up) experiments corresponds to the process obtained in the O₂-jump (down) experiments. The results are discussed in terms of a state with intermediate affinity in O₂-binding and with the possible existence of a slow step in O₂-binding.Abbreviations Hr hemerythrin - Hb hemoglobin - CM carboxymethyl - T-jump temperature-jump Offprint requests to: H. Kihara     

Purification of a mouse nuclear factor that binds to both the A and B cores of the polyomavirus enhancer.

We have previously identified a protein factor, PEBP2 (polyomavirus enhancer-binding protein), in the nuclear extract from mouse NIH 3T3 cells which binds to the sequence motif, PEA2, located within the polyomavirus enhancer A element. Upon cellular transformation with activated oncogene c-Ha-ras, this factor frequently undergoes drastic molecular modifications into an altered form having a considerably reduced molecular size. In this study, the altered form, PEBP3, was purified to near homogeneity. The purified PEBP3 comprised two sets of families of polypeptides, alpha-1 to alpha-4 and beta-1 to beta-2, which were 30 to 35 kilodaltons and 20 to 25 kilodaltons in size, respectively. Both kinds of polypeptides possessed DNA-binding activities with exactly the same sequence specificity. Individual alpha or beta polypeptides complexed with DNA showed faster gel mobilities than did PEBP3. However, the original gel retardation pattern was restored when alpha and beta polypeptides were mixed together in any arbitrary pair. These observation along with the results of UV- and chemical-cross-linking studies led us to conclude that PEBP3 is a heterodimer of alpha and beta subunits, potentially having a divalent DNA-binding activity. Furthermore, PEBP3 was found to bind a second, hitherto-unnoticed site of the polyomavirus enhancer that is located within the B element and coincides with the sequence previously known as the simian virus 40 enhancer core homology. From comparison of this and the original binding sites, the consensus sequence for PEBP3 was defined to be PuACCPuCA. These findings provided new insights into the biological significance of PEBP3 and PEBP2.     

Maintenance of mitochondrial DNA by the Caenorhabditis elegans ATR checkpoint protein ATL-1

Here we show that inactivation of the ATR-related kinase ATL-1 results in a significant reduction in mitochondrial DNA (mtDNA) copy numbers in Caenorhabditis elegans. Although ribonucleotide reductase (RNR) expression and the ATP/dATP ratio remained unaltered in atl-1 deletion mutants, inhibition of RNR by RNAi or hydroxyurea treatment caused further reductions in mtDNA copy number. These results suggest that ATL-1 functions to maintain mtDNA independently of RNR.     

Transient shielding of intimin and the type III secretion system of enterohemorrhagic and enteropathogenic Escherichia coli by a group 4 capsule

Enterohemorrhagic and enteropathogenic Escherichia coli (EHEC and EPEC, respectively) strains represent a major global health problem. Their virulence is mediated by the concerted activity of an array of virulence factors including toxins, a type III protein secretion system (TTSS), pili, and others. We previously showed that EPEC O127 forms a group 4 capsule (G4C), and in this report we show that EHEC O157 also produces a G4C, whose assembly is dependent on the etp, etk, and wzy genes. We further show that at early time points postinfection, these G4Cs appear to mask surface structures including intimin and the TTSS. This masking inhibited the attachment of EPEC and EHEC to tissue-cultured epithelial cells, diminished their capacity to induce the formation of actin pedestals, and attenuated TTSS-mediated protein translocation into host cells. Importantly, we found that Ler, a positive regulator of intimin and TTSS genes, represses the expression of the capsule-related genes, including etp and etk. Thus, the expression of TTSS and G4C is conversely regulated and capsule production is diminished upon TTSS expression. Indeed, at later time points postinfection, the diminishing capsule no longer interferes with the activities of intimin and the TTSS. Notably, by using the rabbit infant model, we found that the EHEC G4C is required for efficient colonization of the rabbit large intestine. Taken together, our results suggest that temporal expression of the capsule, which is coordinated with that of the TTSS, is required for optimal EHEC colonization of the host intestine.