Eukaryotic DNA topoisomerase and in vitro recombination between circular supercoiled plasmid DNA's in an in vitro system

Eukaryotic DNA topoisomerase II was shown to catalyze recombination between circular supercoiled plasmid DNAs in vitro. The results obtained are discussed with regard to the organization of eukaryotic chromosomes in the form of topologically independent domains (loops).     

Computed tomographic angiography of the thoracic vessels: the method, initial experience and outlook for its use

The paper presents the data available in the literature on computed tomographic angiography and the first experience with it to study thoracic vessels. It details the principles of spiral computed tomography and CT angiography. Practical aspects of their implementation, as well as basic concepts are outlined. It is concluded that CT angiography is promising in studying thoracic vessels in various abnormalities.     

Age changes in cells in Streptococcus diacetilactis cultures

The activity of metabolism changed in the cultures of Streptococcus diacetilactis, strain Bogdan, with aging. The number of viable cells decreased as well as the ability to evolve oxygen, to produce C-4 compounds, and to react to pH changes by liberating acids upon alkalifying or neutral products upon acidifying. After the culture had been grown during 72 hours, 2,3-butyleneglycol was found in the cultural broth, and the number of viable cells was low. As was revealed by electron microscopy, the cells were intact during 48 hours; they started to disintegrate by 72 hours. After 120 hours all cells were disintegrated. First, the cells produced no more mesosomes, became less electron dense; then the cell wall was decomposed and the contents of the cell poured through disruptions in the cytoplasmic membrane.     

Postresuscitation changes in brain free radical-mediated processes and nitric oxide synthase activity in rats: effects of individual behavior in "emotional resonance" test

Effects of 7-min cardiac arrest and individual behavior on free radical-mediated processes and nitric oxide synthase (NOS) activity was evaluated in brains of male Wistar rats one hour and one week after resuscitation. "Emotional resonance" test was used for the behavioral selection of rats. The test includes factors of significance for rats: the choice between large and lighted or small and dark space as well as signals of pain of another rat. Free radical generation (using chemiluminescence method), superoxide scavenging/generating activity, substances reacting with 2-thiobarbituric acid and NOS activity (by measuring mononitrosyl iron complex of NO with diethyl dithiocarbamate and endogenous brain Fe2+ by electron spin resonance spectroscopy) were determined in cerebral cortex, cerebellum and hippocampus. Cardiac arrest induced oxidative stress accompanied by the loss of NOS activity, as well as compensatory changes of free radical-mediated processes in cerebral cortex. Oxidative stress was also evident in cerebellum and, to a lesser extent, in hippocampus. Most of neurochemical differences between behavioral groups were induced by cardiac arrest. These differences were global, related to a specific brain region or became apparent in cerebral lateralization of biochemical indices.     

Effect of flurenizide on adaptive reactions in patients with chronic obstructive pulmonary disease

Studies on the wake-sleep cycle with participation of eight female volunteers were performed before, during, and after a 120-day bed rest with the head-end of bed tilted down at 6 degrees (HDT). Methods of polysomnography and actography were applied. The test-subjects were assigned into 2 groups. Group A was prescribed to use countermeasures throughout the experiment; no countermeasures was administered by group B. Evidence of significant alteration in sleep structure at different time points in HDT is given in comparison with data about females of the control group under the conditions of everyday activity. Sleep deviations in the subjects were reordered at each of the three points of investigation and differed from those in control. HDT was shown to modify the sleep structure in experimental groups A and B. A supposition is made that under these conditions the dynamics of physical activity during night sleep had an adaptive character.     

Simultaneous bleaching of vital teeth and an open-chamber nonvital tooth with 10% carbamide peroxide

OBJECTIVE: The purpose of this study was to evaluate the effectiveness of bleaching a nonvital tooth with an open pulp chamber while simultaneously bleaching the other vital teeth with 10% carbamide peroxide. METHOD AND MATERIALS: Ten discolored nonvital teeth were treated. Each nonvital tooth was prepared as in the conventional "walking bleaching" fashion, so that the gutta-percha was sealed from the pulp chamber. The 10% carbamide peroxide was injected into the chamber of the nonvital tooth and loaded into the custom-fitted tray for all teeth. The nonvital teeth were bleached from both the inside and the outside. The patient closed the orifice with a cotton pellet during the day and changed the cotton pellet after meals. The patient applied fresh solution nightly. RESULTS: All teeth were successfully lightened. The time required to lighten the nonvital tooth was related to the duration of the discoloration. CONCLUSION: With proper patient selection and education, this technique can provide an effective way to lighten nonvital and vital teeth simultaneously, especially where extended treatment time may be required for difficult discolorations.     

Perturbation of the internal water chain in cytochrome f of oxygenic photosynthesis: loss of the concerted reduction of cytochromes f and b6

The 1.96 A structure of turnip cytochrome f revealed a linear internal chain of H2O molecules with the oxygen atoms of the chain having occupancies and "B" factors comparable to those of neighboring atoms [Martinez et al. (1996) Protein Sci. 5, 1081-1092. ]. Four waters extend 11 A from the heme toward Lys66 on the cytochrome surface. All residues that contribute an atom to the 15 H-bonds of five internal H2O molecules are essentially conserved in 23 cytochrome sequences. With only Gln and Asn side chains involved in H-bonding, the water chain resembles a "proton wire". The function of the conserved H2O chain was tested through site-directed mutagenesis of these Asn and Gln residues. Four of the five conserved Asn/Gln residues were changed in six mutants generated in the green alga, Chlamydomonas reinhardtii. Except for the N168F mutant, all grew photosynthetically. Although the rates of oxidation of cyt f oxidation and of reduction of cyt b6 (5-6 ms in the wild type) were not significantly affected, the rates of cyt f reduction and generation of the slow electrochromic band shift (Deltapsis) were markedly decreased, the half-times increasing to as much as 38 and 18 ms, respectively. Thus, in these mutants, reduction of cyt b6 reduction clearly precedes that of cyt f. Retardation of Deltapsis in the absence of an observable change in the rate of cyt b6 reduction implied that the rate of H+ translocation decreased in the mutants, and electron transfer was concomitantly retarded, most likely between the ISP and cyt f. The following was concluded: (i) proton and electron transfer are coupled in reduction of cyt f, and the cyt f water chain functions in H+ transfer; (ii) reduction of the high- and low-potential chains in the b6f complex is not concerted in the water chain mutants; and (iii) quinol deprotonation and electron transfer from reduced quinone are initiated by an early event, probably the movement of the ISP triggered by oxidation of cyt f.     

Radiotherapy, toxicity and dosimetry of copper-64-TETA-octreotide in tumor-bearing rats

The efficacy of 64Cu [T1/2 = 12.7 hr; beta+ (0.655 MeV; 19%); beta- (0.573 MeV; 40%)] as a radioisotope for radiotherapy has been recently established. Here we demonstrate that 64Cu-1,4,8,11 -tetraazacyclotetradecane-N,N',N",N'-tetraacetic acid (TETA)-octreotide, a somatostatin receptor ligand, inhibits the growth of CA20948 rat pancreatic tumors in Lewis rats at doses that cause minimal toxicity. METHODS: Tumor-bearing rats were administered a single 15 mCi (555 MBq) dose, a fractionated dose of 15 mCi given in 2-3 doses over 2-8 days, or control agents of buffer, unlabeled octreotide or 64Cu-labeled TETA. In certain experiments, blood was removed at times from 4-23 days post-treatment, and a complete blood count along with blood chemistry analyses were obtained. RESULTS: Tumor-growth inhibition was significantly greater in rats injected with a single 15 mCi dose than in rats injected with control agents (p < 0.05). Dose fractionation in two doses, either 1 or 2 days apart, induced significantly increased tumor-growth inhibition compared with rats given a single dose (p < 0.05). The only toxicity observed in treated rats was a decrease in the white blood cell count. This drop was more pronounced in rats treated with a single dose compared with those treated with a fractionated dose. Human absorbed doses of 64Cu-TETA-octreotide to normal organs were estimated from biodistribution data in Lewis rats, and these data indicate that radiotherapy with 64Cu-TETA-octreotide in humans would be feasible. CONCLUSION: Copper-64-TETA-octreotide is a promising radiopharmaceutical for targeted radiotherapy of somatostatin receptor-positive tumors.     

Molecular modelling and site-directed mutagenesis of the active site of endothelin-converting enzyme

Mammalian endothelin-converting enzyme is a membrane-bound metalloprotease; its C-terminal domain contains sequence motifs characteristic of zinc metalloproteases. We examined residues expected from molecular modelling to be important for substrate binding using selectively mutated recombinant rat ECE-1alpha expressed in CHO cells. A conserved N-A-Ar-Ar (Ar = aromatic) motif is likely to be important for substrate binding. Mutating N550 to Gln or Y552 to Phe reduces Vmax/Km by 8- and 18-fold, respectively. The equivalent residue to Y553 in thermolysin binds the inhibitor through its NH group. Removing this putative interaction by mutating Tyr to Pro destroys activity, but mutating it to Ala or Phe also removes most activity. Mutating G583 (in a conserved GGI motif N-terminal of the zinc-binding helix) to Ala has no measurable effect, but mutating G584 to Ala destroys activity. Changing V583 in the zinc-binding helix to Met, to mimic the sequence pattern in bovine ECE-2, increases Vmax/Km to 1.7-fold that of the wild- type. Assays of phosphoramidon binding follow the pattern of those of substrate binding, but the IC50 of the more potent ECE inhibitor CGS 26303 was not significantly altered by any of these mutations, suggesting that this compound may bind to ECE in a different mode from phosphoramidon.