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Distribution of organic carbon, selected stable elements and artificial radionuclides among dissolved, colloidal and particulate phases in the Rhône River (France): preliminary results.

The behaviour of radionuclides discharged from nuclear facilities in the Rhône River depends on their distribution among the dissolved, colloidal and particulate phases. A large water sample was fractionated using sequential ultrafiltration. Size distributions of organic carbon, Fe, Al, Si, Ca, Mg, Cu, Zn, 137Cs, 60Co and 106Ru were obtained. Our results show that organic colloids account for 11% of the total organic carbon content. Approximately 20% of the dissolved (< 450 nm) Fe and Al are in colloidal classes. 137Cs is not significantly transferred by the colloidal phase while 25% of 60Co or 106Ru is associated with organic and inorganic colloids.

Carbon↗

Thioether complexes of palladium(II) and platinum(II) as artificial peptidases. residue-selective peptide cleavage by a palladium(II) complex.

We report the synthesis and characterization of perchlorate salts containing the following three novel complex cations each with a bidentate thioether ligand: binuclear cis-[Pt(CH3SCH2CH2CH2SCH3)(mu-OH)]22+, mononuclear cis-[Pt(CH3SCH2CH2CH2SCH3)(H2O)2]2+, and mononuclear cis-[Pd(CH3SCH2CH2CH2SCH3)(H2O)2]2+. Despite their analogous compositions, the mononuclear Pt(II) and Pd(II) complexes differ in the selectivity with which they promote the hydrolysis of polypeptides. The complex cis-[Pt(CH3SCH2CH2CH2SCH3)(H2O)2]2+ promotes slow but selective cleavage of Met-Pro peptide bonds at pH 2.0. The selectivity of the complex cis-[Pd(CH3SCH2CH2CH2SCH3)(H2O)2]2+ is pH-dependent. At pH 2.0, this Pd(II) complex promotes residue-selective hydrolysis of the X-Y bond in X-Y-Met and X-Y-His sequences; the rate is enhanced when residue Y is proline. At pH 7.0, this kinetic preference becomes sequence-selective in that the Pd(II) complex exclusively cleaves the X-Pro bond in X-Pro-Met and X-Pro-His sequences. The enhanced reactivity of the X-Pro amide group is attributed to the high basicity of its carbonyl oxygen atom. Binding of the metal(II) atom enhances the electrophilicity of the carbonyl carbon atom and promotes nucleophilic attack by a solvent water molecule. The bidentate thioether ligand disfavors the formation of hydrolytically unreactive complexes, allowing the Pd(II) complex to promote the cleavage reaction.

Chromatography, High Pressure Liquid↗

Design of artificial transcription factors to selectively regulate the pro-apoptotic bax gene.

The tumor suppressor p53 is the most commonly mutated gene in human cancers. Active p53 is able to stimulate the transcription of a variety of genes including the pro-apoptotic gene bax, as well as p21, a cell cycle regulator. In this study we produced novel zinc finger transcription factors that would selectively increase the expression of bax, but not of other p53 targets. Reporter gene assays in p53-negative Saos-2 cells showed that the novel zinc finger proteins stimulated transcription driven by a minimal bax promoter, but not that driven by a minimal p21 promoter. Moreover, electromobility shift assays demonstrated that the novel transcription factors could bind the bax promoter sequence with high affinity and selectivity. Expression of a five zinc finger protein (5ZFAV) in COS-7 cells resulted in an increase in Bax protein levels, indicating that this novel transcription factor could act on endogenous gene expression. Expression of 5ZFAV also drastically reduced Saos-2 cell survival; this effect could be reversed by the general caspase inhibitor B-D-FMK. These data suggest that 5ZFAV is able to induce apoptosis through increased Bax expression. Further, while expression of 5ZFAV in p53-deficient Saos-2 cells reduced cell survival, there was little effect on U-2 OS cells which have wild-type p53. Thus the selective induction of the pro-apoptotic bax gene may be a valuable adjunct to cancer chemotherapy by diminishing survival of p53-deficient tumor cells.

Animals↗

Synthesis and evaluation of oligonucleotides incorporating novel artificial nucleobases for the selective formation of non-natural type triplexes.

Regulation of specific gene expression by the sequence specific triplex formation has been expected as a new biological tool or a new therapeutic method in the so-called antigene strategy. As native nucleobases form triplexes only within homopurine-homopyrimidine duplexes, expansion of the target sequences to general ones has been desired. We have previously reported the selective formation of base triplets by the non-native nucleobases (1, 2) with a CG or a TA base pair, respectively, in the organic solvent. In this paper, we wish to report the synthesis of the new non-native bases (3, 4, 9-11) and their incorporation into the triplex-forming oligonucleotides (TFOs), as well as their ability of triplex formation.

Base Composition↗

Artificial neural network-based channel selection and loudness mapping.

We present in this paper artificial neural network techniques for implementing loudness mapping and "smart" channel selection for cochlear implant systems. For loudness mapping, a multilayer perceptron (MLP) is trained to perform the mapping for each channel according to threshold and comfort levels. It is shown that good accuracy mapping can be performed by a very simple MLP architecture. For channel selection, we propose a neural network-based method that can make "smart" selection. We describe and report results for the case in which 6 channels are to be selected from 18. The neural network-based selection system is trained on a multispeaker labeled speech database and tested on a database of different speakers and spoken sentences. Compared with methods used by leading cochlear implant systems, our approach produces significantly better results, and it is easy to implement in the speech processor of the cochlear implant system.

Cochlear Implants↗

Growth of methylaminotrophic, acetotrophic and hydrogenotrophic methanogenic bacteria on artificial supports.

The efficiency of organic matter degradation in attached biomass reactors depends on the suitable selection of artificial support for the retention of bacterial communities. We have studied the growth on glass and clay beads of methylaminotrophic, acetotrophic and hydrogenotrophic methanogenic bacterial communities isolated from anaerobic reactors. Bacterial counts were performed by the standard MPN technique. Experiments were performed in 50 ml vials for 12 days at 35 degrees C. Increase in the counts of methylaminotrophic and hydrogenotrophic methanogens occurred on both glass and clay beads. The latter support material also stimulated the growth rate of methylaminotrophic methanogens.

Aluminum Silicates↗

Using weather indices to predict survival of winter wheat in a cool temperate environment.

Seven years of winter survival data for winter wheat ( Triticum aestivum L.) were collected on a loam soil located on the Central Experimental Farm at Ottawa, Ontario (45 degrees 23'N, 75 degrees 43'W). The site was low-lying and subject to frequent winter flooding and ice-sheet formation. Two cultivars, a soft white and a hard red winter wheat, were planted in September. Crop establishment was measured in late fall and the percentage survival was measured in April of the following year. Meteorological data, which were available from the nearby weather site, were used to develop a large set of monthly weather indices that were felt to be important for winter survival. The objective of the study was to use genetic selection algorithms and artificial neural networks to select a subset of critical weather factors and topographic features and to model winter survival. The six weather indices selected were the total rain depth for December (mm), the total rain depth for February (mm), the number of days of the month with snow on the ground for January, the extreme minimum observed daily air temperature for March ( degrees C), the number of days of the month with snow on the ground for March, and the number of days of April with a daily maximum air temperature greater than 0 degrees C. It was found 89% of the variation in winter survival could be explained by these six weather indices, the cultivar, elevation and plot location.

Forecasting↗

Artificial metalloprotease with active site comprising aldehyde group and Cu(II)cyclen complex.

To design artificial proteases that cleave peptide backbones of a wide range of proteins at selected sites, artificial active sites comprising the Cu(II) complex of cyclen (Cu(II)Cyc) and aldehyde group were synthesized on a cross-linked polystyrene. The aldehyde group was employed as the binding site in view of its ability of reversible formation of imine bonds with epsilon-amino groups of Lys residues exposed on the surface of proteins and Cu(II)Cyc as the catalytic group for peptide hydrolysis. The two polymeric artificial metalloproteases synthesized in the present study cleaved all of the protein substrates examined (myoglobin, gamma-globulin, bovine serum albumin, human serum albumin, lysozyme, and ovalbumin), manifesting saturation kinetic behavior. At 50 degrees C and pH 9.0 or 9.5, K(m) was (1.3-22) x 10(-)(4) M, comparable to those of natural proteases, and k(cat) was (6.0-25) x 10(-)(4) s(-)(1), corresponding to half-lives of 4.6-19 min. Intermediacy of the imine complexes formed between the aldehyde group of the catalyst and the epsilon-amino groups of Lys residues of the substrates was confirmed by the trapping experiment with NaB(OAc)(3)H. MALDI-TOF MS of the proteolytic reaction mixtures revealed formation of various cleavage products. Structures of some of the cleavage products were determined by using carboxypeptidase A and trypsin. Among various cleavage sites thus identified, Gln(91)-Ser(92) and Ala(94)-Thr(95) were the major initial cleavage sites in the degradation of myoglobin by the two catalysts. The selective cleavage of Gln(91)-Ser(92) and Ala(94)-Thr(95) was attributed to general acid assistance in peptide cleavage by Tyr(146) located in proximity to the two peptide bonds. Broad substrate selectivity, high cleavage-site selectivity, and high proteolytic rate are achieved, therefore, by positioning the aldehyde group in proximity to Cu(II)Cyc attached to a cross-linked polystyrene.

Aldehydes↗

Monoiodo-D-tyrosine, an artificial amino acid radiopharmaceutical for selective measurement of membrane amino acid transport in the pancreas.

Using 11C-labeled natural amino acids, the functional diagnosis of tissue metabolism has been actively studied. Our interest has been focused on developing a clinically available 123I-labeled artificial amino acid with a single metabolic function. For this study, [123I]3-iodo-D-tyrosine ([123I]D-MIT) was selected. In vitro and in vivo studies using 125I-labeled D-MIT indicated that it showed a high pancreatic accumulation, selective affinity for membrane active transport systems, and was stable against enzymatic deiodination. A canine scintigraphic study using 123I-labeled D-MIT and kinetic analysis showed that it behaved as an "artificial amino acid" radiopharmaceutical with selective membrane amino acid transport affinity in the pancreas.

Amino Acids↗

[Urea sensor for the continuous ammonium-selective enzymatic process control of the artificial kidney].

Presented is a flow-through method for continuous ammonium-selective enzymatic monitoring of the artificial kidney by means of a bioelectrochemical urea electrode. The urea is converted in an enzyme membrane by covalently bound urease and the ammonium ions are detected by a Nonactin-PVC-membrane. In addition to detailed data from the oxygen-independent solid-state contact sensor, curves are obtained on-line from the patient during the haemodialysis session. The advantages of the method are described in detail. Furthermore, the urea sensor can be used for measurements in heparinized blood.

Anti-Bacterial Agents↗

[The regulation of re-selection form in the aesthetic factor of upper anterior artificial teeth].

Sixty one cases of selection error due to the esthetic factors of artificial tooth forms were investigated out of the 72 cases of selection error which were committed at the department of Prodosthontics in Nihon University school of dentistry at Matsudo for 17 years from 1971 to 1988. The following were the results of investigation, 1. The cases of S-type were changed to those of ST-type, which emphasized the Tapering type factors. 2. The those of ST-type were changed to those of SO-type and S-type which emphasized the Ovoid type factors and got rid of the Tapering type factors. 3. The those of SO-type were changed to those of STO-type which emphasized the Tapering type factors. 4. The those of T-type were changed to those of TO-type which emphasized the Ovoid type factors. 5. The those of O-type were changed to those of TO-type and STO-type which emphasized the Tapering and Square type factors. The regulations of re-selection showed the tendency of changing from single form to complex form, emphasizing the Tapering type factors.

Denture Design↗

Absence of tight junction formation in an allogeneic graft cell line used for developing an engineered artificial salivary gland.

An essential structural feature of fluid-secreting epithelial tissues is the presence of tight junctions. To develop a tissue-engineered organ capable of fluid secretion, the cellular component must establish these structures. As part of efforts to create an engineered artificial salivary gland, we have examined the ability of a candidate allogeneic graft cell line, HSG, to produce several key tight junction proteins, as well as to exhibit functional activities consistent with effective tight junction strand formation. In contrast to results obtained with a control kidney cell line, MDCK-II, HSG cells were unable to synthesize four important tight junction-associated proteins: ZO-1, occludin, claudin-1, and claudin-2. In addition, unlike MDCK-II cells, HSG cell monolayers could not restrict paracellular permeability. HSG cells were, thus, unable to generate significant transepithelial electrical resistance or serve as an effective barrier to osmotically imposed fluid movement. Furthermore, these two functional activities could not be reconstituted via the stable transfection of HSG cells with cDNAs encoding either claudin-1 or claudin-2. We conclude that because of their inability to form tight junctions, HSG cells are unsuitable for use as an allogeneic graft cell in an artificial salivary fluid secretory device. These studies also emphasize the importance of graft cell selection in artificial organ development, as certain required characteristics may be difficult to reengineer.

Cell Line↗

Sex-linked expression of a sexually selected trait in the stalk-eyed fly, Cyrtodiopsis dalmanni.

Recent theoretical and empirical work has suggested that the X chromosome may play a special role in the evolution of sexually dimorphic traits. We tested this idea by quantifying sex chromosome influence on male relative eyespan, a dramatically sexually selected trait in the stalk-eyed fly, Cyrtodiopsis dalmanni. After 31 generations of artificial sexual selection on eyespan:body length ratio, we reciprocally crossed high- with low-line flies and found no evidence for maternal effects; the relative eyespan of F1 females from high- and low-line dams did not differ. However, F1 male progeny from high-line dams had longer relative eyespan than male progeny from low-line dams, indicating X-linkage. Comparison of progeny from a backcross involving reciprocal F1 males and control line females confirmed X-linked inheritance and indicated no effect of the Y chromosome on relative eyespan. We estimated that the X chromosome accounts for 25% (SE = 6%) of the change in selected lines, using the average difference between reciprocal F1 males divided by the difference between parental males, or 34%, using estimates of the number of effective factors obtained from reciprocal crosses between a high and low line. These estimates exceed the relative size of the X in the diploid genome of a male, 11.9% (SE = 0.3%), as measured from mitotic chromosome lengths. However, they match expectations if X-linked genes in males exhibit dosage compensation by twofold hyperactivation, as has been observed in other flies. Therefore, sex-linked expression of relative eyespan is likely to be commensurate with the size of the X chromosome in this dramatically dimorphic species.

Animals↗