Formation of negative and positive ions in the radiosensitizer nimorazole upon low-energy electron collisions.

A comprehensive investigation of low-energy electron attachment and electron ionization of the nimorazole radiosensitizer used in cancer radiation therapy is reported by means of a gas-phase crossed beam experiment in an electron energy range from 0 eV to 70 eV. Regarding negative ion formation, we discuss the formation of fifteen fragment anions in the electron energy range of 0 eV-10 eV, where the most intense signal is assigned to the nitrogen dioxide anion NO2 -. The other fragment anions have been assigned to form predominantly from a common temporary negative ion state close to 3 eV of the nitroimidazole moiety, while the morpholine moiety seems to act only as a spectator in the dissociative electron attachment event to nimorazole. Quantum chemical calculations have been performed to help interpreting the experimental data with thermochemical thresholds, electron affinities, and geometries of some of the neutral molecules. As far as positive ion formation is concerned, the mass spectrum at the electron energy of 70 eV shows a weakly abundant parent ion and C5H10NO+ as the most abundant fragment cation. We report appearance energy (AE) measurements for six cations. For the intact nimorazole molecular cation, the AE of 8.16 ± 0.05 eV was obtained, which is near the presently calculated adiabatic ionization energy.

Impact of the Conformational Variability of Oligopeptides on the Computational Prediction of Their CD Spectra.

Although successful in the structural determination of ordered biomolecules, the spectroscopic investigation of oligopeptides in solution is hindered by their complex and rapidly changing conformational ensemble. The measured circular dichroism (CD) spectrum of an oligopeptide is an ensemble average over all microstates, severely limiting its interpretation, in contrast to ordered biomolecules. Spectral deconvolution methods to estimate the secondary structure contributions in the ensemble are still mostly based on databases of larger ordered proteins. Here, we establish how the interpretation of CD spectra of oligopeptides can be enhanced by the ability to compute the same observable from a set of atomic coordinates. Focusing on two representative oligopeptides featuring a known propensity toward an α-helical and ß-hairpin motif, respectively, we compare and cross-validate the structural information coming from deconvolution of the experimental CD spectra, sequence-based de novo structure prediction, and molecular dynamics simulations based on enhanced sampling methods. We find that small conformational variations can give rise to significant changes in the CD signals. While for the simpler conformational landscape of the α-helical peptide de novo structure prediction can already give reasonable agreement with the experiment, an extended ensemble of conformers needs to be considered for the ß-hairpin sequence.

Low-energy electron-induced decomposition of 5-trifluoromethanesulfonyl-uracil: A potential radiosensitizer.

5-trifluoromethanesulfonyl-uracil (OTfU), a recently proposed radiosensitizer, is decomposed in the gas-phase by attachment of low-energy electrons. OTfU is a derivative of uracil with a triflate (OTf) group at the C5-position, which substantially increases its ability to undergo effective electron-induced dissociation. We report a rich assortment of fragments formed upon dissociative electron attachment (DEA), mostly by simple bond cleavages (e.g., dehydrogenation or formation of OTf-). The most favorable DEA channel corresponds to the formation of the triflate anion alongside with the reactive uracil-5-yl radical through the cleavage of the O-C5 bond, particularly at about 0 eV. Unlike for halouracils, the parent anion was not detected in our experiments. The experimental findings are accounted by a comprehensive theoretical study carried out at the M06-2X/aug-cc-pVTZ level. The latter comprises the thermodynamic thresholds for the formation of the observed anions calculated under the experimental conditions (383.15 K and 3 × 10-11 atm). The energy-resolved ion yield of the dehydrogenated parent anion, (OTfU-H)-, is discussed in terms of vibrational Feshbach resonances arising from the coupling between the dipole bound state and vibrational levels of the transient negative ion. We also report the mass spectrum of the cations obtained through ionization of OTfU by electrons with a kinetic energy of 70 eV. The current study endorses OTfU as a potential radiosensitizer agent with possible applications in radio-chemotherapy.

The situation of sanitary systems in rural areas in the Miyun catchment, China.

The Miyun Reservoir provides most of Beijing's drinking water. Despite its importance, the Miyun reservoir suffers from decreasing water quality caused by uncontrolled wastewater discharges, inadequate land use and over fertilization, which increase the pressure on soil and water resources. The major pollutants are nitrogen and phosphorus which emanate to some extent from untreated sewage. So far there is little data about the existing wastewater quantity and quality in rural settlements in northern China. This study was conducted in typical villages situated along upstream rivers in the catchment of the Miyun Reservoir. The main objective was to determine the current situation and efficiency of the wastewater treatment system in rural settlements.

Strategies to mitigate diffuse phosphorus pollution during rewetting of fen peat soils.

The cultivation of fen peat soils (Eutric Histosols) for agricultural purposes, started in Europe about 250 years ago, resulting in decreased soil fertility, increased oxidation of peat and corresponding greenhouse gas fluxes to the atmosphere, nutrient transfer to aquatic ecosystems and losses in total area of the former native wetlands. To prevent these negative environmental effects set-aside programs and rewetting measures were promoted in recent years. Literature results and practical experiences showed that large scale rewetting of intensively used agricultural Histosols may result in mobilisation of phosphorus (P), its transport to adjacent surface waters and an accelerated eutrophication. The paper summarises results from an international European Community sponsored research project and demonstrates how results obtained at different scales and from different scientific disciplines were compiled to derive a strategy to carry out rewetting measures. Based on this findings a simple decision support system (DSS) for a hydrologically sensitive area in the Droemling catchment in north-eastern Germany was developed and since 2005 practically used to prevent freshwater resources from non point P pollution.

The impact of pulse pressure on the accuracy of wrist blood pressure measurement.

There is an increasing number of wrist blood pressure measurement devices that successfully passed the validation procedures of the British Hypertension Society (BHS) and the European Society of Hypertension (ESH). It remains unknown, however, whether pulse pressure as a marker of arterial stiffness and vascular ageing affects the accuracy of these devices. An ESH protocol validated wrist device was compared with the upper arm mercury sphygmomanometry in a study population (33 patients, 99 measurements) including a relevant number of subjects with pulse pressure >50 mm Hg (84.8%) and isolated systolic hypertension (27.3%). Mean systolic bias was 10.2 mm Hg with 95% limits of agreement of -13.1 and 33.6 mm Hg, mean diastolic bias was 4.8 mm Hg with limits of agreement of -11.0 and 20.7 mm Hg. The impact of body mass index, age, systolic blood pressure and pulse pressure on the absolute value of blood pressure bias was tested by stepwise multiple regression analysis. The systolic bias significantly depended on pulse pressure, whereas there was no significant effect of the independent variables on the diastolic bias. Separate correlation analysis showed a significant correlation between pulse pressure and both absolute systolic bias (Pearson r=0.48, P<0.001) and relative systolic bias (systolic bias divided by systolic blood pressure, Pearson r=0.29, P=0.003). Even well-validated wrist blood pressure devices can show a clinically relevant bias in patients with elevated pulse pressure. Increased arterial stiffness may impair the accuracy of oscillometric blood pressure measurement at the wrist.

Evaluation of the polymorphisms in the exons 2 to 4 of the TP53 in cervical carcinoma patients from a Brazilian population.

The majority of TP53 polymorphisms and cervical cancer association studies have only analyzed codon 72 polymorphism. Eight polymorphisms were reported in the region encompassing exon 2 to 4 of TP53 that codify the aminoterminal p53 region containing domains involved in the transcription transactivation and apoptosis induction. We investigated if the polymorphisms present in this region were associated with cervical cancer risk. A total of 140 samples (83 from Brazilian patients with cervical carcinoma and 57 from Brazilian healthy women) were analyzed by PCR and DNA sequencing. Only three from the eight TP53 polymorphisms described in the analyzed region were polymorphic within our samples: the 11827 base from intron 2, the 16bp duplication in the intron3 and the codon 72 (Arg>Pro) from exon 4. No statistically significant association was observed between polymorphisms from intron 2 and the 16bp duplication from intron 3 with cervical cancer. No statistically significant difference in the frequency of homozygotes for Arg in relation to other genotypes was found when comparing patient and healthy groups (OR=0.70; 95% CI= 0.31-1.56; p= 0.222). However, Arg/Pro heterozygotes were more frequent within HPV positive cancer patients than in healthy women (p=0.023; OR (Arg/Pro:Pro/Pro)= 5.82; 95% CI: 1.22-30.78; p=0.024).

Differences in the absorption, metabolism and biliary excretion of a diastereomeric pair of alphavbeta3-antagonists in rat: limited role of P-glycoprotein.

1. The study investigated mechanisms underlying the pharmacokinetic differences of two zwitterionic diastereomers ((3S)-3-[(3R or 3S)-2-oxo-3-[3-(5,6,7,8-tetrahydro-1,8-naphthyridin-2-yl)propyl]pyrrolidin-1-yl]-3-quinolin-3-ylpropanoic acid) with different lipophilicities using a combination of in vivo and in vitro approaches. 2. In rat, both isomers possessed comparable plasma clearances (CL). However, the more lipophilic diastereomer I exhibited a higher metabolic clearance (>2-fold higher than II), whereas the hydrophilic zwitterion II exhibited a higher biliary clearance (approximately 5-fold higher than I). Following oral administration, the bioavailability (F) of I (17%) was much higher than that of II (1%). 3. Consistent with these in vivo observations and the expectation based on their lipophilicity differences, the metabolism in rat liver microsomes was faster and the permeability in Caco-2 and LLC-PK1 cells and in situ rat intestinal loop was better for I than for II. 4. Only the absorption of the more lipophilic diastereomer I was subjected to an efflux system in the Caco-2 and in situ rat intestinal loop models. I was a good substrate for P-glycoprotein (P-gp) in both the human MDR1 and mouse mdr1a transfected cell lines, and in the wild-type mdr1a (-/-) mouse when compared with the P-gp-deficient mdr1a (-/-) mouse. Concomitant administration of I with verapamil in rat caused significant increases in oral AUC, F and Cmax of I without affecting its CL, further supporting the effect of P-gp in limiting the intestinal absorption of I in vivo in this animal model. 5. Since the findings that the lipophilic diastereomer I, but not II, was a good P-gp substrate were not in line with the observations that I was excreted to bile much slower than II and that I was absorbed better than II, the results suggested that P-gp played a minor role to the observed differences in the biliary excretion and intestinal absorption of the diastereomers I and II in rat.

MYB61 is required for mucilage deposition and extrusion in the Arabidopsis seed coat.

We have undertaken a systematic reverse genetic approach to understand R2R3-MYB gene function in Arabidopsis. Here, we report the functional characterization of MYB61 based on the phenotype of three independent insertion alleles. Wide-ranging phenotype screens indicated that MYB61 mutants were deficient in seed mucilage extrusion upon imbibition. This phenotype was expressed in the sporophytic tissues of the seed. Deposition and extrusion of the principal component of the mucilage, a relatively unbranched rhamnogalacturonan, were reduced in the MYB61 mutant seed coats. Additional defects in the maturation of the testa epidermal cells suggested a potential deficiency in extracellular secretion in myb61 lines. Consistent with a proposed role in testa development, reverse transcription-polymerase chain reaction analysis showed the highest MYB61 expression in siliques, which was localized to the seed coat by a beta-glucuronidase (GUS) reporter gene fusion. Lower levels of GUS expression were detected in developing vascular tissue. Parallel analysis of the ttg1-1 mutant phenotype indicated that this mutant showed more severe developmental defects than myb61 and suggested that MYB61 may function in a genetic pathway distinct from that of TTG1. The transient nature of seed epidermal characteristics in the ttg1-1 mutant suggested that TTG1 was required for maintenance rather than initiation of testa epidermal differentiation. Germination and seedling establishment were compromised in the myb61 and ttg1-1 mutants under conditions of reduced water potential, suggesting a function for Arabidopsis seed mucilage during germination in dry conditions.

Technical advance: a high throughput system for transposon tagging and promoter trapping in tomato.

We describe new tools for functional analysis of the tomato genome based on insertional mutagenesis with the maize Ac/Ds transposable elements in the background of the miniature cultivar Micro-Tom. 2932 F3 families, in which Ds elements transposed and were stabilized, were screened for phenotypic mutations. Out of 10 families that had a clear mutant phenotype, only one mutant was Ds-tagged. In addition, we developed promoter trapping using the firefly luciferase reporter gene and enhancer trapping, using beta-glucuronidase (GUS). We show that luciferase can be used as a non-invasive reporter to identify, isolate and regenerate somatic sectors, to study the time course of mutant expression, and to identify inducible genes. Out of 108 families screened for luciferase activity 55% showed expression in the flower, 11% in the fruit and 4% in seedlings, suggesting a high rate of Ds insertion into genes. Preferential insertion into genes was supported by the analysis of Ds flanking sequences: 28 out of 50 sequenced Ds insertion sites were similar to known genes or to ESTs. In summary, the 2932 lines described here contain 2-3 Ds inserts per line, representing a collection of approximately 7500 Ds insertions. This collection has potential for use in high-throughput functional analysis of genes and promoter isolation in tomato.

[Rotavirus detection in feces of children with acute diarrhea] / Detecção de rotavírus nas fezes de crianças com diarréia aguda.

OBJECTIVE: To determine the prevalence of rotavirus in the etiology of acute diarrhea in children from Natal city, RN, Brazil and investigate the existence or not of a seasonal distribution of this pathogen in our environment.METHODS: Fecal samples from 1,903 children (boys and girls) with ages ranging from 1 month to 10 years, living in Natal-RN, who presented acute diarrhea episodes in a period from January 1996 to December 1998, were analyzed. We searched viral particles directly in the feces by a passive agglutination reaction using anti-rotavirus specific-group monoclonal antibodies coated latex particles.RESULTS: 151 children (7.9%) of the studied population presented a positive reaction, revealing the presence of rotavirus particles in feces. Considering, however, only the children (ages from 1 to 24 months) who are more susceptible to rotavirus infection, we verified that from 1,065 examined children, 136 of them (12.8%) presented positive reaction for rotavirus, and the great majority of all children with positive reaction (96.3%) had ages ranging from 6 to 24 months. Analysis of the distribution of the cases of rotavirus infection in the three year revealed that the incidence of infection was higher in July, August, and September.CONCLUSIONS: The results indicated rotavirus infections have an important role in the etiology of acute diarrhea cases. The majority of the acute diarrhea cases positive for rotavirus occurred during the first two years of life, reaching mainly children from 6 to 24 months, with the highest incidence of infection during the months of July, August, and September.

Function search in a large transcription factor gene family in Arabidopsis: assessing the potential of reverse genetics to identify insertional mutations in R2R3 MYB genes.

More than 92 genes encoding MYB transcription factors of the R2R3 class have been described in Arabidopsis. The functions of a few members of this large gene family have been described, indicating important roles for R2R3 MYB transcription factors in the regulation of secondary metabolism, cell shape, and disease resistance, and in responses to growth regulators and stresses. For the majority of the genes in this family, however, little functional information is available. As the first step to characterizing these genes functionally, the sequences of >90 family members, and the map positions and expression profiles of >60 members, have been determined previously. An important second step in the functional analysis of the MYB family, through a process of reverse genetics that entails the isolation of insertion mutants, is described here. For this purpose, a variety of gene disruption resources has been used, including T-DNA-insertion populations and three distinct populations that harbor transposon insertions. We report the isolation of 47 insertions into 36 distinct MYB genes by screening a total of 73 genes. These defined insertion lines will provide the foundation for subsequent detailed functional analyses for the assignment of specific functions to individual members of the R2R3 MYB gene family.

Nonpeptide glycoprotein IIB/IIIA inhibitors. 19. A new design paradigm employing linearly minimized, centrally constrained, exosite inhibitors.

A new series of potent, linearly-minimized, orally active, selective GPIIb/IIIa inhibitors is identified. Thus 15 (L-750,034) achieves interaction via a constrained, non-turned conformation that maintains the proper distance between its charged termini and full sulfonamide exosite interaction. The diminutive stature and the proposed linear conformation of L-750,034 define a new paradigm for the conceptualization of RGD mimics.

Towards functional characterisation of the members of the R2R3-MYB gene family from Arabidopsis thaliana.

Transcription factors containing a conserved DNA-binding domain similar to that of the proto-oncogene c-myb have been identified in nearly all eukaryotes. MYB-related proteins from plants generally contain two related helix-turn-helix motifs, the R2 and R3 repeats. It was estimated that Arabidopsis thaliana contains more than 100 R2R3-MYB genes. The few cases where functional data are available suggest an important role of these genes in the regulation of secondary metabolism, the control of cell shape, disease resistance, and hormone responses. To determine the full regulatory potential of this large family of regulatory genes, a systematic search for the function of all genes of this family was initiated. Sequence data for more than 90 different A. thaliana R2R3-MYB genes have been obtained. Sequence comparison revealed conserved amino acid motifs shared by subgroups of R2R3-MYB genes in addition to the characteristic DNA-binding domain. No significant clustering of the genes was detected, although they are not uniformly distributed throughout the A. thaliana genome.

A polymorphism in exon b2 of the major breakpoint cluster region (M-bcr) identified in chronic myeloid leukaemia patients.

The BCR/ABL junctional region and the b3 exon from chronic myeloid leukaemia (CML) patients were sequenced. In all 21 samples analysed the junctional region, as well as the b3 exon of 8 b3a2 mRNA molecules, presented no differences to the already described sequences. However, we identified a polymorphic base in the b2 exon in two out of seven b3a2 samples, four out of 10 b2a2 samples and all four b3a2/b2a2 samples analysed. In the eighth position before the junctional region of BCR/ABL cDNA, a cytosine replaces thymine in these cases. The polymorphism described here could be a useful marker for the differentiation of normal and rearranged BCR alleles in heterozygotes.

Isolation and characterisation of a diverse family of Arabidopsis two and three-fingered C2H2 zinc finger protein genes and cDNAs.

In animal systems the C2H2 zinc finger protein (ZFP) gene family is the largest group of regulatory proteins and its members have a wide and important role in growth and development. It is likely that this family of ZFP transcription factors will also be important in plants. We have used a PCR approach employing highly degenerate oligonucleotide primers to isolate several Arabidopsis genomic and cDNA clones encoding potential ZFPs. In addition we have used the sequence information from these clones to identify two ESTs as members of this family. Five two-fingered and one three-fingered ZFPs have been identified. Outside of the zinc finger regions, there is considerable sequence diversity, including the sequence and length of the interfinger region; the expression pattern of each gene is different. This is the first report of the isolation of a three-fingered plant C2H2 ZFP gene and we discuss its possible evolutionary origin from two-fingered ZFP genes.

Partially Molten Middle Crust Beneath Southern Tibet: Synthesis of Project INDEPTH Results

INDEPTH geophysical and geological observations imply that a partially molten midcrustal layer exists beneath southern Tibet. This partially molten layer has been produced by crustal thickening and behaves as a fluid on the time scale of Himalayan deformation. It is confined on the south by the structurally imbricated Indian crust underlying the Tethyan and High Himalaya and is underlain, apparently, by a stiff Indian mantle lid. The results suggest that during Neogene time the underthrusting Indian crust has acted as a plunger, displacing the molten middle crust to the north while at the same time contributing to this layer by melting and ductile flow. Viewed broadly, the Neogene evolution of the Himalaya is essentially a record of the southward extrusion of the partially molten middle crust underlying southern Tibet.

Autoencapsulation through intermolecular forces: a synthetic self-assembling spherical complex.

The synthesis and characterization of a system for the study of molecular recognition phenomena are described. The system involves a tetraurea molecule that is capable of assembly into various associated states through hydrogen bonding. In organic solvents, the dynamic transition between a low-ordered (aggregate) state and a highly ordered dimeric assembly can be induced by the introduction of smaller molecules of appropriate size and shape. These smaller molecules, such as benzene, adamantanes, and ferrocenes, act as guests that occupy the pseudospherical capsule formed by the dimeric host. Among various guests, those that best fill the cavity and offer chemical complementarity to the host are preferentially encapsulated.

Isolation and characterization of the tomato homeobox gene THOM1.

We have screened a cDNA library prepared from tomato (Lycopersicon esculentum Mill. cv. VFNT Cherry) shoot tips for homeobox-containing clones. The isolated cDNA clone THOM1 (tomato homeobox gene 1) contains a homeobox, a leucine zipper motif and and an acidic region. These features support the hypothesis that the THOM1 gene product acts as a transcriptional activator. THOM1 is highly expressed in the vegetative shoot apical meristem, the floral meristem and axillary meristems. Young derivatives of these meristems show similar levels of THOM1 transcripts which decrease with increasing age of the respective tissue. The THOM1 gene is located in the centromere region of chromosome 1 and belongs to a small gene family of the tomato genome.