SECURITIES CLASS ACTIONS MAY HAVE HIT THEIR PEAK
After 3 years of poignant expansion, sovereign bonds category actions forsaken off in 2009, according to total from dual consulting as well as investigate firms -- a pointer which a flurry of wake up spurred by a credit predicament has died down. Credit crisis-related filings were roughly twice as usual during a initial half of a year than during a second, upon condition which serve justification which those sorts of actions have been upon a approach out.
CELLULAR AND MOLECULAR MECHANISMS FOR THE SYNERGISTIC CYTOTOXICITY ELICITED BY OXALIPLATIN AND PEMETREXED IN COLON CANCER CELL LINES.
Cancer Chemother Pharmacol. 2009 Dec 18;
Nannizzi S, Veal GJ, Giovannetti E, Mey V, Ricciardi S, Ottley CJ, Del Tacca M, Danesi R
PURPOSE: Oxaliplatin effect in the treatment of colorectal cancer is improved upon combination with thymidylate synthase (TS) inhibitors. Pemetrexed is polyglutamated by the folylpolyglutamate synthase (FPGS) and blocks folate metabolism and DNA synthesis by inhibiting TS, dihydrofolate reductase (DHFR) and glycinamide ribonucleotide formyltransferase (GARFT). The present study evaluates the pharmacological interaction between oxaliplatin and pemetrexed in colorectal cancer cells. METHODS: Human HT29, WiDr, SW620 and LS174T cells were treated with oxaliplatin and pemetrexed. Drug interaction was studied using the combination index method, while cell cycle was investigated with flow cytometry. The effects of drugs on Akt phosphorylation and apoptosis were studied with ELISA and fluorescence microscopy, respectively. RT-PCR analysis was performed to assess whether drugs modulated the expression of pemetrexed targets and of genes involved in DNA repair (ERCC1 and ERCC2). Finally, platinum-DNA adduct levels were detected by ultra-sensitive multi-collector inductively coupled plasma mass spectrometry (ICP-MS). RESULTS: A dose-dependent inhibition of cell growth was observed after drug exposure, while a synergistic interaction was detected preferentially with sequential combinations. Oxaliplatin enhanced cellular population in the S-phase. Drug combinations increased apoptotic indices with respect to single agents, and both drugs inhibited Akt phosphorylation. RT-PCR analysis showed a correlation between the FPGS/(TS x DHFR x GARFT) ratio and pemetrexed sensitivity, as well as a downregulation of ERCC1, ERCC2, TS, DHFR and GARFT after drug exposure. In addition, pretreatment with pemetrexed resulted in an increase of oxaliplatin-DNA adducts. CONCLUSION: These data demonstrate that oxaliplatin and pemetrexed synergistically interact against colon cancer cells, through modulation of cell cycle, inhibition of Akt phosphorylation, induction of apoptosis and modulation of gene expression.
ASSOCIATION OF MELAMINE EXPOSURE WITH URINARY STONE AND OXIDATIVE DNA DAMAGE IN INFANTS.
Arch Toxicol. 2009 Dec 18;
Ke Y, Duan X, Wen F, Xu X, Tao G, Zhou L, Zhang R, Qiu B
There is evidence in experimental animals for the urolithiasis and carcinogenicity of melamine, but no evidence for melamine in humans. To evaluate any association between melamine-contaminated powdered formula (MCPF) feeding and urolithiasis, and further the MCPF feeding and oxidative damage to DNA in infants. A cross-sectional study was carried out to assess urolithiasis and urinary 8-hydroxy-2'-deoxyguanosine (8-OHdG) in four groups of infants according to the type of feeding: (1) Infants receiving over 90% of their intake as MCPF. (2) Infants receiving 50-90% of their intake as MCPF. (3) Infants receiving less than 50% of their intake as MCPF. (4) Infants receiving over 90% of their intake as imported milk powdered formula free of melamine contamination. Groups 1 to 3 are the observation groups, and Group 4 is the reference group. There is a significant correlation between urolithiasis and percentage of MCPF intake. The mean urinary 8-OHdG concentrations for Groups 1, 2, 3, and 4 were: 2.03 +/- 0.52, 1.67 +/- 0.28, 1.90 +/- 0.39, and 1.85 +/- 0.47 micromoles per mole of creatinine, respectively. There were no significant differences in the mean urinary 8-OHdG concentrations among the observation and control groups. There were also no correlation between mean urinary 8-OHdG excretions and percentage of MCPF intake. Our data suggested that melamine exposure were associated with urolithiasis, but it might not cause any increase in oxidative damage of DNA in infants.
MUTATION IN THE LYSA GENE IMPAIRS THE SYMBIOTIC PROPERTIES OF MESORHIZOBIUM CICERI.
Arch Microbiol. 2009 Dec 18;
Das SK, Gautam US, Sandhu KV, Bandyopadhyay S, Chakrabartty PK, Singh A
A Tn5-induced mutant of Mesorhizobium ciceri, TL28, requiring the amino acid lysine for growth on minimal medium was isolated and characterized. The Tn5 insertion in the mutant strain TL28 was located on a 6.8-kb EcoRI fragment of the chromosomal DNA. Complementation analysis with cloned DNA indicated that 1.269 kb of DNA of the 6.8-kb EcoRI fragment restored the wild-type phenotype of the lysine-requiring mutant. This region was further characterized by DNA sequence analysis and was shown to contain a coding sequence homologous to lysA gene of different bacteria. The lys (-) mutant TL28 was unable to elicit development of effective nodules on the roots of Cicer arietinum L. There was no detectable level of lysine in the root exudates of chickpea. However, addition of lysine to the plant growth medium restored the ability of the mutant to produce effective nodules with nitrogen fixation ability on the roots of C. arietinum.
MONITORING CELLULAR BEHAVIOUR USING RAMAN SPECTROSCOPY FOR TISSUE ENGINEERING AND REGENERATIVE MEDICINE APPLICATIONS.
J Mater Sci Mater Med. 2009 Dec 18;
Boyd AR, Burke GA, Meenan BJ
Raman spectroscopy has been used to determine the chemical composition of materials for over 70 years. Recent spectacular advances in laser and CCD camera technology creating instruments with higher sensitivity and lower cost have initiated a strong resurgence in the technique, ranging from fundamental research to process control methodology. One such area of increased potential is in tissue engineering and regenerative medicine (TERM), where autologous cell culture, stem cell biology and growth of human cells on biomaterial scaffolds are of high importance. Traditional techniques for the in vitro analysis of biochemical cell processes involves cell techniques such as fixation, lysis or the use of radioactive or chemical labels which are time consuming and can involve the perpetuation of artefacts. Several studies have already shown the potential of Raman spectroscopy to provide useful information on key biochemical markers within cells, however, many of these studies have utilised micro- or confocal Raman to do this, which are not suited to the rapid and non-invasive monitoring of cells. For this study a versatile fit-for-purpose Raman spectrometer was used, employing a macro-sampling optical platform (laser spot size 100 mum at focus on the sample) to discriminate between different TERM relevant cell types and viable and non-viable cells. The results clearly show that the technique is capable of obtaining Raman spectra from live cells in a non-destructive, rapid and non-invasive manner, however, in these experiments it was not possible to discriminate between different cell lines. Despite this, notable differences were observed in the spectra obtained from viable and non-viable cells, showing significant changes in the spectral profiles of protein, DNA/RNA and lipid cell constituents after cell death. It is evident that the method employed here shows significant potential for further utilisation in TERM, providing data directly from live cells that fits within a quality assurance framework and provides the opportunity to analyse cells in a non-destructive manner.
CHLOROPLAST DNA PHYLOGEOGRAPHY OF BETULA MAXIMOWICZIANA, A LONG-LIVED PIONEER TREE SPECIES AND NOBLE HARDWOOD IN JAPAN.
J Plant Res. 2009 Dec 18;
Tsuda Y, Ide Y
Betula maximowicziana is an ecologically and economically important tree species in Japan. In order to examine the phylogeographical pattern of the species in detail, maternally inherited chloroplast (cp) DNA variations of 25 natural populations of Betula maximowicziana and a total of 12 populations of three related species were evaluated by PCR-RFLP analysis. Two main haplotypic groups of B. maximowicziana populations (northern and southern) were detected, with the main boundary passing through the Tohoku region in northeastern Japan; in addition there was high genetic differentiation among the 25 populations studied (G (ST) = 0.950, [Formula: see text]). The phylogeographical pattern exhibited by B. maximowicziana was much more similar to that of alpine plants than to that of beech and oak. Comparison of the patterns of genetic structure obtained from the cpDNA with previously and newly acquired data on bi-parentally inherited nuclear DNA indicates that the nuclear genome was transferred via pollen from the northern haplotypic group to the southern group more frequently than it moved in the opposite direction. Although common haplotypes were detected among B. maximowicziana and the two related species examined, these haplotypes were not shared sympatrically, suggesting very rare hybridization among the species currently occurring in their natural populations.
INITIAL TEMPORAL FIELD DEFECT IN LEBER HEREDITARY OPTIC NEUROPATHY.
Jpn J Ophthalmol. 2009 Nov; 53(6): 603-7
Wakakura M, Fujie W, Emoto Y
PURPOSE: To determine the site of the initial field defect in patients with Leber hereditary optic neuropathy (LHON). METHODS: We studied nine eyes of nine consecutive LHON patients with the 11778 mitochondrial DNA mutation who had no visual loss (four eyes) or only minimal visual loss (five eyes). When unilateral visual loss was observed, Humphrey field analysis (HFA) (HFA 30-2 program and sometimes the HFA 10-2 program) was immediately and repeatedly performed on the better eye. RESULTS: For the 12 centralmost points in the visual field, a loss of sensitivity (P < 0.02) was initially found in the upper temporal field of nine eyes and in the lower temporal field of three eyes. These results indicate that it was possible to detect the initial site of sensitivity loss in the centralmost temporal test points in all nine cases. The HFA 10-2 program confirmed the sensitivity loss in the temporal field in two cases. CONCLUSIONS: The centralmost temporal visual field appears to be the most susceptible site in eyes of LHON patients. This suggests that the most susceptible cells during the early stages of LHON are the retinal ganglion cells located in the corresponding region of the retina.
NMR ASSIGNMENTS OF THE DNA BINDING DOMAIN OF ML4 PROTEIN FROM MESORHIZOBIUM LOTI.
Biomol NMR Assign. 2009 Dec 19;
Russo L, Palmieri M, Baglivo I, Esposito S, Isernia C, Malgieri G, Pedone PV, Fattorusso R
Ml4 protein from Mesorhizobium loti has a 58% sequence identity with the Ros protein from Agrobacterium tumefaciens that contains a prokaryotic Cys(2)His(2) zinc finger domain. Interestingly, Ml4 is a zinc-lacking protein that does not contain the Cys(2)His(2) motif and is able to bind the Ros DNA target sequence with high affinity. Here we report the (1)H, (15)N and (13)C NMR assignments of the Ml4 protein DNA binding domain (residue 52-151), as an important step toward elucidating at a molecular level how this prokaryotic domain can overcome the metal requirement for proper folding and DNA-binding activity.
COMPARATIVE PROTEIN MODELING, PREDICTION OF CONSERVED RESIDUE AND ACTIVE SITES IN COLD RESISTANT PROTEIN ISOLATED FROM CRPF(1), A COLD TOLERANT MUTANT OF PSEUDOMONAS FLUORESCENS.
Curr Microbiol. 2009 Dec 18;
Khan M, Kumar A, Goel R
Proteins interacting with the biological information molecules DNA and RNA play important cellular roles in all organisms. One widespread super family of proteins implicated in such function(s) is cold shock protein (CSP) that contains the cold shock domain (CSD). This work is planned to study the three-dimensional structure, conserved residues, and different active sites in the structure of cold resistant protein (CRP) from CRPF(1), cold tolerant mutant of Pseudomonas fluorescence by comparative homology modeling. Here we tried to identify crucial residues that are involved in active sites or functional sites of the protein. The study reveals that CRP represent the prototype of the CSD and share a highly similar overall fold consisting of five antiparallel beta-sheets forming a beta-barrel structure with surface exposed aromatic and basic residues that were responsible for nucleic acid binding properties of variable binding affinities and sequence selectivity and harbors the nucleic acid binding motifs RNP1 and RNP2 that is highly conserved in CSP family.
ISOLATION AND GENETIC MANIPULATION OF THE ANTIBIOTIC DOWN-REGULATORY GENE, WBLA ORTHOLOG FOR DOXORUBICIN-PRODUCING STREPTOMYCES STRAIN IMPROVEMENT.
Appl Microbiol Biotechnol. 2009 Dec 18;
Noh JH, Kim SH, Lee HN, Lee SY, Kim ES
Cross-genome comparative transcriptome analyses were previously conducted using the sequenced Streptomyces coelicolor genome microarrays to understand the genetic nature of doxorubicin (DXR) and daunorubicin (DNR) overproducing industrial mutant (OIM) of Streptomyces peucetius. In this previous work, a whiB-like putative transcription factor (wblA ( sco )) was identified as a global antibiotic down-regulator in S. coelicolor (Kang et al., J Bacteriol 189:4315-4319, 2007). In this study, a total genomic DNA library of a DXR/DNR-overproducing S. peucetius OIM was constructed and screened using wblA ( sco ) as a probe, resulting in the isolation of a wblA ortholog (wblA ( spe )) that had 95% amino acid identity to wblA ( sco ). Gene disruption of wblA ( spe ) from the S. peucetius OIM resulted in an approximately 70% increase in DXR/DNR productivity, implying that the DXR/DNR production in the S. peucetius OIM could be further improved via comparative transcriptomics-guided target gene manipulation. Furthermore, several putative wblA ( spe ) -dependent genes were also identified using S. coelicolor interspecies DNA microarray analysis between the S. peucetius OIM and wblA ( spe )-disrupted S. peucetius OIM. Among the genes whose expressions were significantly stimulated in the absence of wblA ( spe ), the overexpression of a conserved hypothetical protein (SCO4967) further stimulated the total production of DXR/DNR/akavinone by 1.3-fold in the wblA ( spe )-disrupted S. peucetius OIM, implying that the sequential genetic manipulation of target genes identified from interspecies comparative microarray analysis could provide an efficient and rational strategy for Streptomyces strain improvement.
