High-performance liquid chromatography of platinum complexes on solvent generated anion exchangers : III. Application to the analysis of cisplatin in urine using automated column switching

Platinum complexes are retained on solvent generated anion exchangers, prepared by coating reversed-phase (C-18) supports with a monolayer of hexadecyltrimethylammonium bromide. The retention mechanism is described in terms of ion—dipole interactions in the stationary phase, reinforced by a hydrophobic effect. The high degree of ligand selectivity exhibited by these systems arises from the use of purely aqueous mobile phases which maximize the differences in solute dipole and hydrophobic surface area. By using stationary phases of different surface characteristics and the application of automated column switching, the technique is applicable to the clinical analysis of cisplatin in urine. After chromatography, the purified cisplatin fractions are determined by atomic absorption spectrophotometry. The recovery of cisplatin from urine is 101.1% with a relative standard deviation of 3.6% and the limit of detection is 2 μg/ml.     

Cellular immunotherapy with multiple infusions of in vitro-expanded haploidentical natural killer cells after autologous transplantation for patients with plasma cell myeloma

Background aimsTo investigate the feasibility and safety of haploidentical natural killer (NK) cell infusions as consolidation immunotherapy after autologous stem cell transplant (ASCT) in patients with plasma cell myeloma.MethodsTen patients (median age, 59 years) received induction treatment followed by high-dose melphalan (200 mg/m²) at day –1, ASCT at day 0 and increasing NK cell doses (1.5 × 10⁶, 1.5 × 10⁷ and multiple doses of 1.0 × 10⁸ cells/kg body weight) from day +1 to day +30 after ASCT. NK cells were harvested and purified from peripheral blood of haploidentical donors and expanded for 19 days with interleukin (IL)-2 and IL-15 under Good Manufacturing Practice conditions.ResultsNK cell numbers increased 56.0-fold (37.4- to 75.5-fold). Patients received a median of 3.8 × 10⁸ (0.9–5.7 × 10⁸) NK cells/kg body weight in six (three to eight) infusions. Multiparametric mass cytometry analysis demonstrated an altered surface receptor repertoire of expanded NK cells with increased degranulation and cytokine production activities but diminished expression of perforin. Donor NK cells were detectable in the peripheral blood, peaking 1 h after each dose (up to 90% donor NK cells). The treatment was safe and well tolerated, without evidence of graft-versus-host disease. Comparison with a control patient population receiving ASCT without NK cell infusions showed no significant difference in relapse, progression-free survival and overall survival.ConclusionsThis study demonstrates reliable manufacturing of high numbers of activated NK cells for multiple-dose infusions and safe administration of these cellular products. The trial was registered at ClinicalTrials.gov (identifier no. NCT01040026).     

The Biological Connection Markup Language: a SBGN-compliant format for visualization, filtering and analysis of biological pathways

MOTIVATION: Many models and analysis of signaling pathways have been proposed. However, neither of them takes into account that a biological pathway is not a fixed system, but instead it depends on the organism, tissue and cell type as well as on physiological, pathological and experimental conditions. RESULTS: The Biological Connection Markup Language (BCML) is a format to describe, annotate and visualize pathways. BCML is able to store multiple information, permitting a selective view of the pathway as it exists and/or behave in specific organisms, tissues and cells. Furthermore, BCML can be automatically converted into data formats suitable for analysis and into a fully SBGN-compliant graphical representation, making it an important tool that can be used by both computational biologists and 'wet lab' scientists. Availability and implementation: The XML schema and the BCML software suite are freely available under the LGPL for download at http://bcml.dc-atlas.net. They are implemented in Java and supported on MS Windows, Linux and OS X.     

RF hydrazine plasma modification of chitosan for antibacterial activity and nanofiber applications

Chitosan nano powders were modified using RF hydrazine plasma produced at low pressure (26.66 Pa) with 13.56 MHz frequency at a power of 100 W for 30 min. Characterization and investigation of the properties of plasma-modified chitosan (PMCh) and non-modified chitosan (Ch) were carried out using an optical monochromator, FTIR, florescence analysis, TGA, SEM, and X-ray techniques. FTIR spectra of PMCh indicated a band broadening at 3436 cm⁻¹ that confirmed increasing functional groups based on H-bonding. The number of NH₂ groups was determined from fluorescence analysis. TGA analysis shows that the moisture absorption is three times higher in the PMCh structure. Ch and PMCh in PVA solutions were used to produce nanofibers by the electrospinning method; average fiber diameters were 480 and 280 nm for Ch and PMCh, respectively. It was found that the antibacterial effect of PMCh is better than the Ch for Gram-positive strains.     

ABCG9, ABCG11 and ABCG14 ABC transporters are required for vascular development in Arabidopsis

In order to obtain insights into the regulatory pathways controlling phloem development, we characterized three genes encoding membrane proteins from the G sub‐family of ABC transporters (ABCG9, ABCG11 and ABCG14), whose expression in the phloem has been confirmed. Mutations in the genes encoding these dimerizing ‘half transporters’ are semi‐dominant and result in vascular patterning defects in cotyledons and the floral stem. Co‐immunoprecipitation and bimolecular fluorescence complementation experiments demonstrated that these proteins dimerize, either by flexible pairing (ABCG11 and ABCG9) or by forming strict heterodimers (ABCG14). In addition, metabolome analyses and measurement of sterol ester contents in the mutants suggested that ABCG9, ABCG11 and ABCG14 are involved in lipid/sterol homeostasis regulation. Our results show that these three ABCG genes are required for proper vascular development in Arabidopsis thaliana.     

Gene expression profile during the life cycle of the urochordate Ciona intestinalis

Recent whole-genome studies and in-depth expressed sequence tag (EST) analyses have identified most of the developmentally relevant genes in the urochordate, Ciona intestinalis. In this study, we made use of a large-scale oligo-DNA microarray to further investigate and identify genes with specific or correlated expression profiles, and we report global gene expression profiles for about 66% of all the C. intestinalis genes that are expressed during its life cycle. We succeeded in categorizing the data set into 5 large clusters and 49 sub-clusters based on the expression profile of each gene. This revealed the higher order of gene expression profiles during the developmental and aging stages. Furthermore, a combined analysis of microarray data with the EST database revealed the gene groups that were expressed at a specific stage or in a specific organ of the adult. This study provides insights into the complex structure of ascidian gene expression, identifies co-expressed gene groups and marker genes and makes predictions for the biological roles of many uncharacterized genes. This large-scale oligo-DNA microarray for C. intestinalis should facilitate the understanding of global gene expression and gene networks during the development and aging of a basal chordate.     

Oncofetal H19 RNA promotes tumor metastasis

The oncofetal H19 gene transcribes a long non-coding RNA(lncRNA) that is essential for tumor growth. Here we found that numerous established inducers of epithelial to mesenchymal transition(EMT) also induced H19/miR-675 expression. Both TGF-β and hypoxia concomitantly induced H19 and miR-675 with the induction of EMT markers. We identified the PI3K/AKT pathway mediating the inductions of Slug, H19 RNA and miR-675 in response to TGF-β treatment, while Slug induction depended on H19 RNA. In the EMT induced multidrug resistance model, H19 level was also induced. In a mouse breast cancer model, H19 expression was tightly correlated with metastatic potential. In patients, we detected high H19 expression in all common metastatic sites tested, regardless of tumor primary origin. H19 RNA suppressed the expression of E-cadherin protein. H19 up-regulated Slug expression concomitant with the suppression of E-cadherin protein through a mechanism that involved miR-675. Slug also up-regulated H19 expression and activated its promoter. Altogether, these results may support the existence of a positive feedback loop between Slug and H19/miR-675, that regulates E-cadherin expression. H19 RNA enhanced the invasive potential of cancer cells in vitro and enhanced tumor metastasis in vivo. Additionally, H19 knockdown attenuated the scattering and tumorigenic effects of HGF/SF. Our results present novel mechanistic insights into a critical role for H19 RNA in tumor progression and indicate a previously unknown link between H19/miR-675, Slug and E-cadherin in the regulation of cancer cell EMT programs.