The human proteome project: current state and future direction

After the successful completion of the Human Genome Project, the Human Proteome Organization has recently officially launched a global Human Proteome Project (HPP), which is designed to map the entire human protein set. Given the lack of protein-level evidence for about 30% of the estimated 20,300 protein-coding genes, a systematic global effort will be necessary to achieve this goal with respect to protein abundance, distribution, subcellular localization, interaction with other biomolecules, and functions at specific time points. As a general experimental strategy, HPP research groups will use the three working pillars for HPP: mass spectrometry, antibody capture, and bioinformatics tools and knowledge bases. The HPP participants will take advantage of the output and cross-analyses from the ongoing Human Proteome Organization initiatives and a chromosome-centric protein mapping strategy, termed C-HPP, with which many national teams are currently engaged. In addition, numerous biologically driven and disease-oriented projects will be stimulated and facilitated by the HPP. Timely planning with proper governance of HPP will deliver a protein parts list, reagents, and tools for protein studies and analyses, and a stronger basis for personalized medicine. The Human Proteome Organization urges each national research funding agency and the scientific community at large to identify their preferred pathways to participate in aspects of this highly promising project in a HPP consortium of funders and investigators.     

Anti-Inflammatory Activity of S. Marianum and N. Sativa Extracts on Macrophages

Background:Nigella sativa (N. sativa) and Silybum marianum (S. marianum) are used to regulate macrophage polarization in lipopolysaccharide-induced RAW 264.7 cells and thioglycollate-elicited peritoneal inflammation.Methods:Cytotoxicity assays and acute toxicity tests were performed to investigate the safe dose and toxicity of the prepared extracts. Also, nitric oxide production was determined by Griess assay on RAW264.7 and peritoneal macrophage supernatants. After RNA extraction from macrophages, real-time PCR was performed to measure the relative gene expression of tumor necrosis factor (TNF)-α, interleukin (IL)-6, transforming growth factor (TGF)-β, and IL-10. Finally, regulatory T cells (Treg cells) were counted by flow cytometry.Results:S. marianum methanolic extract (SME), N. sativa ethanolic extract (NEE), and their mixture (SME+NEE) decreased NO levels significantly in RAW264.7 and peritoneal murine macrophages. N. sativa ethanolic extract significantly increased IL-10 gene expression and significantly decreased IL-6 and TNF-α expression in RAW264.7 cells. In mixture-treated peritoneal macrophages, IL-10 and TGF-β expression were significantly increased, while IL-6 and TNF-α were significantly decreased. Also, the percentage of Treg cells was significantly greater in the mixture-treated cells than in controls.Conclusion:These results suggest that an SME and NEE mixture has anti-inflammatory and immunomodulatory activities and may be useful in the treatment of diseases of immunopathologic origin characterized by macrophage hyperactivation.Key Words: Cytokine, Inflammation, Nigella sativa, Nitric oxide, Silybum marianum     

Prevalence and detection of metallo-β-lactamase (MBL)-producingPseudomonas aeruginosa strains from clinical isolates in Iran

Of the 126 isolates obtained from clinical specimens, seventy strains were selected because of resistance or reduced susceptibilities to imipenem and/or ceftazidime. Screening for detection of MBL-producing strains was performed in latter isolates by the Etest MBL strips. Isolates that were positive to Etest MBL strips were analysed by PCR. PCR was performed with specific DNA probes for detection of genes coding IMP or VIM enzymes and positive controls (MBL-producing strains). Finally, eight isolates ofPseudomonas aeruginosa were detected to carry a bla_VIM gene.     

Dictyostelium myosin II G680V suppressors exhibit overlapping spectra of biochemical phenotypes including facilitated phosphate release.

We have biochemically characterized 13 intragenic suppressors of the G680V mutation of Dictyostelium myosin II. In the absence of the G680V mutation, the suppressors result in a number of deviant behaviors, most commonly an increase in the basal (actin-independent) ATPase of the motor. This phenotype is complementary to that of the G680V mutant and supports our proposal that the latter impairs phosphate release. Different subsets of the mutants also suffer from poor ATPase enhancement by 1 mg/ml actin, failure to release from actin in the presence of ATPgammaS (or ADP and salt), and excessive release from actin in the presence of ADP. The patterns of suppressor behaviors suggest that, in general, they are facilitating P(i)-releasing state(s) of the motor, but that different individual suppressors may secondarily perturb other states or actions of the motor.     

Tomato (Solanum lycopersicum) growth and fruit quality affected by organic fertilization and ozonated water

Protoplasma - The use of modern and safe techniques to increase plant growth and yield is of significance. There is little data, to our knowledge, on the use of organic fertilization and ozonated...     

A new RAPD marker for beet necrotic yellow vein virus resistance gene in Beta vulgaris

RAPD markers linked to beet necrotic yellow vein virus (BNYVV) resistance genes were identified in two Beta vulgaris accessions Holly-1-4 and WB42 using bulked segregant analysis. The polymorphism revealed by the RAPD markers in the F₂ generations of WB42 was higher than that of Holly-1-4. The segregation distortion at marker loci was slightly lower in the B. vulgaris × B. maritima cross than in the B. vulgaris × B. vulgaris cross. For Holly-1-4, a RAPD marker was identified in a long distance from the resistance gene of Rz ₁ . However, a RAPD marker tightly linked with Rz ₂ gene in repulsion phase was detected with an approximate distance of 0.036 rf. This marker was not generation specific and showed high repeatability. The distance between Rz ₁ and Rz ₂ genes was estimated as 0.464 rf. After the Rz ₂ Rz ₂ and Rz ₂ rz ₂ genotypes were identified using ELISA values and repulsion phase RAPD markers, comparison of their ELISA means revealed lack of the gene dosage effects. Nevertheless, under the field or severe infection conditions, the difference between ELISA mean values of the Rz ₂ Rz ₂ and Rz ₂ rz ₂ genotypes might be more than that observed in this study and the gene dosage effects of Rz ₂ allele might be important.     

A new on-line, in-tube pre-column derivatization technique for high performance liquid chromatographic determination of azithromycin in human serum

Pre-column derivatization methods for high performance liquid chromatographic assay of specific pharmaceutical agents using 9-fluorenylmethyl chloroformate (FMOC-Cl) have received special attention because highly fluorescent and stable adducts are provided by these methods. However, unlike the post-column on-line techniques, long derivatization time is needed and the reaction cannot be well controlled. A new, sensitive and fast pre-column on-line derivatization technique coupled with high-performance liquid chromatography using FMOC-Cl as labeling agent is described and validated for determination of azithromycin in human serum. After extraction of the drug from serum, the residue was reconstituted in mixture of acetonitrile-phosphate buffer (3:1, v/v; pH 8.5) and directly injected onto the chromatographic system. Continuous on-line derivatization and analysis of the compounds were successfully performed using in-tube elution of FMOC-Cl. The total time needed for derivatization and chromatographic analysis of the drug was 13 min. The assay was reliable and reproducible, with limit of quantification of 10 ng/ml. The described technique may offer significant advantages over existing off-line derivatization methods using FMOC-Cl.