Evaluation of the potential of Raman microspectroscopy for prediction of chemotherapeutic response to cisplatin in lung adenocarcinoma

The study of the interaction of anticancer drugs with mammalian cells in vitro is important to elucidate the mechanisms of action of the drug on its biological targets. In this context, Raman spectroscopy is a potential candidate for high throughput, non-invasive analysis. To explore this potential, the interaction of cis-diamminedichloroplatinum(II) (cisplatin) with a human lung adenocarcinoma cell line (A549) was investigated using Raman microspectroscopy. The results were correlated with parallel measurements from the MTT cytotoxicity assay, which yielded an IC(50) value of 1.2 ± 0.2 μM. To further confirm the spectral results, Raman spectra were also acquired from DNA extracted from A549 cells exposed to cisplatin and from unexposed controls. Partial least squares (PLS) multivariate regression and PLS Jackknifing were employed to highlight spectral regions which varied in a statistically significant manner with exposure to cisplatin and with the resultant changes in cellular physiology measured by the MTT assay. The results demonstrate the potential of the cellular Raman spectrum to non-invasively elucidate spectral changes that have their origin either in the biochemical interaction of external agents with the cell or its physiological response, allowing the prediction of the cellular response and the identification of the origin of the chemotherapeutic response at a molecular level in the cell.     

Synthesis of 3,5-diaryl-4-chlorophthalates by [4+2] cycloaddition of 1-ethoxy-2-chloro-1,3-bis(trimethylsilyloxy)-1,3-diene with dimethyl acetylenedicarboxylate and subsequent site-selective Suzuki-Miyaura reactions

The [4+2] cycloaddition of 1-ethoxy-2-chloro-1,3-bis(trimethylsilyloxy)-1,3-diene with dimethyl acetylenedicarboxylate (DMAD) afforded dimethyl 4-chloro-3,5-dihydroxyphthalate. Site-selective Suzuki-Miyaura reactions of its bis(triflate) provide a convenient approach to 3,5-diaryl-4-chlorophthalates containing two different aryl groups.     

Oral feeding of pomegranate fruit extract inhibits early biomarkers of UVB radiation-induced carcinogenesis in SKH-1 hairless mouse epidermis

Pomegranate from the plant Punica granatum L. possesses strong antioxidant and anti-inflammatory properties. Recently, we have demonstrated that treatment of normal human epidermal keratinocytes with pomegranate fruit extract (PFE) inhibited UVB-mediated activation of nuclear factor kappa B (NF-κB) and mitogen activated protein kinases pathways. Here, we evaluated the effect of PFE on early biomarkers of photocarcinogenesis employing SKH-1 hairless mice. PFE was provided in drinking water (0.2%, wt/vol) to SKH-1 hairless mice for 14 days before a single UVB (180 mJ cm⁻²) irradiation. We found that oral feeding of PFE inhibited UVB-induced: (1) skin edema; (2) hyperplasia; (3) infiltration of leukocytes; (4) lipid peroxidation; (5) hydrogen peroxide generation; (6) ornithine decarboxylase (ODC) activity; and (7) ODC, cyclooxygenase-2 and proliferating cell nuclear antigen protein expression. Oral feeding of PFE enhanced repair of UVB-mediated formation of cyclobutane pyrimidine dimers (CPDs) and 8-oxo-7,8-dihydro-2′-deoxyguanosine (8-oxodG). Importantly, PFE treatment further enhanced UVB-mediated increase in tumor suppressor p53 and cyclin kinase inhibitor p21. Furthermore, oral feeding of PFE inhibited UVB-mediated: (1) nuclear translocation of NF-κB; (2) activation of IKKα; and (3) phosphorylation and degradation of IκBα. Taken together, we provide evidence that oral feeding of PFE to mice affords substantial protection from the adverse effects of UVB radiation via modulation in early biomarkers of photocarcinogenesis and provide suggestion for its photochemopreventive potential.     

A Novel Approach for Mining High‐Utility Sequential Patterns in Sequence Databases

Mining sequential patterns is an important research issue in data mining and knowledge discovery with broad applications. However, the existing sequential pattern mining approaches consider only binary frequency values of items in sequences and equal importance/significance values of distinct items. Therefore, they are not applicable to actually represent many real‐world scenarios. In this paper, we propose a novel framework for mining high‐utility sequential patterns for more real‐life applicable information extraction from sequence databases with non‐binary frequency values of items in sequences and different importance/significance values for distinct items. Moreover, for mining high‐utility sequential patterns, we propose two new algorithms: UtilityLevel is a high‐utility sequential pattern mining with a level‐wise candidate generation approach, and UtilitySpan is a high‐utility sequential pattern mining with a pattern growth approach. Extensive performance analyses show that our algorithms are very efficient and scalable for mining high‐utility sequential patterns.     

An application of univariate marginal distribution algorithm in MIMO communication systems

The paper discusses a sequence detector based on univariate marginal distribution algorithm (UMDA) that jointly estimates the symbols transmitted in a multiple input multiple output (MIMO) communication system. While an optimal maximum likelihood detection using an exhaustive search method is prohibitively complex, it has been shown that sphere decoder (SD) achieves the optimal bit error rate (BER) performance with polynomial time complexity for smaller array sizes. However, the worst‐case complexity of SD is exponential in the problem dimensions, this brings in question its practical implementation for larger number of spatial layers and for higher‐order signal constellation. The proposed detector shows promising results for this overly difficult and complicated operating environment, confirmed through simulation results. A performance comparison of the UMDA detector with SD is presented for higher‐order complex MIMO architectures with limited average transmit power. The proposed detector achieves substantial performance gain for higher‐order systems attaining a near optimal BER performance with reduced computational complexity as compared with SD. Copyright © 2009 John Wiley & Sons, Ltd.     

Two-step on-particle ionization/enrichment via a washing- and separation-free approach: multifunctional TiO2 nanoparticles as desalting, accelerating, and affinity probes for microwave-assisted tryptic digestion of phosphoproteins in ESI-MS and MALDI-MS: comparison with microscale TiO2

We introduce a simplified sample preparation method using bare TiO₂ nanoparticles (NPs) to serve as multifunctional nanoprobes (desalting, accelerating, and affinity probes) for effective enrichment of phosphopeptides from microwave-assisted tryptic digestion of phosphoproteins (α-casein, β-casein and milk) in Electrospray Ionization Mass Spectrometry (ESI-MS) and Matrix Assisted Laser Desorption Ionization Mass Spectrometry (MALDI-MS). The results demonstrate that TiO₂ NPs can effectively enrich and accelerate the digestion reactions of phosphoproteins in aqueous solutions and also from complex real samples. After the microwave experiments, we directly injected the resulting solutions into the ESI-MS and MALDI-MS systems for analysis, and excellent sensitivity was achieved without the need for any washing procedure or separation process. The reasons are attributed to the high binding affinity and selectivity of TiO₂ NPs toward phosphopeptides. Thus, phosphopeptides can be adsorbed onto the TiO₂ NP surface. The digested or partially digested phosphoproteins can be concentrated onto the TiO₂ NP surface. This results in the effective or complete digestion of phosphoproteins in a short period of time (45 s). In addition, high sensitivity and sequence coverage of phosphopeptide can be obtained using TiO₂ NPs as microwave absorbers and affinity probes in MALDI-MS and ESI-MS. This is due to the photocatalytic nature of the TiO₂ NPs because the absorption of microwave radiation that can accelerate the activation of trypsin for efficient digestion of phosphoproteins and enhances the ionization of phosphopeptides. The lowest concentrations detected for ESI-MS and MALDI-MS were 0.1 μM and 10 fmol, respectively, for α-casein. Comparing the two-step approach of TiO₂ NPs with microscale TiO₂ particles, the microscale TiO₂ particles shows no effect on the microwave-assisted tryptic digestion of phosphoproteins. The current approach offers multiple advantages, such as great simplicity, high sensitivity and selectivity, straightforward and separation/washing-free technique for phosphorpeptide enrichment analysis.     

Modification of tricine–SDS–PAGE for online and offline analysis of phosphoproteins by ICP-MS

This study describes a modification to tricine sodium dodecyl sulphate polyacrylamide gel electrophoresis to make it more effective for the separation of low molecular mass proteins and for coupling with inductively coupled plasma mass spectrometry (ICP-MS). The modified method employs low-percentage polyacrylamide gels (7–10%) (w/v) and low reagent concentrations that provide efficient separations, good quantitation and low matrix levels that are compatible with ICP-MS. Using phosphopeptides as a model system, and offline analysis, we obtained recoveries of 73% (w/v) in a 9% gel compared with 55% in a conventional 16% gel. Online coupling was achieved by modification of a standard commercially available gel electroelution apparatus and casting of the gel into a 7.3-cm-long tube. Online separation of a digest of β-casein was demonstrated with recovery of the mono- and tetraphosphopeptides, which were identified by comparison with peptide standards. A mass balance study with the standards yielded recoveries of 95% for tetraphosphopeptides and 48% for monophosphopeptides. The factors affecting the separations and recoveries are discussed in detail. The detection limits for 10-μL samples of the mono- and tetraphosphopeptides were 0.7 μM (7 pmol) and 0.2 μM (2 pmol) respectively.     

Simple gas chromatographic method for the quantification of total cholesterol in fish meats

Six fish species-Cirrhinus mrigala (Morakhi), Labeo rohita (Danbhro), Catla catla (Thalli), Wallago attu (Jarko), Ctenopharyngodon idella (Grass), and Cyprimus carpio (Gulfam)--commonly found in the Pakistani Indus river, were selected for fat and cholesterol evaluation. Fat content was quantified by the Folch method and cholesterol content was determined by a simple GC method. The application of the proposed method to quantify cholesterol content revealed variations among the six breeds evaluated. The breeds studied contained 0.80-1.95% fat, indicating that all investigated fishes were lean; significant variation was found in cholesterol content, and ranged between 72 and 392 mg/100 g. Standard addition analyses showed that the method was accurate, as the recovery of cholesterol varied from 95 to 103%, with a coefficient of variance not more than 5.6%. The results showed that, after the development of suitable calibration, within 11 min, the cholesterol could be accurately determined by GC.     

Ionic studies of sodium alginate isolated from Sargassum terrarium (brown algea) karachi coast with 2,1-electrolyte

Physical and chemical analysis of the polysaccharide isolated from Sargassum Terarrium (brown algae) of Karachi coast showed characteristics of the sodium alginate. Optical rotations and sulphated ash content were found and FTIR spectra showed a sharp and strong absorption band at 1600 cm^?1 representing carboxylate ion which conforms high uronic acid content of the product. The viscosities of aqueous 0.1% sodium alginate solution were measured in the presence of copper II chloride (CuCl₂). The viscosities were found to be increased with the increase in the concentration of electrolyte. Viscosities were also found affected with temperature. ‘A’ and ‘B’ coefficients of Jones–Dole equation were evaluated. The increase in positive values of ‘B’ coefficient with the rise of temperature led to conclusion that given electrolyte in 0.1% aqueous sodium alginate solution behaves as structure maker. Thermodynamic parameters regarding to activated state like energy of activation E_η, change in free energy of activation ΔG_η and change of entropy of activation ΔS_η were also evaluated. Straight-line plots of log η versus 1/T observed with positive slopes show the effect of temperature on the viscosities of solutions. Energy of activation (E_η) was found to be decreased with the rise of temperature. Change in free energy of activation (ΔG_η) was also found to be increased with increase in concentrations of electrolyte and also with rise of temperature. The values of change in entropy of activation (ΔS_η) were also calculated. Negative values of ΔS_η were found to be increased with increase in concentration of electrolyte and also with rise of temperature.