Improved in vitro anti-tumoral activity, intracellular uptake and apoptotic induction of gemcitabine-loaded pegylated unilamellar liposomes

Anaplastic thyroid carcinoma is one of the most aggressive and lethal solid carcinomas affecting humans. A major limit of the chemotherapeutic agents is represented by their low therapeutic index. In this work, we investigated the possibility of improving the anti-tumoral activity of gemcitabine by using pegylated unilamellar liposomes. Liposomes were made up of 1,2-dipalmitoyl-sn-glycero-3-phospocholine monohydrate/cholesterol/N-(carbonyl-methoxypolyethylene glycol-2000)-1, 2-distearoyl-sn-glycero-3-phosphoethanolamine (6:3:1 molar ratio) and they were prepared with a pH gradient to improve the gemcitabine loading capacity. The anti-tumoral efficacy of the liposomal formulation was tested in vitro on human anaplastic thyroid carcinoma cells (ARO) in culture, comparing the effects with those of the free drug. Gemcitabine-loaded unilamellar liposomes had a mean size approximately 200 nm with a zeta potential approximately -2 mV. The liposomal carrier noticeably improved the anti-tumoral activity of gemcitabine against ARO cells in terms of both dose-dependent cytotoxic effect and of drug exposition effect. Namely, gemcitabine-loaded liposomes showed a cytotoxic effect (58.2% increase of cell mortality at 1 microM with respect to free drug) after 12 h incubation, while the free drug showed a significant activity only after 72 h incubation. Moreover, a significant effect on the cell mortality appeared at 0.1 microM and 100% mortality was detected at a concentration of 1 microM of gemcitabine-loaded liposomes, while the free drug elicited the same effect at a concentration of 100 microM. The improved anti-tumoral activity of gemcitabine determined by the liposomal carrier was due to a greater intracellular uptake. The intracellular gemcitabine levels as a function of time showed a sinusoidal profile with peaks after 2 h, 6 h and 11 h, related to the cellular cycle of ARO. PARP cleavage and DNA fragmentation analysis provided clear evidence of the apoptosis induction in ARO cells by treatment with liposomally entrapped gemcitabine after 72 h incubation. Thus, gemcitabine-loaded liposomes may have a potential therapeutic relevance for the treatment of anaplastic thyroid carcinoma.     

Cytogenetic profile of lymphoma of follicle mantle lineage: correlation with clinicobiologic features

Conventional chromosome analysis (CCA) and interphase fluorescence in situ hybridization (FISH) was performed in 42 patients with mantle-cell lymphoma (MCL), with BCL1 rearrangement. The t(11;14)(q13;q32) or 11q abnormalities were detected by CCA in 34 cases, 20 of which had additional aberrations. A normal karyotype was observed in 8 cases. Probes detecting the chromosome aberrations that were observed in at least 3 cases by CCA, ie, +12, 13q14 deletion, and 17p deletion, were used for interphase FISH analysis. FISH detected total or partial +12, 13q14 deletion and 17p- in 28.5%, 52.4%, and 26% of the cases, respectively. The presence of these anomalies was not a function of karyotype complexity. Based on the results of CCA/FISH, three groups of increasing karyotype complexity were recognized: group 1, including 11 patients without detectable aberrations in addition to BCL1 rearrangement; group 2, including 14 patients with 1 to 2 additional anomalies; and group 3, including 17 patients with three or more additional anomalies. Clinical parameters associated with shorter survival were male sex (P =.006) and primary lymph-node involvement compared with primary bone marrow involvement (P =.015). Trisomy 12 was the only single cytogenetic parameter predictive of a poor prognosis (P =.006) and the best prognostic indicator was the derived measure of karyotype complexity (P <.0001), which maintained statistical significance in multivariate analysis (P<.0001). We arrived at the following conclusions: 13q14 deletion occurs at a high incidence in MCL; 17p deletion and total/partial +12 are relatively frequent events in MCL, the latter aberration being associated with a shorter survival; and the degree of karyotype complexity has a strong impact on prognosis in this neoplasia.     

Migration analysis,antioxidant, and mechanical characterization of polypropylene-based active food packaging films loaded with BHA,BHT, and TBHQ

Polypropylene (PP) based active composite films were prepared by adding butylated hydroxy anisole (BHA), butylated hydroxytoluene (BHT), and tertiary butylated hydroquinone (TBHQ) antioxidants using the extrusion molding process. All concentrations of BHT, 2% to 3% BHA, and 3% TBHQ significantly increased the tensile strength (TS) of the composite films compared with control films. Increasing antioxidant concentration decreased TS values for BHT films, whereas an opposite trend was observed for BHA and TBHQ films. BHA at < 2%, BHT at > 2%, and TBHQ at all added concentrations significantly reduced elongation at break (E_b) of the composite films compared to control films. Water vapor permeability (WVP) of 1% BHT film was not significantly different from control. However, other antioxidants especially at increased concentrations significantly increased WVP values. TBHQ films with 300% to 662% increase had the highest WVP and BHT films with 5% to 81% increase had the lowest WVP among composite films. All three antioxidants had a negative effect on the transparency of the films; however the effect of BHA at higher concentrations was greater. The antioxidants did not change the color attributes of the films. Films containing all antioxidants showed 2,2-diphenyl-1-picrylhydrazyl radical-scavenging activity, which increased with increase in their concentration, especially for those containing 3 wt.% BHT and TBHQ. Overall, incorporating BHA and BHT into a PP matrix improved mechanical, barrier, antioxidant properties, and film appearance and consequently were proposed for the development of antioxidant active PP films. TBHQ film is not recommended for food packaging because of its weak mechanical properties (lower E_b and TS values, higher WVP, and greater migration).     

Simplified automatic fault detection in wind turbine induction generators

This paper presents a simplified automated fault detection scheme for wind turbine induction generators with rotor electrical asymmetries. Fault indicators developed in previous works have made use of the presence of significant spectral peaks in the upper sidebands of the supply frequency harmonics; however, the specific location of these peaks may shift depending on the wind turbine speed. As wind turbines tend to operate under variable speed conditions, it may be difficult to predict where these fault‐related peaks will occur. To accommodate for variable speeds and resulting shifting frequency peak locations, previous works have introduced methods to identify or track the relevant frequencies, which necessitates an additional set of processing algorithms to locate these fault‐related peaks prior to any fault analysis. In this work, a simplified method is proposed to instead bypass the issue of variable speed (and shifting frequency peaks) by introducing a set of bandpass filters that encompass the ranges in which the peaks are expected to occur. These filters are designed to capture the fault‐related spectral information to train a classifier for automatic fault detection, regardless of the specific location of the peaks. Initial experimental results show that this approach is robust against variable speeds and further shows good generalizability in being able to detect faults at speeds and conditions that were not presented during training. After training and tuning the proposed fault detection system, the system was tested on “unseen” data and yielded a high classification accuracy of 97.4%, demonstrating the efficacy of the proposed approach.     

LC with Evaporative Light-Scattering Detection for Quantitative Analysis of Organic Acids in Juices

In this work, a performing LC method with evaporative light-scattering detector was described for the analysis of tartaric, malic, ascorbic, citric and succinic acids in fruit juices. The method was optimized and validated in comparison with LC-UV, in terms of accuracy, LODs, LOQs and precision. The successive application to ACE, orange, pear, peach, mulberry and apple juices allowed separation and quantitative determination of organic acids in about 20 min just diluting and filtering the sample before LC determination. In the analysed juices, citric, malic and ascorbic acids were always present, and in all the samples, citric acid was the prevailing acid followed by malic acid except for peach and apple juices where an opposite trend was found. Tartaric and succinic acids were detected at lower concentrations and more rarely, except for peach juices, all containing tartaric acid, and ACE and orange juices, all containing succinic acid.     

Profiling of hydrophilic proteins from Olea europaea olive pollen by MALDI TOF mass spectrometry

The antigenic profile of Olea europaea pollen from different Mediterranean cultivars was obtained by MALDI mass spectrometry. A simple procedure of chemical fractionation of the whole antigen extract was developed, whereby less complex, or pure, fractions of antigen candidate were obtained prior to mass spectrometric analysis. Some of the features of protein structure and distribution probably depend on cultivar adaptation to the environment. The profilings of pollen proteins thus obtained allow the distinction of the analyzed cultivars into three distinct groups: (i) those characterized by a low Ole e 1 content; (ii) those over-enriched in Ole e 1 and (iii) that containing Ole e 3 and Ole e 7 only. The latter consists of at least four isoforms differing by the degree of glycosilation. These results demonstrate that the proposed experimental procedure, can supply valuable information on the antigens' micro heterogeneity.     

Contamination of UHT milk by aflatoxin M1 in Iran

In this study the levels of aflatoxin M₁ (AFM₁) in UHT milk samples were determined in May, August and November and February. Two hundred and ten UHT milk samples were obtained from supermarkets in Tehran, Iran. The occurrence and concentration range of AFM₁ in the samples were investigated by competitive enzyme-linked immunoabsorbent assay (ELISA) method. AFM₁ was found in 116 (55.2%) of 210 UHT milk samples examined. The levels of AFM₁ in 70 (33.3%) samples were higher than the maximum tolerance limit (0.05 μg/l) accepted by some European countries while none of the samples exceeded the prescribed limit of US regulations. The highest mean concentration of AFM₁ was recorded in February (0.087 μg/l). The lowest mean concentration of AFM₁ was recorded in August (0.021 μg/l). Statistical evaluation showed that there were significant difference (P < 0.01) between the mean concentrations of AFM₁ of UHT milk samples taken in February with May and August. AFM₁ contents of milk samples taken in February were not higher than UHT milk samples taken in November (P < 0.01). The AFM₁ incidence of exceeding legal limit in UHT milk samples (33.3%) was relatively much higher than some other countries. It was therefore concluded that, the levels of AFM₁ in UHT milk samples consumed in Iran were high and seemed to pose a threat to public health.     

A comparative analysis of a modified picture frame test for characterization of woven fabrics

An experimental, finite‐element analysis framework is utilized to estimate the deformation state in a modified version of the picture frame test. During the analysis, the effect of fiber misalignment and the deformation heterogeneity in the tested fabric, a 2 × 2 PP/E‐Glass twill, is accounted for and a force prediction model is presented. Using an equivalent stress–strain normalization scheme, the comparison of the modified test with the conventional (original) picture frame and bias‐extension tests is also made, and results reveal similarities and differences that should receive attention in the identification of constitutive models of woven fabrics using these basic tests. Ideally, the trellising behavior should not change from one test to another but results show that in the presence of fiber misalignment, the modified picture frame test yields a behavior closer to that of the bias‐extension test, while the general form of the test's repeatability, measured by a signal‐to‐noise metric, remains similar to the original picture frame test. POLYM. COMPOS., 2010. © 2009 Society of Plastics Engineers