Adhesion and proliferation of fibroblasts on cluster-assembled nanostructured carbon films: the role of surface morphology

We have investigated the influence on adhesion and proliferation of NIH 3T3 fibroblasts of the surface morphology of cluster assembled carbon films deposited by Supersonic Cluster Beam Deposition. Nanostructured carbon films exhibit a multi-scale morphology, which resembles the surface structure of the extracellular matrix, and possess a high specific area, while being relatively smooth at all scales. Correlations between measured morphological parameters and adaptive cell response have been brought out. High specific area and smoothness appear to conceivably favour both the early attachment of plated cells and the long-term survival of adherent cells. Moreover, nano-structured carbon films affect the cells morphology as well as the extension and the number of the focal contacts.     

Torsades de pointes: a case with multiple variables

PURPOSE: The human medulloblastoma cell line D283 Med (4-HCR), a line resistant to 4-hydroperoxycyclophosphamide (4-HC), displays enhanced repair of DNA interstrand crosslinks induced by phosphoramide mustard. D283 Med (4-HCR) cells are cross-resistant to 1,3-bis(2-chloroethyl)- -nitrosourea, but partial sensitivity is restored after elevated levels of O6-alkylguanine-DNA alkyltransferase (AGT) are depleted by O6-benzylguanine (O6-BG). Studies were conducted to define the activity of 4-HC and 4-hydroperoxydidechlorocyclophosphamide against D283 Med (4-HCR) after AGT is depleted by O6-BG. METHODS: Limiting dilution and xenograft studies were conducted to define the activity of 4-HC and 4-hydroperoxydidechlorocyclophosphamide with or without O6-BG. RESULTS: The activity of 4-HC and 4-hydroperoxydidechlorocyclophosphamide against D283 Med (4-HCR) was increased after AGT depletion by O6-BG preincubation. Similar studies with Chinese hamster ovary cells, with or without stable transfection with a plasmid expressing the human AGT protein, revealed that the AGT-expressing cells were significantly less sensitive to 4-HC and 4-hydroperoxydidechlorocyclophosphamide. Reaction of DNA with 4-HC, phosphoramide mustard, or acrolein revealed that only 4-HC and acrolein caused a decrease in AGT levels. CONCLUSIONS: We propose that a small but potentially significant part of the cellular toxicity of cyclophosphamide in these cells is due to acrolein, and that this toxicity is abrogated by removal of the acrolein adduct from DNA by AGT.     

Application of micromagnetics to a boundless plate

The case of magnetization reversal in a boundless plate of magnetically homogeneous material is treated in the framework of micromagnetics. It is assumed that the magnetocrystalline symmetry axis and the applied field lie along the normal of the plate surface. The mathematical analysis performed facilitates the determination of the nucleation field in an exact analytical way, leading to the conclusion that nucleation takes place by spin rotation in unison. Some indirect experimental evidences are discussed.     

Modulating D-amino acid oxidase substrate specificity: production of an enzyme for analytical determination of all D-amino acids by directed evolution

Recent research on the flavoenzyme D-amino acid oxidase from Rhodotorula gracilis (RgDAAO) has revealed new, intriguing properties of this catalyst and offers novel biotechnological applications. Among them, the reaction of RgDAAO has been exploited in the analytical determination of the D-amino acid content in biological samples. However, because the enzyme does not oxidize acidic D-amino acids, it cannot be used to detect the total amount of D-amino acids. We now present the results obtained using a random mutagenesis approach to produce RgDAAO mutants with a broader substrate specificity. The libraries of RgDAAO mutants were generated by error-prone PCR, expressed in BL21(DE3)pLysS Escherichia coli cells and screened for their ability to oxidize different substrates by means of an activity assay. Five random mutants that have a 'modified' substrate specificity, more useful for the analytical determination of the entire content of D-amino acids than wild-type RgDAAO, have been isolated. With the only exception of Y223 and G199, none of the effective amino acid substitutions lie in segments predicted to interact directly with the bound substrate. The substitutions appear to cluster on the protein surface: it would not have been possible to predict that these substitutions would enhance DAAO activity. We can only conclude that these substitutions synergistically generate small structural changes that affect the dynamics and/or stability of the protein in a way that enhances substrate binding or subsequently catalytic turnover.     

p53 expression in B-cell chronic lymphocytic leukemia: a marker of disease progression and poor prognosis

We have analyzed by immunocytochemistry (ICC) the frequency of p53 protein expression in 181 cases of B-cell chronic lymphocytic leukemia (CLL) followed at a single institution to assess the relationship between p53 and the clinical and morphological features of the disease, as well as the possible involvement of this protein in the pathogenesis of the more aggressive forms of CLL. The overall frequency of p53 protein positivity in CLL was 15% (27 of 181 cases). There were no significant differences in age, sex, absolute lymphocyte count, or lymphocyte doubling time between p53-positive and -negative patients. By contrast, p53-positive patients had a significantly higher percentage of prolymphocytes (P = .002) and a significantly lower percentage of residual CD3-positive T lymphocytes (P = .0001). No correlation was found between the percentage of p53-positive cells and the percentage of cells in cycle assessed by the monoclonal antibody Ki-67. When the percentage of p53 positivity was correlated with the clinical stage of the disease, the proportion of p53-positive cases increased significantly from Binet's stage A (8 of 108; 7.4%), to stage B (12 of 49; 24.4%) and C (7 of 24; 29.2%) (P = .002). p53 positivity correlated also with the phase of the disease, showing a low expression at diagnosis (8 of 112; 7.1%) and a significantly higher expression in patients studied during the course of the disease (7 of 35; 20%) and, to a further extent, with disease progression (12 of 34; 35.3%) (P = .0001). The association of p53 protein expression with mutations in the gene was confirmed by direct sequence of the entire cDNA in 15 of the 17 ICC positive cases tested (88%). A significantly shorter treatment-free interval from diagnosis (P = .003) and a poorer response to therapy (P = .007) was observed in p53-positive compared with p53-negative patients. Overall survival from the time of diagnosis, as well as from the time of p53 protein analysis, was significantly shorter in patients with p53 protein expression (P = .03 and .0001, respectively). Moreover, in multivariate analysis, p53 expression and stage C were independently associated with a short survival. The results of this study indicate that in CLL the expression of the p53 protein, analyzed by a simple and reliable immunocytochemical method, is strongly associated with p53 gene mutations, a morphological variant (CLL with >10% prolymphocytes), advanced clinical stage, progressive disease, poor response to therapy, and short survival.     

Data mining with Temporal Abstractions: learning rules from time series

A large volume of research in temporal data mining is focusing on discovering temporal rules from time-stamped data. The majority of the methods proposed so far have been mainly devoted to the mining of temporal rules which describe relationships between data sequences or instantaneous events and do not consider the presence of complex temporal patterns into the dataset. Such complex patterns, such as trends or up and down behaviors, are often very interesting for the users. In this paper we propose a new kind of temporal association rule and the related extraction algorithm; the learned rules involve complex temporal patterns in both their antecedent and consequent. Within our proposed approach, the user defines a set of complex patterns of interest that constitute the basis for the construction of the temporal rule; such complex patterns are represented and retrieved in the data through the formalism of knowledge-based Temporal Abstractions. An Apriori-like algorithm looks then for meaningful temporal relationships (in particular, precedence temporal relationships) among the complex patterns of interest. The paper presents the results obtained by the rule extraction algorithm on a simulated dataset and on two different datasets related to biomedical applications: the first one concerns the analysis of time series coming from the monitoring of different clinical variables during hemodialysis sessions, while the other one deals with the biological problem of inferring relationships between genes from DNA microarray data.     

Collision modification functions: Incorporating changes over time

Collision modification factors (CMFs) are considered the primary tools for estimating the effectiveness of safety treatments at road sites. Three main techniques are commonly used to estimate CMFs: the empirical Bayes (EB) method, the comparison-group (CG) method, and a combination of the EB and CG methods. CMF estimates from these techniques are usually provided with a measure of uncertainty of the estimate, in the form of standard error and confidence interval.     

Healthy Virgin Olive Oil: A Matter of Bitterness

Virgin olive oil (VOO) is the pillar fat of Mediterranean diet. It is made from olive fruits and obtained by squeezing olives without any solvent extraction. Respect to the seed oils, an unique polar polyphenol-rich fraction gives VOO a bitter and pungent taste.

The recent substantiation by European Food Safety Authority (EFSA) of a health claim for VOO polyphenols may represent an efficient stimulus to get the maximum health benefit from one of the most valuable traditional product of Mediterranean countries educating consumers to the relationship between the VOO bitterness and its health effect.

Agronomical practices and new processing technology to avoid phenolic oxidation and hydrolysis and to enhance the aromatic components of the VOO have been developed and they can be used to modulate taste and flavor to diversify the products on the market. VOOs having high concentration of phenol compounds are bitter and pungent therefore many people do not consume them, thus loosing the health benefits related to their intake.

In this paper, the chemist's and nutritionist's point of view has been considered to address possible strategies to overcome the existing gap between the quality perceived by consumer and that established by expert tasters. Educational campaigns emphasizing the bitter-health link for olive oils should be developed.