Generation of Functional Immortalized Human Corneal Stromal Stem Cells

In addition to their therapeutic potential in regenerative medicine, human corneal stromal stem cells (CSSCs) could serve as a powerful tool for drug discovery and development. Variations from different donors, their isolation method, and their limited life span in culture hinder the utility of primary human CSSCs. To address these limitations, this study aims to establish and characterize immortalized CSSC lines (imCSSC) generated from primary human CSSCs. Primary CSSCs (pCSSC), isolated from human adult corneoscleral tissue, were transduced with ectopic expression of hTERT, c-MYC, or the large T antigen of the Simian virus 40 (SV40T) to generate imCSSC. Cellular morphology, proliferation capacity, and expression of CSSCs specific surface markers were investigated in all cell lines, including TNFAIP6 gene expression levels in vitro, a known biomarker of in vivo anti-inflammatory efficacy. SV40T-overexpressing imCSSC successfully extended the lifespan of pCSSC while retaining a similar morphology, proliferative capacity, multilineage differentiation potential, and anti-inflammatory properties. The current study serves as a proof-of-concept that immortalization of CSSCs could enable a large-scale source of CSSC for use in regenerative medicine.     

Studying the Impact of Persistent Organic Pollutants Exposure on Human Health by Proteomic Analysis: A Systematic Review

Persistent organic pollutants (POPs) are organic chemical substances that are widely distributed in environments around the globe. POPs accumulate in living organisms and are found at high concentrations in the food chain. Humans are thus continuously exposed to these chemical substances, in which they exert hepatic, reproductive, developmental, behavioral, neurologic, endocrine, cardiovascular, and immunologic adverse health effects. However, considerable information is unknown regarding the mechanism by which POPs exert their adverse effects in humans, as well as the molecular and cellular responses involved. Data are notably lacking concerning the consequences of acute and chronic POP exposure on changes in gene expression, protein profile, and metabolic pathways. We conducted a systematic review to provide a synthesis of knowledge of POPs arising from proteomics-based research. The data source used for this review was PubMed. This study was carried out following the PRISMA guidelines. Of the 742 items originally identified, 89 were considered in the review. This review presents a comprehensive overview of the most recent research and available solutions to explore proteomics datasets to identify new features relevant to human health. Future perspectives in proteomics studies are discussed.     

Synthesis of an Anti-CD7 Recombinant Immunotoxin Based on PE24 in CHO and E. coli Cell-Free Systems

Recombinant immunotoxins (RITs) are an effective class of agents for targeted therapy in cancer treatment. In this article, we demonstrate the straight-forward production and testing of an anti-CD7 RIT based on PE24 in a prokaryotic and a eukaryotic cell-free system. The prokaryotic cell-free system was derived from Escherichia coli BL21 Star^TM (DE3) cells transformed with a plasmid encoding the chaperones groEL/groES. The eukaryotic cell-free system was prepared from Chinese hamster ovary (CHO) cells that leave intact endoplasmic reticulum-derived microsomes in the cell-free reaction mix from which the RIT was extracted. The investigated RIT was built by fusing an anti-CD7 single-chain variable fragment (scFv) with the toxin domain PE24, a shortened variant of Pseudomonas Exotoxin A. The RIT was produced in both cell-free systems and tested for antigen binding against CD7 and cell killing on CD7-positive Jurkat, HSB-2, and ALL-SIL cells. CD7-positive cells were effectively killed by the anti-CD7 scFv-PE24 RIT with an IC₅₀ value of 15 pM to 40 pM for CHO and 42 pM to 156 pM for E. coli cell-free-produced RIT. CD7-negative Raji cells were unaffected by the RIT. Toxin and antibody domain alone did not show cytotoxic effects on either CD7-positive or CD7-negative cells. To our knowledge, this report describes the production of an active RIT in E. coli and CHO cell-free systems for the first time. We provide the proof-of-concept that cell-free protein synthesis allows for on-demand testing of antibody–toxin conjugate activity in a time-efficient workflow without cell lysis or purification required.     

Reactivity to visual signals and the cerebellar vermis: Evidence from a rare case with rhombencephalosynapsis.

The modulatory role of the cerebellum was investigated in a case with rhombencephalosynapsis (RS), a rare dysplasia characterized by the absence of the cerebellar vermis. The visual psychophysical task involved localizing a target and ignoring a distractor appearing either before, at the same time as, or after the target. It allowed us to assess reactivity to warning signals, distraction, and reactivity to signals appearing during attentive processing. Compared with a control sample, the patient exhibited greater reactivity to warning signals and difficulties interrupting attentive processing, whereas distraction was not increased. Complementary analyses showed that these deficits did not reflect just delayed development of attentional processes. These results suggest that the cerebellum modulates responsiveness and commands attentional/exploratory flexibility in response to sensory signals. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Isolation and characterization of a psychrotolerant toxin producer, Bacillus weihenstephanensis, in liquid egg products

A psychrotolerant bacteria of the Bacillus cereus group was found responsible for the spoilage of whole liquid egg products. By sequencing a 16S rRNA region and performing a PCR amplification of specific 16S rRNA and cspA signatures, a Bacillus weihenstephanensis was identified. Characterization of this strain shows its ability to grow in defined medium as well as in whole liquid egg at refrigerated temperatures. The strain isolated possesses genes encoding for hemolysin BL, nonhemolytic enterotoxin, and B. cereus enterotoxins and produces enterotoxins with cytotoxic activity in whole liquid egg, even at refrigerated temperatures. The isolate exhibits a clear ability to stick and form biofilms on stainless steel, the most common material used in egg breaking factories, as well as on model hydrophilic (glass) and hydrophobic (polytetrafluoroethylene) materials. These findings show the necessity to monitor for Bacillus contamination in egg products that are often used in the composition of particularly susceptible finished products such as cream, dessert, dairy, meat, and seafood.     

Thick lead-free ferroelectric films with high Curie temperatures through nanocomposite-induced strain

Ferroelectric materials are used in applications ranging from energy harvesting to high-power electronic transducers. However, industry-standard ferroelectric materials contain lead, which is toxic and environmentally unfriendly. The preferred alternative, BaTiO(3), is non-toxic and has excellent ferroelectric properties, but its Curie temperature of ～130 °C is too low to be practical. Strain has been used to enhance the Curie temperature of BaTiO(3) (ref. 4) and SrTiO(3) (ref. 5) films, but only for thicknesses of tens of nanometres, which is not thick enough for many device applications. Here, we increase the Curie temperature of micrometre-thick films of BaTiO(3) to at least 330 °C, and the tetragonal-to-cubic structural transition temperature to beyond 800 °C, by interspersing stiff, self-assembled vertical columns of Sm(2)O(3) throughout the film thickness. The columns, which are 10 nm in diameter, strain the BaTiO(3) matrix by 2.35%, forcing it to maintain its tetragonal structure and resulting in the highest BaTiO(3) transition temperatures so far.     

Surface-immobilized peptide aptamers as probe molecules for protein detection

We demonstrate the use of surface-immobilized, oriented peptide aptamers for the detection of specific target proteins from complex biological solutions. These peptide aptamers are target-specific peptides expressed within a protein scaffold engineered from the human protease inhibitor stefin A. The scaffold provides stability to the inserted peptides and increases their binding affinity owing to the resulting three-dimensional constraints. A unique cysteine residue was introduced into the protein scaffold to allow orientation-specific surface immobilization of the peptide aptamer and to ensure exposure of the binding site to the target solution. Using dual-polarization interferometry, we demonstrate a strong relationship between binding affinity and aptamer orientation and determine the affinity constant KD for the interaction between an oriented peptide aptamer ST(cys+)_(pep9) and the target protein CDK2. Further, we demonstrate the high selectivity of the peptide aptamer STM_(pep9) by exposing surface-immobilized ST(cys+)_(pep9) to a complex biological solution containing small concentrations of the target protein CDK2.     

Transgenerational effects of the holocaust: Externalization of aggression in second generation of holocaust survivors.

Examined the psychological characteristics of the children of holocaust survivors. 38 males and females in their late 20's were interviewed; the parents of 19 Ss were holocaust survivors, and the parents of the other 19 Ss were in Israel during World War II and did not suffer directly. It was hypothesized that reactions of the children of the holocaust survivors would include guilt rather than external aggression and a tendency not to externalize aggressive impulses in reaction to frustrating events. The Ss' reactions to a projective test were analyzed, and the responses provide support for the major hypothesis. Results show that sons and daughters of holocaust survivors were less likely to externalize aggression than were those in a control group. This finding is supported by the content analysis of the structured interview conducted with the Ss in both groups. (20 ref) (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Infiltration and Top Seed Growth-Textured YBCO Bulks With Multiple Holes

The recently reported hole-patterned YBa₂Cu₃O _y (Y123) bulks with improved superconducting properties are highly interesting from material quality and application variety points of view. It is well known that the core of plain bulk superconductors needs to be fully oxygenated and some defects like cracks, pores, and voids must be suppressed in order that the material can trap a high magnetic field or carry a high current density. To minimize the above defects, we have used a combination of standard superconducting ceramic processing and an infiltration technique to prepare regularly perforated YBa₂Cu₃O _y (Y123) bulk superconductors. This process leads to negligible shrinkage upon annealing and a uniform distribution of Y211 inclusions. Texture was evidenced by neutron pole figure measurements. Flux mapping was used to verify the superconducting homogeneity of the samples and to investigate the field-trapping ability. In addition, the textured drilled samples were reinforced using resin or metal impregnation and the influence of the different processing steps on the hardness of the materials has been investigated.     

Synthesis of polymer materials for use as cell culture substrates

Up to today, several techniques have been used to maintain cells in culture for studying many aspects of cell biology and physiology. More often, cell culture is dependent on proper anchorage of cells to the growth surface. Thus, poly-l-lysine, fibronectin or laminin are the most commonly used substrates. In this study, electrosynthesized biocompatible polymer films are proposed as an alternative to these standard substrates. The electrosynthesized polymers tested were polyethylenimine, polypropylenimine and polypyrrole. Then, the adhesion, proliferation and morphology of rat neuronal cell lines were investigated on these polymer substrates in an attempt to develop new and efficient polymer materials for cell culture. During their growth on the polymers, the evolution of the cell morphology was monitored using both confocal microscopy and immunohistochemistry, leading to the conclusion of a normal development. An estimation of the adhesion and proliferation rates of rat neuronal cell cultures indicated that polyethylenimine and polypropylenimine were the best substrates for culturing olfactory neuronal cells. A method to favour the differentiation of the neuronal cells was also developed since the final aim of this work is to develop a biosensor for odour detection using differentiated neuronal cells as transducers. Consequently, a biosensor was microfabricated using silicon technology. This microsystem allowed us to culture the cells on a silicon wafer and to position the cells on certain parts of the silicon wafer.