Chickpea protein hydrolysate as a substitute for serum in cell culture

The growth of mammalian cells in vitro requires the use of rich culture media that are prepared by combining serum with specific nutrient formulations. Serum, the most expensive component of culture media, provides a complex mixture of growth factors and nutrients. Protein hydrolysates that can support in vitro cell growth and eliminate or reduce the need to use serum have been obtained from different sources. Here we describe the use of two food grade proteases to produce a chickpea protein hydrolysate that has been added to cell culture medium in order to determine whether it can be used as a substitute for serum. Medium containing the hydrolysate has been tested using two human cells lines: the monocytic THP-1 cell line which grows in suspension, and the epithelial Caco-2 cell line which grows as a monolayer. The chickpea protein hydrolysate was a good substitute for serum in the first case, but did not allow growth of Caco-2 cells. Supplementation of culture media with this inexpensive and safe hydrolysate would greatly reduce the cost of cell culture.     

Efficacy and safety of pregabalin 600 mg/d for treating painful diabetic peripheral neuropathy: A double-blind placebo-controlled trial

Background  

Recent consensus guidelines recommend pregabalin as a first-tier treatment for painful diabetic peripheral neuropathy (DPN). We evaluated the efficacy of pregabalin 600 mg/d (300 mg dosed BID) versus placebo for relieving DPN-associated neuropathic pain, and assessed its safety using objective measures of nerve conduction (NC).     

Intrinsically antibacterial materials based on polymeric derivatives of eugenol for biomedical applications

Infections are the most common cause of biomaterial implant failure representing a constant challenge to the more widespread application of medical implants. This study reports on the preparation and characterization of novel hydrophilic copolymeric systems provided with antibacterial properties coming from eugenol residues anchored to the macromolecular chains. Thus, high conversion copolymers were prepared from the hydrophilic monomer 2-hydroxyethyl methacrylate (HEMA) and different eugenol monomeric derivatives, eugenyl methacrylate (EgMA) and ethoxyeugenyl methacrylate (EEgMA), by bulk polymerization reaction. Thermal evaluation revealed glass transition temperature values in the range 95-58 degrees C following the order HEMA-co-EgMA > PHEMA > HEMA-co-EEgMA and a clear increase in thermal stability with the presence of any eugenyl monomer in the system. In vitro wettability studies showed a reduction of water sorption capacity and surface free energy values with increasing the content of eugenol residues in the copolymer. The antimicrobial activity of copolymeric discs was evaluated by determining their capacity to reduce or inhibit colony formation by different bacterial species. All eugenyl containing materials showed bacteria growth inhibition, this one being higher for the EEgMA derivative copolymers.     

Algorithm for identification of fusion proteins via mass spectrometry

Identification of fusion proteins has contributed significantly to our understanding of cancer progression, yielding important predictive markers and therapeutic targets. While fusion proteins can be potentially identified by mass spectrometry, all previously found fusion proteins were identified using genomic (rather than mass spectrometry) technologies. This lack of MS/MS applications in studies of fusion proteins is caused by the lack of computational tools that are able to interpret mass spectra from peptides covering unknown fusion breakpoints (fusion peptides). Indeed, the number of potential fusion peptides is so large that the existing MS/MS database search tools become impractical even in the case of small genomes. We explore computational approaches to identifying fusion peptides, propose an algorithm for solving the fusion peptide identification problem, and analyze the performance of this algorithm on simulated data. We further illustrate how this approach can be modified for human exons prediction.     

Association of human mitochondrial DNA variants with plasma LDL levels

Increasing evidence supports the relationship between mitochondrial DNA variability and differences in energy metabolism, which may have pathophenotypic consequences. MtDNA pathological mutation has been also described to be associated with hypercholesterolemia. The target of this work consisted in studying the possible existence of an association between the mitochondrial DNA variability and plasma cholesterol levels. For this, two populations of 61 sedentary and 83 sportsmen were used to estimate the association of the lipidemic levels with the mitochondrial DNA variant harboured by them. Triglycerides, HDL-c, LDL-c and cholesterol/HDL-c were essayed, and mitochondrial DNA polymorphisms were assessed by HVR I sequencing and PCR/RFLP analysis. Major Caucasian mtDNA clades (HV, JT, U and IWX) did not associate with lipidemic levels in the sedentary population. However, in the case of a more disciplined population in term of nutritional habits and life style as sportsmen are, a significantly higher and lower level of LDL-c was associated with HV and JT clade, respectively. This observation could have relevant significance for metabolic distress diseases affecting plasma cholesterol levels.     

A combined proteome and ultrastructural localization analysis of 14-3-3 proteins in transformed human amnion (AMA) cells: definition of a framework to study isoform-specific differences

The 14-3-3 proteins constitute a family of highly conserved and broadly expressed multifunctional polypeptides that are involved in a variety of important cellular processes that include cell cycle progression, growth, differentiation, and apoptosis. Although the exact cellular function(s) of 14-3-3 proteins is not fully elucidated, as a rule these proteins act by binding to protein ligands, thus regulating their activity; so far more than 300 cellular proteins have been reported to interact with 14-3-3 proteins. Binding to cognate interacting partners is isoform-specific, but redundancy also exists as several binding peptides can be recognized by all isoforms, and some functions can be carried out by any isoform indistinctly. Moreover by interacting with different ligands in a spatially and temporally regulated fashion the same isoform can play multiple possibly even opposing roles where the resultant cellular outcome will be determined by the integration of the various effects. Although there is a large body of literature on specific aspects of 14-3-3 biology, not much is known on the coordinated aspects of 14-3-3 isoform expression, post-translational modifications, and subcellular localization. To address the question of isoform-specific differences, we carried out a comparative analysis of the patterns of expression, phosphorylation, and subcellular localization of the 14-3-3 beta, epsilon, sigma, tau, and zeta protein isoforms in transformed human amnion (AMA) cells. To validate as well as broaden our observations we analyzed the occurrence of the various isoforms in a large number of established cell lines and mammary and urothelial tissue specimens. Given the systematic approach we undertook and our application of isoform-discriminating technologies to the analysis of various cellular systems, we expect the data presented in this study to serve as an enabling resource for researchers working with 14-3-3 proteins.     

Flexible informatics for linking experimental data to mathematical models via DataRail

MOTIVATION: Linking experimental data to mathematical models in biology is impeded by the lack of suitable software to manage and transform data. Model calibration would be facilitated and models would increase in value were it possible to preserve links to training data along with a record of all normalization, scaling, and fusion routines used to assemble the training data from primary results. RESULTS: We describe the implementation of DataRail, an open source MATLAB-based toolbox that stores experimental data in flexible multi-dimensional arrays, transforms arrays so as to maximize information content, and then constructs models using internal or external tools. Data integrity is maintained via a containment hierarchy for arrays, imposition of a metadata standard based on a newly proposed MIDAS format, assignment of semantically typed universal identifiers, and implementation of a procedure for storing the history of all transformations with the array. We illustrate the utility of DataRail by processing a newly collected set of approximately 22 000 measurements of protein activities obtained from cytokine-stimulated primary and transformed human liver cells. AVAILABILITY: DataRail is distributed under the GNU General Public License and available at http://code.google.com/p/sbpipeline/     

Antifouling activity of sessile bacilli derived from marine surfaces

Marine biofilms are a virtually untapped source of bioactive molecules that may find application as novel antifoulants in the marine paint industry. This study aimed at determining the potential of marine biofilm bacteria to produce novel biomolecules with potential application as natural antifoulants. Nine representative strains were isolated from a range of surfaces and were grown in YEB medium and harvested during the late exponential growth phase. Bacterial biomass and spent culture medium were extracted with ethanol and ethyl acetate, respectively. Extracts were assayed for their antifouling activity using two tests: (1) antimicrobial well diffusion test against a common fouling bacterium, Halomonas marina, and (2) anti-crustacean activity test using Artemia salina. Our results showed that none of the ethanolic extracts (bacterial biomass) were active in either test. In contrast, most of the organic extracts had antimicrobial activity (88%) and were toxic towards A. salina (67%). Sequencing of full 16 S ribosomal DNA analysis showed that the isolates were related to Bacillus mojavensis and Bacillus firmus. Matrix-assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF-MS) profiling of ethyl acetate extracts of culture supernatants showed that these species produce the bioactive lipopeptides surfactin A, mycosubtilin and bacillomycin D.