Plant-Based Seafood Analogs

There is a growing global need to shift from animal- towards plant-based diets. The main motivations are environmental/sustainability-, human health- and animal welfare concerns. The aim is to replace traditional animal-based food with various alternatives, predominantly plant-based analogs. The elevated consumption of fish and seafood, leads to negative impacts on the ecosystem, due to dwindling biodiversity, environmental damage and fish diseases related to large-scale marine farming, and increased intake of toxic substances, particularly heavy metals, which accumulate in fish due to water pollution. While these facts lead to increased awareness and rising dietary shifts towards vegetarian and vegan lifestyles, still the majority of seafood consumers seek traditional products. This encourages the development of plant-based analogs for fish and seafood, mimicking the texture and sensorial properties of fish-meat, seafood, or processed fish products. Mimicking the internal structure and texture of fish or seafood requires simulating their nanometric fibrous-gel structure. Common techniques of structuring plant-based proteins into such textures include hydrospinning, electrospinning, extrusion, and 3D printing. The conditions required in each technique, the physicochemical and functional properties of the proteins, along with the use of other non-protein functional ingredients are reviewed. Trends and possible future developments are discussed.     

Gene Expression Profile of Prostate Cancer Patients by Chemiluminescent Analysis

Abstract

The heterogeneous and multifactorial nature of prostate cancer that generates differential gene expression patterns in tumor cells leads us to investigate the molecular mRNA profiling of 14 genes through streptavidin-alkaline phosphatase–labeled RNA probes from tissue samples with prostate cancer and benign prostatic hyperplasia. Hybridizations were performed using cDNA amplification for each gene spotted onto positively charged nylon membranes and densitometry readings. The constitutive gene GAPDH was used to normalize the data. The methods developed in this study may be applicable to the prostate cancer diagnosis using AR, CEACAM-1, DD3 (also called PCA3), OPN-1, and PSMA significant differential expression.     

Synthesis of Optically Active 1,4-Dioxane Nucleotide Analogs

Optically active nucleotide analogs were prepared that were composed of a 1,4-dioxane ring as the sugar analog to which either uracil or adenine attached together with two carboxylic ester groups, to be used as vehicles for formation of oligomers. The chiral 1,4-dioxane moiety was constructed from dimethyl L-tartrate via the corresponding (2R,3R)-dimethyl 2-O-allyl-tartrate.     

Very efficient search for nucleotide alignments

We describe a very efficient search for nucleotide alignments, which is analogous to the novel very efficient search for protein alignment. Just as it has been the case with the alignment of proteins, based on 20 × 20 adjacency matrices for amino acids, obtained from a superposition of labeled amino acids adjacency matrices for the proteins considered, one can construct labeled matrices of size 4 × 4, listing adjacencies of nucleotides in DNA sequence. The matrix elements correspond to 16 pairs of adjacent nucleotides. To obtain DNA alignments, one combines information in the corresponding matrices for a pair of DNA nucleotides. Matrices are obtained by insertion of the sequential labels for pairs of nucleotides in the corresponding cells of the 4 × 4 tables. When two such matrices are superimposed, one can identify all segments in two DNA sequences, which are shifted relative to one another by the same amount in either direction, without using trial‐and‐error displacements of the two sequences one relative to the other to find local nucleotide alignments. © 2012 Wiley Periodicals, Inc.     

Accurate pKa Calculation of the Conjugate Acids of Alkanolamines,Alkaloids and Nucleotide Bases by Quantum Chemical Methods

The pK_a of the conjugate acids of alkanolamines, neurotransmitters, alkaloid drugs and nucleotide bases are calculated with density functional methods (B3LYP, M08‐HX and M11‐L) and ab initio methods (SCS‐MP2, G3). Implicit solvent effects are included with a conductor‐like polarizable continuum model (CPCM) and universal solvation models (SMD, SM8). G3, SCS‐MP2 and M11‐L methods coupled with SMD and SM8 solvation models perform well for alkanolamines with mean unsigned errors below 0.20 pK_a units, in all cases. Extending this method to the pK_a calculation of 35 nitrogen‐containing compounds spanning 12 pK_a units showed an excellent correlation between experimental and computational pK_a values of these 35 amines with the computationally low‐cost SM8/M11‐L density functional approach.     

Contribution and limits of a non‐intrusive Raman spectroscopic method compared with HPLC for routine application to pre‐delivery analytical control of two major camptothecin analogs: irinotecan and topotecan

The purpose of this study was to develop a Raman spectroscopy (RS) method as an effective tool for the non‐intrusive pre‐delivery analytical quality control (AQC) of two camptothecin analogs, i.e. irinotecan (IRI) and topotecan (TPT), which are prescribed and compounded at the hospital. Following a phase of analytical pre‐validation, based on the actual conditions of use of the analogs, the protocol was validated and compared with the reference high‐performance liquid chromatography (HPLC) method. For IRI, AQC by RS has been validated in ranges from 0.94 to 3.27 mg/ml in saline solutions and from 0.89 to 3.30 mg/ml in dextrose solutions. These ranges recover the entire therapeutic concentrations encountered in clinical practice, i.e. 1.08–2.8 mg/ml. The RS and HPLC methods were validated by calculating the accuracy profile and provided excellent results for the analytical validation key criteria. The Spearman and Kendall correlation tests (p‐value < 1.10⁻¹¹) and the statistical studies performed on the Bland and Altman graphs confirm a strong correlation between RS and HPLC results. However, we show that a routine apparatus is unable to quantify TPT therapeutic concentrations ranging between 25 and 50 µg/ml but that a sufficiently powerful RS bench is able to detect and quantify TPT. Overall, these results confirm the potential of the RS option for future innovative applications. Owing to its analytical and practical qualities, this promising method contributes to the improvement of the safety of the medication circuit at the hospital and to the protection of caregivers and their working environment. Copyright © 2015 John Wiley & Sons, Ltd.     

Discriminative detection of low‐abundance point mutations using a PCR/ligase detection reaction/capillary gel electrophoresis method and fluorescence dual‐channel monitoring

We applied a facile LIF dual‐channel monitoring system recently developed and reported by our group to the polymerase chain reaction/ligase detection reaction/CGE method for detecting low‐abundance point mutations present in a wild‐type sequence‐dominated population. Mutation discrimination limits and signaling fidelity of the analytical system were evaluated using three mutant variations in codon 12 of the K‐ras oncogene that have high diagnostic value for colorectal cancer. We demonstrated the high sensitivity of the present method by detecting rare mutations present among an excess of wild‐type alleles (one mutation among ～100 normal sequences). This method also simultaneously interrogated the allelic compositions of the test samples with high specificity through spectral discrimination of the dye‐tagged ligase detection reaction products using the dual‐channel monitoring system.     

Novel 1,2,3-Triazole Derivatives as Mimics of Steroidal System—Synthesis,Crystal Structures Determination,Hirshfeld Surfaces Analysis and Molecular Docking

Herein, we present the synthesis and crystal structures determination of five 4-(1-phenyl-1H-1,2,3-triazol-4-yl)phenol derivatives containing halogen atoms, 6a–e, which may be used as an excellent mimic of steroids in the drug development process. Good quality crystals obtained for all of the synthesized compounds allowed the analysis of their molecular structures. Subsequently, the determined crystal structures were used to calculate the Hirshfeld surfaces for each of the synthesized compounds. Furthermore, results of our docking studies indicated that synthesized derivatives are able to bind effectively to the active sites of selected enzymes and receptors involved in the hormone biosynthesis and signaling pathways, analogously to the native steroids.     

Computational analyses prioritize and reveal the deleterious nsSNPs in human angiotensinogen gene

Angiotensinogen (AGT) is a key component of renin-angiotensin-aldosterone system (RAAS), which plays central role in blood pressure homeostasis. Association of AGT polymorphisms have been investigated in different ethnic populations in variety of cardiovascular and non-cardiovascular conditions. In this study, 354 non-synonymous SNPs (nsSNPs) of AGT were evaluated to predict damaging and structurally important variants. Majority of the deleterious nsSNPs occurred in the evolutionary conserved regions. Several of these nsSNPs were found to affect post-translational modifications like methylation, glycosylation, phosphorylation, ubiquitination etc. Structural evaluations predicted 19 variants as destabilizing and some of them were also predicted to destabilize the renin-AGT interaction. Therefore, the present computational investigation predicted pathogenic and functionally important variants of human AGT gene. The study has also shown that AGT deregulation is associated with survival outcome in patients with gastric and breast cancer, using microarray gene expression profile. Furthermore, the computationally screened nsSNPs can be analyzed in population based genotyping studies and may help futuristic drug development in the area of AGT pharmacogenomics.