Quantitative assessment of specific carbonic anhydrase inhibitors effect on hypoxic cells using electrical impedance assays

Carbonic anhydrase IX (CA IX) is an important orchestrator of hypoxic tumour environment, associated with tumour progression, high incidence of metastasis and poor response to therapy. Due to its tumour specificity and involvement in associated pathological processes: tumourigenesis, angiogenesis, inhibiting CA IX enzymatic activity has become a valid therapeutic option. Dynamic cell-based biosensing platforms can complement cell-free and end-point analyses and supports the process of design and selection of potent and selective inhibitors. In this context, we assess the effectiveness of recently emerged CA IX inhibitors (sulphonamides and sulphocoumarins) and their antitumour potential using an electrical impedance spectroscopy biosensing platform. The analysis allows discriminating between the inhibitory capacities of the compounds and their inhibition mechanisms. Microscopy and biochemical assays complemented the analysis and validated impedance findings establishing a powerful biosensing tool for the evaluation of carbonic anhydrase inhibitors potency, effective for the screening and design of anticancer pharmacological agents.     

Chemical evidence for the mechanism of the biodecoloration of Amaranth by <Emphasis Type="Italic">Trametes versicolor</Emphasis>

The white-rot fungus Trametes versicolor decolorized Amaranth. The hypothesis that the carbon structure of Amaranth was broken down in smaller mass fragments was investigated analyzing the products of decoloration. FTIR spectroscopy, ion chromatography, sulfite and ammonia analysis were used to compare the culture filtrate before dye addition, with the pure dye, the culture filtrate after dye addition, and the culture filtrate during the treatment. The hypothesis of polymerization of the decoloration products was tested by spectrophotometric analysis of dialysates of the pure dye, the culture filtrate before dye addition, and the culture filtrates after dye addition and decoloration. FTIR showed that the signals typical for the azo group disappeared after decoloration, while new peaks appeared that were characteristic of substituted naphthalenic or benzenic compounds. Ion chromatography showed that the level of sulfate in the treatment increased when compared with the level of the sulfate in control, suggesting that the sulfonic groups were being stripped from Amaranth’s structure and metabolized to sulfate. Sulfite measurements for the treatment and controls showed no significant difference, and were well below the saturation concentration for sulfite in water, confirming that the medium was aerobic. Ammonia concentration did not change with the decoloration. Absorbance scans after dialysis of decolorized samples showed no new peaks, suggesting that the decoloration products were not polymerized. These observations suggests that the decoloration mechanism starts with the azo link removal, followed by desulfonation, naphthalene ring opening, and the formation of smaller mass fragments, similar to fungal metabolites.     

Identifying dynamical persistent biomarker structures for rare events using modern integrative machine learning approach

The evolution of omics and computational competency has accelerated discoveries of the underlying biological processes in an unprecedented way. High throughput methodologies, such as flow cytometry, can reveal deeper insights into cell processes, thereby allowing opportunities for scientific discoveries related to health and diseases. However, working with cytometry data often imposes complex computational challenges due to high-dimensionality, large size, and nonlinearity of the data structure. In addition, cytometry data frequently exhibit diverse patterns across biomarkers and suffer from substantial class imbalances which can further complicate the problem. The existing methods of cytometry data analysis either predict cell population or perform feature selection. Through this study, we propose a “wisdom of the crowd” approach to simultaneously predict rare cell populations and perform feature selection by integrating a pool of modern machine learning (ML) algorithms. Given that our approach integrates superior performing ML models across different normalization techniques based on entropy and rank, our method can detect diverse patterns existing across the model features. Furthermore, the method identifies a dynamic biomarker structure that divides the features into persistently selected, unselected, and fluctuating assemblies indicating the role of each biomarker in rare cell prediction, which can subsequently aid in studies of disease progression.     

Aquatain AMF efficacy on juvenile mosquito stages in control of Culex pipiens complex and Aedes albopictus

Aquatain^® mosquito formulation (AMF) is a silicone-based monomolecular film, which has recently been approved for use in the European Union. The physical mode of action based on lowering water surface tension prevents mosquito larvae/pupae respiration. Additionally, AMF disables gravid females from landing on the water surface and obstructs the natural oviposition process. Due to multistage effects on mosquitoes, AMF could be a product of choice for defined water body and container breeders such as Culex pipiens L. complex, principal vector of West Nile virus in Europe, and the invasive Aedes albopictus (Skuse) (both Diptera: Culicidae), vector of dengue and chikungunya viruses. The primary objectives of this study were to evaluate the efficacy of AMF, to determine the susceptibility of the immature forms of C. pipiens and A. albopictus, and the persistence/longevity of the product to suppress the eclosion of adults. AMF achieved high mortality rates of juvenile A. albopictus and C. pipiens under laboratory conditions. However, in the field C. pipiens larvae showed higher susceptibility to AMF than A. albopictus. Pupae of the two mosquito species were highly susceptible to the presence of AMF. When C. pipiens juveniles were exposed to AMF in the wild, effects lasted for 21 days in densely covered water bodies and 56 days in water recipients with less vegetation. In both breeding sites, natural habitat and artificial water recipient, the two mosquito species with high impact on public health in Europe could successfully be suppressed by application of AMF (1 ml m⁻²).     

Improved determination of plasmid copy number using quantitative real-time PCR for monitoring fermentation processes

Background  

Recombinant protein production in Escherichia coli cells is a complex process, where among other parameters, plasmid copy number, structural and segregational stability of plasmid have an important impact on the success of productivity. It was recognised that a method for accurate and rapid quantification of plasmid copy number is necessary for optimization and better understanding of this process. Lately, qPCR is becoming the method of choice for this purpose. In the presented work, an improved qPCR method adopted for PCN determination in various fermentation processes was developed.     

Coming to Grips with Evolvability

To explain the evolution of complex organisms by random mutation, drift, and selection is not a trivial task. This becomes obvious if we imagine an organism in which most genes affect most traits and all mutations are immediately expressed in the phenotype. Most of the mutations will be deleterious. Computer programmers experienced a similar problem when trying to evolve computer programs by introducing random changes to a conventional computer code, realizing that almost all random changes are “lethal.” Everyone who has done any programming knows that conventional computer languages are very brittle! Real organisms are not organized in this way but rather involve mediation between the genes and the phenotypic traits, namely development, also sometimes called the genotype–phenotype map. This map of genetic effects is structured in a way that enables evolvability, that is, enhances the probability that mutations will improve the performance of the organism. Here we outline two properties of organismal development, namely modularity and robustness. Modularity refers to the situation in which genes affect a restricted number of functionally related phenotypic characters. Robustness describes a situation in which cryptic mutations can accumulate without effect on fitness but can become visible to selection in a new environment or genetic background. We discuss recent empirical evidence in support of both phenomena and their effect on evolvability and also briefly address their evolution.     

Automatic detection of basal cell carcinoma by hyperspectral imaging

The purpose of this study was to test the ability of hyperspectral imaging (HSI) combined with unsupervised anomaly detectors to automatically differentiate basal cell carcinoma (BCC) from normal skin. Hyperspectral images of the face of a female patient with a BCC of the lower lip were acquired using a visible/near-infrared HSI system and two anomaly detection algorithms (Reed-Xiaoli and Reed-Xiaoli/Uniform Target hybrid anomaly detectors) were used to detect pathological tissue from normal skin. The results revealed that the receiver operating characteristic curve of the Reed-Xiaoli/Uniform Target hybrid detector was higher than that of the Reed-Xiaoli detector in the range of false positive rates between 0 and 0.8. The area under curve values were good (0.7074 and 0.8607, respectively) with Reed-Xiaoli/Uniform Target hybrid detector performing better. In conclusion, HSI combined with either of two anomaly detectors can play a promising role in the automated screening of BCC.