Biosafety and Proteome Profiles of Different Heat Inactivation Methods for Mycobacterium tuberculosis

Inactivation of Mycobacterium tuberculosis is an important step to guarantee biosafety for subsequent M. tuberculosis identification and related research, notably in areas of endemicity with minimal resources. However, certain biomolecules might be denatured or hydrolyzed because of the harsh inactivation process, and a standardized protocol is yet to be determined.

Mineral Inactivation of Zinc in Polluted Soil—Sustainability of Zeolite, Bentonite and Blends

The study outlines a novel and traceable procedure for inactivating zinc polluted soil (an Anthrosols) adjacent to a former zinc (Zn) ore mine “Orzel Biały” in Bytom (Poland), where the total content of Zn amounted to 3988.0 mg kg−1. This pollution level initiated an inactivation process involving two natural mineral sorbents, i.e., zeolite (Z) and bentonite (B), as well as their five blends (ZeoBen) expressed as ZB: (1) ZB15/85, (2) ZB30/70, (3) ZB50/50, (4) ZB70/30 and (5) ZB85/15. Next, phosphorus (P) as triple superphosphate (TSP, 46% P2O5) was added to individual ZB at rates: 0.25%, 0.5%, 1.0% and 2.0%. All sorbents were added to the Zn polluted soil at 0%, 0.25%, 0.5%, 1.0% and 2.0% (dry weight basis). Treatments (1.0 kg of Zn-polluted soil with ZB sorbents) were aged for 115 days. Data revealed that ZB85/15 with prevailing zeolite caused a Znact inactivation of 66–71%, while zeolite induced 54% and 47% for bentonite. Reactive zinc (Znreac) decreased much more (20%) when zeolite was incorporated at the rate 2.5 g·kg−1 soil, and bentonite was (10%) at the same rate. The application of the sorbent ZB50/50 enriched with triple superphosphate (TSP) raised the stabilization degree for both Zn fractions. The efficiency was significant at the TSP rate of 2.0% of the sorbent and at least the sorbent +TSP of 10 g·kg−1 soil. The cation exchange capacity (CEC) of about 2 cmol(+)·kg−1 controlled the activity −0.50 mmol·dm−3 of either γZnreac or γZnact, hence a very low zinc ionic activity. The use of mineral blends with higher sharing of zeolite is promising for remediating metal-polluted lands in the case of zinc.

Computational Investigation of Protein Photoinactivation by Molecular Hyperthermia

To precisely control protein activity in a living system is a challenging yet long-pursued objective in biomedical sciences. Recently, we have developed a new approach named molecular hyperthermia (MH) to photoinactivate protein activity of interest without genetic modification. MH utilizes nanosecond laser pulse to create nanoscale heating around plasmonic nanoparticles to inactivate adjacent protein in live cells. Here we use a numerical model to study important parameters and conditions for MH to efficiently inactivate proteins in nanoscale. To quantify the protein inactivation process, the impact zone is defined as the range where proteins are inactivated by the nanoparticle localized heating. Factors that reduce the MH impact zone include the laser pulse duration, temperature-dependent thermal conductivity (versus constant properties), and nonspherical nanoparticle geometry. In contrast, the impact zone is insensitive to temperature-dependent material density and specific heat, as well as thermal interface resistance based on reported data in the literature. The low thermal conductivity of cytoplasm increases the impact zone. Different proteins with various Arrhenius kinetic parameters have significantly different impact zones. This study provides guidelines to design the protein inactivation process by MH.

Computational investigation of protein photoinactivation by molecular hyperthermia

To precisely control protein activity in a living system is a challenging yet long-pursued objective in biomedical sciences. Recently we have developed a new approach named molecular hyperthermia (MH) to photoinactivate protein activity of interest without genetic modification. MH utilizes nanosecond laser pulse to create nanoscale heating around plasmonic nanoparticles to inactivate adjacent protein in live cells. Here we use a numerical model to study important parameters and conditions for MH to efficiently inactivate proteins in nanoscale. To quantify the protein inactivation process, the impact zone is defined as the range where proteins will be inactivated by nanoparticle localized heating. Factors that reduce the MH impact zone include stretching the laser pulse duration, temperature-dependent thermal conductivity (versus constant properties), and non-spherical nanoparticle geometry. In contrast, the impact zone is insensitive to temperature-dependent material density and specific heat, as well as thermal interface resistance based on reported data. The low thermal conductivity of cytoplasm increases the impact zone. Different proteins with various Arrhenius kinetic parameters have significantly different impact zones. This study provides guidelines to design the protein inactivation process in MH.

Ultra-Deep DNA Methylation Analysis of X-Linked Genes: GLA and AR as Model Genes

Recessive X-linked disorders may occasionally evolve in clinical manifestations of variable severity also in female carriers. For some of such diseases, the frequency of the symptoms’ appearance during women’s life may be particularly relevant. This phenomenon has been largely attributed to the potential skewness of the X-inactivation process leading to variable phenotypes. Nonetheless, in many cases, no correlation with X-inactivation unbalance was demonstrated. However, methods for analyzing skewness have been mainly limited to Human Androgen Receptor methylation analysis (HUMARA). Recently, the X-inactivation process has been largely revisited, highlighting the heterogeneity existing among loci in the epigenetic state within inactive and, possibly, active X-chromosomes. We reasoned that gene-specific and ultra-deep DNA methylation analyses could greatly help to unravel details of the X-inactivation process and the roles of specific X genes inactivation in disease manifestations. We recently provided evidence that studying DNA methylation at specific autosomic loci at a single-molecule resolution (epiallele distribution analysis) allows one to analyze cell-to-cell methylation differences in a given cell population. We here apply the epiallele analysis at two X-linked loci to investigate whether females show allele-specific epiallelic patterns. Due to the high potential of this approach, the method allows us to obtain clearly distinct allele-specific epiallele profiles.