ChIP — Does it work correctly? The optimization steps of chromatin immunoprecipitation

The proteins interaction with DNA is one of the key regulatory elements of many biological processes; including gene transcription, epigenetic modification or cell differentiation. Immunoprecipitation of chromatin; ChIP; is a method used to assess the interaction of the protein with a DNA sequence, and determines the localization of specific locus in the genome. The main steps of this method are fixation, sonication, immunoprecipitation and analysis of DNA. Although the immunoprecipitation assay is a multipurpose tool applied in biochemistry and biotechnology, it requires optimization. This paper describes several critical parameters that should be taken into account when immunoprecipitation assay is applied

TiO 2

One-step TiO2 nanoparticle synthesis based on the interaction between thiourea and metatitanic acid is applied for sulfur and carbon anatase codoping. The synthesis of the doped TiO2 has been monitored by means of differential thermal analysis and thermogravimetric analysis (DTA-TG), which allows determining the optimal thermal conditions for the process. Electron microscopy showed micrometer-sized (5–15 μm) randomly distributed crystal aggregates, consisting of many 15–40-nm TiO2 nanoparticles. The obtained phase composition and chemical states of the doping elements are analyzed by means of X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), infrared (IR) and Raman spectroscopies, and electron paramagnetic resonance (EPR). XRD displays in both samples (doped and pristine) the existence of only one crystalline phase—the tetragonal modification of TiO2—anatase. Further data assessment by means of Rietveld refinement allowed detection of a slight c lattice parameter and volume increase related to incorporation of the doping elements. XPS demonstrated the presence of carbon and sulfur as doping elements in the material. It was confirmed that carbon is in elemental form and also present in oxygen-containing compounds, which are adsorbed on the particle surface. The binding energy for sulfur electron core shell corresponds to the established data for sulfate compounds, where sulfur is in 6+ oxidation state. The synthesized S- and C-codoped TiO2 showed excellent photocatalytic performance during the degradation of organic dyes (rhodamine B, methylene blue), gas-phase oxidation of ethanol under visible light, and photocatalytic hydrogen generation from ethanol under ultraviolet light

Treasure on the Earth—Gold Nanoparticles and Their Biomedical Applications

Recent advances in the synthesis of metal nanoparticles (NPs) have led to tremendous expansion of their potential applications in different fields, ranging from healthcare research to microelectronics and food packaging. Among the approaches for exploiting nanotechnology in medicine, gold nanomaterials in particular have been found as the most promising due to their unique advantages, such as in sensing, image enhancement, and as delivery agents. Although, the first scientific article on gold nanoparticles was presented in 1857 by Faraday, during the last few years, the progress in manufacturing these nanomaterials has taken an enormous step forward. Due to the nanoscale counterparts of gold, which exhibit distinct properties and functionality compared to bulk material, gold nanoparticles stand out, in particular, in therapy, imaging, detection, diagnostics, and precise drug delivery. This review summarizes the current state-of-the-art knowledge in terms of biomedical applications of gold nanoparticles. The application of AuNPs in the following aspects are discussed: (i) imaging and diagnosing of specific target; (ii) treatment and therapies using AuNPs; and (iii) drug delivery systems with gold nanomaterials as a carrier. Among the different approaches in medical imaging, here we either consider AuNPs as a contrast agent in computed tomography (CT), or as a particle used in optical imaging, instead of fluorophores. Moreover, their nontoxic feature, compared to the gadolinium-based contrast agents used in magnetic resonance imaging, are shown. The tunable size, shape, and functionality of gold nanoparticles make them great carriers for targeted delivery. Therefore, here, we summarize gold-based nanodrugs that are FDA approved. Finally, various approaches to treat the specific diseases using AuNPs are discussed, i.e., photothermal or photodynamic therapy, and immunotherapy

Biocompatibility and Cytotoxicity of Gold Nanoparticles: Recent Advances in Methodologies and Regulations

Recent advances in the synthesis of metal nanoparticles (MeNPs), and more specifically gold nanoparticles (AuNPs), have led to tremendous expansion of their potential applications in different fields, ranging from healthcare research to microelectronics and food packaging. The properties of functionalised MeNPs can be fine-tuned depending on their final application, and subsequently, these properties can strongly modulate their biological effects. In this review, we will firstly focus on the impact of MeNP characteristics (particularly of gold nanoparticles, AuNPs) such as shape, size, and aggregation on their biological activities. Moreover, we will detail different in vitro and in vivo assays to be performed when cytotoxicity and biocompatibility must be assessed. Due to the complex nature of nanomaterials, conflicting studies have led to different views on their safety, and it is clear that the definition of a standard biosafety label for AuNPs is difficult. In fact, AuNPs’ biocompatibility is strongly affected by the nanoparticles’ intrinsic characteristics, biological target, and methodology employed to evaluate their toxicity. In the last part of this review, the current legislation and requirements established by regulatory authorities, defining the main guidelines and standards to characterise new nanomaterials, will also be discussed, as this aspect has not been reviewed recently. It is clear that the lack of well-established safety regulations based on reliable, robust, and universal methodologies has hampered the development of MeNP applications in the healthcare field. Henceforth, the international community must make an effort to adopt specific and standard protocols for characterisation of these products

Higher thermal and ethanol tolerance of a yeast strain isolated from oral cavity

798-803Efficient bioethanol producing microorganisms must be endowed with peculiar physiological and technological traits, such as, higher thermal and ethanol tolerance. This encompasses a strain thermotolerance and an ability to grow at elevated sugar and ethanol concentration or ability to sustain dehydration process such as freeze drying. In this study, we characterized a thermotolerant yeast strain isolated from the human oral cavity regarding the above mentioned parameters. Such an uncommon niche was considered as the great potential reservoir, to isolate strains endowed with metabolic aptitudes requested for ethanol production. In the process, we have defined the YTerm-1strain ability to sustain high sugar and ethanol concentration that are two technological constrains in bioethanol production. Finally, we highlighted that the strain YTerm-1 was able to accumulate, at a high level, trehalose and β-glucan, two compounds conferring the cells an increased resistance to freeze drying process and osmotic stress. Our results suggest that the Yterm-1 strain showed a better growth ability and higher ethanol yield as compared to the industrial strain. Other metabolic traits, such as resistance to dehydration stress, tolerance to ethanol, accumulation of intracellular trehalose or membrane β-glucan confer to that isolate all the characteristics requested in industrial production of ethanol

Biomaterial composed of chitosan, riboflavin, and hydroxyapatite for bone tissue regeneration

Abstract Biomaterial engineering approaches involve using a combination of miscellaneous bioactive molecules which may promote cell proliferation and, thus, form a scaffold with the environment that favors the regeneration process. Chitosan, a naturally occurring biodegradable polymer, possess some essential features, i.e., biodegradability, biocompatibility, and in the solid phase good porosity, which may contribute to promote cell adhesion. Moreover, doping of the materials with other biocompounds will create a unique and multifunctional scaffold that will be useful in regenerative medicine. This study is focused on the manufacturing and characterization of composite materials based on chitosan, hydroxyapatite, and riboflavin. The resulting films were fabricated by the casting/solvent evaporation method. Morphological and spectroscopy analyses of the films revealed a porous structure and an interconnection between chitosan and apatite. The composite material showed an inhibitory effect on Staphylococcus aureus and exhibited higher antioxidant activity compared to pure chitosan. In vitro studies on riboflavin showed increased cell proliferation and migration of fibroblasts and osteosarcoma cells, thus demonstrating their potential for bone tissue engineering applications