Rapid determination of uracil in biological fluids at mercury thin film electrode for early detection of potential 5-fluorouracil toxicity due to dihydropyrimidine dehydrogenase deficiency.

Determination of plasma uracil was reported as a method for evaluation of Dihydropyrimidine dehydrogenase (DPD) activity that is highly demanded to ensure the safe administration of 5-fluorouracil (5-FU)-based therapies to cancer patients. This work reports the development of a simple electroanalytical method based on adsorptive stripping square wave voltammetry (AdSWV) at mercury film-coated glassy carbon electrode (MF/GCE) for the highly sensitive determination of uracil in biological fluids that can be used for diagnosis of decreased DPD activity. Due to the formation of the HgII-Uracil complex at the electrode surface, the accuracy of the measurement was not affected by the complicated matrices in biological fluids including human serum, plasma, and urine. The high sensitivity of the developed method results in a low limit of detection (≈1.3 nM) in human plasma samples, falling below the practical cut-off level of 15 ng mL-1 (≈0.14 µM). This threshold concentration is crucial for predicting 5-FU toxicity, as reported in buffer, and ≤1.15% in biological samples), and accuracy (recovery percentage close to 100%).

Enhancing the substrate selectivity of enzyme mimetics in biosensing and bioassay: Novel approaches.

A substantial development in nanoscale materials possessing catalytic activities comparable with natural enzymes has been accomplished. Their advantages were owing to the excellent sturdiness in an extreme environment, possibilities of their large-scale production resulting in higher profitability, and easy manipulation for modification. Despite these advantages, the main challenge for artificial enzyme mimetics is the lack of substrate selectivity where natural enzymes flourish. This review addresses this vital problem by introducing substrate selectivity strategies to three classes of artificial enzymes: molecularly imprinted polymers, nanozymes (NZs), and DNAzymes. These rationally designed strategies enhance the substrate selectivity and are discussed and exemplified throughout the review. Various functional mechanisms associated with applying enzyme mimetics in biosensing and bioassays are also given. Eventually, future directives toward enhancing the substrate selectivity of biomimetics and related challenges are discussed and evaluated based on their efficiency and convenience in biosensing and bioassays.

Exploring the potential nutritional benefits of Arthrospira maxima and Chlorella vulgaris: A focus on vitamin B12, amino acids, and micronutrients.

Arthrospira (Limnospira) maxima (A. maxima) and Chlorella vulgaris (Ch. vulgaris) are among the approved microalgae and cyanobacteria (MaC) in the food industry that are known to be safe for consumption. However, both organisms are controversial regarding their vitamin B12 content, due to the possible occurrence of pseudo-cobalamin. Concurrently, their nutrition profiles remain understudied. The main purpose of the present study was to identify their nutrition profiles, focusing mainly on vitamin B12, amino acids, and micronutrients under iron-induced hormesis (10 mg/L Fe in treated samples). Our findings indicate a higher B12 content in A. maxima compared to Ch. vulgaris (both control and treated samples). Using liquid chromatography with tandem mass spectrometry (LC-MS/MS), the cyanocobalamin content was determined as 0.42 ± 0.09 µg/g dried weight (DW) in the A. maxima control and 0.55 ± 0.02 µg/g DW in treated A. maxima, resulting in an insignificant difference. In addition, the iron-enriched medium increased the amount of iron in both tested biomasses (p < 0.01). However, a more pronounced (approximately 100×) boost was observed in Ch. vulgaris, indicating a better absorption capacity (control Ch. vulgaris 0.16 ± 0.01 mg/g Fe, treated Ch. vulgaris 15.40 ± 0.34 mg/g Fe). Additionally, Ch. vulgaris also showed a higher micronutrient content. Using both tested microalgae, meeting the sufficient recommended daily mineral allowance for an adult is possible. By combining biomass from A. maxima and Ch. vulgaris in a ratio of 6:1, we can fulfill the recommended daily allowance of vitamin B12 and iron by consuming 6 tablets/6 g. Importantly, iron hormesis stimulated amino acid composition in both organisms. The profile of amino acids may suggest these biomasses as promising potential nutrition sources.

2D graphene-based advanced nanoarchitectonics for electrochemical biosensors: Applications in cancer biomarker detection.

Low-cost, rapid, and easy-to-use biosensors for various cancer biomarkers are of utmost importance in detecting cancer biomarkers for early-stage metastasis control and efficient diagnosis. The molecular complexity of cancer biomarkers is overwhelming, thus, the repeatability and reproducibility of measurements by biosensors are critical factors. Electrochemical biosensors are attractive alternatives in cancer diagnosis due to their low cost, simple operation, and promising analytical figures of merit. Recently graphene-derived nanostructures have been used extensively for the fabrication of electrochemical biosensors because of their unique physicochemical properties, including the high electrical conductivity, adsorption capacity, low cost and ease of mass production, presence of oxygen-containing functional groups that facilitate the bioreceptor immobilization, increased flexibility and mechanical strength, low cellular toxicity. Indeed, these properties make them advantageous compared to other alternatives. However, some drawbacks must be overcome to extend their use, such as poor and uncontrollable deposition on the substrate due to the low dispersity of some graphene materials and irreproducibility of the results because of the differences in various batches of the produced graphene materials. This review has documented the most recently developed strategies for electrochemical sensor fabrication. It differs in the categorization method compared to published works to draw greater attention to the wide opportunities of graphene nanomaterials for biological applications. Limitations and future scopes are discussed to advance the integration of novel technologies such as artificial intelligence, the internet of medical things, and triboelectric nanogenerators to eventually increase efficacy and efficiency.

Short-term autophagy inhibition by autophinib or SAR405 does not alter the effect of cisplatin on ATP production in prostate cancer cells.

OBJECTIVES: Cisplatin is a widely used anticancer drug for the treatment of many solid cancers. DNA damage is thought to be the key mechanism of cisplatin's anticancer activity. However, cisplatin may also affect cellular metabolism. The aim of this study was to determine the effect of cisplatin on the types of ATP production (OXPHOS versus glycolysis) and their rate in prostate cancer cells and to determine the potentially protective effect of autophagy and amino acids during cisplatin treatment. We also wanted to investigate the potential synergy between the metabolic effects of cisplatin on ATP production and the inhibition of autophagy. METHODS: Cisplatin treatment can significantly affect the metabolism of cancer cells. Important metabolic pathways can be altered, leading to changes in energy production and nutrient utilization. Autophagy and amino acid pool modulations can serve as protective mechanisms significantly affecting tumor cell survival under metabolic stress caused by anticancer treatment. By enabling the recycling of amino acids, autophagy helps cancer cells maintain cellular homeostasis and overcome nutrient limitations. Thus, inhibition of autophagy could have a supportive effect on the metabolic effects of cisplatin. RESULTS: After cisplatin treatment, ATP production by way of OXPHOS was significantly decreased in 22Rv1 and PC-3 cells. On the other hand, ATP production by glycolysis was not significantly affected in 22Rv1 cells. DU145 cells with dysfunctional autophagy were the most sensitive to cisplatin treatment and showed the lowest ATP production. However, short-term autophagy inhibition (24h) by autophinib or SAR405 in 22Rv1 and PC-3 cells did not alter the effect of cisplatin on ATP production. Levels of some amino acids (arginine, methionine) significantly affected the fitness of cancer cells. CONCLUSION: Persistent defects of autophagy can affect the metabolic sensitivity of cancer cells due to interference with arginine metabolism. Amino acids contained in the culture medium had an impact on the overall effect of cisplatin (Fig. 3, Ref. 38).

Physiological and transcriptome profiling of Chlorella sorokiniana: A study on azo dye wastewater decolorization.

Over decades, synthetic dyes have become increasingly dominated by azo dyes posing a significant environmental risk due to their toxicity. Microalgae-based systems may offer an alternative for treatment of azo dye effluents to conventional physical-chemical methods. Here, microalgae were tested to decolorize industrial azo dye wastewater (ADW). Chlorella sorokiniana showed the highest decolorization efficiency in a preliminary screening test. Subsequently, the optimization of the experimental design resulted in 70% decolorization in a photobioreactor. Tolerance of this strain was evidenced using multiple approaches (growth and chlorophyll content assays, scanning electron microscopy (SEM), and antioxidant level measurements). Raman microspectroscopy was employed for the quantification of ADW-specific compounds accumulated by the microalgal biomass. Finally, RNA-seq revealed the transcriptome profile of C. sorokiniana exposed to ADW for 72 h. Activated DNA repair and primary metabolism provided sufficient energy for microalgal growth to overcome the adverse toxic conditions. Furthermore, several transporter genes, oxidoreductases-, and glycosyltransferases-encoding genes were upregulated to effectively sequestrate and detoxify the ADW. This work demonstrates the potential utilization of C. sorokiniana as a tolerant strain for industrial wastewater treatment, emphasizing the regulation of its molecular mechanisms to cope with unfavorable growth conditions.

Surface modification strategies and the functional mechanisms of gold nanozyme in biosensing and bioassay.

Gold nanozymes (GNZs) have been widely used in biosensing and bioassay due to their interesting catalytic activities that enable the substitution of natural enzyme. This review explains different catalytic activities of GNZs that can be achieved by applying different modifications to their surface. The role of Gold nanoparticles (GNPs) in mimicking oxidoreductase, helicase, phosphatase were introduced. Moreover, the effect of surface properties and modifications on each catalytic activity was thoroughly discussed. The application of GNZs in biosensing and bioassay was classified in five categories based on the combination of the enzyme like activities and enhancing/inhibition of the catalytic activities in presence of the target analyte/s that is realized by proper surface modification engineering. These categories include catalytic activity enhancer, reversible catalytic activity inhibitor, binding selectivity enhancer, agglomeration base, and multienzyme like activity, which are explained and exemplified in this review. It also gives examples of those modifications that enable the application of GNZs for in vivo biosensing and bioassays.

New insight into the biocompatibility/toxicity of graphene oxides and their reduced forms on Chlamydomonas reinhardtii.

Graphene oxides (GOs) and their reduced forms are often discussed both positively and negatively due to the lack of information about their chemistry and structure. This study utilized GOs with two sheet sizes that were further reduced by two reducing agents (sodium borohydride and hydrazine) to obtain two different degrees of reduction. The synthesized nanomaterials were characterized using scanning electron microscopy (SEM), atomic force microscopy (AFM), X-ray photoelectron spectroscopy (XPS), elemental analysis (EA), Fourier transform infrared (FTIR) spectroscopy, and Raman spectroscopy (RA) to understand their chemistry and structure. The second focus of our investigation included in vitro testing of the biocompatibility/toxicity of these materials on a model organism, the freshwater microalga Chlamydomonas reinhardtii. The effects were studied on the basis of biological endpoints complemented by biomass investigation (FTIR spectroscopy, EA, and atomic absorption spectrometry (AAS)). The results showed that the biocompatibility/toxicity of GOs is dependent on their chemistry and structure and that it is impossible to generalize the toxicity of graphene-based nanomaterials.

Not so dangerous? PET microplastics toxicity on freshwater microalgae and cyanobacteria.

Microalgae and cyanobacteria are among the most important primary producers and are responsible for the production of 50-80% of the oxygen on Earth. They can be significantly affected by plastic pollution, as the vast majority of plastic waste ends up in rivers and then the oceans. This research focuses on green microalgae Chlorella vulgaris (C. vulgaris), Chlamydomonas reinhardtii (C. reinhardtii), filamentous cyanobacterium Limnospira (Arthrospira) maxima (L.(A.) maxima) and how they are affected by environmentally relevant PET-MPs (polyethylene-terephtalate microplastics). Manufactured PET-MPs have asymmetric shape, size between 3 and 7 µm and were used in concentrations ranging from 5 mg/L to 80 mg/L. The highest inhibitory rate of growth was found in C. reinhardtii (-24%). Concentration-dependent changes in chlorophyll a composition were found in C. vulgaris and C. reinhardtii, not in L. (A.) maxima. Furthermore, cell damage was detected in all three organisms by CRYO-SEM (shriveling, cell wall disruption), but the cyanobacterium was the least damaged. A PET-fingerprint was detected on the surface of all tested organisms using FTIR, indicating the adherence of PET-MPs. The highest rate of PET-MPs adsorption was detected in L. (A.) maxima. Specifically, characteristic spectra were observed at ∼721, 850, 1100, 1275, 1342, and 1715 cm-1 which are specific for functional groups of PET-MPs. Nitrogen and carbon content significantly increased in L. (A.) maxima under exposure to 80 mg/L due to the PET-MPs adherence and mechanical stress. In all three tested organisms, weak exposure-related ROS generation was detected. In general, cyanobacteria seem to be more resistant to the effects of MPs. However, organisms in the aquatic environment are exposed to MPs over a longer time scale, so it is important to use the present findings for further longer-term experiments on environmentally relevant organisms.

The combined effect of graphene oxide and elemental nano-sulfur on soil biological properties and lettuce plant biomass.

The impact of graphene oxide (GO) nanocarbon on soil properties is mixed, with both negative and positive effects. Although it decreases the viability of some microbes, there are few studies on how its single amendment to soil or in combination with nanosized sulfur benefits soil microorganisms and nutrient transformation. Therefore, an eight-week pot experiment was carried out under controlled conditions (growth chamber with artificial light) in soil seeded with lettuce (Lactuca sativa) and amended with GO or nano-sulfur on their own or their several combinations. The following variants were tested: (I) Control, (II) GO, (III) Low nano-S + GO, (IV) High nano-S + GO, (V) Low nano-S, (VI) High nano-S. Results revealed no significant differences in soil pH, dry plant aboveground, and root biomass among all five amended variants and the control group. The greatest positive effect on soil respiration was observed when GO was used alone, and this effect remained significant even when it was combined with high nano-S. Low nano-S plus a GO dose negatively affected some of the soil respiration types: NAG_SIR, Tre_SIR, Ala_SIR, and Arg_SIR. Single GO application was found to enhance arylsulfatase activity, while the combination of high nano-S and GO not only enhanced arylsulfatase but also urease and phosphatase activity in the soil. The elemental nano-S probably counteracted the GO-mediated effect on organic carbon oxidation. We partially proved the hypothesis that GO-enhanced nano-S oxidation increases phosphatase activity.