Cross sectional study on exposure to BPA and its analogues and semen parameters in Czech men.

Exposure to bisphenols has been found to have adverse effects on male reproductive function in animals. Human exposure to bisphenols is widespread. Bisphenol A (BPA) and its analogues, including bisphenol S (BPS), bisphenol F (BPF), and bisphenol AF (BPAF) are utilized in various consumer products such as food contact materials and dental resins. The effects of these compounds on male fertility and spermatogenesis are unclear and findings from human studies are inconsistent. In this cross-sectional study, we evaluated the influence of BPA, BPS, BPF, BPAF (BPs) measured in semen on number of spermatozoa, total motility, progressive motility, morphology, and DNA fragmentation. We also examined the association of bisphenols (BPs) exposure with patients' occupation. A total of 358 patients aged 17-62 years with BMI 18-42 were included in the study from 2019 to 2021. BPs were extracted using solvent extraction followed by preconcentration step and determined by high-performance liquid chromatography and tandem mass spectrometry (LC/MSMS). Bisphenols were detected in 343 from 349 analysed samples (98.3% of all the samples). In 6 samples, the concentration of all BPs was under the limit of detection and in 20 samples under the limit of quantification. We did not find a statistically significant relationship between occupation and BPs. However, we observed significant correlations between the concentration of BPA and a lower motility and normal morphology. For BPS, a significant correlation with a lower ejaculate volume and a lower total sperm count was found. BPF and BPAF were detected only in 14.3% and 23.9% of samples, respectively. For BPF and BPAF, no significant correlations with spermiogram parameters were observed. Our results show that BPs are widespread in the male population (more than 90% of analysed samples), independently of an occupation and in case of BPA and BPS having a negative impact on spermiogram parameters.

Elevated erythroferrone distinguishes erythrocytosis with inherited defects in oxygen-sensing pathway from primary familial and congenital polycythaemia.

Congenital erythrocytoses represent a heterogenous group of rare defects of erythropoiesis characterized by elevated erythrocyte mass. We performed molecular-genetic analysis of 21 Czech patients with congenital erythrocytosis and assessed the mutual link between chronic erythrocyte overproduction and iron homoeostasis. Causative mutations in erythropoietin receptor (EPOR), hypoxia-inducible factor 2 alpha (HIF2A) or Von Hippel-Lindau (VHL) genes were detected in nine patients, including a novel p.A421Cfs*4 EPOR and a homozygous intronic c.340+770T>C VHL mutation. The association and possible cooperation of five identified missense germline EPOR or Janus kinase 2 (JAK2) variants with other genetic/non-genetic factors in erythrocytosis manifestation may involve variants of Piezo-type mechanosensitive ion channel component 1 (PIEZO1) or Ten-eleven translocation 2 (TET2), but this requires further research. In two families, hepcidin levels appeared to prevent or promote phenotypic expression of the disease. No major contribution of heterozygous haemochromatosis gene (HFE) mutations to the erythrocytic phenotype or hepcidin levels was observed in our cohort. VHL- and HIF2A-mutant erythrocytosis showed increased erythroferrone and suppressed hepcidin, whereas no overproduction of erythroferrone was detected in other patients regardless of molecular defect, age or therapy. Understanding the interplay between iron metabolism and erythropoiesis in different subgroups of congenital erythrocytosis may improve current treatment options.

Current possibilities and risks of donating some tissues, cells, and organs from living donors.

Transfusion, transplantation, and regenerative medicine are rapidly developing fields. The authors of the text want to inform about upcoming legislative changes at the EU level and briefly describe and compare the difficulty of some donation procedures from the point of view of a living donor, as well as their risks, including psychosocial risks. The study is based on a qualitative expert investigation. Comparing the complexity of procedures from the perspective of donors is important, for example, for setting compensations. The tables show that the current compensations are disproportionate.

The exposure of Czech firefighters to perfluoroalkyl substances and polycyclic aromatic hydrocarbons: CELSPAC - FIREexpo case-control human biomonitoring study.

The CELSPAC - FIREexpo biomonitoring study investigates the long-term effects of chemical exposure on firefighters' wellness and fitness. It aims to provide science-based measures to minimize the health risks of the firefighting occupation. Here, we present the study design, cohort profile, and first results with respect to internal per- and polyfluoroalkyl substances (PFAS) and polycyclic aromatic hydrocarbons (PAH) levels in study participants. Participants (n = 166) were divided into three subcohorts: i) newly recruited firefighters, ii) professional firefighters with several years' experience, and iii) the control group. Participants underwent physical performance tests, provided information on their lifestyle and diet, and urine and blood samples 1-4 times within an 11-week period. 12 serum PFAS and 10 urinary hydroxylated PAH (OH-PAH) levels were determined using HPLC-MS/MS and compared between subcohorts and samplings. The association of internal exposure with reported lifestyles and occupational factors was investigated using Spearman's correlation, principal component analysis, and multivariate regression analysis. ΣPFAS levels in firefighters were significantly higher than in the control group and were mostly associated with the length of firefighting career, age, blood donation, and population size. 10.9 % and 7.6 % of measurements exceeded the HBM-I or HBM-II value for PFOS and PFOA, respectively. Urinary ΣPAH levels increased significantly after training with burning wooden pallets, but none of them exceeded the no observed genotoxic effect level. Firefighters' occupational exposure, its sources, and pathways, need to be systematically monitored and investigated on a long-term and individual basis. The CELSPAC - FIREexpo study helps to clarify the degree of occupational exposure to the given compounds and the subsequent risks to firefighters.

Effect of Bisphenol S on testicular tissue after low-dose lactation exposure.

Exposure to endocrine disruptors such as bisphenols, can lead to and be the explanation for idiopathic infertility. In our study, we assessed the effect of exposure to bisphenol S (BPS) via breast milk on the testicular tissue health of adult male mice. Lactating dams were exposed to BPS through drinking water (0.216 ng g bw/day and 21.6 ng g bw/day) from post-natal day 0-15. Although there was no significant difference in testicular histopathology between the control and experimental groups, we observed an increase in the number of tight and gap junctions in the blood-testis barrier (BTB) of adult mice after lactation BPS exposure. Moreover, there was an increase in oxidative stress markers in adult testicular tissue of mice exposed via breast milk. Our lactation model indicates that breast milk is a route of exposure to an endocrine disruptor that can be responsible for idiopathic male infertility through the damage of the BTB and weakening of oxidative stress resistance in adulthood.

The Bisphenols Found in the Ejaculate of Men Does Not Pass through the Testes.

Exposure to bisphenols is related to negative effects on male reproduction. The bisphenols exposure is associated with several modes of action including negative impact on the blood-testis barrier (BTB) in testes or direct effect on spermatozoa. Bisphenols have been detected in human seminal plasma, but the possible mechanism of seminal transfer of bisphenols is not clear. Some authors consider the transfer through the blood-testis barrier to be crucial. Therefore, in this work, we compared normozoospermic men and men after vasectomy who have interrupted vas deferens and their ejaculate does not contain testicular products. We measured the concentration of bisphenol A (BPA), bisphenol S (BPS) and bisphenol F (BPF) in the urine and seminal plasma of these men using liquid chromatography tandem mass spectrometry (LC/MSMS). We found that the ratio of urinary and seminal plasma content of bisphenols did not differ in normozoospermic men or men after vasectomy. From the obtained data, it can be concluded that the pathways of transport of bisphenols into seminal plasma are not primarily through the testicular tissue, but this pathway is applied similarly to other routes of transmission by a corresponding ejaculate volume ratio. To a much greater extent than through testicular tissue, bisphenols enter the seminal plasma mainly as part of the secretions of the accessory glands.

Overview of the Mechanisms of Action of Selected Bisphenols and Perfluoroalkyl Chemicals on the Male Reproductive Axes.

Male fertility has been deteriorating worldwide for considerable time, with the greatest deterioration recorded mainly in the United States, Europe countries, and Australia. That is, especially in countries where an abundance of chemicals called endocrine disruptors has repeatedly been reported, both in the environment and in human matrices. Human exposure to persistent and non-persistent chemicals is ubiquitous and associated with endocrine-disrupting effects. This group of endocrine disrupting chemicals (EDC) can act as agonists or antagonists of hormone receptors and can thus significantly affect a number of physiological processes. It can even negatively affect human reproduction with an impact on the development of gonads and gametogenesis, fertilization, and the subsequent development of embryos. The negative effects of endocrine disruptors on sperm gametogenesis and male fertility in general have been investigated and repeatedly demonstrated in experimental and epidemiological studies. Male reproduction is affected by endocrine disruptors via their effect on testicular development, impact on estrogen and androgen receptors, potential epigenetic effect, production of reactive oxygen species or direct effect on spermatozoa and other cells of testicular tissue. Emerging scientific evidence suggests that the increasing incidence of male infertility is associated with the exposure to persistent and non-persistent endocrine-disrupting chemicals such as bisphenols and perfluoroalkyl chemicals (PFAS). These chemicals may impact men's fertility through various mechanisms. This study provides an overview of the mechanisms of action common to persistent (PFAS) and nonpersistent (bisphenols) EDC on male fertility.

The role of new technologies in myeloproliferative neoplasms: Application of next-generation sequencing in myelofibrosis.

INTRODUCTION: Driver mutations in Philadelphia chromosome-negative myeloproliferative neoplasms are well known. In the past, whole-genome sequencing identified nondriver mutations in other genes, potentially contributing to evolution of malignant clones. METHODS: Next-generation sequencing was used to assess the presence of any mutations in 14 candidate genes at the point of diagnosis and the resultant impact on the clinical course of the disease. RESULTS: The study analysed 63 patients with myelofibrosis (MF). Nondriver mutations were detected in 44% of them. The most frequently affected genes were ASXL1 (27%), TET2 (11%) and SF3B1 (6%). The frequency of such mutations was highest in primary MF (59%) and lowest in the prefibrotic phase of primary MF (21%). Patients with prognostically unfavourable sequence variants in genes had significantly worse overall survival (53 vs 71 months; HR = 2.77; 95% CI 1.17-6.56; P = .017). CONCLUSION: In our study, multivariate analysis proved DIPSS to be the only significant factor to predict patient survival. DIPSS contains all of the important clinical and laboratory factors except genetic changes. Stratification of patients according to DIPSS is still beneficial although there are newer and improved scoring systems like GIPSS or MIPSS70. Assessing subclonal mutations in candidate genes during diagnosis may aid in the identification of high-risk MF patients and is therefore relevant for making a prediction for overall survival more accurate.

In vitro intestinal toxicity of commercially available spray disinfectant products advertised to contain colloidal silver.

The use of colloidal silver-containing products as dietary supplements, immune boosters and surface disinfectants has increased in recent years which has elevated the potential for human exposure to silver nanoparticles and ions. Product mislabeling and long-term use of these products may put consumers at risk for adverse health outcomes including argyria. This study assessed several physical and chemical characteristics of five commercial products as well as their cytotoxicity using a rat intestinal epithelial cell (IEC-6) model. Concentrations of silver were determined for both the soluble and particulate fractions of the products. Primary particle size distribution and elemental composition were determined by transmission electron microscopy (TEM) and energy-dispersive X-ray spectroscopy (EDS), respectively. Hydrodynamic diameters were measured using nanoparticle tracking analysis (NTA) and dynamic light scattering (DLS). The effect of gastrointestinal (GI) simulation on the colloidal silver products was determined using two systems. First, physical and chemical changes of the silver nanoparticles in these products was assessed after exposure to Synthetic Stomach Fluid (SSF) resulting in particle agglomeration, and the appearance of AgCl on the surfaces and between particles. IEC-6 cells were exposed for 24 h to dilutions of the products and assessed for cell viability. The products were also treated with a three-stage simulated GI system (stomach and intestinal fluids) prior to exposure of the IEC-6 cells to the isolated silver nanoparticles. Cell viability was affected by each of the consumer products. Based on the silver nitrate and commercial silver nanoparticle dose response, the cytotoxicity for each of the colloidal silver products was attributed to the particulate silver, soluble silver or non­silver matrix constituents.

Transformation of Silver Nanoparticle Consumer Products during Simulated Usage and Disposal.

Twenty-two silver nanoparticle (AgNP) consumer products (CPs) were analyzed with respect to their silver speciation. Three CPs and three lab-synthesized particles were selected to simulate environmental fate and transport by simulating their intended usage and disposal methods. Since many of these products are meant for ingestion, we simulated their usage by exposing them to human synthetic stomach fluid followed by exposure to wastewater sludge. We found that during the products individual exposure to wastewater sludge, the conversion rate of silver to AgCl and Ag2S was affected by both the amount of silver ion present and the properties of the AgNP. The rates of conversion of metallic silver to silver sulfide was heavily dependent on the particle size for the lab-synthesized particles, with 90 nm PVP-capped particles reacting to a much lesser extent than the 15 nm PVP-capped or the citrate-capped particles. We observed similar sulfidation rates on two of the tested CPs with the 15 nm lab-synthesized particles despite containing silver nanoparticles >5 times larger, indicating the presence of other influencing factors. Pre-treatment with synthetic stomach fluid modified the rates of Ag2S formation. Due to the variable composition of CPs and the conditions they are exposed to between manufacture, sale, use, and disposal, their final composition may be somewhat unpredictable in the environment. In the present study, we have achieved a more accurate approximation of the expected interactions between silver nanoparticle-containing CPs and environmental media by utilizing real CPs and evaluating them with solid phase and aqueous phase analytical techniques.

In vitro intestinal toxicity of copper oxide nanoparticles in rat and human cell models.

Human oral exposure to copper oxide nanoparticles (NPs) may occur following ingestion, hand-to-mouth activity, or mucociliary transport following inhalation. This study assessed the cytotoxicity of Cupric (II) oxide (CuO) and Cu2O-polyvinylpyrrolidone (PVP) coated NPs and copper ions in rat (intestine epithelial cells; IEC-6) and human intestinal cells, two- and three-dimensional models, respectively. The effect of pretreatment of CuO NPs with simulated gastrointestinal (GI) fluids on IEC-6 cell cytotoxicity was also investigated. Both dose- and time-dependent decreases in viability of rat and human cells with CuO and Cu2O-PVP NPs and Cu2+ ions was observed. In the rat cells, CuO NPs had greater cytotoxicity. The rat cells were also more sensitive to CuO NPs than the human cells. Concentrations of H2O2 and glutathione increased and decreased, respectively, in IEC-6 cells after a 4-h exposure to CuO NPs, suggesting the formation of reactive oxygen species (ROS). These ROS may have damaged the mitochondrial membrane of the IEC-6 cells causing a depolarization, as a dose-related loss of a fluorescent mitochondrial marker was observed following a 4-h exposure to CuO NPs. Dissolution studies showed that Cu2O-PVP NPs formed soluble Cu whereas CuO NPs essentially remained intact. For GI fluid-treated CuO NPs, there was a slight increase in cytotoxicity at low doses relative to non-treated NPs. In summary, copper oxide NPs were cytotoxic to rat and human intestinal cells in a dose- and time-dependent manner. The data suggests Cu2O-PVP NPs are toxic due to their dissolution to Cu ions, whereas CuO NPs have inherent cytotoxicity, without dissolving to form Cu ions.

Determination of Silver Nanoparticle Dose in vitro.

An important issue for interpreting in vitro nanomaterial testing is quantifying the dose delivered to target cells. Considerations include the concentration added to the culture, the proportion of the applied dose that interacts with the target cells, and the amount that is eventually absorbed by the target cells. Rapid and efficient techniques are needed to determine delivered doses. Previously, we demonstrated that TiO2 and silver nanoparticles (AgNP) were absorbed by cells in a dose dependent manner between 1 µg/ml and 30 µg/ml and were detected in cells by light scatter using a flow cytometer. Here, we compare four potential indices of the dose of AgNP to cells, including: inductively coupled plasma - mass spectrometry (ICP-MS); flow cytometry side scatter (SSC); and amount of silver deposited to the cell layer as estimated with both an integrated Volumetric Centrifugation Method - In Vitro Sedimentation, Diffusion and Dosimetry Model (VCM-ISDD) and a Distorted Grid (DG) model. A retinal pigment epithelial cell line was exposed to 20 nm or 75 nm citrate-coated AgNP for 24 hr. The relationships between particle sizes and internalized doses varied according to the dose metric. Twenty-four hours after exposure, the cell layer contained a greater mass of silver when treated with 75 nm AgNP than with 20 nm AgNP. When the dose was expressed as the number of particles or as the total surface area of absorbed particles, however, the reverse was true; the dose to the cells was higher after exposure to 20 than 75 nm AgNP. Flow cytometry SSC increased with dose for both sizes of AgNP, and was correlated with Ag in cells measured by ICP-MS. The rate of SSC increase was greater for 75 than for 20 nm AgNP, suggesting it could be used as an indicator of cellular dose after accounting for particle size and composition. Silver was detected by ICP-MS in re-suspended supernates of the isolated cell layer suggested that not all the silver deposited to the cell layer was absorbed by the cells. Both the VCM-ISDD and DG models estimated the proportion of Ag deposited to the cellular layer, which in both cases was greater than the amount of silver in the cells measured by ICP-MS. Modeled deposition more closely compared to the total Ag deposition by ICP-MS, i.e. mass of silver in the cells plus the resuspended, unabsorbed Ag from the cell layer. ICP-MS indicated the mass of silver in cells from AgNP treatment, but not whether the Ag was in the form of particles or dissolved ions. Deposition models predicted the amount of AgNP deposited to the cell layer, but not cellular uptake. Flow cytometry SSC was correlated to cellular uptake of particle-form AgNP and could be calibrated against ICP-MS to indicate mass of cellular uptake. Therefore, a combination of approaches may be required to accurately understand cellular dosimetry of in vitro nanotoxicology experiments. In summary, cellular dosimetry is an important consideration for nanotoxicology experiments, and not necessarily related to the applied dose.

Characterization of engineered nanoparticles in commercially available spray disinfectant products advertised to contain colloidal silver.

Given the potential for human exposure to silver nanoparticles from spray disinfectants and dietary supplements, we characterized the silver-containing nanoparticles in 22 commercial products that advertised the use of silver or colloidal silver as the active ingredient. Characterization parameters included: total silver, fractionated silver (particulate and dissolved), primary particle size distribution, hydrodynamic diameter, particle number, and plasmon resonance absorbance. A high degree of variability between claimed and measured values for total silver was observed. Only 7 of the products showed total silver concentrations within 20% of their nominally reported values. In addition, significant variations in the relative percentages of particulate vs. soluble silver were also measured in many of these products reporting to be colloidal. Primary silver particle size distributions by transmission electron microscopy (TEM) showed two populations of particles - smaller particles (<5nm) and larger particles between 20 and 40nm. Hydrodynamic diameter measurements using nanoparticle tracking analysis (NTA) correlated well with TEM analysis for the larger particles. Z-average (Z-Avg) values measured using dynamic light scattering (DLS); however, were typically larger than both NTA or TEM particle diameters. Plasmon resonance absorbance signatures (peak absorbance at around 400nm indicative of metallic silver nanoparticles) were only noted in 4 of the 9 yellow-brown colored suspensions. Although the total silver concentrations were variable among products, ranging from 0.54mg/L to 960mg/L, silver containing nanoparticles were identified in all of the product suspensions by TEM.

Transformations of Nanoenabled Copper Formulations Govern Release, Antifungal Effectiveness, and Sustainability throughout the Wood Protection Lifecycle.

Here we compare the standard European benchmark of wood treatment by molecularly dissolved copper amine (Cu-amine), also referred to as aqueous copper amine (ACA), against two nanoenabled formulations: copper(II)oxide nanoparticles (CuO NPs) in an acrylic paint to concentrate Cu as a barrier on the wood surface, and a suspension of micronized basic copper carbonate (CuCO3·Cu(OH)2) for wood pressure treatment. After characterizing the properties of the (nano)materials and their formulations, we assessed their effects in vitro against three fungal species: Coniophora puteana, Gloeophyllum trabeum, and Trametes versicolor, finding them to be mediated only partially by ionic transformation. To assess the use phase, we quantify both release rate and form. Cu leaching rates for the two types of impregnated wood (conventional and nanoenabled) are not significantly different at 172 ± 6 mg/m2, with Cu being released predominantly in ionic form. Various simulations of outdoor aging with release sampling by runoff, during condensation, by different levels of mechanical shear, all resulted in comparable form and rate of release from the nanoenabled or the molecular impregnated woods. Because of dissolving transformations, the nanoenabled impregnation does not introduce additional concern over and above that associated with the traditional impregnation. In contrast, Cu released from wood coated with the CuO acrylate contained particles, but the rate was at least 100-fold lower. In the same ranking, the effectiveness to protect against the wood-decaying basidiomycete Coniophora puteana was significant with both impregnation technologies but remained insignificant for untreated wood and wood coated by the acrylic CuO. Accordingly, a lifecycle-based sustainability analysis indicates that the CuO acrylic coating is less sustainable than the technological alternatives, and should not be developed into a commercial product.

Nanoscale Coloristic Pigments: Upper Limits on Releases from Pigmented Plastic during Environmental Aging, In Food Contact, and by Leaching.

The life cycle of nanoscale pigments in plastics may cause environmental or human exposure by various release scenarios. We investigated spontaneous and induced release with mechanical stress during/after simulated sunlight and rain degradation of polyethylene (PE) with organic and inorganic pigments. Additionally, primary leaching in food contact and secondary leaching from nanocomposite fragments with an increased surface into environmental media was examined. Standardized protocols/methods for release sampling, detection, and characterization of release rate and form were applied: Transformation of the bulk material was analyzed by Scanning Electron Microscopy (SEM), X-ray-tomography and Fourier-Transform Infrared spectroscopy (FTIR); releases were quantified by Inductively Coupled Plasma Mass Spectrometry (ICP-MS), single-particle-ICP-MS (sp-ICP-MS), Transmission Electron Microscopy (TEM), Analytical Ultracentrifugation (AUC), and UV/Vis spectroscopy. In all scenarios, the detectable particulate releases were attributed primarily to contaminations from handling and machining of the plastics, and were not identified with the pigments, although the contamination of 4 mg/kg (Fe) was dwarfed by the intentional content of 5800 mg/kg (Fe as Fe2O3 pigment). We observed modulations (which were at least partially preventable by UV stabilizers) when comparing as-produced and aged nanocomposites, but no significant increase of releases. Release of pigments was negligible within the experimental error for all investigated scenarios, with upper limits of 10 mg/m2 or 1600 particles/mL. This is the first holistic confirmation that pigment nanomaterials remain strongly contained in a plastic that has low diffusion and high persistence such as the polyolefin High Density Polyethylene (HDPE).

Meeting the Needs for Released Nanomaterials Required for Further Testing-The SUN Approach.

The analysis of the potential risks of engineered nanomaterials (ENM) has so far been almost exclusively focused on the pristine, as-produced particles. However, when considering a life-cycle perspective, it is clear that ENM released from genuine products during manufacturing, use, and disposal is far more relevant. Research on the release of materials from nanoproducts is growing and the next necessary step is to investigate the behavior and effects of these released materials in the environment and on humans. Therefore, sufficient amounts of released materials need to be available for further testing. In addition, ENM-free reference materials are needed since many processes not only release ENM but also nanosized fragments from the ENM-containing matrix that may interfere with further tests. The SUN consortium (Project on "Sustainable Nanotechnologies", EU seventh Framework funding) uses methods to characterize and quantify nanomaterials released from composite samples that are exposed to environmental stressors. Here we describe an approach to provide materials in hundreds of gram quantities mimicking actual released materials from coatings and polymer nanocomposites by producing what is called "fragmented products" (FP). These FP can further be exposed to environmental conditions (e.g., humidity, light) to produce "weathered fragmented products" (WFP) or can be subjected to a further size fractionation to isolate "sieved fragmented products" (SFP) that are representative for inhalation studies. In this perspective we describe the approach, and the used methods to obtain released materials in amounts large enough to be suitable for further fate and (eco)toxicity testing. We present a case study (nanoparticulate organic pigment in polypropylene) to show exemplarily the procedures used to produce the FP. We present some characterization data of the FP and discuss critically the further potential and the usefulness of the approach we developed.

Detection of Engineered Copper Nanoparticles in Soil Using Single Particle ICP-MS.

Regulatory efforts rely on nanometrology for the development and implementation of laws regarding the incorporation of engineered nanomaterials (ENMs) into industrial and consumer products. Copper is currently one of the most common metals used in the constantly developing and expanding sector of nanotechnology. The use of copper nanoparticles in products, such as agricultural biocides, cosmetics and paints, is increasing. Copper based ENMs will eventually be released to the environment through the use and disposal of nano-enabled products, however, the detection of copper ENMs in environmental samples is a challenging task. Single particle inductively coupled plasma mass spectroscopy (spICP-MS) has been suggested as a powerful tool for routine nanometrology efforts. In this work, we apply a spICP-MS method for the detection of engineered copper nanomaterials in colloidal extracts from natural soil samples. Overall, copper nanoparticles were successfully detected in the soil colloidal extracts and the importance of dwell time, background removal, and sample dilution for method optimization and recovery maximization is highlighted.

Novel arylazopyrazole inhibitors of cyclin-dependent kinases.

Here, we describe new 4-arylazo-3,5-diamino-1H-pyrazole derivatives developed from CAN508, one of the first inhibitors to show preference for transcriptional regulator cyclin-dependent kinase 9. By substituting nitrogen in the pyrazole ring and employing a heteroatom in the 4-aryl ring, we obtained more potent derivatives differing in their CDK-selectivity profiles. The antiproliferative and anti-CDK kinase activities of the novel arylazopyrazoles were examined. The cellular effect of compound IVc was studied on MCF-7 cells synchronized by various methods and compared with other selective CDK inhibitors. The results demonstrated that IVc shows a preference for CDK4 and CDK1. In contrast to cytostatic effects induced by IVc in MCF-7 and K562 cells, we observed apoptotic activities in the RPMI-8226 cell line, which were confirmed by detecting active caspases by different biochemical assays.

In-house validation of a method for determination of silver nanoparticles in chicken meat based on asymmetric flow field-flow fractionation and inductively coupled plasma mass spectrometric detection.

Nanomaterials are increasingly used in food production and packaging, and validated methods for detection of nanoparticles (NPs) in foodstuffs need to be developed both for regulatory purposes and product development. Asymmetric flow field-flow fractionation with inductively coupled plasma mass spectrometric detection (AF(4)-ICP-MS) was applied for quantitative analysis of silver nanoparticles (AgNPs) in a chicken meat matrix following enzymatic sample preparation. For the first time an analytical validation of nanoparticle detection in a food matrix by AF(4)-ICP-MS has been carried out and the results showed repeatable and intermediately reproducible determination of AgNP mass fraction and size. The findings demonstrated the potential of AF(4)-ICP-MS for quantitative analysis of NPs in complex food matrices for use in food monitoring and control. The accurate determination of AgNP size distribution remained challenging due to the lack of certified size standards.

Arylazopyrazole AAP1742 inhibits CDKs and induces apoptosis in multiple myeloma cells via Mcl-1 downregulation.

Inhibitors of cyclin-dependent kinases 9 have been developed as potential anticancer drugs for the treatment of multiple myeloma. We have previously prepared a library of arylazo-3,5-diaminopyrazole inhibitors of CDKs. Here, we describe a novel member, AAP1742 (CDK9 inhibition with IC(50) = 0.28 µm), that reduces the viability of multiple myeloma cell lines in low micromolar concentrations. Consistent with inhibition of CDK9, AAP1742 decreases the phosphorylation of RNA polymerase II and inhibits mRNA synthesis of anti-apoptotic proteins Mcl-1, Bcl-2, and XIAP, followed by apoptosis in the RPMI-8226 cell line in a dose- and a time-dependent manner. These results are consistent with the biochemical profile of AAP1742 and further suggest cellular inhibition of CDK9 as a possible target for anticancer drugs.