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Katarína Poláková,1 Aurélie Fauger,2 Michèle Sayag,2 Eric Jourdan2 1Saint Elisabeth´s Oncological Institute, Bratislava, Slovakia; 2Laboratoire Bioderma, Lyon, France Background: Acne vulgaris is an inflammatory disorder of the pilosebaceous unit. Aim: To confirm that BGM (bakuchiol, Ginkgo biloba extract, and mannitol) complex increases the established clinical efficacy of adapalene 0.1% gel in patients with acne. Methods: A clinical trial was conducted in acne patients. A total of 111 subjects received adapalene 0.1% gel and BGM complex or vehicle cream for 2 months. Assessments comprised Investigator Global Assessment (IGA), global efficacy, seborrhea intensity, inflammatory and non-inflammatory lesions, and subject perception, as well as overall safety and local tolerance and quality of life. Results: At the end of the trial, inflammatory and non-inflammatory lesions, IGA, global efficacy, and seborrhea intensity had significantly improved in both treatment groups. Differences were statistically significant (P

The field of molecular biology aims to explain the fundamentals governing living processes at molecular level by identifying mechanisms that produce such processes. Central discoveries describing the innate systems of living nature helped derive molecular technologies that work in vitro. The application of these methods has not only improved understanding of molecular biology but has also helped to improve prevention, diagnosis and treatment of human diseases. This review focuses on the key technologies of molecular genetics and their development over the past 25 years. The application of these methods in the diagnosis and monitoring of haemato(oncolo)gical patients that have helped significantly to improve prognosis and treatment approaches, is described. [ABSTRACT FROM AUTHOR]

Human leukocyte antigen G (HLA-G) is a non-classical HLA class I protein with various immunosuppressive functions. Besides its profound effect to induce fetal tolerance, HLA-G has been also found to enhance graft acceptance. The aim of the study was to analyse the association between HLA-G 14 bp insertion/deletion polymorphism, soluble HLA-G level and kidney graft outcome in the Slovak population. We investigated 69 kidney transplant recipients aged 27–65 years. Out of this group, 37 recipients developed acute rejection, confirmed by biopsy, and 32 patients had stable allograft function. Plasma was obtained from recipients at 1 day before transplantation and analyzed by ELISA. Genotyping of HLA-G polymorphism was performed by PCR. Significantly higher pre-transplantation levels of sHLA-G were found in the group with stable allograft function in comparison to group with acute rejection (P = 0.0409). In the homozygous −14/−14 recipients with stable allograft function, significantly higher values of sHLA-G were determined in comparison to the recipients with acute rejection (P = 0.0052). The study revealed an association between 14 bp deletion polymorphism and soluble HLA-G level that is proportional to kidney graft acceptance. It is suggested that pre-transplantation levels of soluble HLA-G should be monitored as additional marker to predict kidney graft outcome.

Katerina Tomankova,1 Katerina Polakova,2 Klara Pizova,1 Svatopluk Binder,1 Marketa Havrdova,2 Mary Kolarova,2 Eva Kriegova,3 Jana Zapletalova,1 Lukas Malina,1 Jana Horakova,1 Jakub Malohlava,1 Argiris Kolokithas-Ntoukas,4 Aristides Bakandritsos,4 Hana Kolarova,1 Radek Zboril2 1Department of Medical Biophysics, Institute of Translation Medicine, Faculty of Medicine and Dentistry, 2Regional Centre of Advanced Technologies and Materials, Departments of Physical Chemistry and Experimental Physics, Faculty of Science, 3Department of Immunology, Faculty of Medicine and Dentistry, Palacky University, Olomouc, Czech Republic; 4Department of Materials Science, University of Patras, Patras, Greece Abstract: One of the promising strategies for improvement of cancer treatment is based on magnetic drug delivery systems, thus avoiding side effects of standard chemotherapies. Superparamagnetic iron oxide (SPIO) nanoparticles have ideal properties to become a targeted magnetic drug delivery contrast probes, named theranostics. We worked with SPIO condensed colloidal nanocrystal clusters (MagAlg) prepared through a new soft biomineralization route in the presence of alginate as the polymeric shell and loaded with doxorubicin (DOX). The aim of this work was to study the in vitro cytotoxicity of these new MagAlg–DOX systems on mouse fibroblast and breast carcinoma cell lines. For proper analysis and understanding of cell behavior after administration of MagAlg–DOX compared with free DOX, a complex set of in vitro tests, including production of reactive oxygen species, comet assay, cell cycle determination, gene expression, and cellular uptake, were utilized. It was found that the cytotoxic effect of MagAlg–DOX system is delayed compared to free DOX in both cell lines. This was attributed to the different mechanism of internalization of DOX and MagAlg–DOX into the cells, together with the fact that the drug is strongly bound on the drug nanocarriers. We discovered that nanoparticles can attenuate or even inhibit the effect of DOX, particularly in the tumor MCF7 cell line. This is a first comprehensive study on the cytotoxic effect of DOX-loaded SPIO compared with free DOX on healthy and cancer cell lines, as well as on the induced changes in gene expression. Keywords: DOX/SPIO nanocarriers, superparamagnetic iron oxide nanoparticles, doxorubicin, in vitro cytotoxicity

Josef Skopalik,1 Katerina Polakova,2 Marketa Havrdova,2 Ivan Justan,1 Massimiliano Magro,3 David Milde,2 Lucia Knopfova,4 Jan Smarda,4 Helena Polakova,1 Eva Gabrielova,5 Fabio Vianello,2,3 Jaroslav Michalek,1 Radek Zboril21Department of Pharmacology, Masaryk University, Brno, Czech Republic; 2Regional Centre of Advanced Technologies and Materials, Department of Physical Chemistry and Analytical Chemistry, Faculty of Science, Palacky University, Olomouc, Czech Republic; 3Department of Comparative Biomedicine and Food Science, University of Padua, Padova, Italy; 4Department of Experimental Biology, Faculty of Science, Masaryk University, Brno, Czech Republic; 5Department of Medical Chemistry and Biochemistry, Faculty of Medicine, Palacky University, Olomouc, Czech RepublicObjective: Cell therapies have emerged as a promising approach in medicine. The basis of each therapy is the injection of 1–100×106 cells with regenerative potential into some part of the body. Mesenchymal stromal cells (MSCs) are the most used cell type in the cell therapy nowadays, but no gold standard for the labeling of the MSCs for magnetic resonance imaging (MRI) is available yet. This work evaluates our newly synthesized uncoated superparamagnetic maghemite nanoparticles (surface-active maghemite nanoparticles – SAMNs) as an MRI contrast intracellular probe usable in a clinical 1.5 T MRI system.Methods: MSCs from rat and human donors were isolated, and then incubated at different concentrations (10–200 µg/mL) of SAMN maghemite nanoparticles for 48 hours. Viability, proliferation, and nanoparticle uptake efficiency were tested (using fluorescence microscopy, xCELLigence analysis, atomic absorption spectroscopy, and advanced microscopy techniques). Migration capacity, cluster of differentiation markers, effect of nanoparticles on long-term viability, contrast properties in MRI, and cocultivation of labeled cells with myocytes were also studied.Results: SAMNs do not affect MSC viability if the concentration does not exceed 100 µg ferumoxide/mL, and this concentration does not alter their cell phenotype and long-term proliferation profile. After 48 hours of incubation, MSCs labeled with SAMNs show more than double the amount of iron per cell compared to Resovist-labeled cells, which correlates well with the better contrast properties of the SAMN cell sample in T2-weighted MRI. SAMN-labeled MSCs display strong adherence and excellent elasticity in a beating myocyte culture for a minimum of 7 days.Conclusion: Detailed in vitro tests and phantom tests on ex vivo tissue show that the new SAMNs are efficient MRI contrast agent probes with exclusive intracellular uptake and high biological safety.Keywords: mesenchymal stromal cells, stem cell tracking, magnetic resonance imaging, superparamagnetic iron oxide nanoparticles, stem cell labelling

Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

Background: Although emergency medical service is focused on providing acute prehospital treatment, it is often used by terminally ill patients and their informal caregivers during the last days of patient's life. Little is known about why they decide to use the emergency medical services., Study Objective: The aim was to explore informal caregivers' motivation and decision-making process for calling emergency medical services for their terminally ill loved ones., Methods: This study used a qualitative design. Data were collected by semi-structured interviews with 31 relatives of 30 patients who used the emergency medical services. Data were analyzed with NVivo software by utilizing principles of thematic analysis., Results: Through the analysis, four distinct themes emerged: (1) limited availability of support from health care services; (2) insufficient planning of care; (3) decline in the health of the patient and (4) being lost and desperate., Conclusion: For informal caregivers, emergency medical services represented an important source of support while caring for their terminally ill loved ones due to the limited availability of other sources of help, including a lack of specialist palliative care providers. Additionally, informal caregivers had limited knowledge of the dying process and used emergency medical services for professional advice., (Copyright © 2024 Wolters Kluwer Health, Inc. All rights reserved.)

Clinical trials frequently include multiple end points that mature at different times. The initial report, typically based on the primary end point, may be published when key planned co-primary or secondary analyses are not yet available. Clinical Trial Updates provide an opportunity to disseminate additional results from studies, published in JCO or elsewhere, for which the primary end point has already been reported. The European Stop Kinase Inhibitors (EURO-SKI) study is the largest clinical trial for investigating the cessation of tyrosine kinase inhibitors (TKIs) in patients with chronic myeloid leukemia in stable deep molecular remission (DMR). Among 728 patients, 434 patients (61%; 95% CI, 57 to 64) remained in major molecular response (MMR) at 6 months and 309 patients of 678 (46%; 95% CI, 42 to 49) at 36 months. Duration of TKI treatment and DMR before TKI stop were confirmed as significant factors for the prediction of MMR loss at 6 months. In addition, the type of BCR::ABL1 transcript was identified as a prognostic factor. For late MMR losses after 6 months, TKI treatment duration, percentage of blasts in peripheral blood, and platelet count at diagnosis were significant factors in multivariate analysis. For the entire study period of 36 months, multiple logistic regression models confirmed duration of treatment, blasts, and transcript type as independent factors for MMR maintenance. In addition to the duration of treatment, transcript type as well as blasts in peripheral blood at diagnosis should be considered as important factors to predict treatment-free remission.

The phylogenetic and taxonomic affinities of lineages currently assigned to the non-monophyletic ciliate order Loxocephalida Jankowski (1980) within subclass Scuticociliatia Small (1967) remain unresolved. In the current study, we redescribe the morphology of the type species, Loxocephalus luridus Eberhard (1862) based on two Czech populations and include the first scanning and transmission electron microscopy images of the species. We provide the first 18S rRNA gene sequences for L. luridus and consider its phylogenetic position. Our results support the separation of Dexiotricha from Loxocephalus; however, the former genus is recovered as non-monophyletic. The monophyly of genus Dexiotricha and that of Loxocephalus + Dexiotricha is rejected. Loxocephalus luridus, together with Dexiotricha species, nests within a fully supported clade with Conchophthirus species, long presumed to belong to the Pleuronematida. Haptophrya is recovered as sister to this clade. The monophyly of the Astomatia Schewiakoff (1896) including Haptophrya is rejected. No clear morphologic synapomorphy is identified for the fully supported clade consisting of Haptophrya, Dexiotricha, Loxocephalus, and Conchophthirus., (© 2024 International Society of Protistologists.)

Ciliates are a diverse group of protists known for their ability to establish various partnerships and thrive in a wide variety of oxygen-depleted environments. Most anaerobic ciliates harbor methanogens, one of the few known archaea living intracellularly. These methanogens increase the metabolic efficiency of host fermentation via syntrophic use of host end-product in methanogenesis. Despite the ubiquity of these symbioses in anoxic habitats, patterns of symbiont specificity and fidelity are not well known. We surveyed two unrelated, commonly found groups of anaerobic ciliates, the Plagiopylea and Metopida, isolated from anoxic marine sediments. We sequenced host 18S rRNA and symbiont 16S rRNA marker genes as well as the symbiont internal transcribed spacer region from our cultured ciliates to identify hosts and their associated methanogenic symbionts. We found that marine ciliates from both of these co-occurring, divergent groups harbor closely related yet distinct intracellular archaea within the Methanocorpusculum genus. The symbionts appear to be stable at the host species level, but at higher taxonomic levels, there is evidence that symbiont replacements have occurred. Gaining insight into this unique association will deepen our understanding of the complex transmission modes of marine microbial symbionts, and the mutualistic microbial interactions occurring across domains of life., (© The Author(s) 2024. Published by Oxford University Press on behalf of the International Society for Microbial Ecology.)

GO is a 2D nanomaterial that has attracted attention in many industries in recent years, such as the chemical industry, electronics or medicine. Due to its unique properties such as strength, hydrophilicity and large specific surface area with the possibility of functionalization, GO is a particularly attractive material in biomedicine as a candidate for use in targeted drug delivery. In such a case, we need information on whether graphene oxide penetrates into cells and whether we are able to detect and monitor GO in these cells during and also after the treatment to evaluate possible degradation process of GO and its interaction within the cell compartements. This work introduces the Raman spectroscopy as label-free detection method showing the advantages of combining Raman spectroscopy with MCR (Multivariate Curve Resolution) analysis for advanced detection of GO in cervical cancer (HeLa) cells. Our synthesized GO is characterized firstly by AFM, SEM and Raman spectroscopy and then MCR-Raman spectroscopy is used to detect internalized GO in individual HeLa cells. Moreover, by using our methodology, distribution of GO as well as its chemical stability inside the cell for up to six months is investigated without using any additional labeling or tracing the GO. Thus, MCR-Raman spectroscopy may become a new analytical tool in preclinical and clinical applications of graphene-based nanotheranostics.

The research on anaerobic ciliates, to date, has mainly been focused on representatives of obligately anaerobic classes such as Armophorea or Plagiopylea. In this study, we focus on the anaerobic representatives of the subclass Scuticociliatia, members of the class Oligohymenophorea, which is mainly composed of aerobic ciliates. Until now, only a single anaerobic species, Cyclidium porcatum (here transferred to the genus Anaerocyclidium gen. nov.), has been described both molecularly and morphologically. Our broad sampling of anoxic sediments together with cultivation and single cell sequencing approaches have shown that scuticociliates are common and diversified in anoxic environments. Our results show that anaerobic scuticociliates represent a distinctive evolutionary lineage not closely related to the family Cyclidiidae (order Pleuronematida), as previously suggested. However, the phylogenetic position of the newly recognized lineage within the subclass Scuticociliatia remains unresolved. Based on molecular and morphological data, we establish the family Anaerocyclidiidae fam. nov. to accommodate members of this clade. We further provide detailed morphological descriptions and 18S rRNA gene sequences for six new Anaerocyclidium species and significantly broaden the described diversity of anaerobic scuticociliates., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier GmbH. All rights reserved.)

Coffee processing is a major contributor to the creation of food and product waste. Using coffee co-products can play an essential role in addressing environmental problems and issues with nutritionally unbalanced foods, population growth, and food-related diseases. This research aimed to determine the quality and sensory parameters (aw, pH, dry matter, TAC, TPC, fat, fatty acids profile, fiber, caffeine, chlorogenic acids, color, and sensory analysis) of different botanical origins of cascara (coffee husks) and silverskin (thin layer). The results of this study show that silverskin and cascara are a good source of TAC (1S 58.17 ± 1.28%, 2S 46.65 ± 1.20%, 1C 36.54 ± 1.84%, 2C 41.12 ± 2.11%). Cascara showed the presence of polyphenols (2C 49.135 g GAE·kg -1 ). Coffee co-products are good sources of fiber. Silverskin had higher values of caffeine than cascara. Palmitic, stearic, oleic, linoleic, and arachidic acids were the most represented acids in the samples. Given the obtained results, cascara can be considered "low-fat" (1C 4.240 g·kg -1 and 2C 5.4 g·kg -1 ). Based on the sensory evaluation, no sample reached the acceptable index value of 70%. Understanding the link between the character, identification properties, and composition of coffee co-products of different botanical origins can enable their application in the food industry.

Free-living anaerobic ciliates are of considerable interest from an ecological and an evolutionary standpoint. Extraordinary tentacle-bearing predatory lineages have evolved independently several times within the phylum Ciliophora, including two rarely encountered anaerobic litostomatean genera, Legendrea and Dactylochlamys . In this study, we significantly extend the morphological and phylogenetic characterization of these two poorly known groups of predatory ciliates. We provide the first phylogenetic analysis of the monotypic genus Dactylochlamys and the three valid species of Legendrea based on the 18S rRNA gene and ITS-28S rRNA gene sequences. Prior to this study, neither group had been studied using silver impregnation methods. We provide the first protargol-stained material and also a unique video material including documentation, for the first time, of the hunting and feeding behavior of a Legendrea species. We briefly discuss the identity of methanogenic archaeal and bacterial endosymbionts of both genera based on 16S rRNA gene sequences, and the importance of citizen science for ciliatology from a historical and contemporary perspective.

On-chip vascular microfluidic models provide a great tool to study aspects of cardiovascular diseases in vitro. To produce such models, polydimethylsiloxane (PDMS) has been the most widely used material. For biological applications, its hydrophobic surface has to be modified. The major approach has been plasma-based surface oxidation, which has been very challenging in the case of channels enclosed within a microfluidic chip. The preparation of the chip combined a 3D-printed mold with soft lithography and commonly available materials. We have introduced the high-frequency low-pressure air-plasma surface modification of seamless channels enclosed within a PDMS microfluidic chip. The plasma treatment modified the luminal surface more uniformly than in previous works. Such a setup enabled a higher degree of design freedom and a possibility of rapid prototyping. Further, plasma treatment in combination with collagen IV coating created a biomimetic surface for efficient adhesion of vascular endothelial cells as well as promoted long-term cell culture stability under flow. The cells within the channels were highly viable and showed physiological behavior, confirming the benefit of the presented surface modification.

Nowadays, there is an increased interest in coffee derivatives (green beans, roasted beans, and coffee by-products (Cascara and Silverskin)) due to their particular chemical composition. This study aimed to compare the content of dry matter, total fat, fatty acids, and fiber (ADF, NDF) of coffee by-products (Cascara and Silverskin) and coffee beans (green and roasted under different conditions). Coffee beans and their by-products were obtained from 100% C. arabica coffee cherries from Panama by dry process. The lowest concentrations of fat corresponded to Cascara 4.24 g·kg -1 and Silverskin 23.70 g·kg -1 , respectively. The major fatty acids detected in all samples were palmitic, stearic, oleic, and linoleic acids, the latter two being essential fatty acids. LDA showed that 89.01% of the variability between beans and by-products was explained by lignoceric, myristic, behenic, tricosanoic, arachidic, and heneicosanoic acids. Silverskin appeared to be a good source of lignoceric, myristic, and behenic acids and had a higher concentration of dietary fiber (314.95 g·kg -1 ) than Cascara (160.03 g·kg -1 ). Coffee by-products (Silverskin and Cascara) are low-fat products enriched in dietary fiber. Their incorporation, after adjustment, into the global diet may contribute to nutrition security, the sustainability of the coffee sector, and human health.

Fluorescent carbon dots (CDs) are potential tools for the labeling of cells with many advantages such as photostability, multicolor emission, small size, rapid uptake, biocompatibility, and easy preparation. Affinity towards organelles can be influenced by the surface properties of CDs which affect the interaction with the cell and cytoplasmic distribution. Organelle targeting by carbon dots is promising for anticancer treatment; thus, intracellular trafficking and cytotoxicity of cationic CDs was investigated. Based on our previous study, we used quaternized carbon dots (QCDs) for treatment and monitoring the behavior of two human cancer cell MCF-7 and HeLa lines. We found similarities between human cancer cells and mouse fibroblasts in the case of QCDs uptake. Time lapse microscopy of QCDs-labeled MCF-7 cells showed that cells are dying during the first two hours, faster at lower doses than at higher ones. QCDs at a concentration of 100 µg/mL entered into the nucleus before cellular death; however, at a dose of 200 µg/mL, blebbing of the cellular membrane occurred, with a subsequent penetration of QCDs into the nuclear area. In the case of HeLa cells, the dose-depended effect did not happen; however, the labeled cells were also dying in mitosis and genotoxicity occurred nearly at all doses. Moreover, contrasted intracellular compartments, probably mitochondria, were obvious after 24 h incubation with 100 µg/mL of QCDs. The levels of reactive oxygen species (ROS) slightly increased after 24 h, depending on the concentration, thus the genotoxicity was likely evoked by the nanomaterial. A decrease in viability did not reach IC 50 as the DNA damage was probably partly repaired in the prolonged G0/G1 phase of the cell cycle. Thus, the defects in the G2/M phase may have allowed a damaged cell to enter mitosis and undergo apoptosis. The anticancer effect in both cell lines was manifested mainly through genotoxicity.

Caffeine content is a crucial attribute of coffee. Its concentration and thus maximum cups of Coffea arabica from Africa, Asia, Central America, and South America from different altitudes of growing areas, altitude, and process using different post-harvest processing (dry, wet, and pulped natural). Our results suggest that geographical origin might affect the alkaloid concentration in C. arabica . The caffeine concentration pattern in green samples was as follows: Central America > South America > Asia > Africa. Altitude affected the concentrations, lowlands > midlands > highlands, however, not significantly. Given caffeine is thermostable, the medium roasting process did not affect the concentration of caffeine directly, but a small increase was observed. Scientific opinion on the safety of habitual caffeine consumption of up to 400 mg per day does not raise safety concerns for non-pregnant adults. A cup (7 g coffee in 120 mL of water) was used for recalculation. Results suggest that mostly highlands and midlands coffee from Africa reached levels of caffeine that might be consumed in more than 5.5 cups a day.

Objectives: Despite a number of studies on effectiveness of palliative care, there is a lack of complex updated review of the impact of in-hospital palliative care consult service. The objective is to update information on the impact of palliative care consult service in inpatient hospital setting., Methods: This study was a systematic literature review, following the standard protocols (Preferred Reporting Items for Systematic Reviews and Meta-Analyses, Joanna Briggs Institute tools) to ensure the transparent and robust review procedure. The effect of palliative care consult service was classified as being associated with improvement, no difference, deterioration or mixed results in specific outcomes. PubMed, Scopus, Academic Search Ultimate and SocINDEX were systematically searched up to February 2020. Studies were included if they focused on the impact of palliative care consult service caring for adult palliative care patients and their families in inpatient hospital setting., Results: After removing duplicates, 959 citations were screened of which 49 full-text articles were retained. A total of 28 different outcome variables were extracted. 18 of them showed positive effects within patient, family, staff and healthcare system domains. No difference was observed in patient survival and depression. Inconclusive results represented patient social support and staff satisfaction with care., Conclusions: Palliative care consult service has a number of positive effects for patients, families, staff and healthcare system. More research is needed on factors such as patient spiritual well-being, social support, performance, family understanding of patient diagnosis or staff stress., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2021. No commercial re-use. See rights and permissions. Published by BMJ.)

Recent experience with the COVID-19 pandemic should be a lesson learnt with respect to the effort we have to invest in the development of new strategies for the treatment of viral diseases, along with their cheap, easy, sensitive, and selective detection. Since we live in a globalized world where just hours can play a crucial role in the spread of a virus, its detection must be as quick as possible. Thanks to their chemical stability, photostability, and superior biocompatibility, carbon dots are a kind of nanomaterial showing great potential in both the detection of various virus strains and a broad-spectrum antiviral therapy. The biosensing and antiviral properties of carbon dots can be tuned by the selection of synthesis precursors as well as by easy post-synthetic functionalization. In this review, we will first summarize current options of virus detection utilizing carbon dots by either electrochemical or optical biosensing approaches. Secondly, we will cover and share the up-to-date knowledge of carbon dots' antiviral properties, which showed promising activity against various types of viruses including SARS-CoV-2. The mechanisms of their antiviral actions will be further adressed as well. Finally, we will discuss the advantages and distadvantages of the use of carbon dots in the tangled battle against viral infections in order to provide valuable informations for further research and development of new virus biosensors and antiviral therapeutics., (© 2021. The Author(s).)

Background: From 2012 to 2013, there was a mass methanol poisoning outbreak in the Czech Republic. Methanol metabolites can cause specific lesions in the basal ganglia, subcortical white matter, and optic nerve. However, long-term sequelae of methanol poisoning on cognitive functioning have not yet been explored. The current study aimed to delineate the cognitive changes observed in methanol poisoning survivors in the seven years since 2012., Methods: We conducted longitudinal research with repeated measurements in 2013, 2015, 2017 and 2019 to evaluate the development of cognitive changes after acute methanol poisoning. A complex neuropsychological battery consisted of tests of global cognitive performance, auditory and visual attention, executive functioning, learning and memory, working memory and language. Motor performance measures and depression scale were also included., Results: Repeated measures ANOVA of four measurements with post-hoc tests showed a significant decline in the Mini-Mental State Examination (p = 0.007); however, other parameters were not significantly decreasing. In comparison to normative values, the z-scores for each test measure, in the memory domain, in particular, ranged from 43 to 60 % of participants below 1.5 SD. Mild to severe depression levels from the onset of poisoning improved during the seven years, returning to normal in up to 27 % of participants., Conclusion: In the longitudinal perspective, methanol poisoning survivors manifest progressive global cognitive decline and overall persistent below-average cognitive performance with some improvements in the frequency of depressive symptoms., Competing Interests: Declaration of Competing Interest The authors report no declarations of interest., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Objective: The aim of this study was to analyse longitudinal development of prognostic awareness in advanced cancer patients and their families., Methods: This was a longitudinal cohort study, involving 134 adult cancer patients, 91 primary family caregivers and 21 treating oncologists. Key eligibility criterion for patients was life expectancy less than 1 year (estimated by their oncologists using the 12-month surprised question). Structured interviews, including tools to measure prognostic awareness, health information needs, and demographics were conducted face to face or via phone three times over 9 months. Forty-four patients completed all three phases of data collection., Results: Only 16% of patients reported accurate prognostic awareness, 58% being partially aware. Prognostic awareness of both patients and family caregivers remained stable over the course of the study, with only small non-significant changes. Gender, education, type of cancer, spirituality or health information needs were not associated with the level of prognostic awareness. Family caregivers reported more accurate prognostic awareness, which was not associated with patients' own prognostic awareness (agreement rate 59%, weighted kappa 0.348, CI = 0.185-0.510)., Conclusions: Prognostic awareness appears to be a stable concept over the course of the illness. Clinicians must focus on the initial patients' understanding of the disease and be able to communicate the prognostic information effectively from the early stages of patients' trajectory., (© 2021 John Wiley & Sons Ltd.)

Tea ( Camellia sinensis) is widely sought for beverages worldwide. Heavy metals are often the main aims of the survey of teas, given that the use of agricultural fertilization is very frequent. Some of these may affect the content of bioactive compounds. Therefore, in this study, we analyzed fermented and non-fermented teas of a single plant origin from Japan, Nepal, Korea, and China, and described mutual correlations and changes in the total antioxidant capacity (TAC), and the content of polyphenols (TPC), caffeine, and heavy metals in tea leaves, in relation to the origin and fermentation process. Using UV-VIS spectrophotometry and HPLC-DAD, we determined variations in bioactive compounds' content in relation to the fermentation process and origin and observed negative correlations between TAC and TPC. Heavy metal content followed this order: Mn > Fe > Cu > Zn > Ni > Cr > Pb > Co > Cd > Hg. Given the homogenous content of these elements in relation to fermentation, this paper also describes the possibility of using heavy metals as determinants of geographical origin. Linear Discriminant Analysis showed an accuracy of 75% for Ni, Co, Cd, Hg, and Pb, explaining 95.19% of the variability between geographical regions.

Loxocephalids represent a nonmonophyletic assemblage of oligohymenophorean ciliates with morphological features common to both scuticociliates (mode of ontogenesis) and hymenostomes (morphology of the oral structures). In phylogenetic analyses that include both nuclear and mitochondrial molecular markers, relationships among loxocephalid ciliates are still largely unresolved. With the aim to clarify the phylogeny of the controversial order Loxocephalida Jankowski, 1980, we provide the first 18S rRNA gene sequences of three morphologically well-described ciliates currently included in the order, namely Cinetochilum margaritaceum, Zitheron hovassei, and Zitheron muscorum comb. nov. We also provide the first 18S rRNA gene sequence for the monotypic genus Urozona. Here we present improved diagnoses for these species based on updated morphologic data. Our molecular data circumscribe the family Cinetochilidae Perty, 1852 and support the exclusion of genus Cinetochilides from family Cinetochilidae; therefore, we establish a family Cinetochilididae fam. nov. We consider Urozona as incertae sedis in Oligohymenophorea at the current state of knowledge., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier GmbH. All rights reserved.)

The scope of application of carbon nanomaterials in biomedical, environmental and industrial fields is recently substantially increasing. Since in vitro toxicity testing is the first essential step for any commercial usage, it is crucial to have a reliable method to analyze the potentially harmful effects of carbon nanomaterials. Even though researchers already reported the interference of carbon nanomaterials with common toxicity assays, there is still, unfortunately, a large number of studies that neglect this fact. In this study, we investigated interference of four bio-promising carbon nanomaterials (graphene acid (GA), cyanographene (GCN), graphitic carbon nitride (g-C 3 N 4 ) and carbon dots (QCDs)) in commonly used LIVE/DEAD assay. When a standard procedure was applied, materials caused various types of interference. While positively charged g-C 3 N 4 and QCDs induced false results through the creation of free agglomerates and intrinsic fluorescence properties, negatively charged GA and GCN led to false signals due to the complex quenching effect of the fluorescent dye of a LIVE/DEAD kit. Thus, we developed a new approach using a specific gating strategy based on additional controls that successfully overcame all types of interference and lead to reliable results in LIVE/DEAD assay. We suggest that the newly developed procedure should be a mandatory tool for all in vitro flow cytometry assays of any class of carbon nanomaterials.