University of Rzeszów

In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, research on this topic has continued to accelerate, and many new scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Accordingly, it is important to update these guidelines for monitoring autophagy in different organisms. Various reviews have described the range of assays that have been used for this purpose. Nevertheless, there continues to be confusion regarding acceptable methods to measure autophagy, especially in multicellular eukaryotes. For example, a key point that needs to be emphasized is thatthere is a difference between measurements that monitor the numbers or volume of autophagic elements (e.g., autophagosomes or autolysosomes) at any stage of the autophagic process versus those that measure flux through the autophagy pathway (i.e., the completeprocess including the amount and rate of cargo sequestered and degraded). In particular, a block in macroautophagy that results in autophagosome accumulation must be differentiated from stimuli that increase autophagic activity, defined as increasedautophagy induction coupled with increased delivery to, and degradation within, lysosomes (inmost higher eukaryotes and some protists such as Dictyostelium) or the vacuole (in plants and fungi). In other words, it is especially important that investigators new to the field understand that the appearance of more autophagosomes does not necessarily equate with more autophagy. In fact, in manycases, autophagosomes accumulate because of a block in trafficking to lysosomes without a concomitant change in autophagosome biogenesis, whereas an increase in autolysosomes may reflect a reduction in degradative activity. It is worth emphasizing here that lysosomal digestion is a stage of autophagy and evaluating its competence is a crucial part of the evaluation of autophagic flux, or complete autophagy. Here, we present a set of guidelines for the selection and interpretation of methods for use by investigators who aim to examine macroautophagy and related processes, as well as forreviewers who need to provide realistic and reasonable critiques of papers that are focused on these processes. These guidelines are not meant to be a formulaic set of rules, because the appropriate assays depend in part on the question being asked and the system being used. In addition, we emphasize that no individual assay is guaranteed to be the most appropriate one in every situation, and we strongly recommend the use of multipleassays to monitor autophagy. Along these lines, because of the potential for pleiotropic effects due to blocking autophagy through genetic manipulation, it is imperative to target by gene knockout or RNA interference more than one autophagyrelated protein. In addition, some individual Atg proteins, or groups of proteins, are involved in other cellular pathways implying that not all Atg proteins can be used as a specific marker for an autophagic process. In these guidelines, we consider these various methods of assessing autophagy and what information can, or cannot, be obtained from them. Finally, by discussing the merits and limits of particular assays, we hope to encourage technical innovation in the field.

autophagic responses. Here, we critically discuss current methods of assessing autophagy and the information they can, or cannot, provide. Our ultimate goal is to encourage intellectual and technical innovation in the field.

Climate warming is causing a shift in biological communities in favor of warm-affinity species (i.e., thermophilization). Species responses often lag behind climate warming, but the reasons for such lags remain largely unknown. Here, we analyzed multidecadal understory microclimate dynamics in European forests and show that thermophilization and the climatic lag in forest plant communities are primarily controlled by microclimate. Increasing tree canopy cover reduces warming rates inside forests, but loss of canopy cover leads to increased local heat that exacerbates the disequilibrium between community responses and climate change. Reciprocal effects between plants and microclimates are key to understanding the response of forest biodiversity and functioning to climate and land-use changes.

Abstract Body-mass index (BMI) has increased steadily in most countries in parallel with a rise in the proportion of the population who live in cities 1,2 . This has led to a widely reported view that urbanization is one of the most important drivers of the global rise in obesity 3–6 . Here we use 2,009 population-based studies, with measurements of height and weight in more than 112 million adults, to report national, regional and global trends in mean BMI segregated by place of residence (a rural or urban area) from 1985 to 2017. We show that, contrary to the dominant paradigm, more than 55% of the global rise in mean BMI from 1985 to 2017—and more than 80% in some low- and middle-income regions—was due to increases in BMI in rural areas. This large contribution stems from the fact that, with the exception of women in sub-Saharan Africa, BMI is increasing at the same rate or faster in rural areas than in cities in low- and middle-income regions. These trends have in turn resulted in a closing—and in some countries reversal—of the gap in BMI between urban and rural areas in low- and middle-income countries, especially for women. In high-income and industrialized countries, we noted a persistently higher rural BMI, especially for women. There is an urgent need for an integrated approach to rural nutrition that enhances financial and physical access to healthy foods, to avoid replacing the rural undernutrition disadvantage in poor countries with a more general malnutrition disadvantage that entails excessive consumption of low-quality calories.

The aim of this review is to present an overview of changes in the contemporary use of wild food plants in Europe, mainly using the examples of our home countries: Poland, Italy, Spain, Estonia and Sweden. We set the scene referring to the nutrition of 19th century peasants, involving many famine and emergency foods. Later we discuss such issues as children's wild snacks, the association between the decline of plant knowledge and the disappearance of plant use, the effects of over-exploitation, the decrease of the availability of plants due to ecosystem changes, land access rights for foragers and intoxication dangers. We also describe the 20th and 21st century vogues in wild plant use, particularly their shift into the domain of haute-cuisine.

BACKGROUND: Neuraminidase inhibitors were widely used during the 2009-10 influenza A H1N1 pandemic, but evidence for their effectiveness in reducing mortality is uncertain. We did a meta-analysis of individual participant data to investigate the association between use of neuraminidase inhibitors and mortality in patients admitted to hospital with pandemic influenza A H1N1pdm09 virus infection. METHODS: We assembled data for patients (all ages) admitted to hospital worldwide with laboratory confirmed or clinically diagnosed pandemic influenza A H1N1pdm09 virus infection. We identified potential data contributors from an earlier systematic review of reported studies addressing the same research question. In our systematic review, eligible studies were done between March 1, 2009 (Mexico), or April 1, 2009 (rest of the world), until the WHO declaration of the end of the pandemic (Aug 10, 2010); however, we continued to receive data up to March 14, 2011, from ongoing studies. We did a meta-analysis of individual participant data to assess the association between neuraminidase inhibitor treatment and mortality (primary outcome), adjusting for both treatment propensity and potential confounders, using generalised linear mixed modelling. We assessed the association with time to treatment using time-dependent Cox regression shared frailty modelling. FINDINGS: We included data for 29,234 patients from 78 studies of patients admitted to hospital between Jan 2, 2009, and March 14, 2011. Compared with no treatment, neuraminidase inhibitor treatment (irrespective of timing) was associated with a reduction in mortality risk (adjusted odds ratio [OR] 0·81; 95% CI 0·70-0·93; p=0·0024). Compared with later treatment, early treatment (within 2 days of symptom onset) was associated with a reduction in mortality risk (adjusted OR 0·48; 95% CI 0·41-0·56; p<0·0001). Early treatment versus no treatment was also associated with a reduction in mortality (adjusted OR 0·50; 95% CI 0·37-0·67; p<0·0001). These associations with reduced mortality risk were less pronounced and not significant in children. There was an increase in the mortality hazard rate with each day's delay in initiation of treatment up to day 5 as compared with treatment initiated within 2 days of symptom onset (adjusted hazard ratio [HR 1·23] [95% CI 1·18-1·28]; p<0·0001 for the increasing HR with each day's delay). INTERPRETATION: We advocate early instigation of neuraminidase inhibitor treatment in adults admitted to hospital with suspected or proven influenza infection. FUNDING: F Hoffmann-La Roche.

The presence of heavy metals in food is a threat to human health. Exposure to heavy metals as a result of consumption of contaminated vegetables, as well as their toxicity, is a serious problem. Different branches of industry and the road traffic have a significant impact on environmental pollution with heavy metals. Municipal and industrial sewage also is an important source of those substances. Furthermore, the mineral content of vegetables depends on factors such as the natural content of trace elements in the environment, their levels in mineral fertilizers, and fertilizer doses. In the soil, a natural source of these metals is bedrock. In soils used for agricultural purposes, some quantities of metals are introduced together with fertilizers, both organic and mineral. Additionally, another sources of the metals are plant protection products. Heavy metal dynamics in the soil and their uptake by plants are influenced by soil properties, which play a key role in the bioavailability of these metals. Metal mobility and assimilation are also influenced by the addition of organic and inorganic matter. A significant body of evidence also suggest that the age of the soil plays an important role in modulation of metal bioavailability to plants. Apart from being influenced by the soil-related factors, absorption of metals differs in different types of plants. A significant variation in metal concentrations was also found depending on their location in plant tissues, on plant species, or even on varieties of the same species.

AIM: The article addresses selected determinants of the nursing shortage in Poland and other countries in the face of employee ageing. BACKGROUND: Global demographic changes have led to a systematic increase in the elderly population and a decreasing number of births, which have impacted health policy and healthcare systems in various countries. Both processes necessitate transitions in global health care. Nursing care, which has faced a human resources crisis, is a strategic area within this context. SOURCES OF EVIDENCE: This study is based on national listings and strategic documents for nursing policy in Poland, including Increasing average age of nurses and midwives prepared by the Polish Main Council of Nurses and Midwives, the incorporation of big data, international reports and a literature review on nursing and healthcare challenges. DISCUSSION/CONCLUSIONS: This paper argues that the causes of the nursing shortage are multifaceted with no single global or local measure of its nature. An overview of the problem indicates ineffective planning and use of available nursing resources, poor recruitment or an undersupply of a new staff, and global demographic conditions. The overview highlights the fact that nursing shortages have reached a critical point for healthcare services on both the local and global levels. CONCLUSIONS FOR NURSING AND HEALTH POLICY: The general recommendations for nursing policy include the need to prepare and implement national social security agendas into services provided by nurses. Such a programme would include general issues: improving working and employment conditions, implementing mechanisms regulating salary and providing the possibility of lifelong learning with the incorporation of mobile and technological innovations as a sustainable solution.

We we present some algorithms, based on rough set theory, that can be used for the problem of new cases classification. Most of the algorithms were implemented and included in Rosetta system [43]. We present several methods for computation of decision rules based on reducts. We discuss the problem of real value attribute discretization for increasing the performance of algorithms and quality of decision rules. Finally we deal with a problem of resolving conflicts between decision rules classifying a new case to different categories (classes). Keywords: knowledge discovery, rough sets, classification algorithms, reducts, decision rules, real value attribute discretization

Awareness is an uncommon complication of anaesthesia, affecting 0.1-0.2% of all surgical patients. Bispectral index (BIS) monitoring measures the depth of anaesthesia and facilitates anaesthetic titration. In this trial we determined whether BIS-guided anaesthesia reduced the incidence of awareness during surgery in adults.We did a prospective, randomised, double-blind, multicentre trial. Adult patients at high risk of awareness were randomly allocated to BIS-guided anaesthesia or routine care. Patients were assessed by a blinded observer for awareness at 2-6 h, 24-36 h, and 30 days after surgery. An independent committee, blinded to group identity, assessed every report of awareness. The primary outcome measure was confirmed awareness under anaesthesia at any time.Of 2463 eligible and consenting patients, 1225 were assigned to the BIS group and 1238 to the routine care group. There were two reports of awareness in the BIS-guided group and 11 reports in the routine care group (p=0.022). BIS-guided anaesthesia reduced the risk of awareness by 82% (95% CI 17-98%).BIS-guided anaesthesia reduces the risk of awareness in at-risk adult surgical patients undergoing relaxant general anaesthesia. With a cost of routine BIS monitoring at US16 dollars per use in Australia and a number needed to treat of 138, the cost of preventing one case of awareness in high-risk patients is about 2200 dollars.

Riboflavin [7,8-dimethyl-10-(1'-d-ribityl)isoalloxazine, vitamin B₂] is an obligatory component of human and animal diets, as it serves as the precursor of flavin coenzymes, flavin mononucleotide, and flavin adenine dinucleotide, which are involved in oxidative metabolism and other processes. Commercially produced riboflavin is used in agriculture, medicine, and the food industry. Riboflavin synthesis starts from GTP and ribulose-5-phosphate and proceeds through pyrimidine and pteridine intermediates. Flavin nucleotides are synthesized in two consecutive reactions from riboflavin. Some microorganisms and all animal cells are capable of riboflavin uptake, whereas many microorganisms have distinct systems for riboflavin excretion to the medium. Regulation of riboflavin synthesis in bacteria occurs by repression at the transcriptional level by flavin mononucleotide, which binds to nascent noncoding mRNA and blocks further transcription (named the riboswitch). In flavinogenic molds, riboflavin overproduction starts at the stationary phase and is accompanied by derepression of enzymes involved in riboflavin synthesis, sporulation, and mycelial lysis. In flavinogenic yeasts, transcriptional repression of riboflavin synthesis is exerted by iron ions and not by flavins. The putative transcription factor encoded by SEF1 is somehow involved in this regulation. Most commercial riboflavin is currently produced or was produced earlier by microbial synthesis using special selected strains of Bacillus subtilis, Ashbya gossypii, and Candida famata. Whereas earlier RF overproducers were isolated by classical selection, current producers of riboflavin and flavin nucleotides have been developed using modern approaches of metabolic engineering that involve overexpression of structural and regulatory genes of the RF biosynthetic pathway as well as genes involved in the overproduction of the purine precursor of riboflavin, GTP.

Ascomycete yeasts are metabolically diverse, with great potential for biotechnology. Here, we report the comparative genome analysis of 29 taxonomically and biotechnologically important yeasts, including 16 newly sequenced. We identify a genetic code change, CUG-Ala, in Pachysolen tannophilus in the clade sister to the known CUG-Ser clade. Our well-resolved yeast phylogeny shows that some traits, such as methylotrophy, are restricted to single clades, whereas others, such as l-rhamnose utilization, have patchy phylogenetic distributions. Gene clusters, with variable organization and distribution, encode many pathways of interest. Genomics can predict some biochemical traits precisely, but the genomic basis of others, such as xylose utilization, remains unresolved. Our data also provide insight into early evolution of ascomycetes. We document the loss of H3K9me2/3 heterochromatin, the origin of ascomycete mating-type switching, and panascomycete synteny at the MAT locus. These data and analyses will facilitate the engineering of efficient biosynthetic and degradative pathways and gateways for genomic manipulation.

Obesity has become a major epidemic in the 21st century. It increases the risk of dyslipidemia, hypertension, and type 2 diabetes, which are known cardiometabolic risk factors and components of the metabolic syndrome. Although overt cardiovascular (CV) diseases such as stroke or myocardial infarction are the domain of adulthood, it is evident that the CV continuum begins very early in life. Recognition of risk factors and early stages of CV damage, at a time when these processes are still reversible, and the development of prevention strategies are major pillars in reducing CV morbidity and mortality in the general population. In this review, we will discuss the role of well-known but also novel risk factors linking obesity and increased CV risk from prenatal age to adulthood, including the role of perinatal factors, diet, nutrigenomics, and nutri-epigenetics, hyperuricemia, dyslipidemia, hypertension, and cardiorespiratory fitness. The importance of 'tracking' of these risk factors on adult CV health is highlighted and the economic impact of childhood obesity as well as preventive strategies are discussed.

This paper is an ethnobotanical review of wild edible plants gathered for consumption from the 19th century to the present day, within the present borders of Slovakia. Twenty-four sources (mainly ethnographic) documenting the culinary use of wild plants were analysed. The use of 106 species (over 3% of the Slovak flora) has been recorded. Nowadays most of them are no longer used, or used rarely, apart from a few species of wild fruits. The most frequently used plants include the fruits of &lt;em&gt;Rubus idaeus&lt;/em&gt;, &lt;em&gt;Fragaria &lt;/em&gt;spp., &lt;em&gt;Rubus &lt;/em&gt;subgenus &lt;em&gt;Rubus&lt;/em&gt;, &lt;em&gt;Vaccinium myrtillus&lt;/em&gt;, &lt;em&gt;V. vitis-idaea&lt;/em&gt;, &lt;em&gt;Fagus sylvatica&lt;/em&gt;, &lt;em&gt;Corylus avellana&lt;/em&gt;, &lt;em&gt;Prunus spinosa&lt;/em&gt;, &lt;em&gt;Pyrus &lt;/em&gt;spp., &lt;em&gt;Malus &lt;/em&gt;spp., &lt;em&gt;Crataegus &lt;/em&gt;spp. and the leaves of &lt;em&gt;Urtica dioica&lt;/em&gt;, &lt;em&gt;Rumex acetosa&lt;/em&gt;, Chenopodiaceae species, &lt;em&gt;Cardamine amara&lt;/em&gt;, &lt;em&gt;Glechoma &lt;/em&gt;spp., &lt;em&gt;Taraxacum &lt;/em&gt;spp. and &lt;em&gt;Oxalis acetosella&lt;/em&gt;. The most commonly used wild food taxa are nearly identical to those used in Poland, and the same negative association of wild vegetables with famine exists in Slovakia, resulting in their near complete disappearance from the present-day diet.

Four spectrophotometric methods of determination of antioxidant capacity: a method based on the scavenging of the l,1-diphenyl-2-picrylhydrazyl (DPPH) free radical, the "ferric-reducing ability of plasma" (FRAP), a method based on reduction of the 2.2'-azinobis (3-ethylbenzthiazolinesulfonate) free radical (ABTS.+) and a kinetic method based on the oxidation of dihydro-2,7-dichlorofluorescein by 2,2'-azobis(2-amidopropane) (ABAP) were compared with respect to standard antioxidants (ascorbate, glutathione, Trolox and urate) and human blood plasma. Various reactivities of standard antioxidants in different tests were found. glutathione showing a low reactivity in the FRAP assay. Kinetic measurements show that the reduction of indicators, especially by blood plasma, may not be complete at recommended times of the assays and the time of measurement is an important parameter when comparing the results.

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.

There has been a significant increase in the development of edible films and coatings in recent times, and this is expected to have a significant impact on the quality of fruit and vegetables in the coming years. Consumers expect fresh fruit and vegetables free from pesticide residues, with high quality, nutritional value and an extended shelf life. The application of coatings and edible films to fruits and vegetables represents an environmentally friendly approach to an innovative solution to this problem. Coatings and edible films can act as ecological and biodegradable packaging. The coating strategy involves a combination of natural biopolymers and appropriate preservation methods. The article presents the applicability, trends and perspectives of polysaccharide coatings and edible films and their impact on the quality of fruit and vegetables, providing an understanding of their main functions and benefits. Numerous studies show that natural polysaccharides are well suited for use as packaging material for fresh fruit and vegetables and can often be an important alternative to synthetic compounds. Natural polymer materials are a good barrier to oxygen and carbon dioxide; however, they are characterised by excessive solubility in the water environment, water vapour permeability and low extensibility. The properties of edible coatings can be modified by the addition of plasticisers, surfactants, cross-linkers, antimicrobial agents, functional additives, nanosilver particles or fruit and vegetable residues. The use of an electric field is also a promising technology here. The use of polysaccharides for the preparation of edible films and coatings is justified not only by the possibility of reducing the consumption of packaging made of synthetic polymer materials but also by the fact that the production of some natural polymers can be made using waste products generated during the processing of food raw materials.

Betalains are water-soluble pigments present in vacuoles of plants of the order Caryophyllales and in mushrooms of the genera Amanita, Hygrocybe and Hygrophorus. Betalamic acid is a constituent of all betalains. The type of betalamic acid substituent determines the class of betalains. The betacyanins (reddish to violet) contain a cyclo-3,4-dihydroxyphenylalanine (cyclo-DOPA) residue while the betaxanthins (yellow to orange) contain different amino acid or amine residues. The most common betacyanin is betanin (Beetroot Red), present in red beets Beta vulgaris, which is a glucoside of betanidin. The structure of this comprehensive review is as follows: Occurrence of Betalains; Structure of Betalains; Spectroscopic and Fluorescent Properties; Stability; Antioxidant Activity; Bioavailability, Health Benefits; Betalains as Food Colorants; Food Safety of Betalains; Other Applications of Betalains; and Environmental Role and Fate of Betalains.

BACKGROUND & AIMS: Cholangiocarcinoma (CCA) is a rare and heterogeneous biliary cancer, whose incidence and related mortality is increasing. This study investigates the clinical course of CCA and subtypes (intrahepatic [iCCA], perihilar [pCCA], and distal [dCCA]) in a pan-European cohort. METHODS: The ENSCCA Registry is a multicenter observational study. Patients were included if they had a histologically proven diagnosis of CCA between 2010-2019. Demographic, histomorphological, biochemical, and clinical studies were performed. RESULTS: Overall, 2,234 patients were enrolled (male/female=1.29). iCCA (n = 1,243) was associated with overweight/obesity and chronic liver diseases involving cirrhosis and/or viral hepatitis; pCCA (n = 592) with primary sclerosing cholangitis; and dCCA (n = 399) with choledocholithiasis. At diagnosis, 42.2% of patients had local disease, 29.4% locally advanced disease (LAD), and 28.4% metastatic disease (MD). Serum CEA and CA19-9 showed low diagnostic sensitivity, but their concomitant elevation was associated with increased risk of presenting with LAD (odds ratio 2.16; 95% CI 1.43-3.27) or MD (odds ratio 5.88; 95% CI 3.69-9.25). Patients undergoing resection (50.3%) had the best outcomes, particularly with negative-resection margin (R0) (median overall survival [mOS] = 45.1 months); however, margin involvement (R1) (hazard ratio 1.92; 95% CI 1.53-2.41; mOS = 24.7 months) and lymph node invasion (hazard ratio 2.13; 95% CI 1.55-2.94; mOS = 23.3 months) compromised prognosis. Among patients with unresectable disease (49.6%), the mOS was 10.6 months for those receiving active palliative therapies, mostly chemotherapy (26.2%), and 4.0 months for those receiving best supportive care (20.6%). iCCAs were associated with worse outcomes than p/dCCAs. ECOG performance status, MD and CA19-9 were independent prognostic factors. CONCLUSION: CCA is frequently diagnosed at an advanced stage, a proportion of patients fail to receive cancer-specific therapies, and prognosis remains dismal. Identification of preventable risk factors and implementation of surveillance in high-risk populations are required to decrease cancer-related mortality. LAY SUMMARY: Edition (ICD-11) (i.e., intrahepatic [2C12], perihilar [2C18], or distal [2C15] affected bile ducts), which come into effect in 2022. General and tumor-type specific features at diagnosis, risk factors, biomarker accuracy, as well as patient management and outcomes, are presented and compared, outlining the current clinical state of cholangiocarcinoma in Europe.

Humans the world over have depended on wild-growing plants in their diets for hundreds of thousands of years, and many people continue to rely on these species to meet at least part of their daily nutritional needs. Wild harvested plant foods include: roots and other underground parts; shoots and leafy greens; berries and other fleshy fruits; grains, nuts and seeds; and mushrooms, lichens, algae and other species. Use of any of these species requires special cultural knowledge regarding harvesting, preparation, cooking and other forms of processing. Many were, and are, prepared and served in mixtures or combinations. In most cases, too, the species are managed, tended or manipulated in some way to increase their productivity and availability. Many of the most widely used species are categorized as weeds—species that grow and reproduce readily in disturbed or cleared land, and are common around human settlements and agricultural areas. This paper presents case examples of edible wild plant use and the roles of these species in agroecosystems from different parts of the world and discusses similarities and differences in use across different cultures and segments of society.