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Extracellular vesicles (EVs), through their complex cargo, can reflect the state of their cell of origin and change the functions and phenotypes of other cells. These features indicate strong biomarker and therapeutic potential and have generated broad interest, as evidenced by the steady year-on-year increase in the numbers of scientific publications about EVs. Important advances have been made in EV metrology and in understanding and applying EV biology. However, hurdles remain to realising the potential of EVs in domains ranging from basic biology to clinical applications due to challenges in EV nomenclature, separation from non-vesicular extracellular particles, characterisation and functional studies. To address the challenges and opportunities in this rapidly evolving field, the International Society for Extracellular Vesicles (ISEV) updates its Minimal Information for Studies of Extracellular Vesicles, which was first published in 2014 and then in 2018 as MISEV2014 and MISEV2018, respectively. The goal of the current document, MISEV2023, is to provide researchers with an updated snapshot of available approaches and their advantages and limitations for production, separation and characterisation of EVs from multiple sources, including cell culture, body fluids and solid tissues. In addition to presenting the latest state of the art in basic principles of EV research, this document also covers advanced techniques and approaches that are currently expanding the boundaries of the field. MISEV2023 also includes new sections on EV release and uptake and a brief discussion of in vivo approaches to study EVs. Compiling feedback from ISEV expert task forces and more than 1000 researchers, this document conveys the current state of EV research to facilitate robust scientific discoveries and move the field forward even more rapidly.

Abstract Extracellular vesicles (EVs) are lipid-membrane enclosed structures that are associated with several diseases, including those of genitourinary tract. Urine contains EVs derived from urinary tract cells. Owing to its non-invasive collection, urine represents a promising source of biomarkers for genitourinary disorders, including cancer. The most used method for urinary EVs separation is differential ultracentrifugation (UC), but current protocols lead to a significant loss of EVs hampering its efficiency. Moreover, UC protocols are labor-intensive, further limiting clinical application. Herein, we sought to optimize an UC protocol, reducing the time spent and improving small EVs (SEVs) yield. By testing different ultracentrifugation times at 200,000g to pellet SEVs, we found that 48 min and 60 min enabled increased SEVs recovery compared to 25 min. A step for pelleting large EVs (LEVs) was also evaluated and compared with filtering of the urine supernatant. We found that urine supernatant filtering resulted in a 1.7-fold increase on SEVs recovery, whereas washing steps resulted in a 0.5 fold-decrease on SEVs yield. Globally, the optimized UC protocol was shown to be more time efficient, recovering higher numbers of SEVs than Exoquick-TC (EXO). Furthermore, the optimized UC protocol preserved RNA quality and quantity, while reducing SEVs separation time.

Summary: During DNA repair, ATM-induced H2AX histone phosphorylation and MDC1 recruitment spread megabases beyond the damage site. While loop extrusion has been suggested to drive this spread, the underlying mechanism remains unclear. Herein, we provide two lines of evidence that loop extrusion is not the only driver of damage-induced γH2AX spread. First, cohesin loader NIPBL and cohesin subunit RAD21 accumulate considerably later than the phosphorylation of H2AX and MDC1 recruitment at micro-IR-induced damage. Second, auxin-induced RAD21 depletion does not affect γH2AX/MDC1 spread following micro-irradiation or DSB induction by zeocin. To determine if diffusion of activated ATM could account for the observed behavior, we measured the exchange rate and diffusion constants of ATM and MDC1 within damaged and unperturbed chromatin. Using these measurements, we introduced a quantitative model in which the freely diffusing activated ATM phosphorylates H2AX. This model faithfully describes the dynamics of ATM and subsequent γH2AX/MDC1 spread at complex DNA lesions.

The etiology of Hashimoto's thyroiditis is still unclear. Genetic predisposition and the effects of various environmental factors are discussed. The present article is a systematic review of publications on the occurrence and development of this disease in the presence of impaired functions of the gastrointestinal tract and changes in the microbiota. Increased intestinal permeability, as well as changes in the microbiota, can disrupt the normal functioning of the immune system and can lead to the development of an autoimmune disease. Normalization of the diversity of the microbiota before an autoimmune process occurs can be achieved both with the use of pro-biotic strains and with appropriate dietary changes.

Cytokines are the key mediators which control and regulate immune and inflammatory responses via complex networks and serve as biomarkers of many diseases. Quantitative determination of cytokines is helpful in assessing immune status and adjusting therapy for various inflammatory diseases, such as sepsis and pneumonia. Since community-acquired pneumonia remains a common cause of childhood morbidity and mortality. At the present stage, prognosis of the disease severity in children is an urgent problem. The aim of our study was to identify associations between cytokine levels in healthy children and in children with community-acquired pneumonia (CAP), depending on the age of patients and severity of the pathological process. The work was carried out at the Department of Microbiology, Virology and Immunology, Department of Propaedeutics of Childhood Diseases and Pediatrics, and at the Research Institute of Immunology at the South Ural State Medical University. The study included 117 children aged 1 to 18 years with radiologically confirmed diagnosis of CAP, either severe or mild degree. The comparison group was composed from 28 healthy children who did not have community-acquired pneumonia, as well as other signs of acute respiratory viral infection at the time of examination, being observed for any chronic disorders. The levels of IL-1β, IFNγ, IL-6, IL-4, IL-10, IL-2, TNFα, IFNλ2 (IL-28A), IFNλ3 (IL-28B), IL-8, MCP-1, IL-17AF, GM-CSF were determined in blood serum by means of ELISA test systems (a “sandwich” technique with peroxidase as an indicator enzyme). For statistical analysis, a multidimensional method was used, i.e., non-linear analysis of the principal components using the CATPCA algorithm. Among the children with CAP, our study revealed a consistent increase of IL-1ß, IL-4, IL-10, IL-2, TNFα, IFNλ2 (IL-28A), IFNλ3 (IL-28B), IL-8, MCP-1, IL-17AF, GM-CSF in blood serum. The highest correlation with severity was shown for IFNλ2 (IL-28A), IFNλ3 (IL-28B) and MCP-1, which may be considered additional biomarkers of the CAP severity. There was also a significant variability of the cytokine profile in healthy children and its significant skewing in pneumonia, especially in severe cases.

Alzheimer’s disease (AD) has long proven to be a complex neurodegenerative disorder, with cholinergic dysfunction, oxidative stress, and neuroinflammation being just a few of its pathological features. The complexity of the disease requires a multitargeted treatment covering its many aspects. In the present investigation, an arylhydrazone derivative of 5-methoxyindole-2-carboxylic acid (5MeO), with in vitro strong antioxidant, neuroprotective and monoamine oxidase B-inhibiting effects, was studied in a scopolamine-induced Alzheimer-type dementia in rats. Using behavioral and biochemical methods, we evaluated the effects of 5MeO on learning and memory, and elucidated the mechanisms of these effects. Our experiments demonstrated that 5MeO had a beneficial effect on different types of memory as assessed by the step-through and the Barnes maze tasks. It efficiently restored the decreased by scopolamine brain-derived neurotrophic factor and acetylcholine levels and normalized the increased by scopolamine acetylcholine esterase activity in hippocampus. Most effective 5MeO was in counteracting the induced by scopolamine oxidative stress by decreasing the increased by scopolamine levels of lipid peroxidation and by increasing the reduced by scopolamine catalase activity. Blood biochemical analyses demonstrated a favorable safety profile of 5MeO, prompting further pharmacological studies suggesting 5MeO as a safe and efficient candidate in a multitargeted treatment of AD.

Introduction: Metal ions, released from dental alloys due to corrosion, come in contact with the cells of the surrounding tissues and may spread throughout the body via the gastrointestinal system, thus inducing dose-dependent cytopathological effects. This study aimed to assess and compare the salivary cobalt and chromium concentrations in individuals aged 18–65 years with and without dental restorations containing metal alloys. Methods: Participants were divided into two main groups according to the existence of metal alloys in the oral cavity—18 patients had fixed prosthetic restorations made of metal alloys, and 17 patients had no metal objects in their oral cavity. Each main group was subdivided into two subgroups according to the type of saliva sample—with or without additional stimulation. Salivary cobalt and chromium concentrations were measured by inductively coupled plasma mass spectrometry. A non-parametric Mann–Whitney test and Spearman’s rank correlation coefficient were applied, and the level of significance was set to p < 0.05. Results: The results showed that the chromium level in non-stimulated saliva was higher in the group of patients with metal dental restorations. No statistical difference was found in cobalt levels. There was no statistical difference in Co or Cr concentrations in stimulated saliva between the studied groups. A positive correlation was found between Cr and Co concentrations in non-stimulated saliva and between cobalt concentrations in stimulated and non-stimulated saliva. Conclusions: Metal alloys in the oral cavity induced elevated chromium levels in non-stimulated saliva, and a correlation between chromium and cobalt ion concentration was found. A detailed examination of patients and their medical history prior to prosthetic treatment is advisable in order to avoid any undesired health effects.

Colorectal cancer (CRC) is a leading cause of mortality worldwide. Its incidence holds a major position among the most common life-threatening diseases. Hence, the early identification and precise characterization of disease activity based on proper biomarkers are of utmost importance for therapeutic strategy and patient survival. The identification of new biomarkers for colorectal cancer or disease-specific levels/combinations of biomarkers will significantly contribute to precise diagnosis and improved personalized treatment of patients. Therefore, the present study aims to identify colorectal cancer-specific immunological biomarkers. The plasma levels of several cytokines (interleukin-1β /IL-1β/, IL-2, IL-4, IL-10, IL-12, IL-15, TGFβ and IFNγ) of 20 patients with colorectal cancer and 21 healthy individuals were determined by ELISA. The expression of several types of glycoproteins on the surface of peripheral blood leukocytes isolated from CRC patients and healthy volunteers was evaluated by flow cytometry. Correlations between cytokine levels and cell surface glycoprotein expression were analyzed. The obtained results demonstrated significantly elevated levels of CD80, CD86, CD279 and CD274 expressing leukocyte populations in the cancer patient group, while the numbers of NK cells and CD8- and CD25-positive cells were decreased. Based on these data and the correlations with cytokine levels, it can be concluded that CD25, CD80, CD86, CD274 and CD279 glycoproteins combined with specific plasma levels of IL-1β, IL-2, IL-15 and TGFβ could represent potential biomarkers for colorectal cancer.

BackgroundRespiratory syncytial virus (RSV) is a common cause of upper and lower respiratory tract infections. This study aimed to explore the prevalence of respiratory syncytial virus (RSV) and other respiratory viruses in Bulgaria, characterize the genetic diversity of RSV strains, and perform amino acid sequence analyses of RSV surface and internal proteins.MethodsClinical and epidemiological data and nasopharyngeal swabs were prospectively collected from patients with acute respiratory infections between October 2020 and May 2023. Real-time PCR for 13 respiratory viruses, whole-genome sequencing, phylogenetic, and amino acid analyses were performed.ResultsThis study included three epidemic seasons (2020–2021, 2021–2022, and 2022–2023) from week 40 of the previous year to week 20 of the following year. Of the 3,047 patients examined, 1,813 (59.5%) tested positive for at least one viral respiratory pathogen. RSV was the second most detected virus (10.9%) after SARS-CoV-2 (22%). Coinfections between RSV and other respiratory viruses were detected in 68 cases, including 14 with SARS-CoV-2. After two seasons of low circulation, RSV activity increased significantly during the 2022–2023 season. The detection rates of RSV were 3.2, 6.6, and 13.7% in the first, second, and third seasons, respectively. RSV was the most common virus found in children under 5 years old with bronchiolitis (40%) and pneumonia (24.5%). RSV-B drove the 2022–2023 epidemic. Phylogenetic analysis indicated that the sequenced RSV-B strains belonged to the GB5.0.5a and GB5.0.6a genotypes. Amino acid substitutions in the surface and internal proteins, including the F protein antigenic sites were identified compared to the BA prototype strain.ConclusionThis study revealed a strong resurgence of RSV in the autumn of 2022 after the lifting of anti-COVID-19 measures, the leading role of RSV as a causative agent of serious respiratory illnesses in early childhood, and relatively low genetic diversity in circulating RSV strains.

Agriculture is an important sector in Bulgaria, especially in the rural areas where it is considered an important source of income and employment. Bulgarian agriculture has changed substantially over the past ten years, which has led to a transformation in production, farm, organizational and sectorial structures. The aim of the paper is to observe the regional differences in Bulgarian farm structure and outline main prospects for the next programming period, 2023-2027. The survey reveals a significant decline in the number of holdings in parallel with the decreased role of small farms. By contrast, the importance of large holdings is increasing. There are different patterns of agricultural transformation in the regions of Bulgaria. While large extensive producers dominate North Bulgaria, there is more balanced agricultural development in the South part of the country. The new CAP 2023-2027 is directed to more ambitious objectives related to climate, innovations and convergence. The subsidiarity principles could help Bulgaria to overcome some of these challenges if the agricultural policy is better targeted and oriented towards intensive sectors and more balanced and sustainable rural development.

Bioeconomy, as an evolving concept, gained serious interest in the past two decades. It is considered a key driver in resolving a number of global challenges and is integrated into a broader framework with circularity, green transition and sustainable development. At this point, many international, national and regional strategic documents are linked to bioeconomy and directed to its strategic objectives. The aim of the paper is to present comparative analysis of bioeconomy strategies at global and national levels in order to observe the main concepts, vision and priorities and highlighted future agenda, prospects and recommendations for better integration and coordination. The analysis of the published strategic documents shows that there are no global framework and internationally accepted definition of bioeconomy. There are diverse understandings and differences in key priorities and goals between Europe, Asia, and South and North America. The bioeconomy remains a political concept, with a significant number of countries engaged in implementing its guiding principles. Until 2030, the ambitions in bioeconomy strategies and policies will be tested. In this regard, significant support is needed in the form of specific measures and financial instruments to realize investments in this field.

Aim: The purpose of this study was to compare the mechanical properties of materials used for orthodontic retainers made by direct 3D printing and thermoforming. Materials and methods: Twenty-one specimens (n=7) from 3 different materials (Formlabs Dental LT Clear V2 - Formlabs Inc., Somerville, Massachusetts, USA; NextDent Ortho Flex - Vertex-Dental B.V., Soesterberg, The Netherlands, and Erkodent Erkodur - ERKODENT, Germany) were manufactured and their mechanical properties were evaluated. Two of the specimen groups were 3D printed and the other one was fabricated using a material for thermoforming. The statistical methods we applied were descriptive statistics, the Kruskal-Wallis and Dunn’s post-hoc tests. Results: With respect to Young’s modulus (E), the Kruskal-Wallis test (df=2, χ2=17.121, p=0.0002) showed a significant difference between the materials for direct 3D printing of orthodontic retainers (E=2762.4 MPa±115.16 MPa for group 1 and 2393.05 MPa±158.13 MPa for group 2) and thermoforming foils (group 3, E=1939.4 MPa±74.18 MPa). Statistically significant differences were also found between the flexural strength (FS) (Kruskal-Wallis test, df=2, χ2=17.818, p=0.0001) and F(max) (Kruskal-Wallis test, df=2, χ2=17.818, p=0.0001). Conclusions: The materials tested in the current study showed statistically significant differences in their Young’s modulus, flexural strength, and F(max).

The efficiency of virus internalization into target cells is a major determinant of infectivity. SARS-CoV-2 internalization occurs via S-protein-mediated cell binding followed either by direct fusion with the plasma membrane or endocytosis and subsequent fusion with the endosomal membrane. Despite the crucial role of virus internalization, the precise kinetics of the processes involved remains elusive. We developed a pipeline, which combines live-cell microscopy and advanced image analysis, for measuring the rates of multiple internalization-associated molecular events of single SARS-CoV-2-virus-like particles (VLPs), including endosome ingression and pH change. Our live-cell imaging experiments demonstrate that only a few minutes after binding to the plasma membrane, VLPs ingress into RAP5-negative endosomes via dynamin-dependent scission. Less than two minutes later, VLP speed increases in parallel with a pH drop below 5, yet these two events are not interrelated. By co-imaging fluorescently labeled nucleocapsid proteins, we show that nucleocapsid release occurs with similar kinetics to VLP acidification. Neither Omicron mutations nor abrogation of the S protein polybasic cleavage site affected the rate of VLP internalization, indicating that they do not confer any significant advantages or disadvantages during this process. Finally, we observe that VLP internalization occurs two to three times faster in VeroE6 than in A549 cells, which may contribute to the greater susceptibility of the former cell line to SARS-CoV-2 infection. Taken together, our precise measurements of the kinetics of VLP internalization-associated processes shed light on their contribution to the effectiveness of SARS-CoV-2 propagation in cells.

Abstract Climate change has been associated with both latitudinal and elevational shifts in species’ ranges. The extent, however, to which climate change has driven recent range shifts alongside other putative drivers remains uncertain. Here, we use the changing distributions of 378 European breeding bird species over 30 years to explore the putative drivers of recent range dynamics, considering the effects of climate, land cover, other environmental variables, and species’ traits on the probability of local colonisation and extinction. On average, species shifted their ranges by 2.4 km/year. These shifts, however, were significantly different from expectations due to changing climate and land cover. We found that local colonisation and extinction events were influenced primarily by initial climate conditions and by species’ range traits. By contrast, changes in climate suitability over the period were less important. This highlights the limitations of using only climate and land cover when projecting future changes in species’ ranges and emphasises the need for integrative, multi-predictor approaches for more robust forecasting.

Abstract Extracellular vesicles (EVs), through their complex cargo, can reflect the state of their cell of origin and change the functions and phenotypes of other cells. These features indicate strong biomarker and therapeutic potential and have generated broad interest, as evidenced by the steady year‐on‐year increase in the numbers of scientific publications about EVs. Important advances have been made in EV metrology and in understanding and applying EV biology. However, hurdles remain to realising the potential of EVs in domains ranging from basic biology to clinical applications due to challenges in EV nomenclature, separation from non‐vesicular extracellular particles, characterisation and functional studies. To address the challenges and opportunities in this rapidly evolving field, the International Society for Extracellular Vesicles (ISEV) updates its ‘Minimal Information for Studies of Extracellular Vesicles’, which was first published in 2014 and then in 2018 as MISEV2014 and MISEV2018, respectively. The goal of the current document, MISEV2023, is to provide researchers with an updated snapshot of available approaches and their advantages and limitations for production, separation and characterisation of EVs from multiple sources, including cell culture, body fluids and solid tissues. In addition to presenting the latest state of the art in basic principles of EV research, this document also covers advanced techniques and approaches that are currently expanding the boundaries of the field. MISEV2023 also includes new sections on EV release and uptake and a brief discussion of in vivo approaches to study EVs. Compiling feedback from ISEV expert task forces and more than 1000 researchers, this document conveys the current state of EV research to facilitate robust scientific discoveries and move the field forward even more rapidly.

Ribose 2′O-methylation (Nm, ribomethylation) is the most abundant RNA modification present in rRNA. It has been shown that alterations in ribosomal 2′O-methylation at individual Nm sites likely reflect regulated cellular processes. Although several analytical approaches for Nm detection and profiling have been developed, a simple and affordable method for the screening and measurement of individual Nm sites in large numbers of tissue samples is required to examine their potential for clinical translation. Here, we describe a new quantitative reverse transcription PCR-based method that can sensitively assess ribomethylation levels at specific rRNA sites at single-nucleotide resolution in low input amounts of total RNA.

Introduction: Adipose tissue is immunologically and hormonally active, and these effects are mediated largely by adipocytokines. Thyroid hormones regulate metabolism and organ function, and Hashimoto’s thyroiditis (HT) is the most common autoimmune disease affecting thyroid function.Aim: To evaluate the levels of the adipocytokines leptin and adiponectin in patients with autoimmune HT, and to perform a comparative intragroup analysis in patients with different stages of gland functional activity, and in a control group.Materials and methods: Ninety-five patients with HT and 21 healthy controls were enrolled in the study. Venous blood was taken without anticoagulants after at least 12 hours of fasting, and serum samples were frozen at −70°C until analysis. Serum levels of leptin and adiponectin were determined by an enzyme-linked immunosorbent assay (ELISA).Results: Serum levels of leptin in HT patients were higher than those in the control group (4.5±5.2 ng/mL vs. 1.9±1.3 ng/mL). The hypothyroid patient’s group showed significantly higher levels of leptin than those of the healthy controls (5.1±5.2 ng/mL vs. 1.9±1.3 ng/mL), (p=0.031). Leptin levels correlated positively with body mass index (r=0.533, p

The progress of information and communication technologies in the era of COVID-19 created an unprecedented opportunity for medicine to adapt to new models of care. Telemedicine and telehealth have enabled medical care at a distance in various fields, including ophthalmology. The aim of this article is to review the current state and the opportunities for telemedicine in ophthalmology.

Introduction: Prostate cancer (PCa), one of the most prevalent malignancies affecting men worldwide, presents significant challenges in terms of early detection, risk stratification, and active surveillance. In recent years, liquid biopsies have emerged as a promising non-invasive approach to complement or even replace traditional tissue biopsies. Extracellular vesicles (EVs), nanosized membranous structures released by various cells into body fluids, have gained substantial attention as a source of cancer biomarkers due to their ability to encapsulate and transport a wide range of biological molecules, including RNA. In this study, we aimed to validate 15 potential RNA biomarkers, identified in a previous EV RNA sequencing study, using droplet digital PCR.Methods: The candidate biomarkers were tested in plasma and urinary EVs collected before and after radical prostatectomy from 30 PCa patients and their diagnostic potential was evaluated in a test cohort consisting of 20 benign prostate hyperplasia (BPH) and 20 PCa patients’ plasma and urinary EVs. Next, the results were validated in an independent cohort of plasma EVs from 31 PCa and 31 BPH patients.Results: We found that the levels of NKX3-1 (p = 0.0008) in plasma EVs, and tRF-Phe-GAA-3b (p < 0.0001) tRF-Lys-CTT-5c (p < 0.0327), piR-28004 (p = 0.0081) and miR-375-3p (p < 0.0001) in urinary EVs significantly decreased after radical prostatectomy suggesting that the main tissue source of these RNAs is prostate and/or PCa. Two mRNA biomarkers—GLO1 and NKX3-1 showed promising diagnostic potential in distinguishing between PCa and BPH with AUC of 0.68 and 0.82, respectively, in the test cohort and AUC of 0.73 and 0.65, respectively, in the validation cohort, when tested in plasma EVs. Combining these markers in a biomarker model yielded AUC of 0.85 and 0.71 in the test and validation cohorts, respectively. Although the PSA levels in the blood could not distinguish PCa from BPH in our cohort, adding PSA to the mRNA biomarker model increased AUC from 0.71 to 0.76.Conclusion: This study identified two novel EV-enclosed RNA biomarkers–NKX3-1 and GLO1–for the detection of PCa, and highlights the complementary nature of GLO1, NKX3-1 and PSA as combined biomarkers in liquid biopsies of PCa.

Community-acquired pneumonia (CAP) is one of the most common acute infectious diseases. To date, the incidence of CAP among children was decreased significantly worldwide, mainly due to increasing use of effective preventive measures. Nevertheless, CAP remains a common cause of childhood morbidity and mortality. Pneumonia may develop at any age, but most often it occurs in young children, who are more likely to have a more severe course of pneumonia. Currently, early diagnosis and prognosis of the disease severity in children is an urgent issue. It was found that, in most cases, a panel of conventional biomarkers, including the number of leukocytes, procalcitonin, CRP is not sufficient for the diagnosis of pediatric CAP. There is a demand for new biological markers which, along with clinical evaluation, may significantly improve diagnostics and management of CAP in children, thus reducing the risk of adverse outcomes associated with this disease. Such markers could be found among the cytokines, which are active participants in the CAP pathogenesis. The aim of this study was to determine the level of several cytokines in blood serum of children with CAP and to assess changes in the cytokine profile depending on the patient’s age and severity of the disease. The study included 117 children aged 1 to 18 years with a diagnosis of CAP confirmed by X-ray examination. The comparison group included 28 healthy children who did not have CAP or other signs of acute respiratory viral infection at the time of examination, being free of any chronic pathology requiring outpatient observation. A number of cytokines were determined quantitatively in blood serum, i.e., IL-1β, IFNγ, IL-2, IL-4, IL-6, IL-10, TNFα, IFNλ2 (IL-28A), IFNλ3 (IL-28B), IL-8, MCP-1, IL-17AF, GM-CSF using test systems based on the “sandwich” method of solid-phase ELISA using peroxidase labeling. As a result, it was found that the content of IL-6, IL-17AF, IL-1β, IFNγ, MCP-1, IFNλ2 (IL-28A), IFNλ3 (IL-28B), GM-CSF was significantly higher in the group of children with severe community-acquired pneumonia. The levels of certain cytokines, e.g., IL-6, IFNλ2 (IL-28A), IFNλ3 (IL-28B), GM-CSF varied depending on the age of patients, thus, probably, reflecting the degree of immune system activation in the children of different age groups.

Electronic decision-support tools are becoming an essential component of government strategies to tackle non-native species invasions. This study describes the development and application of a multilingual electronic decision-support tool for screening terrestrial animals under current and future climate conditions: the Terrestrial Animal Species Invasiveness Screening Kit (TAS-ISK). As an adaptation of the widely employed Aquatic Species Invasiveness Screening Kit (AS-ISK), the TAS-ISK question template inherits from the original Weed Risk Assessment (WRA) and related WRA-type toolkits and complies with the ‘minimum requirements’ for use with the recent European Regulation on invasive alien species of concern. The TAS-ISK consists of 49 basic questions on the species’ biogeographical/historical traits and its biological/ecological interactions, and of 6 additional questions to predict how climate change is likely to influence the risks of introduction, establishment, dispersal and impact of the screened species. Following a description of the main features of this decision-support tool as a turnkey software application and of its graphical user interface with support for 32 languages, sample screenings are provided in different risk assessment areas for one representative species of each of the main taxonomic groups of terrestrial animals supported by the toolkit: mammals, birds, reptiles, amphibians, annelids, insects, molluscs, nematodes, and platyhelminths. The highest-scoring species were the red earthworm Lumbricus rubellus for the Aegean region of Turkey and the New Zealand flatworm Arthurdendyus triangulatus for Croatia. It is anticipated that adoption of this toolkit will mirror that of the worldwide employed AS-ISK, hence allowing to share information and inform decisions for the prevention of entry and/or dispersal of (high-risk) non-native terrestrial animal species – a crucial step to implement early-stage control and eradication measures as part of rapid-response strategies to counteract biological invasions.

The aim of the present study was to investigate the changes in the fatty acid profile and acid value of milkfat during ripening of Kashkaval cheese with reduced sodium chloride content. Three batches of Kashkaval cheese (with low 0.7%; medium 1.5% and standard 3.1% NaCl content) were produced. Analyses were carried out on days 1 and 45 of ripening. Based on the results obtained, it was established that lipolysis in Kashkaval cheese during ripening was significantly delayed (p

CRISPR elements in the human genome are expressed in both healthy tissues and tumors but with distinct patterns, representing a potential biomarker for cancer.

Alzheimer’s disease manifests as a complex pathological condition, with neuroinflammation, oxidative stress and cholinergic dysfunction being a few of the many pathological changes. Due to the complexity of the disease, current therapeutic strategies aim at a multitargeted approach, often relying on a combination of substances with versatile and complementary effects. In the present study, a unique combination of α-lipoic acid, citicoline, extracts of leaves from olive tree and green tea, vitamin D3, selenium and an immune-supporting complex was tested in scopolamine-induced dementia in rats. Using behavioral and biochemical methods, we assessed the effects of the combination on learning and memory, and elucidated the mechanisms of these effects. Our results showed that, compared to its components, the experimental combination was most efficient in improving short- and long-term memory as assessed by the step-through method as well as spatial memory as assessed by T-maze and Barnes maze underlined by decreases in AChE activity (p < 0.05) and LPO (p < 0.001), increases in SOD activity in the cortex (p < 0.05) and increases in catalase (p < 0.05) and GPx (p < 0.01) activities and BDNF (p < 0.001) and pCREB (p < 0.05) levels in the hippocampus. No significant histopathological changes or blood parameter changes were detected, making the experimental combination an effective and safe candidate in a multitargeted treatment of AD.

The recent expansion of the round goby (Neogobius melanostomus) in tributaries of the lower Danube (Bulgaria) prompted assessing potential negative effects on local fish fauna such as competition for food resources. Diet overlap between the round goby and native fishes was analyzed using samples of fishes and macroinvertebrates collected monthly for a one-year period. Significant dietary overlap between N. melanostomus and Barbus petenyi, Perca fluviatilis, Gobio gobio, Vimba vimba, and Neogobius fluviatilis was observed. Dietary overlap in the Iskar River was found to be considerable mainly in the spring months. Conversely, overlap in the Vit River was observed during summer and late fall. The main reason for the overlap noted was the preference for Chironomidae and Hydropsychidae larvae and Gammaridae. The results suggest that there is potential for competition between the round goby and native fish species inhabiting the tributaries of the Danube. Potentially harmful effects on rare, protected fish species in the area are discussed.

Introduction: Recent studies have shown that the intestinal microbiota can modulate certain systemic metabolic and immune responses, including liver graft function and the development of complications in patients after liver transplantation (LT). Akkermansia muciniphila (AKM) and Faecalibacterium prausnitzii (FAEP) are two of the most abundant gut commensal bacteria, with mucosa-protective and anti-inflammatory effects that are important for maintaining normal intestinal homeostasis and gut barrier function. Our objective was to quantify levels of Akkermansia muciniphila and Faecalibacterium prausnitzii in immunosuppressed patients with LT. Materials and methods: Fecal samples from 23 liver transplant patients (15 adults and 8 children) and 9 non-LT controls were examined. Bacterial DNA was isolated from the samples using the stool DNA isolation kit and the obtained DNA was analyzed with commercially available qPCR kit for AKM and FAEP. Results: We found a statistically significant decrease in the amount of AKM and FAEP compared to the control group. The median values were: for AKM 8.75 for patients and 10.25 for the control group (p = 0.030), and for FAEP 9.72 and 10.47, p = 0.003, respectively. In children after LT, this difference was also statistically significant: AKM (p = 0.051) and FAEP (p = 0.014). In contrast no statistically significant differences were found between adult patients and controls, AKM (p = 0.283) and FAEP (p = 0.056), although the amount of both bacteria showed tendency for reduction. Conclusions: In this pilot study, we found a reduction in the total amount of the two studied bacteria in transplanted patients compared to the control healthy group.

The composition of grape seeds oil from a mixture of two varieties of red grape (Sangiovese and Pinot noir) from Varna region was investigated. Different analytical techniques have been used to determine the content of biologically active compounds, fatty acids, sterols and elemental compositions. It has been found that the oxidation stability of the oil is relatively low - about 4.5 hours. The fluorescence spectrum analyzed at an excitation wavelength of 350 nm contains peaks that are attributed to pigments, oxidizing products and vitamins. Transmission spectra were used to determine the content of β-carotene, chlorophyll and color characteristics in the CIELab colorimetric system. Infrared spectroscopic experiments (ATR and permeability) were used for confirmation of the fatty acid profile of the analyzed oil. The results show that this oil is a good source of various healthy nutrients

The European Commission presented the 2018 Bioeconomy strategy, which developed an action plan for a resource-efficient, competitive and sustainable economy. The bioeconomy sectors are also linked to the European Green Deal, Stepping up Europe’s 2030 climate ambition Communication, Farm to Folk Strategy and other innovation strategies. In the transition to a low carbon world, biomass plays a crucial role as an alternative to fossil resources. In the EU, agriculture is the main source of biomass with 68% of the total supply. The paper analyses European Union agricultural biomass potential, supply and use. Agricultural biomass is part of the core bioeconomy sectors and its demand is increasing. However, the potential of biomass and its alternative uses are a major concern. Biomass is a renewable but limited resource and, on that basis, it is important to outline the balance and to pay attention to the relationship between the nutritional and industrial needs of biomass in terms of food and energy security. The policy framework in this regard has to be complex and well-targeted. The biomass use could lead to a number of benefits associated with resolving global issues. On the other hand, if sustainability is not taken into account, the opportunities for inclusive growth and development will not be achieved.

The topic was to investigate the anatomical features of the rabbit liver by two- and three-dimensional ultrasonography. Eighteen sexually mature healthy clinically New Zealand rabbits aged eight months were studied. Two-dimensional ultarsonographic anatomical image of the rabbit liver presented it in the cranial abdominal region as a relatively hypoechoic finding. Its contours were regular and in close contact with the hyperechoic diaphragm. Liver parenchyma was heterogeneous. The gall bladder was visualized as an oval soft tissue structure, filled with anechoic content. Its walls were hypoechoic. Two-dimensional ultarsonographic anatomical image of left hepatic lobe was sharply distinguished to right hepatic lobe’s outlines. In three–dimensional ultrasonogarphic anatomical study, the organ image was in three orthogonal planes. Its relief was regular and uninterrupted. Left hepatic lobe was found on the left and the right hepatic lobe was a soft tissue point for gall bladder position. Left and lateral was left lateral hepatic lobe. It was covered partly by left medial hepatic lobe. The right hepatic lobe was visualized as a single structure. The gall bladder was an oval finding. Its walls were hyperechoic and regular, without roughness. The results could be used as a base for modern interpretation of rabbit liver anatomy.