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Motivation The gut microbiota is the human body’s largest population of microorganisms that interact with human intestinal cells. They use ingested nutrients for fundamental biological processes and have important impacts on human physiology, immunity and metabolome in the gastrointestinal tract. Results Here, we present M2R, a Python add-on to cobrapy that allows incorporating information about the gut microbiota metabolism models to human genome-scale metabolic models (GEMs) like RECON3D. The idea behind the software is to modify the lower bounds of the exchange reactions in the model using aggregated in- and out-fluxes from selected microbes. M2R enables users to quickly and easily modify the pool of the metabolites that enter and leave the GEM, which is particularly important for those looking into an analysis of the metabolic interaction between the gut microbiota and human cells and its dysregulation. Availability and implementation M2R is freely available under an MIT License at https://github.com/e-weglarz-tomczak/m2r. Supplementary information Supplementary data are available at Bioinformatics online.

The purpose of the study was to assess the variability between 16 wild thyme populations introduced into ex situ conditions, in terms of morphological traits and chemical profiles. The observations were carried out on 2-year-old plants. Total content of essential oil and phenolic compounds (flavonoids, phenolic acids and tannins) was determined according to Polish Pharmacopoeia 6th. The composition of essential oil was carried out by GC/MS and GC/FID. Populations differed in respect of examined traits, i.e.: plant’s height (from 50 to 90 mm), plant diameter (from 21.3 to 53.0 cm), length of stems (from 8.1 to 20.5 cm). Fresh mass of herb per plant ranged from 36.0 to 307.0 g, while dry mass from 15.63 to 73.11 g (CV 55.0 and 47.5%, respectively). Total content of essential oil varied from 0.23 to 0.97%. Three chemotypes were distinguished within investigated populations: geranyl acetate + β-terpineol + β-myrcene, geranyl acetate + β-terpineol + borneol and pure linalool type. The total content of phenolic compounds ranged, as following: flavonoids (0.17–0.26%), tannins (0.57–1.32%), phenolic acids (0.28–0.61%). To sum up, investigated populations differed significantly in respect of developmental features as well as content and composition of identified compounds. Obtained results may be used in future works on selection and breeding of wild thyme, providing a high-quality raw material characterized by specific chemical composition and pharmacological activity.

Cancer cell metabolism is dependent on cell-intrinsic factors, such as genetics, and cell-extrinsic factors, such nutrient availability. In this context, understanding how these two aspects interact and how diet influences cellular metabolism is important for developing personalized treatment. In order to achieve this goal, genome-scale metabolic models (GEMs) are used, however, genetics and nutrient availability are rarely considered together. Here, we propose integrated metabolic profiling, a framework that allows enriching GEMs with metabolic gene expression data and information about nutrients. First, the RNA-seq is converted into Reaction Activity Score (RAS) to further scale reaction bounds. Second, nutrient availability is converted to Maximal Uptake Rate (MUR) to modify exchange reactions in a GEM. We applied our framework to the human osteosarcoma cell line (U2OS). Osteosarcoma is a common and primary malignant form of bone cancer with poor prognosis, and, as indicated in our study, a glutamine-dependent type of cancer.

Dropwort is a wild-growing plant, long used in traditional European medicine. The species is a rich source of phenolic compounds. The plant raw materials are: flowers, herb, and underground organs (rhizomes with tuberous roots). They have been used in the treatment of difficult-to-heal wounds, cold, rheumatism, and kidney problems. Dropwort extracts reveal potential in the prevention of neurodegenerative disorders, as well. Due to dynamic changes in the use of agricultural lands, natural sites of this species gradually disappear. Introduction into the cultivation seems to be a chance to preserve its natural resources and to provide standardized raw materials for the industry. In this chapter, we present the results of our investigations concerning the factors affecting the quality of different raw materials obtained from cultivated dropwort plants.

The purpose of the study was to determine the differences between two subspecies: O. vulgare L. subsp. hirtum (Link) Ietswaart (Greek oregano) and O. vulgare L. subsp. vulgare (common oregano) growing in cultivation conditions within temperate climate of Central Europe. The characteristic of the subspecies was undertaken in terms of selected morphological parameters and the quality of the raw material. The herb of both subspecies was evaluated on the content and composition of essential oil by hydrodistillation followed by GC-MS and GC-FID (gas chromatography coupled with mass spectrometry and flame ionization detector), the total content of phenolic acids (according to PP 6th ed.) and the content of rosmarinic acid (by HPLC). The sensory evaluation (QDA) was performed, as well. Greek oregano was distinguished by visibly higher number of glandular trichomes on the leaves (up to 4.85 per 1 mm2) followed by higher content of essential oil in the herb (up to 3.36 g &times, 100 g&minus, 1 DW) in comparison to common oregano. Based on the essential oil composition, Greek oregano was classified as mixed carvacrol/&gamma, terpinene chemotype, while common oregano as mixed sabinyl/cymyl type rich in sesquiterpenes. Greek oregano was also characterized by higher total content of phenolic acids (up to 6.16 g &times, 1 DW) and rosmarinic acid (up to 6787.2 mg &times, 1 DW) than common oregano. Essential oil content reached the maximum at the beginning of blooming (common oregano) and at the full blooming stage (Greek oregano). In turn, the amount of phenolic acids followed by rosmarinic acid was the highest at the beginning of seed-setting stage, in the case of both subspecies. The differences between subspecies concerning chemical composition (especially essential oil) were reflected in the sensory attributes, where both odor and taste notes were found at higher level for Greek oregano. Results of our work indicate that Greek oregano is well adapted to grow in the temperate zone conditions. Such adaptation was reflected mainly in the satisfied yield and maintaining characters typical for the Mediterranean plant, e.g., a high essential oil content followed by high carvacrol share, traits the most important from practice viewpoint.

Common valerian (Valeriana officinalis L.) is one of the most important medicinal plants, with a mild sedative, nervine, antispasmodic and relaxant effect. Despite a substantial number of studies on this species, the genetic diversity and population structure have not yet been analyzed. Here, we use a next-generation sequencing-based Diversity Array Technology sequencing (DArT-seq) technique to analyze Polish gene bank accessions that originated from wild populations and cultivars. The major and, also, the most astounding result of our work is the low level of observed heterozygosity of individual plants from natural populations, despite the fact that the species is widespread in the studied area. Inbreeding in naturally outcrossing species such as valerian decreases reproductive success. The analysis of the population structure showed the potential presence of a metapopulation in the central part of Poland and the formation of a distinct gene pool in the Bieszczady Mountains. The results also indicate the presence of the cultivated gene pool within wild populations in the region where the species is cultivated for the needs of the pharmaceutical industry, and this could lead to structural and genetic imbalances in wild populations.

The phytochemical diversity of Melittis melissophyllum was investigated in terms of seasonal changes and age of plants including plant organs diversity. The content of phenolics, namely: coumarin, 3,4-dihydroxycoumarin, o-coumaric acid 2-O-glucoside, verbascoside, apiin, luteolin-7-O-glucoside, and o-coumaric, p-coumaric, chlorogenic, caffeic, ferulic, cichoric acids, was determined using HPLC-DAD. Among these, luteolin-7-O-glucoside, verbascoside, chlorogenic acid, and coumarin were the dominants. The highest content of flavonoids and phenolic acids was observed in 2-year-old plants, while coumarin in 4-year-old plants (272.06 mg 100 g&ndash, 1 DW). When considering seasonal changes, the highest content of luteolin-7-O-glucoside was observed at the full flowering, whereas verbascoside and chlorogenic acid were observed at the seed-setting stage. Among plant organs, the content of coumarin and phenolic acids was the highest in leaves, whereas verbascoside and luteolin-7-O-glucoside were observed in flowers. The composition of essential oil was determined using GC-MS/GC-FID. In the essential oil from leaves, the dominant was 1-octen-3-ol, whilst from flowers, the dominant was &alpha, pinene.

Since December 2019 a novel a coronavirus identified as SARS-CoV-2 or COV2 has been spreading around the world. On the 16th of May around 4.5 million people got infected and over 300,000 died due to the infection of COV2. The effective treatment remains a challenge. Targeted therapeutics are still under investigation. The papain-like protease (PLPro) from the human SARS-CoV-2 coronavirus is a cysteine protease that plays a critical role in virus replication. Its activity is required to process the viral polyprotein into functional, mature subunits. Moreover, COV2 uses this enzyme to modulate the host’s immune system to its own benefit. Therefore, it represents a highly promising target for the development of antiviral drugs.In this work, we discovered that ebselen, a synthetic organoselenium drug molecule with anti-inflammatory, anti-oxidant and cytoprotective activity in mammalian cells and cytotoxicity in lower organisms, is a highly active inhibitor of PLProCoV2. We proved that ebselen is a covalent, fast-binding inhibitor of PLProCoV2 exhibiting a low micromolar potency. Furthermore, we identified a difference between PLPro from SARS-CoV-1 (the corona virus which caused the 2002–2004 outbreak, SARS) and SARS-CoV-2 that allows to explain the difference in dynamics of the replication, and, thus, the disease progression. Namely, we present that they show differences in the binding affinity of substrates that we observed through kinetics and molecular docking studies. Using a novel Approximate Bayesian Computation method we were able to find kinetic constants for both enzymes. Molecular modeling study on the structure of the active site and binding mode of the ebselen with SARS and COV2 showed also significant differences that could explain our observation that ebselen is less active and slower bounding with SARS than COV2.In conclusion, we show that ebselen inhibits the activity of the essential viral enzyme papain-like protease (PLpro) from SARS-COV-2 in low micromolar range.

Aim Probiotic bacteria administered directly after birth to preterm neonates may improve gastrointestinal function and may reduce the incidence of late-onset sepsis, which is a frequent complication in this group. Purpose The main objective of this study was to evaluate whether a new probiotic bacterial mixture of Lactobacillus rhamnosus KL53A and Bifidobacterium breve PB04 given to preterm, low-birth-weight neonates would influence composition of their gut microbiota and sepsis rates. Patients and methods This study was a multicenter, randomized, double-blind, placebo-controlled trial conducted in clinical centers of neonatal care in Poland. A probiotic or placebo preparation was given twice daily to 181 preterm low-birth-weight neonates who were eligible for enteral feeding between July 2012 and July 2013. The probiotic was given to 90 neonates, while placebo was given to 91 neonates. The gut microbiota was monitored by microbiological analysis of stool samples. Sepsis episodes were detected on the basis of clinical and laboratory findings and confirmed by blood cultures. Results Tested probiotic administration resulted in continuous increase of the Lactobacillus and Bifidobacterium counts in the gut microbiota. The applied tested strains successfully colonized the neonates gut since they were present in over 90% of stool samples, which was confirmed by molecular analysis. Regardless of the study group (probiotic or placebo), B. breve colonization correlated with lower staphylococcal sepsis incidence, which was irrespective of whether probiotics were given. No sepsis case caused by strains included in study probiotic was recorded. Conclusion Appropriately selected and characterized probiotic bacteria may be safely given to preterm neonates to normalize their distorted gut microbiota and may contribute to lower staphylococcal sepsis rates.

The potent transcription inhibitor Actinomycin D is used with several cancers. Here, we report the discovery that this naturally occurring antibiotic inhibits two human neutral aminopeptidases, the cell-surface alanine aminopeptidase and intracellular methionine aminopeptidase type 2. These metallo-containing exopeptidases participate in tumor cell expansion and motility and are targets for anticancer therapies. We show that the peptide portions of Actinomycin D and Actinomycin X2 are not required for effective inhibition, but the loss of these regions changes the mechanism of interaction. Two structurally less complex Actinomycin D analogs containing the phenoxazone chromophores, Questiomycin A and Actinocin, appear to be competitive inhibitors of both aminopeptidases, with potencies similar to the non-competitive macrocyclic parent compound (Ki in the micromolar range). The mode of action for all four compounds and both enzymes was demonstrated by molecular modeling and docking in the corresponding active sites. This knowledge gives new perspectives to Actinomycin D’s action on tumors and suggests new avenues and molecules for medical applications.

The gray short-tailed opossum (Monodelphis domestica) is a small marsupial gaining recognition as a laboratory animal in biomedical research. Despite numerous studies on opossum neuroanatomy, a consistent and comprehensive neuroanatomical reference for this species is still missing. Here we present the first three-dimensional, multimodal atlas of the Monodelphis opossum brain. It is based on four complementary imaging modalities: high resolution ex vivo magnetic resonance images, micro-computed tomography scans of the cranium, images of the face of the cutting block, and series of sections stained with the Nissl method and for myelinated fibers. Individual imaging modalities were reconstructed into a three-dimensional form and then registered to the MR image by means of affine and deformable registration routines. Based on a superimposition of the 3D images, 113 anatomical structures were demarcated and the volumes of individual regions were measured. The stereotaxic coordinate system was defined using a set of cranial landmarks: interaural line, bregma, and lambda, which allows for easy expression of any location within the brain with respect to the skull. The atlas is released under the Creative Commons license and available through various digital atlasing web services. Electronic supplementary material The online version of this article (10.1007/s00429-017-1540-x) contains supplementary material, which is available to authorized users.

BACKGROUND Malformations of cortical development, such as focal cortical dysplasia, are commonly associated with intractable epilepsy. Multiple animal models were created in attempts to recapitulate features of human malformations of cortical development. These manipulations give rise to various focal or diffuse anatomical abnormalities, accompanied by altered susceptibility to epileptic seizures. Both in humans and in models of dysplasia, the question of timing of the initiating insult is important. METHODS Here, we used a rat model of cerebral dysplasia elicited by prenatal irradiation at gestational days (E) 13, 15, 17, or 19. Previous results suggest these animals are characterized by different patterns of dysplasia as well as different reactivity to seizurogenic stimuli in several seizure models. Rats were implanted with telemetric electroencephalography (EEG) transmitters. We compared EEG data from freely moving animals with anatomical observations obtained with MRI, as well as Western blotting and immunohistochemistry. RESULTS We performed spectral analyses of EEG signal, revealing differential regulation of specific bands (including delta, theta, alpha-beta, gamma) in animals with different patterns of dysplasia. Relative contribution of low-frequency activity in delta band is the lowest in E15 and the highest in E19. Conversely, higher frequency bands, corresponding to alpha/beta and gamma components, are reduced in E15 versus E19. This is accompanied by altered expression of glial markers in the E19 group. CONCLUSION To our knowledge, this constitutes the first quantitative description of power spectral properties in this experimental model, providing insight into mechanisms underlying the anatomical and electrophysiological abnormalities associated with brain dysplasia of different types. Birth Defects Research, 2017.© 2017 Wiley Periodicals, Inc.

Inositol hexaphosphate (IP6), a natural dietary component, has been found as an antitumor agent by stimulating apoptosis and inhibiting cancer cell proliferation, their migration, and metastasis in diverse cancers including colon cancer. However, molecular mechanisms of its action have not been well understood. In recent years, microRNAs (miRNAs) have been reported to play important roles in a broad range of biologic processes, such as cell growth, proliferation, apoptosis, or autophagy. These small noncoding molecules regulate post-transcriptional expression of targets genes via degradation of transcript or inhibition of protein synthesis. Aberrant expression and/or dysregulation of miRNAs have been characterized during tumor development and progression, thus, they are potential molecular targets for cancer prevention. The aim of this study was to investigate the effect of IP6 on the miRNAs expression profile in Caco-2 colon cancer cells. 84 miRNAs were analyzed in Caco-2 cells treated with 2.5 mM and 5 mM IP6 by the use of PCR (Polymerase Chain Reaction) array. The effect of 5 mM IP6 on selected potential miR-155 targets was determined by real-time (RT)-qPCR and ELISA (quantitative Polymerase Chain Reaction and Enzyme-Linked Immunosorbent Assay )method. The results indicated alteration in the specific 10 miRNA expression in human colon cancer cells following their treatment with 5 mM IP6. It down-regulated 8 miRNAs (miR-155, miR-210, miR-144, miR-194, miR-26b, miR-126, miR-302c, and miR-29a) and up-regulated 2 miRNAs (miR-223 and miR-196b). In silico analysis revealed that FOXO3a, HIF-1&alpha, and ELK3 mRNAs are those of predicted targets of miR-155. IP6 at the concentration of 5 mM markedly induced FOXO3a and HIF-1a genes&rsquo, expression at both mRNA and protein level and decreased the amount of ELK3 mRNA as well as protein concentration in comparison to the control. In conclusion, the present study indicates that one of the mechanisms of antitumor potential of IP6 is down-regulation of the miR-155 expression in human colon cancer cells. Moreover, the expression of genes that are targeted by miRNA are also modulated by IP6.

How cancer cells utilize nutrients to support their growth and proliferation in complex nutritional systems is still an open question. However, it is certainly determined by both genetics and an environmental-specific context. The interactions between them lead to profound metabolic specialization, such as consuming glucose and glutamine and producing lactate at prodigious rates. To investigate whether and how glucose and glutamine availability impact metabolic specialization, we integrated computational modeling on the genome-scale metabolic reconstruction with an experimental study on cell lines. We used the most comprehensive human metabolic network model to date, Recon3D, to build cell line-specific models. RNA-Seq data was used to specify the activity of genes in each cell line and the uptake rates were quantitatively constrained according to nutrient availability. To integrated both constraints we applied a novel method, named GENSI (Gene Expression and Nutrients Simultaneous Integration), that translates the relative importance of gene expression and nutrient availability data into the metabolic fluxes based on an observed experimental feature(s). We applied GENSI to study hepatocellular carcinoma addiction to glucose/glutamine. We were able to identify that proliferation, and lactate production is associated with the presence of glucose but does not necessarily increase with its concentration when the latter exceeds the physiological concentration. There was no such association with glutamine. We show that the integration of gene expression and nutrient availability data into genome-wide models improves the prediction of metabolic phenotypes.

Background: Valeriana officinalis L. is one of the most important medicinal plant with a mild sedative, nervine, antispasmodic and relaxant effect. Despite a substantial number of studies on this species, population genomics has not yet been analyzed. The main aim of this study was: characterization of genetic variation of natural populations of V. officinalis in Poland and comparison of variation of wild populations and the cultivated form using Next Generation Sequencing based DArTseq technique. We also would like to establish foundations for genetic monitoring of the species in the future and to develop genetic fingerprint profile for samples deposited in gene bank and in natural sites in order to assess the degree of their genetic integrity and population structure preservation in the future.Results: The major and also the most astounding result of our work is the low level of observed heterozygosity of individual plants from natural populations despite the fact that the species is widespread in the studied area. Inbreeding, in naturally outcrossing species such as valerian, decreases the reproductive success. The analysis of the population structure indicated the potential presence of metapopulation in a broad area of Poland and the formation of a distinct gene pool in Bieszczady Mountains. The results also indicate the presence of individuals of the cultivated form in natural populations in the region where the species is cultivated for the needs of the pharmaceutical industry and this could lead to structural and genetic imbalance in wild populations.Conclusions: The DArTseq technology can be applied effectively in genetic studies of V. officinalis. The genetic variability of wild populations is in fact significantly lower than assumed. Individuals from the cultivated population are found in the natural environment and their impact on wild populations should be monitored.

How cancer cells utilize nutrients to support their growth and proliferation in complex nutritional systems is still an open question. However, it is certainly determined by both genetics and an environmental-specific context. The interactions between them lead to profound metabolic specialization, such as consuming glucose and glutamine and producing lactate at prodigious rates. To investigate whether and how glucose and glutamine availability impact metabolic specialization, we integrated computational modeling on the genome-scale metabolic reconstruction with an experimental study on cell lines. We used the most comprehensive human metabolic network model to date, Recon3D, to build cell line-specific models. RNA-Seq data was used to specify the activity of genes in each cell line and the uptake rates were quantitatively constrained according to nutrient availability. To integrated both constraints we applied a novel method, named Gene Expression and Nutrients Simultaneous Integration (GENSI), that translates the relative importance of gene expression and nutrient availability data into the metabolic fluxes based on an observed experimental feature(s). We applied GENSI to study hepatocellular carcinoma addiction to glucose/glutamine. We were able to identify that proliferation, and lactate production is associated with the presence of glucose but does not necessarily increase with its concentration when the latter exceeds the physiological concentration. There was no such association with glutamine. We show that the integration of gene expression and nutrient availability data into genome-wide models improves the prediction of metabolic phenotypes.

Summary Introduction. In the group of plants revealing adaptogenic activity, species belonging to Scutellaria genus are considered to be the most important ones. Common skullcap (Scutellaria galericulata L.) is less known species from this genus, growing wild in Poland in wet habitats. Its herb has been used in the past as a sedative and diuretic agent. Objective. The aim of the study was to determine the variability of common skullcap in terms of the accumulation of flavonoids in the herb. Methods. The objects of the study were 17 common skullcap populations growing wild in eastern Poland. At each natural site, the list of main flowering plant species was carried out, according to Braun-Blanquet scale of plant abundance. The total content of flavonoids was determined according to EP 8th. The analysis of flavonoids was carried out using HPLC, Shimadzu chromatograph. Results. The total content of flavonoids ranged from 0.21 to 0.50% dry matter (DM). Using HPLC, eight compounds were identified within the group, with chrysin-7-O-glucuronide (30.91–589.27 mg × 100 g−1 DM), baicalin (61.90–482.93 mg × 100 g−1 DM) and scutellarin (43.77–213.54 mg × 100 g−1 DM) as dominants. Chrysin was the compound which in the highest degree differentiated investigated populations (CV=0.77). Conclusion. Obtained results indicate that investigated common skullcap populations differed in terms of the total content of flavonoids as well as the content of indentified flavonoids in the herb. However, there was no clear relationship between geographical localization of populations and the content of identified flavonoids in raw material.

Summary Introduction: Purple betony (Stachys officinalis L., Lamiaceae) is a perennial of versatile medicinal usage. Nowadays, in Poland betony herb is collected exclusively from wild growing plants. Decreasing number of its natural sites results in lack of the herb supply and thus, in its limited usage. Objective: The aim of the study was to determine the effects of the age of plant and term of raw material harvest on its yield and quality in cultivation conditions. Methods: The observations were carried out on 2- and 3-year-old plants. During vegetation the herb was collected for four times. The raw material was subjected to chemical analysis. Tannins (as pyrogallol equivalent) were determined according to Polish Pharmacopoeia, phenolic acids and flavonoids – by HPLC. Results: The mass of herb, both in the second and third year, had increased from the beginning of vegetation up to seed setting. The highest content of tannins was found in the herb collected at the vegetative stage of plant development (2.05% in the second and 2.91% in the third year). Four phenolic acids (chlorogenic, ferulic, caffeic and rosmarinic acids) and five flavonoid compounds (orientin, luteolin-7-glucoside, apigenin-7-glucoside, apigenin-3-glucoside, apigenin) were identified in the obtained raw materials. In these groups, the dominant compounds were caffeic acid and apigenin. The highest content of caffeic acid was found at the beginning of plant vegetation, whereas apigenin – at the stage of full blooming and seed setting. Conclusion: In cultivation conditions, purple betony produces high mass of herb which may be used as a valuable raw material in herbal industry.

Greek oregano and common oregano were compared in respect of the antioxidant and antibacterial activity of the corresponding essential oils and hydroethanolic extracts in relation with their chemical profile. The chemical composition of essential oils was determined by GC-MS and GC-FID, while extracts (phenolic acids and flavonoids fractions) were analyzed by HPLC-DAD. Based on given volatiles, the investigated subspecies represented two chemotypes: a carvacrol/γ-terpinene/p-cymene type in the case of Greek oregano and a sabinyl/cymyl type rich in terpinen-4-ol in common oregano. Within non-volatile phenolic compounds, rosmarinic acid appeared to dominate in both subspecies. Lithospermic acid B, chlorogenic acid and isovitexin were present only in Greek oregano extracts. However, the total content of flavonoids was higher in common oregano extracts. The essential oil and extract of Greek oregano revealed visibly stronger antibacterial activity (expressed as MIC and MBC) than common oregano, whereas the antioxidant potential (determined by DPPH, ABTS and FRAP) of these extracts was almost equal for both subspecies. In the case of Origanum plants, the potential application of essential oils and extracts as antiseptic and antioxidant agents in the food industry should be preceded by subspecies identification followed by recognition of their chemotype concerning both terpene and phenolics composition.

Background: There are only a few reports in the literature about translocation of coagulase-negative staphylococci (CoNS) as a primary cause of sepsis in neonates, although CoNS are among a short list of &ldquo, translocating&rdquo, bacteria when present in abundance. Methods: 468 blood samples, 119 stool samples, and 8 catheter tips, from 311 neonates, were tested for presence of microorganisms. CoNS strains isolated from the blood and stool or from blood and catheter tip of the same newborn at approximately the same time were paired and typed with PFGE (Pulse-Field Gel Electrophoresis) method. The strains were then tested for the presence of adherence genes and biofilm formation. Results: The strains with identical PFGE profiles in comparison to those with non-identical profiles differed in terms of the pattern of the virulence genes and showed a lack of the genes related to adherence, but more often presence of IS256, which is related to virulence. They also were phenotypically unable to adhere to intestinal Caco2 cells. Conclusions: A considerable proportion of CoNS strains isolated from bloodstream of VLBW/LWB neonates was identical to the strains isolated from faeces of the same neonates at the same time. These observations may offer indirect evidence indicating that at least some CoNS can translocate from the gastrointestinal tract of the premature neonates into the bloodstream and thus cause generalized infection.

Common thyme is regarded as one of the most important culinary plants. The purpose of the work was to determine the intraspecific variability of common thyme with respect to morphological and chemical characters including the content and composition of essential oil and phenolic compounds in the herb. The objects of the study were 12 clones, vegetatively multiplied from randomly selected individual plants of cultivar &lsquo, Standard Winter&rsquo, The morphological observations and harvest of raw materials were carried out in the first year plants&rsquo, vegetation. The highest differences between clones were on fresh and dry weight of herb (CV = 0.38 and 0.36, respectively), width of leaves (CV = 0.21), and density of glandular trichomes on the abaxial surface of leaves (CV = 0.29). Examined clones were also differentiated as to the chemical features. Essential oil content (performed by hydrodestillation) and composition (by GC-MS and GC-FID) were determined and they ranged from 2.10 to 4.38 g &times, 100 g&minus, 1 DW. Here, thymol, &gamma, terpinen, and p-cymen were the dominant compounds. Clone no 4 was distinctive as to the highest content of essential oil followed by the highest share of thymol (54.59%). The total content of phenolic acids and flavonoids (determined according to PPh 6th) also differed among clones (CV = 0.38 and 0.36, respectively). Using a validated HPLC-DAD method, the following compounds were identified: caffeic, rosmarinic, p-coumaric acids, luteolin 7-O-glucoside, naryngenin, and (-)-epicatechin. Here, rosmarinic acid followed by luteolin 7-O-glucoside were present in the highest amounts (611.47&ndash, 2675.59 and 46.77&ndash, 325.11 mg &times, 1 DW, respectively). The highest differences between clones were the contents of p-coumaric acid (CV = 0.59), luteolin 7-O-glucoside (CV = 0.50) and rosmarinic acid (CV = 0.40). Such a high range of variability can provide problems with raw material standardization. Nevertheless, it opens possibilities for breeders, whereas individual plants/clones may become valuable components for breeding.

The aim of the study was to assess the effects of Melittis melissophyllum shading on its development and accumulation of phenolics. Their content (verbascoside, apiin, luteolin-7-O-glucoside, coumarin, 3,4-dihydroxycoumarin, o-coumaric acid 2-O-glucoside as well as o-coumaric, p-coumaric, chlorogenic, caffeic, ferulic and cichoric acid) was determined in the herb using HPLC-DAD. The results showed that the content of abovementioned flavonoids and phenolic acids was highest in plants grown under full sunlight. On the other hand, a higher content of coumarin was observed in shaded plants, especially after the seed-setting stage. A similar tendency was noted for the amount of chlorophyll a and b. The content of hydrogen peroxide and malondialdehyde, the activity of polyphenol oxidase and catalase and the antioxidant capacity of plant extracts (measured using DPPH, ABTS and FRAP assays) were found to be the highest in the plants grown in full sunlight. However, the plants grown in moderate (30%) shade were found to thrive best.

The aim of the study was to investigate the chemical variability of small-leaved lime ( Tilia cordata Mill.) growing wild in the eastern area of Poland, in terms of content and composition of phenolic compounds (phenolic acids and flavonoids), composition of essential oil and content of mucilage in flowers. Analyzed populations differed significantly when regards the accumulation of biologically active compounds. The total content of phenolic acids ranged from 0.17 to 1.87 g × 100 g −1 , flavonoids from 0.09 to 0.52 g × 100 g −1 , and mucilage from 0.6 to 3.0 g × 100 g −1 . Using validated HPLC-DAD method, the chemical composition of phenolic acids and flavonoids were determined. Among phenolic acids, four compounds were identified with vanillic acid as the dominant one (391.5–1718.0 mg × 100 g −1 ). Among flavonoids, nine compounds were found. Rutoside and (−)-epicatechin (tannin precursor) were present in the highest amounts (1264.1–4654.6 mg × 100 g −1 and 1000.3–4745.6 mg × 100 g −1 , respectively). GC–MS and GC-FID were used to perform the qualitative and quantitative analyze of the essential oil. The major compounds were alkanes: tricosane (5.7–20.7%), pentacosane (3.0–6.1%) and heneicosane (1.1–6.1%), as well as acetophenon (3.5–6.9%), nonanal (1.3–7.8%) and nonanoic acid (3.9–18.2%).

Summary An efficient method for in vitro propagation of bastard balm by enhanced axillary shoot branching has been developed. The material to establish in vitro culture was shoot tips collected from three-year-old plants in May. The shoots obtained from initial explants were placed on MS/B5 medium containing 0.1, 0.5 or 1.0 mg/l BA with 0.01 mg/l NAA or without the auxin. The highest number of shoots per explant was obtained on the medium with 1.0 mg/l BA (3.9 shoots per explant). For the rooting of shoots ½ MS/B5 with IBA (0.25, 0.50 and 1.0 mg/l) medium was used. The medium without plant growth regulators served as a control. The best root regeneration was observed on the medium without IBA (87.1% of cuttings rooted). IBA used in the medium for shoot rooting affected the morphological traits of obtained microcuttings but not affected their weight. Irrespective of auxin concentration in this medium, obtained microcuttings acclimated in ex vitro conditions very well.

The objective of this study was to investigate the composition of polyphenolic compounds of water and ethanol extracts from leather bergenia leaves (LBL) and to determine the antimicrobial properties of pullulan coating enriched with these extracts. The result showed that both extracts consisted mainly of flavonols and phenolic acids. The antimicrobial activity of pullulan coating solution was determined at different concentrations: 0.4, 1, 2, 5, 10, 20% (w/v) of extracts. The incorporation of LBL extracts showed antimicrobial activities of the films against both Gram-positive and Gram-negative bacteria and molds. Pullulan films incorporated with 20% of LBL extracts showed the strongest antimicrobial activity. In the next part of the study pullulan coating solution with a 20% concentration of water LBL extract was chosen and used as coating on pepper and apple surfaces. Non-coated samples of food were also used as controls. Samples coated with the antimicrobial solutions showed advantages in reducing microbial growth by 1 log CFU/g as compared to the control samples. Physico-chemical and organoleptic observations were also evaluated after storage. The application of the pullulan film enabled an extension of the storage time of the peppers and apples as a result of reduced weight losses and helped to maintain their hardness.

Summary Southern sweet-grass (Hierochloë australis /Schrad./ Roem. & Schult.) is a perennial, wild growing tuft grass occurring in North-East Poland, Belarus and Finland. In Poland the species is under the partial legal protection. The raw material harvested from this plant are leaves rich in coumarins, mainly in coumarin responsible for specific sweet aroma of leaves. They are used mostly for the aromatization of alcohol and tobacco products. Due to high demand for the raw material and decrease in the natural resources of the species, it is advisable to introduce the plant into cultivation. In the presented study vegetative planting stock (1-, 2-, and 4-stem cuttings) were used to set the plantation of southern sweet-grass. The influence of the planting stock type on the mass of leaves and their quality in the first and second year of plant vegetation as well as the mass of seeds from two-year-old plants were investigated. The highest number of well rooted plants was obtained from 4-stem cuttings (74.07%) and the least - from 1-stem cuttings (47.53%). Both, on one- and two-year-old plantations the plants from 4-stem cuttings were characterized by the highest mass of leaves (7.73 and 24.65 g ˟ plant-1, respectively). The plants were also characterized by the highest number of generative shoots (40.71 pcs. ˟ plant-1) and mass of seeds (4.62 g ˟ plant-1). The total contents of coumarins and phenolic acids did not depend on the type of planting stock. The contents of these compounds was higher in two-year-old plants than in one-year-old ones, whereas the content of flavonoids was higher in one-year-old plants.

The significant issue for Polish economy is to fulfil the requirements of the EU's climate and energy policies according to the share of renewable energy sources (RES) in total energy production by 2020 should be equal to 15%. At the end of 2016, the share of RES in total energy production in Poland was only 13.54%. The development of Virtual Power Plant (VPP) is the chance to complete the goal set for 2020 and to increase the development of RES. The VPP is an IT structure which integrates different types of distributed energy sources, flexible consumers and energy storage with each other and with other market segments in real time through a smart grid. The VPP has positive effects on the electricity grid, like the improving energy security as well as grid stability and reliability, optimizing energy resources use, lowering the load – demand response and giving the possibility to manage of electricity surplus in small dispersed energy sources. The main aim of the article is the proposal of a new business model for Virtual Power Plant in Polish conditions. The main aim is accomplished by secondary aims such as (i) describing the concept of virtual organizations, (ii) the review of business models and (iii) the presentation of the legal capabilities to create the virtual power plants. First of all, the literature studies made by authors, point out there is a lack of complex analysis related to the establishing the VPP and its business model. The complex analysis means the analysis consists of technical, legal, economic and ecological aspects. The literature review provides information about virtual organizations in the context of the virtual power plant. The VPP is one of the types of virtual organizations, but the definitions of a VPP are primarily focused on its technical aspect and insufficient attention is paid to the aspect of management and, especially, to a business model. Therefore, in this paper, the legal background to establish the VPP has presented as well as the legal opportunities and threats for creating the VPP in Poland. The legal analysis is the starting point for each practical project. There are shown the legal regulations related to prosumers and supporting energy micro-clusters. These issues require the creation of new products, such as VPP. Next, the review of the business model was made to choose an adequate model. Three models emerge from the analysis, which can be used to build a VPP business model to a greater or lesser extent. The first of them emphasize that the business model is mainly the characteristic relations between customers, clients, partners and suppliers. The second of them underlines the significance of the company's resources, which can be expanded and used (new customers – new dispersed energy sources), as well as potential sources of future economic benefits (new products – power market, megawatts). The third of them combines the key resources and the key relations which are important for VPPs, and it is compliant with the systemic approach. This approach was chosen for further analysis. The authors presented a new business model concept, where the attention is paid to a cooperation network between different kinds of energy entities (e.g. VPP’ owners, prosumers, the owners of micro-installations and others). The segmentation of potential clients of the VPP was made and eight potential segments were identified in Polish conditions. Two segmentation criteria were used: (i) the kind of product, (ii) the kind of market. The business model is perceived as a method of increasing and exploiting of company's resources for preparing new products or services for customers in order to obtain the added value (expanding the competitive advantage or increasing profitability). The examples of added values were defined in relating to the identified VPP client segments: (i) a large buyer buying energy under a long-term bilateral contract, (ii) an energy cooperative, (iii) an owner of micro-installation, (iv) a company that owns energy storage, e.g. hybrid vehicle charging stations, storages, (v) energy cluster. Additionally, for all customer segments there are specified (1) relations which connect VPP with the representatives of a specific customer segment and (2) communication channels which significantly affect the final impression on the client. In the knowledge economy, the IT tools are the main kind of communication channel, and IT resources are the key function in the VPP business model. Future research should focus on describing the remaining elements of the business model for each customer segments such as revenue streams, key resources, key activities, key partners and cost structure. In the next step economic capabilities to create the VPP should be analysed with special attention paid to the evaluation of the economic efficiency of the VPP. Different scenarios including consumer segment and value proposition should be taken into consideration. It is necessary to perform an economic analysis of the data coming from a real pilot installation. Значною проблемою польської економіки є виконання вимог кліматичної та енергетичної політик ЄС, у відповідності до яких частка відновлюваних джерел енергії у загальному обсязі виробництва енергії до 2020 року повинна становити 15%. Так, наприкінці 2016 р. частка відновлюваних джерел енергії у Польщі становила лише 13,54%. У рамках даного дослідження, автори розглядають розвиток віртуальних електростанцій як можливість збільшення частки відновлюваних джерел енергії в загальній структурі виробництва електроенергії. Автори визначають віртуальну електростанцію як ІТ-структуру, яка поєднує різні види джерел енергії, споживачів та виконує функції зберігання, обміну або продажу як в самій системі, так із іншими сегментами ринку в режимі реального часу за допомогою смарт-мережі. Позитивний вплив віртуальних електростанцій на електромережу полягає у забезпеченні енергетичної безпеки, стабільності та надійності експлуатації, оптимізації використання енергетичних ресурсів, зниженні навантаження та балансуванні надлишком електроенергії в малих розподільних електромережах. Головна мета статті – розробити нову бізнес-модель віртуальної електростанції в умовах Польщі. Для досягнення поставленої мети було встановлено наступні завдання: (i) опис концепції віртуальних організацій, (ii) огляд наявних бізнес-моделей та (iii) правових можливостей для створення віртуальних електростанцій. Автори досліджують основні нормативно-правові акти, які регулюють права споживачів та особливості функціонування мікрокластерів, що підтримують ідею створення інноваційних продуктів та послуг на енергетичному ринку, таких як віртуальні електростанції. У статті представлена нова концепція бізнес-моделі, в якій увагу приділено мережі співпраці між різними видами енергетичних об'єктів. Нова концепція розглядається як інструмент збільшення та оптимізації використання наявних ресурсів компанії для створення нових продуктів або послуг відповідно до потреб енергетичного ринку з метою отримання додаткової вартості (збільшення конкурентних переваг або підвищення рентабельності).

Seven crystal structures of alanyl aminopeptidase from Neisseria meningitides (the etiological agent of meningitis, NmAPN) complexed with organophosphorus compounds were resolved to determine the optimal inhibitor–enzyme interactions. The enantiomeric phosphonic acid analogs of Leu and hPhe, which correspond to the P1 amino acid residues of well-processed substrates, were used to assess the impact of the absolute configuration and the stereospecific hydrogen bond network formed between the aminophosphonate polar head and the active site residues on the binding affinity. For the hPhe analog, an imperfect stereochemical complementarity could be overcome by incorporating an appropriate P1 side chain. The constitution of P1′-extended structures was rationally designed and the lead, phosphinic dipeptide hPhePψ[CH2]Phe, was modified in a single position. Introducing a heteroatom/heteroatom-based fragment to either the P1 or P1′ residue required new synthetic pathways. The compounds in the refined structure were low nanomolar and subnanomolar inhibitors of N. meningitides, porcine and human APNs, and the reference leucine aminopeptidase (LAP). The unnatural phosphinic dipeptide analogs exhibited a high affinity for monozinc APNs associated with a reasonable selectivity versus dizinc LAP. Another set of crystal structures containing the NmAPN dipeptide ligand were used to verify and to confirm the predicted binding modes; furthermore, novel contacts, which were promising for inhibitor development, were identified, including a π–π stacking interaction between a pyridine ring and Tyr372.

Summary Southern sweet-grass (Hierochloë australis /Schrad./ Roem. & Schult., Poaceae), commonly known as a bison grass, is a perennial tuft grass rarely occurring in mixed forests in Eastern and Northern Europe. The raw material collected from this plant are leaves rich in coumarin compounds (especially coumarin) responsible for sweet, specific aroma of these organs. In Poland, southern sweet-grass leaves are used mainly for alcohol products aromatisation. Growing demand for the raw material results in uncontrolled and excessive collecting of this plant. The best way to solve this problem is to introduce this plant into cultivation. Since southern sweet-grass is allogamous and heterozigotic, strong intraspecific variability of the species is observed. Fourteen clones of selected individuals from one population of southern sweet-grass naturally occurring in East Poland were compared in respect of morphological traits as well as accumulation of biologically active compounds. The clones differed significantly in the weight of leaves (3.76-22.59 g of air-dry weight per plant). The total coumarin content (determined by a spectrophotometric method) in this raw material for investigated clones ranged from 1.49 to 1.94%, flavonoids - from 0.25 to 0.55%, and phenolic acids - from 0.20 to 0.42%. Three coumarin compounds were identified by HPLC, namely coumarin, 3,4-dihydrocoumarin, and o-coumaric acid. The content of coumarin ranged from 84.00 to 310.85, 3,4-dihydrocoumarin: from 17.80 to 168.45, and o-coumaric acid: from 37.50 to 70.00 mg · 100 g-1 dry matter.

The study involved an examination of the antifungal activity on red peppers of pullulan coating (P) and pullulan coating containing either water-ethanol (P + eEMF) or ethanol extract of meadowsweet flowers (P + eEMF). Pullulan was obtained from a culture of Aureobasidium pullulans B-1 mutant. Both non-inoculated peppers and those artificially inoculated with Rhizopus arrhizus were coated and incubated at 24 °C for 5 days. The intensity of the decay caused by Rhizopus arrhizus in the peppers with P and P + eEMF coatings was nearly 3-fold lower, and in the case of P + weEMF 5-fold lower, than that observed in the control peppers. Additionally, the P + weEMF coating decreased, almost two-fold the severity of pepper decay compared to other samples. The influence of coating of pepper postharvest quality was examined after 30 days of storage at 6 °C and 70%–75% RH. All coatings formed a thin and well-attached additional layer of an intensified gloss. During storage, color, total soluble solid content and weight loss of coated peppers were subject to lower changes in comparison with uncoated ones. The results indicate the possibility of the application of pullulan coatings containing MFEs as an alternative to the chemical fungicides used to combat pepper postharvest diseases.

The antimicrobial and antioxidant activities of pullulan films containing sweet basil extract (SBE) at varying concentrations were examined in the paper. Additionally, the content of bioactives in SBE was determined. A strong correlation was noted between SBE concentrations in the film and the obtained film thickness. Pullulan film containing SBE was characterized by a smooth surface as well as a uniform and compact internal structure. The study involved an examination of the effectiveness of pullulan coatings containing 24 mg SBE/cm 2 on the prolongation of the shelf life of “Jonagored” apples. Pullulan coating with SBE was found to offer low antibacterial activity against mesophilic bacteria and good antifungal protection against Rhizopus arrhizus on apple surfaces. This coating also contributed to a reduction in weight losses and lower changes in the color and soluble solids of fruits during storage. Apples with pullulan enriched SBE coating presented better overall preference parameters. Industrial relevance The consumers express an increasing aversion and objections towards an application of chemical means in order to prolong the durability of food products. This prompts food producers to search for new methods of preventing raw materials against harmful microbial activity. Plant extracts, due to their chemical variability, ensure unlimited possibilities of microorganism growth control. An application of natural plant extracts combined with edible coatings for fruit and vegetable durability prolongation may find an acceptance among the consumers. Incorporation of plant extract to edible coating ensures slow release of bioactives on food surface, i.e. at the site of the highest microbiological food changes. Pullulan is a natural polysaccharide, a carrier of small energy amounts, forming glossy, colorless and flavorless films, resistant to oils and impermeable for oxygen. The authors decided to elaborate the procedure concerning an application of edible pullulan coating combined with sweet basil extract for an elongation of apple durability, which will improve the quality of fresh fruits during their storage and in the trade.

This study examined the antimicrobial activity of pullulan films with incorporated meadowsweet flower extracts (EMFs) and the effectiveness of these coatings in the prolonging of apple stability in various storage conditions. Pullulan was obtained from a batch culture of white mutant Aureobasidium pullulans B-1. The content of phenolic components in two extracts-water–ethanol (weEMF) and ethanol one (eEMF)-was compared using the HPLC method. Compared to eEMF, a significantly higher content was noted of phenolic acids and flavonoids in weEMF and its increased activity in inhibiting microorganism growth. Pullulan films with incorporated EMFs were obtained from the solutions to which they were added in concentrations of 3, 6, 9, 12 and 15%. These films were stable and homogenous, and to a higher degree inhibited the growth of tested bacteria than fungi. Apples of the Jonagored variety were coated with a pullulan coating containing 12% EMF. The pullulan coating with EMFs evened and unified the surface of apple skins. Pullulan coating with weEMF more effectively inhibited the growth of natural mesophilic microflora, the amount of which decreased by two logarithmic cycles at the end of storage. EMF addition to the coating significantly limited the growth of the Rhizopus arrhizus mold inoculated to the apples. EMF addition to pullulan coating did not affect color changes nor did apples loss weight during their storage. Pullulan coatings with incorporated EMFs may protect apple quality during their storage and turnover in various temperatures and time conditions.

Sweet basil is one of the most important culinary herbs. Currently, its production is carried out mainly in accordance with conventional agriculture. However, its cultivation in organic systems seems to be better adjusted to consumer demands connected with the lack of pesticide residues in foods and their safety. In the present study, two methods of basil cultivation in organic farming system were applied, i.e., in the open air and under foil tunnels. During the experiment, in central European climatic conditions, it was possible to obtain four successive cuts of herb. The herb was subjected to chemical analysis, including determination of the content of essential oil, phenolic compounds, and chlorophylls. Gas chromatography coupled with mass spectrometry (GC&ndash, MS) and flame ionization detector (GC-FID) analysis of the essential oil was performed, whereas the fresh herb was subjected to sensory analysis. The cumulative mass of fresh herb was distinctly higher in the cultivation under foil tunnels (44.7 kg∙10 m&minus, 2) in comparison to the open field (24.7 kg∙10 m&minus, 2). The content of essential oil, flavonoids, and phenolic acids was also higher in the raw material collected from plants grown under foil tunnels (0.81, 0.36, and 0.78 g&middot, 100g&minus, 1 DW, respectively) than from the open field (0.48, 0.29, and 0.59g&middot, 1 DW, respectively). In turn, the dominant compound of the essential oil, i.e., linalool, was present in higher amounts in the essential oil obtained from plants cultivated in the open field. The sensory and microbiological quality of herb was comparable for both methods of cultivation. The obtained results indicate that, in central European climatic conditions, it is possible to obtain good-quality yield of basil herb. However, for its better productivity, it seems that cultivation under foil tunnels is preferable.

Cyclooxygenase (COX) and lipoxygenase (LOX) are key enzymes of arachidonic acid metabolism. Their products, prostaglandins and leukotrienes, are involved in the pathogenesis of inflammatory bowel diseases and colorectal cancer. The aim of the study was to examine the influence of inositol hexaphosphate (IP6), a naturally occurring phytochemical, on the expression of genes encoding COX and LOX isoforms and synthesis of their products (PGE2 and LTB4) in colon cancer cell line Caco-2 stimulated with pro-inflammatory agents (IL-1β/TNFα). Real-time RT-qPCR was used to validate mRNAs level of examined genes. The concentrations of COX-2 and 5-LOX proteins as well as PGE2 and LTB4 were determined by the ELISA method. Based on these studies it may be concluded that IP6 may limit inflammatory events in the colonic epithelium and prevent colon carcinomas by modulating the expression of genes encoding COX and LOX isoforms at both mRNA and protein levels as well as by affecting the synthesis and secretion of prostaglandins and leukotrienes.

Primula veris L. and Primula elatior (L.) Hill represent medicinal plants used for the production of herbal teas and preparations with antioxidant and expectorant activity. Flowers and roots of both species possess the same biological activity. In the presented study, raw materials of wild growing P. veris and P. elatior were compared in terms of the content and composition of phenolic compounds using a fast and simple HPLC-DAD method. The study showed that flowers of both species were rich in flavonoids. However, P. veris flowers were characterized with a distinctly higher content of isorhamnetin-3-O-glucoside, astragalin, and (+)-catechin, whereas P. elatior occurred to be a richer source of rutoside and isorhamnetin-3-O-rutinoside. Hyperoside was found exclusively in P. elatior flowers. Phenolic glycosides (primverin and primulaverin) were identified only in the roots. Their content was about ten times higher in P. veris in comparison with P. elatior underground organs. The obtained results clearly show that both Primula species differ distinctly in terms of the content and composition of phenolic compounds. The compounds differentiating both species to the highest degree (hyperoside, in flowers, as well as primverin and primulaverin, in the roots) may be useful chemical markers in the identification and evaluation of both species.

Spinal cord injuries are still a serious problem for regenerative medicine. Previous research has demonstrated that activated microglia accumulate in spinal lesions, influencing the injured tissues in various ways. Therefore, transplantation of activated microglia may have a beneficial role in the regeneration of the nervous system. The present study examined the influence of transplanted activated microglial cells in adult rats with injured spinal cords. Rats were randomly divided into an experimental (M) and control (C) group, and were subjected to non-laminectomy focal injury of spinal cord white matter by means of a high-pressured air stream. In group M, activated cultured microglial cells were injected twice into the site of injury. Functional outcome and morphological features of regeneration were analyzed during a 12-week follow-up. The lesions were characterized by means of magnetic resonance imaging (MRI). Neurons in the brain stem and motor cortex were labeled with FluoroGold (FG). A total of 12 weeks after surgery, spinal cords and brains were collected and subjected to histopathological and immunohistochemical examinations. Lesion sizes in the spinal cord were measured and the number of FG-positive neurons was counted. Rats in group M demonstrated significant improvement of locomotor performance when compared with group C (P

The aim of the investigation was to compare 10 populations of great burnet growing wild in Poland in respect of phenolic compounds accumulation. Total content of tannins and phenolic acids were determined according to Polish Pharmacopoeia and qualitative analysis by HPLC. District differences in the content of determined phenolic compounds both between the populations originating from different regions of Poland and between the populations from the same region were observed. Differences in the content of particular phenolic compounds between herb and roots were found too. In herb five phenolic acids were identified, namely: chlorogenic, ellagic, gallic, caffeic, and rosmarinic acid, whereas in underground organs two phenolic acids (ellagic and gallic) and four catechin derivatives (/+/-catechin, /-/-epigallocatechin, /-/-epicatechin, and /-/-epigallocatechin gallate). Dominating phenolic compounds in herb were gallic and chlorogenic acids, whereas in underground organs - gallic acid, catechin and epicatechin.

Introduction The main goal of this study was to examine the patient age and sex dependent expression of KCNQ1 and HERG genes that encode potassium channels responsible for the occurrence of long QT syndrome (LQTS). Material and methods The study enrolled 43 families whose members suffered from LQTS type 1 (LQTS1) or 2 (LQTS2) or were healthy. The study attempted to prove that β-actin is a good endogenous control when determining the expression of the studied genes. Examination of gene expression was achieved with quantitative real-time PCR (QRT-PCR). Expression of the investigated genes was inferred from the analysis of the number of mRNA copies per 1 μg total RNA isolated from whole blood. Results Significantly lower KCNQ1 and KCNH2 mRNA levels in healthy females than healthy males were observed (p = 0.032; p = 0.02). In male patients both transcripts were expressed at a lower level (p = 0.0084; p = 0.035). The comparison of transcriptional activity of KCNQ1 and KCNH2 in healthy adults and children revealed higher KCNQ1 and lower KCNH2 mRNA levels in healthy adults (p = 0.033; p = 0.04), higher KCNQ1 and lower KCNH2 mRNA levels in adult patients below 55 years old than in adults over 55 years old (p=0.036; p = 0.044), and significantly higher KCNQ1 and lower KCNH2 mRNA levels in adult patients (over 55 years) than in paediatric patients (below 15 years) (p=0.047; p = 0.08). Conclusions The results support the hypothesis that KCNQ1 and HERG gene expression is influenced by age and gender in human patients with long QT syndrome and in healthy subjects.

Roseroot (Rhodiola rosea L.) belongs to plants revealing adaptogenic properties, which are attributed to the presence of specific phenolic compounds and are reflected mainly as antioxidant activity. The aim of the present study was to determine the antioxidant and antibacterial activity of various products obtained from R. rosea (underground organs as well as their aqueous and ethanolic dry extracts) in relation to the chemical profiles of phenolic and essential oil compounds. The chemical profiles were determined by High-performance Liquid Chromatography with a diode-array detector (HPLC-DAD) and Gas chromatography-mass spectrometry (GC-MS), antioxidant activity by (1,1-Diphenyl-2-picrylhydrazyl) Scavenging Capacity Assay (DPPH), (2,2&prime, Azino-bis(3-ethylbenzothiazoline-6-sulphonic acid)) Scavenging Capacity Assay (ABTS) and Ferric Reducing Antioxidant Power Assay (FRAP) and antimicrobial properties were expressed as minimum inhibitory concentration (MIC) and minimum bacterial concentration (MBC) values following the broth microdilutions method. The results show that the investigated samples differed in terms of their chemical compositions and biological activities. The extracts were more abundant in phenolic compounds (salidroside, tyrosol, and rosavin derivatives) in comparison to dried underground organs. The content of the determined phenolics in the analyzed extracts was affected by the solvent used for extraction. The ethanolic extract was characterized by the highest content of these substances in comparison to the aqueous one and the dried raw material, especially with regard to rosavin (969.71 mg/100 g). In parallel, this extract showed the strongest antioxidant and antibacterial activity. However, dried R. rosea underground organs also revealed strong antibacterial effects against, for example, Staphylococcus strains.

Eleutherococcus senticosus (Rupr. et Maxim.) Maxim. is a thorny shrub, belonging to the Araliaceae family, occurring primarily in Russia, China, Korea and Japan. In traditional medicine underground parts of this plant are used as a drug with adaptogenic activity. Eleutherosides are the most important biologically active compounds responsible for this action. The content of eleutherosides and polyphenolic acids in rhizomes, roots and stem bark was determined to assess the chemical varibility between ten selected E. senticosus clones. HPLC analyses showed significant differences between clones in the content of eleutherosides B and E. The highest content of these compounds was observed in the stem bark (from 207.2 to 566.0 mg 100 g -1 ), lower in the rhizomes (from 47.5 to 326.2 mg 100 g -1 ) and the lowest in roots (from 38.9 to 228.6 mg 100 g -1 ). In all three investigated plant organs presence of chlorogenic, caffeic, rosmarinic, ferulic, protocatechuic acids and rutoside was determined as well.

The aim of this investigation was to determine the genetic and chemical variability of bennet's herb (Geum urbanum) growing wild in the eastern and central areas of Poland. HPLC was applied for determination of phenolic compounds. Two flavan-3-ols and three polyphenolic acids were found in leaves, while five flavan-3-ols and two polyphenolic acids in underground organs. The populations differed especially in respect of the content of catechin and epicatechin in underground organs. The RAPD analysis indicated genetic variability of investigated populations but there was no clear correlation between this variability and the chemical composition of determined phenolic agents.

Common sage (Salvia officinalis L., Lamiaceae family) is a perennial evergreen subshrub native to the Mediterranean. Leaves collected from this plant are used as an important medicinal raw material with antiseptic, antispasmodic, astringent, carminative, cholagogue and tonic activity. The aim of the research was to evaluate the intraspecific variability of sage with special respect to seed setting and germination. The obtained seeds mass fluctuated from 17.0 to 25.5 g per plant. From a single flower 3 seeds were obtained, on average. A clear relationship between the position of seeds in inflorescence and their germinability was observed. The germinability of seeds obtained from upper whorls fluctuated from 92 to 98% whereas germinability of seeds from the lowest one was 70%. There were no distinct differences between plants in the 1000-grain weight. The mass of 1000 grains ranged from 6.090 to 6.161 g.

Chemical diversity of silverweed (Potentilla anserinal L.) growing at the edges of arable fields The medicinal usage of silverweed herb (Anserinae herba) is primarily related to the presence of phenolic compounds. Three populations of silverweed (Potentilla anserina L.) occurring at the edges of arable fields in Podlasie area (north-eastern Poland) were compared in respect of the content and composition of phenolics. Herb collected from the investigated populations differed in the content of determined compounds. The tannin content was very low (0.81-0.91%). The total flavonoid content ranged from 0.48 to 0.60%. Phenolic acid content was much higher (1.38-2.26%). Three flavan-3-ols: (+)-catechin, (-)-epicatechin, and (-)-epigallocatechin, four flavonol glycosides: rutoside, quercetin-3-O-glucoside, isorhamnetin-3-O-glucoside, and kaempferol-3-O-glucoside, as well as one phenolic acid (ellagic acid) were identified as a result of HPLC analysis of methan-olic extracts from the investigated raw materials. The most considerable differences between populations concerned the content of (+)-catechin (87.0-199.3 mg x 100 g-1), (-)-epigallocatechin (141.9-290.6 mg x 100 g-1), and rutoside (192.3-386.0 mg x 100 g-4) in herb.