How Are the Flower Structure and Nectar Composition of the Generalistic Orchid Neottia ovata Adapted to a Wide Range of Pollinators?

Plant-pollinator interactions significantly influence reproductive success (RS) and drive the evolution of pollination syndromes. In the context of RS, mainly the role of flower morphology is touched. The importance of nectar properties is less studied, despite its significance in pollination effectiveness. Therefore, the aim of this study was to test selection on flower morphology and nectar chemistry in the generalistic orchid Neottia ovata. In 2019–2020, we measured three floral displays and six flower traits, pollinaria removal (PR), female reproductive success (FRS), and determined the soil properties. The sugars and amino acids (AAs) were analyzed using the HPLC method. Data were analyzed using multiple statistical methods (boxplots, ternary plot, one-way ANOVA, Kruskal-Wallis test, and PCA). Variation of flower structure and nectar chemistry and their weak correlation with RS confirms the generalistic character of N. ovata. In particular populations, different traits were under selection. PR was high and similar in all populations in both years, while FRS was lower and varied among populations. Nectar was dominated by glucose, fructose, and included 28 AAs (Ala and Glu have the highest content). Sugars and AAs influenced mainly FRS. Among soil parameters, carbon and carbon:nitrogen ratio seems to be the most important in shaping flower structure and nectar chemistry.     

Effects of quinoa and amaranth on zinc,magnesium and calcium content in beer wort

Pseudocereals such as quinoa and amaranth can be used as partial replacements for malt in the production of new beers. Quinoa and amaranth are of interest due to their high levels of nutrients and micronutrients, including minerals which can significantly improve the performance of brewing yeast and the fermentation rate. In this study, we investigated whether the use of quinoa or amaranth as partial replacements for malt affected the concentration of ions such as zinc and magnesium in beer wort. The use of amaranth, and in particular quinoa, increased the content of both zinc and magnesium ions substantially, even when only 10% of the barley malt was replaced. With 10% quinoa, Zn²⁺ and Mg²⁺ content increased by 41% and 49%, respectively, while the ratio of Mg²⁺ to Ca²⁺ rose from 1.4:1 to 1.9:1. Use of unmalted quinoa and amaranth appears a good way of enriching wort with essential metal ions.     

A Human Cellular Model for Colorectal Anastomotic Repair: The Effect of Localization and Transforming Growth Factor-β1 Treatment on Collagen Deposition and Biomarkers

Anastomotic leakage (AL) is a devastating complication after colorectal surgery, possibly due to the loss of stabilizing collagen fibers in the submucosa. Our aim was to assess the formation of collagen in the colon versus the rectum with or without transforming growth factor (TGF)-β1 exposure in a human cellular model of colorectal repair. Primary fibroblasts were isolated by an explant procedure from clinically resected tissue rings during anastomosis construction in 19 consecutive colorectal patients who underwent laparoscopy. The cells, identified as fibroblasts by morphologic characteristics and flow cytometry analysis (CD90⁺), were cultured for 8 days and in 12 patients in the presence of 1 ng/mL TGF-β1. Total collagen deposition was measured colorimetrically after Sirius red staining of fixed cell layers, and type I, III, and VI collagen biosynthesis and degradation were specifically determined by the biomarkers PINP, PRO-C3, PRO-C6, and C3M in conditioned media by competitive enzyme-linked immunosorbent assays. Total collagen deposition by fibroblasts from the colon and rectum did not significantly differ. TGF-β1 treatment increased PINP, PRO-C6, and total collagen deposition. Mechanistically, TGF-β1 treatment increased COL1A1 and ACTA2 (encoding α-smooth muscle actin), and decreased COL6A1 and MMP2 mRNA levels in colorectal fibroblasts. In conclusion, we found no effect of anatomic localization on collagen production by fibroblasts derived from the large intestine. TGF-β1 represents a potential therapeutic agent for the prevention of AL by increasing type I collagen synthesis and collagen deposition.     

Solvates of New Arylpiperazine Salicylamide Derivative-a Multi-Technique Approach to the Description of 5 HTR Ligand Structure and Interactions

A new ligand for 5-HT1A and 5-HT7 receptors, an arylpiperazine salicylamide derivative with an inflexible spacer, is investigated to identify preferred fragments capable of creating essential intermolecular interactions in different solvates. To fully identify and characterize the obtained crystalline materials, various methods including powder and single-crystal X-ray diffraction, solid-state NMR, and thermal analysis were employed, supplemented by periodic ab initio calculations. The molecular conformation in different solvates, types, and hierarchy of intermolecular interactions as well as the crystal packing were investigated to provide data for future research focused on studying protein–ligand interactions. Based on various methods of crystal structure analysis, including the interaction energy calculation and programs using an artificial neural network, a salicylamide fragment was found to be crucial for intermolecular contacts, mostly of dispersion and electrostatic character. A supramolecular 2D kite-type layer of {4,4} topology was found to form in crystals. The closed voids between layers contain disordered solvents, very weakly interacting with the molecule and the layer. It has been postulated that the separation of the layers might be influenced by an increase in temperature or the size of the solvent; hence, only methanol and ethanol hemi-solvates could be obtained from a series of various alcohols.     

DNA ADDUCTS OF BENZO[A]PYRENE- AND DIBENZO[A,L]PYRENE-DIOL EPOXIDES IN HUMAN LUNG EPITHELIAL CELLS: KINETICS OF ADDUCT REMOVAL,EFFECTS ON CELL CYCLE CHECKPOINTS,AND GENE EXPRESSION

The fjord-region PAH dibenzo[a,l]pyrene (DBP) is considerably more carcinogenic than the bay-region benzo[a]pyrene (BP). This fact can be ascribed to differences in DNA binding efficiency of their ultimate carcinogenic diol epoxide (DEs) intermediates, differences in structural features of the DNA adducts, and differences in DNA adduct recognition and the subsequent lesion removal by nucleotide excision repair (NER). In order to further substantiate previous findings in cell-free human systems (1 Buterin, T., Hess, M. T., Luneva, N., Geacintov, N. E., Amin, S., Kroth, H., Seidel, A. and Naegeli, H. 2000. Unrepaired Fjord Region Polycyclic Aromatic Hydrocarbon-DNA Adducts in Ras Codon 61 Mutational Hot Spots. Cancer Research, 60: 1849–1856.  [Google Scholar]), cultured cells (2 Luch, A., Kudla, K., Seidel, A., Doehmer, J., Greim, H. and Baird, W. M. 1999. The Level of DNA Modification by (+)-syn-(11S,12R,13S,14R)- and (?)-anti-(11R,12S,13S, 14R)-Dihydrodiol Epoxides of Dibenzo[a,l]pyrene Determined the Effect on the Proteins p53 and p21WAF1 in the Human Mammary Carcinoma Cell Line MCF-7. Carcinogenesis, 20: 859–865.  [Google Scholar]), and in rodents (3 Jankowiak, R., Ariese, F., Hewer, A., Luch, A., Zamzow, D., Hughes, N. C., Phillips, D., Seidel, A., Platt, K. L., Oesch, F. and Small, G. J. 1998. Structure, Conformations, and Repair of DNA Adducts from Dibenzo[a,l]pyrene: 32P-Postlabeling and Fluorescence Studies. Chemical Research in Toxicology, 11: 674–685.  [Google Scholar]) we have studied DNA adduct formation of anti-DE of DBP [(±)-anti-DBPDE], in A549 human epithelial lung carcinoma cells and monitored the levels of adducts as a function of time. A high-performance liquid chromatography (HPLC) procedure that allows monitoring of all cis- and trans-nucleoside adducts of dA and dG was used. Circular dichroism and UV-spectroscopy have been employed to gain information on adduct structural features. Incubation of cells with 0.1 μM (±)-anti-DBPDE resulted in rapid formation of adducts, followed by a decline. After 6 h of incubation about 20% of the adducts remained. Repeating the experiment with 0.01 μM (±)-anti-DBPDE resulted in a correspondingly lower adduct level initially, but in this case a larger proportion (35%) of the adducts remained after 6 h of incubation. Notably, at this time point, similar amounts of adducts are observed with both (±)-anti-DBPDE concentrations studied. Independent of diol epoxide concentration and incubation time, the dA/dG adducts were constant in number (～2.8). However, within the different adduct categories, the adduct distribution seemed to be time dependent. Although the data are preliminary, a selection with regard to adduct removal seems to have taken place. In comparative experiments with (+)-anti-BPDE, no significant difference in rates of adduct removal was evident. Preliminary results from global gene expression analysis, with focus on genes involved in DNA maintenance and cell cycle checkpoints, demonstrate interesting differences in the stress response elicited in the cells following exposure to the distorted and flexible nonplanar DBPDE or the rigid and planar BPDE molecule. As expected some major common induction events were clearly related to the activation of p53-dependent cell cycle checkpoint. However, distinct clusters of gene expression alterations were detected which mark one or other treatment specifically, indicating a high degree of subtlety in the stress response to closely related DNA adduct forming species.     

Molecular characterization and antibiotic resistance profiling of Campylobacter isolated from cattle in Polish slaughterhouses

A total of 812 samples from bovine hides and the corresponding carcasses collected at the slaughterhouse level in the eastern part of Poland were examined for the presence of Campylobacter jejuni and Campylobacter coli. Recovered isolates were confirmed using species-specific PCR, characterized by the presence of 11 putative virulence genes and antimicrobial susceptibility was determined using a microbroth dilution method. Furthermore, the genotypic relatedness of the isolates was determined by PFGE profiling and virulence pattern cluster analysis. The prevalence of Campylobacter was 25.6% and 2.7% in bovine hide and carcass samples, respectively. The presence of virulence markers varied between C. jejuni and C. coli species however, the majority of strains possessed the cadF, flhA, flaA genes, irrespective of the bacterial species and origin. The lower number of the strains was positive for the invasive associated markers – virB11 and wlaN. Antibiotic profiling showed that campylobacters were most frequently resistant to quinolones and fluoroquinolones (nalidixic acid and ciprofloxacin, 38.3% of each, respectively) followed by streptomycin (24.3%) and tetracycline (20.9%). Resistance to erythromycin and gentamicin was demonstrated in 4.3% and 2.6% of strains, respectively. Comparisons of the PFGE and virulence marker profiles of the isolates reflected the high genetic diversity of Campylobacter tested. Moreover, a poor correlation between the PFGE type, pathogenic gene marker and antimicrobial resistance patterns was observed.