The Phenolics of the Ornithogalum umbellatum L. (Hyacinthaceae): Phytochemical and Ecological Characterization

Ornithogalum umbellatum L. is a widely distributed species in Europe, that exhibits considerable variability at ecological, morphological, anatomical, and karyological level. Previous reports of the chemical investigations among Ornithogalum species indicate significant diversity of the secondary metabolites, as well. Knowing that environment affects the phenolic composition in plants to a large extent, the main objective of the research was to define relationship between phytochemical and ecological characters. To estimate an environmental influence on these results, plant material was collected at four habitats that differ in ecological factors and belong to two biogeographical regions: the Balkan Peninsula and the Pannonian Plane. Measured phytochemical characters are yield of dry extract, total phenolic and flavonoid contents, the presence of selected phenolic compounds as well as free‐radical scavenging activity (neutralization of DPPH and OH radicals). Results revealed that all analyzed phytochemical parameters differ between investigated O. umbellatum samples. The moisture level of habitat has the highest correlation, either positive or negative, with most of phytochemical characters, and is followed by temperature and soil reaction. Light intensity and nitrogen level have mostly moderate correlation coefficient with phytochemical characters. More complex correlation is revealed between ecological factors and nine phenolic compounds, with three observed patterns of relationship.     

Hypo-Vascular Liver Metastases Treated with Transarterial chemoembolization: Assessment of Early Response by Volumetric Contrast-Enhanced and Diffusion-Weighted Magnetic Resonance Imaging

OBJECTIVE: To evaluate the value of anatomic and volumetric functional magnetic resonance imaging (MRI) in early assessment of response to trans-arterial chemoembolization (TACE) in hypovascular liver metastases. METHODS: This retrospective study included 52 metastatic lesions (42 targeted and 10 non-targeted) in 17 patients who underwent MRI before and early after TACE. Two reviewers reported response by anatomic criteria (Response Evaluation Criteria in Solid Tumor [RECIST], modified RECIST [mRECIST], and European Association for the Study of Liver Disease [EASL]) and functional criteria (volumetric apparent diffusion coefficient and contrast enhancement). Treatment endpoint was RECIST at 6 months. A 2-sample paired t test was used to compare the mean changes after intra-arterial therapy. P < .05 was considered statistically significant. RESULTS: Reduction in mRECIST and EASL at 1 month was significant in the whole cohort as well as in responders by RECIST at 6 months, and the changes fulfilled partial response criteria for both metrics in responders. Responders also had significant changes in volumetric apparent diffusion coefficient (P = .01 and P = .03) and contrast enhancement (P < .0001 and P < .0001) at 1 month for both readers, respectively. CONCLUSION: At 1 month post treatment, responders did not fulfill RECIST criteria but fulfilled mRECIST and EASL criteria. In addition, volumetric contrast-enhanced and diffusion-weighted MRI may be helpful in evaluating early treatment response after TACE in hypovascular liver metastases in patients who have failed to respond to initial chemotherapy.     

Morphology of the human corpus callosum: the shape of its mediosagittal section

The shape of the corpus callosum was studied on mediosagittal sections of 50 human brains. The terms "facies corticalis" and "facies profunda" are proposed for the sides of corpus callosum. Each of 50 brains had a different shape of callosal mediosagittal section and any available classification was not possible. In 94% of cases, the contour of callosal cortical surface exhibited a posterior depression and in 46% also an anterior one. Other details of callosal morphology influencing its shape on mediosagittal section (transverse folds, circumscribed depressions) were described. The junction of fornix and corpus callosum in 78% was in 3rd and in 14% in 4th quarter of callosal length. The possible significance of these findings for further studies of human brain morphology is discussed.     

Continuous internalization of tumor necrosis factor receptors in a human myosarcoma cell line

The cell dynamics of the receptor for tumor necrosis factor (TNF) were examined in TNF-sensitive KYM cells derived from human myosarcoma. With receptor synthesis inhibited by cycloheximide, the half-life of the surface TNF receptor was 2 h in the absence of TNF and 30 min in its presence, suggesting that the TNF receptor is non-recycling and that its internalization is accelerated by TNF. During cell incubation with TNF receptor degradation suppressed by chloroquine, the number of surface TNF receptors remained approximately constant, but the total number of surface and internal TNF receptors increased gradually, at 3 h reaching 1.5 times the initial number, thus suggesting continuous synthesis, externalization, internalization, and degradation of the TNF receptor in the absence of cycloheximide. On cell incubation with 125I-TNF, the intracellular quantity of the pulse-labeled TNF-receptor complex promptly increased, reaching a maximum at 20 min, and then gradually declined, thus confirming that the TNF receptor is internalized as a TNF-receptor complex in the presence of TNF. During incubations with protein synthesis suppressed by cycloheximide following surface TNF receptor digestion by trypsin, TNF receptors reappeared on the cell surface, increasing in number to a peak at 60 min and gradually decreasing, and cells previously exposed to cycloheximide with or without TNF showed no recurrence of surface TNF receptors, suggesting that the TNF receptor is non-recycling. The results of the study thus suggest that the TNF receptor is continuously internalized and degraded intracellularly by lysosomes without being recycled regardless of the presence or absence of TNF and, further, that its internalization is accelerated when it is part of the TNF-receptor complex.     

Enhancement of CRISPR/Cas12a trans-cleavage activity using hairpin DNA reporters

The RNA programmed non-specific (trans) nuclease activity of CRISPR-Cas Type V and VI systems has opened a new era in the field of nucleic acid-based detection. Here, we report on the enhancement of trans-cleavage activity of Cas12a enzymes using hairpin DNA sequences as FRET-based reporters. We discover faster rate of trans-cleavage activity of Cas12a due to its improved affinity (K_m) for hairpin DNA structures, and provide mechanistic insights of our findings through Molecular Dynamics simulations. Using hairpin DNA probes we significantly enhance FRET-based signal transduction compared to the widely used linear single stranded DNA reporters. Our signal transduction enables faster detection of clinically relevant double stranded DNA targets with improved sensitivity and specificity either in the presence or in the absence of an upstream pre-amplification step.     

Indirect estimation of reference intervals for thyroid parameters using advia centaur XP analyzer

BackgroundThe aim of this study was to determine the reference intervals (RIs) for thyroid stimulating hormone (TSH), free thyroxine (FT4), free triiodothyronine (FT3) and FT3/FT4 ratio using indirect methods.MethodsWe analyzed 1256 results TSH, FT4 and FT3 collected from a laboratory information system between 2017 and 2021. All measurements were performed on a Siemens ADVIA Centaur XP analyzer using the chemiluminescent immunoassay. We calculated the values of the 2.5th and 97.5th percentiles as recommended by the IFCC (CLSI C28-A3).ResultsThe RIs derived for TSH, FT4, FT3 and FT3/FT4 ratio were 0.34-4.10 mIU/L, 11.3-20.6 pmol/L, 3.5-6.32 pmol/L and 0.21-0.47, respectively. We found a significant difference between calculated RIs for the TSH and FT4 and those recommended by the manufacturer. Also, FT3 values were significantly higher in the group younger than 30 years relative to the fourth decade (5.26 vs. 5.02, p=0.005), the fifth decade (5.26 vs. 4.94, p=0.001), the sixth decade (5.26 vs. 4.87, p<0.001), the seventh decade (5.26 vs. 4.79, p<0.001) and the group older than 70 years old (5.26 vs. 4.55, p<0.001). Likewise, we found for TSH values and FT3/FT4 ratio a significant difference (p <0.001) between different age groups.ConclusionsThe establishing RIs for the population of the Republic of Srpska were significantly differed from the recommended RIs by the manufacturer for TSH and FT4. Our results encourage other laboratories to develop their own RIs for thyroid parameters by applying CLSI recommendations.     

Antibacterial activity of Punica granatum L. and Areca nut (P.A) combined extracts against some food born pathogenic bacteria

The antibacterial effects of combined extracts of Punica granatum L. and Areca nut (P.A) against resistant bacteria, a gram-positive bacterium, Staphylococcus aureus and three gram-negative bacteria, Escherichia coli, Salmonella, and Enterobacter aerogenes, in individual and biofilm forms was studied. Antibacterial activity was studied using disk diffusion method, microbroth dilution, and microtiter plate methods. Given the disc diffusion test (Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC)), the extracts had inhibitory effects on the individual forms of bacteria. However, the ethanolic extract had greater effectiveness than the methanolic extract. Generally, ethanol efficiently extracts flavonoids and their glycosides, catechol and tannins. This fact due to the ethanol polarity that is equal 0.654. The results indicated that the ability of extracts in inhibiting the formation of biofilms, destruction of biofilms, and prevention of metabolic activity of bacteria had a direct relationship with concentration and the highest inhibitory was seen on Staphylococcus aureus (98.98%), Staphylococcus aureus (94.98%), and Enterobacter aerogenes (88.55%). Based on the results, the P.A. combined extract can be used as an alternative combination with the ability to inhibit antibiotic-resistant bacteria in single and biofilm forms.     

A New Autumn-Flowering Species of Allium (Alliaceae) from Croatia

Here we describe Allium telmatum Bogdanovi?, Brullo, Giusso & Salmeri, a new species of Allium sect. Codonoprasum from North Dalmatia (Croatia). Its chromosome number (2n?=?32), karyotype, leaf anatomy, ecology and taxonomical relationships are examined. Several features, such as phenology (flowering in autumn), occurrence in coastal salt marshes, tetraploid chromosome number, and morphology, indicate that it is most closely related to the Tyrrhenian species Allium savii. On the basis of our herbarium survey, we present here a distribution map of the autumn-flowering species of Allium sect. Codonoprasum in the Mediterranean area.     

Possible relation between a bloom of Distephanus speculum (Silicoflagellata) and anoxia in bottom waters in the Northern Adriatic, 1983

Silicoflagellates, rare planktonic algae, were registered inbloom-like conditions in the 20-m-deep bottom layer of the Gulfof Trieste, Northern Adriatic. The confusing taxonomy and ecologyof these phytoflagellates, as well as their possible contributionto the oxygen depletion of the bottom layer, is discussed.     

Binding site identification of anticancer drug gefitinib to HSA and DNA in the presence of five different probes

This study was carried out to evaluate the binding interaction of gefitinib (GEF) with human serum albumin (HSA) and calf thymus DNA (ct-DNA) using fluorescence, UV–Visible, zeta potential measurements and molecular docking methods in order to understand its pharmacokinetic mechanism. By increasing the temperature, a steady decrease in Stern–Volmer quenching constants was observed for HSA binding properties; this indicates a static type of fluorescence quenching. Negative values were calculated for Gibbs free energy (ΔG), enthalpy (ΔH), and entropy (ΔS) changes, indicating that the reaction is spontaneous and enthalpy-driven. Probe competitive experimental results showed that GEF contains the same binding site as warfarin and are consistent with modeling results. The zeta potential of the HSA increased with increasing GEF, which represents the presence of electrostatic interactions in the system. DNA binding properties were investigated in the presence of three probes. The experimental results showed that by increasing GEF to DNA-AO (acridine-orange) and DNA-MB (methylene-blue) system, the fluorescence intensity and absorbance spectra had no considerable change. Furthermore, with the addition of GEF to DNA, the zeta potential decreased gradually, indicating that the hydrophobic interaction between the GEF and the bases of DNA is the major factor. Thus, GEF can bind to DNA via a groove binding mode. It was also found that GEF entered into the minor groove in the A–T rich region of DNA fragment and bind via van der-Waals forces and three H-bond with double strands of DNA. This is in good agreement with experimental results.