Determination of sotalol, oxprenolol and labetalol in binary mixtures and in spiked human serum by derivative spectrophotometric method

The usefulness of derivative spectrophotometry for the determination of labetalol, sotalol and oxprenolol in binary mixtures and in human spiked serum was checked. To this aim a spectrophotometric analysis of samples in the UV range was carried out and the obtained results revealed that derivative spectrophotometry allows for the fast, accurate and precise determination of the tested substances in spite of their clear interference in the zero-order spectra. For quantitative determinations ìzero-crossingî technique was used to establish wavelengths for zeros of specified component. In a mixture of labetalol and oxprenolol the following wavelengths were established: D1 λ = 245.32 nm and 266.03 nm, D2 λ = 243.30 nm and 301.09 nm. respectively. D3 derivative did not show zeros suitable for quantitative analysis. For the analysis of labetalol and sotalol mixture, D3 derivative spectrophotometry was used at the following wavelengths: λ = 246.03 nm and λ = 249.91 nm, respectively. In this case, the curves of D1 and D2 derivatives showed no zeros that can be used in quantitative analysis. To determine the concentration of the components in a mixture containing oxprenolol and sotalol the following wavelengths were selected: for oxprenolol D1 λ = 245.32 nm, D2 λ = 240.18 nm, D3 λ = 232.05 nm and for sotalol D1 λ = 230.56 nm, D2 λ = 232.65 nm and D3 λ = 238.84 nm, respectively. The developed spectrophotometric method was characterized by high sensitivity and accuracy, LOD determined for sotalol was in the range of 0.21-1.88 μg/mL, for labetalol 1.00-3.43 μg/mL and for oxprenolol 0.16-2.06 μg/mL; LOQ determined for sotalol was in the range of 0.65-5.70 μg/mL, for labetalol 3.11-10.39 μg/mL and for oxprenolol 0.47-6.23 μg/mL, depending on the composition of the tested mixture and the order of the derivative. The recovery of the individual components was within the range of 100 ± 5%. The linearity range was wide and estimated for sotalol in the range of 11.00-38.50 μg/mL, for labetalol 12.80-44.80 μg/mL and for oxprenolol 12.60-44.10 μg/mL with correlation coefficients in the range of 0.9977-0.9999

Fenofibrate interferes with the diapedesis of lung adenocarcinoma cells through the interference with Cx43/EGF-dependent intercellular signaling

Extravasation of circulating cancer cells is regulated by the intercellular/intracellular signaling pathways that locally impair the endothelial barrier function. Co-cultures of human umbilical vein endothelial cells (HUVECs) with lung adenocarcinoma A549 cells enabled us to identify these pathways and to quantify the effect of fenofibrate (FF) on their activity. A549 cells induced the disruption and local activation of endothelial continuum. These events were accompanied by epidermal growth factor (EGF) up-regulation in endothelial cells. Impaired A549 diapedesis and HUVEC activation were seen upon the chemical inhibition of connexin(Cx)43 functions, EGF/ERK1/2-dependent signaling, and RhoA/Rac1 activity. A total of 25 μM FF exerted corresponding effects on Cx43-mediated gap junctional coupling, EGF production, and ERK1/2 activation in HUVEC/A549 co-cultures. It also directly augmented endothelial barrier function via the interference with focal adhesion kinase (FAK)/RhoA/Rac1-regulated endothelial cell adhesion/contractility/motility and prompted the selective transmigration of epithelioid A549 cells. N-acetyl-L-cysteine abrogated FF effects on HUVEC activation, suggesting the involvement of PPARα-independent mechanism(s) in its action. Our data identify a novel Cx43/EGF/ERK1/2/FAK/RhoA/Rac1-dependent signaling axis, which determines the efficiency of lung cancer cell diapedesis. FF interferes with its activity and reduces the susceptibility of endothelial cells to A549 stimuli. These findings provide the rationale for the implementation of FF in the therapy of malignant lung cancers

Evidence for cytoprotective effect of carbon monoxide donor in the development of acute esophagitis leading to acute esophageal epithelium lesions

Exposure to acidic gastric content due to malfunction of lower esophageal sphincter leads to acute reflux esophagitis (RE) leading to disruption of esophageal epithelial cells. Carbon monoxide (CO) produced by heme oxygenase (HMOX) activity or released from its donor, tricarbonyldichlororuthenium (II) dimer (CORM-2) was reported to protect gastric mucosa against acid-dependent non-steroidal anti-inflammatory drug-induced damage. Thus, we aimed to investigate if CO affects RE-induced esophageal epithelium lesions development. RE induced in Wistar rats by the ligation of a junction between pylorus and forestomach were pretreated i.g. with vehicle CORM-2; RuCl3; zinc protoporphyrin IX, or hemin. CORM-2 was combined with NG-nitro-L-arginine (L-NNA), indomethacin, capsazepine, or capsaicin-induced sensory nerve ablation. Esophageal lesion score (ELS), esophageal blood flow (EBF), and mucus production were determined by planimetry, laser flowmetry, histology. Esophageal Nrf-2, HMOXs, COXs, NOSs, TNF-α and its receptor, IL-1 family and IL-1 receptor antagonist (RA), NF-κB, HIF-1α, annexin-A1, suppressor of cytokine signaling (SOCS3), TRPV1, c-Jun, c-Fos mRNA/protein expressions, PGE2, 8-hydroxy-deoxyguanozine (8-OHdG) and serum COHb, TGF-β1, TGF-β2, IL-1β, and IL-6 content were assessed by PCR, immunoblotting, immunohistochemistry, gas chromatography, ELISA or Luminex platform. Hemin or CORM-2 alone or combined with L-NNA or indomethacin decreased ELS. Capsazepine or capsaicin-induced denervation reversed CORM-2 effects. COHb blood content, esophageal HMOX-1, Nrf-2, TRPV1 protein, annexin-A1, HIF-1α, IL-1 family, NF-κB, c-Jun, c-Fos, SOCS3 mRNA expressions, and 8-OHdG levels were elevated while PGE2 concentration was decreased after RE. CO donor-maintained elevated mucosal TRPV1 protein, HIF-1 α, annexin-A1, IL-1RA, SOCS3 mRNA expression, or TGF-β serum content, decreasing 8-OHdG level, and particular inflammatory markers expression/concentration. CORM-2 and Nrf-2/HMOX-1/CO pathway prevent esophageal mucosa against RE-induced lesions, DNA oxidation, and inflammatory response involving HIF-1α, annexin-A1, SOCS3, IL-1RA, TGF-β-modulated pathways. Esophagoprotective and hyperemic CO effects are in part mediated by afferent sensory neurons and TRPV1 receptors activity with questionable COX/PGE2 or NO/NOS systems involvement

The research movement of chloroplasts in the cells of Arabidopsis (Arabidopsis thaliana L.).

Celem niniejszej pracy było zbadanie ruchu chloroplastów podczas reakcji ucieczki wywołanej silnym światłem niebieskim w mezofilu Arabidopsis thaliana. Zastosowano metodę obserwacji mikroskopowej z analizą ilościową otrzymanych obrazów. Analizy ruchu chloroplastów dokonano w komórkach rzodkiewnika pospolitego (Arabidopsis thaliana L.), który jest gatunkiem modelowym w badaniach biologicznych. Zbadano reakcje chloroplastów w ogonkach liściowych, a ilościowej analizie poddano ruchy pojedynczych chloroplastów w czasie reakcji ucieczki w komórkach mezofilu. Szczegółowym celem było określenie trajektorii jaką przebywa pojedynczy chloroplast, a następnie określenie odległości między początkowym, a końcowym położeniem chloroplastu. Otrzymane wyniki pokazują występowanie ruchu chloroplastów w części przyogonkowej liścia rzodkiewnika pospolitego.The aim of this study was to investigate the movement of chloroplasts during the strong blue light-induced avoidance response in Arabidopsis mesophyll. Microscopic observation was followed by quantitative analysis of the obtained images. The analysis of chloroplast movement has been performed in cells of Arabidopsis thaliana L. which is a model species in biological research. First, chloroplast responses in petioles were investigated, followed by quantitative analysis a single chloroplast movements during the avoidance response in mesophyll cells. The specific objective was to determine the trajectory traveled by a single chloroplast, and then to specify the distance between the initial and final position of the chloroplast

The effect of fenofibrate on the invasive properties of lung cancer A549 cells and its interference with their diapedesis

Kaskada metastatyczna jest procesem wieloetapowym, w wyniku którego komórka opuszcza guz pierwotny i przemieszcza się za pomocą układu krążenia do odległych organów, w których następnie tworzy ognisko wtórne choroby. Istotne dla wydajności tego procesu są zarówno właściwości komórek nowotworowych, które warunkują ich potencjał inwazyjny, jak również specyfika i dynamika ich oddziaływań z komórkami śródbłonka podczas penetracji bariery naczyń krwionośnych. Pomimo znaczących postępów w diagnostyce i terapii nowotworów, istnieje potrzeba opracowania nowych wielolekowych terapii alternatywnych dla konwencjonalnych metod leczenia, które będą charakteryzować się szerokim spektrum działania. Lekiem, który może znaleźć takie zastosowanie w terapii nowotworów, jest fenofibrat wykorzystywany w leczeniu hipercholesterolemii, ale również wykazujący właściwości przeciwnowotworowe.W pracy sprawdzono wpływ fenofibratu na komórki raka płuc linii A549. Fenofibrat powodował zmniejszenie aktywności proliferacyjnej komórek A549, a także obniżał wydajność ich migracji, co korelowało ze zmianami organizacji cytoszkieletu aktynowego i wewnątrzkomórkowej lokalizacji białek związanych z przejściem epitelialno-mezenchymalnym (EMT). Stosując model badawczy oparty na ko-hodowli komórek śródbłonka z komórkami nowotworowymi, zaobserwowano interferencję fenofibratu z wydajnością procesu diapedezy komórek A549. Fenofibrat powodował wzmocnienie funkcji barierowej komórek śródbłonka poprzez wpływ na adhezję komórek śródbłonka do podłoża. Wyniki te sugerują możliwość zastosowania fenofibratu jako leku wspomagającego klasyczne strategie terapeutyczne wykorzystywane w leczeniu raka płuc.During the multi-step process of metastatic cascade, invasive cells leave the primary tumor, enter the circulatory system to spread throughout the organism, and to occasionally give rise to secondary tumors. Both the invasive properties of cancer cells and their dynamic interactions with microenvironment are crucial for the efficiency of metastatic cascade. In particular, the interactions of cancer and endothelial cells are crucial for the efficiency of cancer cell diapedesis that directly initiates metastasis formation. Despite the significant progress in the diagnosis and therapy of cancer, new multi-drug therapies are necessary to complement conventional treatment regimens. Fenofibrate, which is conventionally used in the treatment of hypercholesterolemia, may also be used in the treatment of cancers, because it is characterized by a range of anti-cancer properties.Here, the effect of fenofibrate on the invasive potential of lung cancer A549 cells was estimated. An attenuation of A549 cell proliferative and locomotory activity was accompanied by the changes in cell cytoskeleton architecture and in the intracellular localization of proteins associated with epithelial-mesenchymal transition (EMT). An interference of fenofibrate with the process of A549 cell diapedesis was demonstrated by the experimental approach based on the co-cultures of A549 cells with endothelial layer and attributed to the direct effect of fenofibrate on endothelial cells. These data suggest a potential application of fenofibrate for the supplementation of conventional lung cancer chemotherapy