Stark broadening of hydrogen spectral lines with fine structure effects

Formalism and numerical code have been elaborated for calculation of hydrogen line profiles in conditions of plasma in which Stark broadening and fine energy splitting are comparable and it is not possible to neglect either of them. It corresponds to the range of electron densities $10^{11} < N_e~({\rm cm}^{-3}) < 10^{15}$. Lamb shift and spontaneous emission effects have also been included. Computer simulation method was applied in the calculations. Final results have been compared with experimental and theoretical findings by other authors

Hodowla in vitro celomocytow dzdzownic Dendrobaena veneta

In vitro techniques are require to better understand coelomocytes - mediated immune responsed. The main obstacle in the long-term cultures of coelomocytes is the contamination with bacteria and fungi from earthworms exterior and coelomic cavity. To avoid this, coelomocytes are retrieved through a small incision in the body wall from earthworms maintained at 10°C for at least 2 weeks. Culture medium is supplemented with 1% streptomycin/penicylin (Sigma) and 0.001% thiomersal (Merck). Freshly retrieved coelomocytes of Dendrobaena veneta contain granular and non-granular cells in the proportion close to 1 : 1 while in our four-month in vitro culture granular cells predominate.Dla lepszego zrozumienia mechanizmów reakcji odpornościowych mediowanych przez celomocyty dżdżownic niezbędne są hodowle tych komórek w warunkach in vitro. Główną przeszkodą w długoterminowych hodowlach jest ich kontaminacja bakteriami i grzybami pochodzącymi z celomy i powierzchni ciała dżdżownic. Aby tego uniknąć, celomocyty pozyskujemy poprzez rozcięcie ściany ciała dżdżownic przetrzymywanych co najmniej dwa tygodnie w temperaturze 10°C. Pożywkę hodowlaną wzbogacamy 1% roztworem penicyliny/streptomycyny (Sigma) i 0,001% thiomersalem. Świeżo pobrane celomocyty Dendrobaena veneta zawierają komórki ziarniste jak i nieziarniste w proporcji zbliżonej do 1 : 1, podczas gdy w naszych hodowlach czteromiesięcznych przeważają komórki ziarniste

The dynamics of the surface layer of lipid membranes doped by vanadium complex : computer modeling and EPR studies

Penetration of the liposome membranes doped with vanadium complex formed in the liquid-crystalline phase from egg yolk lecithin (EYL) by the TEMPO (2,2,6,6-tetramethylpiperidine-1-oxyl) spin probes has been investigated. The penetration process was followed by 360 hours at 24◦C, using the electron spin resonance (EPR) method. The spectroscopic parameter of the partition (F) of this probe indicated that a maximum rigidity of the membrane was at 3% concentration of the vanadium complex. Computer simulations showed that the increase in the rigidity of the membrane corresponds to the closure of gaps in the surface layer of the membrane, and indicates the essential role of the membrane surface in transport processes

Impact of humic acids on EYL liposome membranes : ESR method

In this paper, the effects of model (commercial) and natural (extracted from peat) humic substances on the membrane of liposomes formed with egg yolk lecithin (EYL) are presented. In our research, mass concentrations of fulvic and humic acids were used, which in relation to lecithin varied from 0% to 13%. To study membrane fluidity, electron spin resonance (EPR) was used with two spin probes, penetrating various regions of the lipid bilayer. The effects of model and natural humic substances (humic acids – HAs and fulvic acids – FAs) on the lipid membrane in different regions were researched: the lipid-water interphase, and in the middle of the lipid bilayer. It was shown that FA and HA impact the fluidity of liposome membranes in different ways. Increased mass concentrations of HAs decreased membrane fluidity in both acids: extracted from peat and the model. However, increased mass concentration of FAs extracted from peat, decreased membrane fluidity in the surface region, at the same time stiffening the central part of the bilayer. Increasing the concentration of FAs extracted from peat had the opposite effect when compared to model FA. This effect may be related to the complexation of xenobiotics present in the soil environment and their impact on biological membranes

Early-phase immunodetection of metallothionein and heat shock proteins in extruded earthworm coelomocytes after dermal exposure to metal ions

This paper provides direct evidence that earthworm immune cells, coelomocytes, are exposed to bio-reactive quantities of metals within 3 days after dermal exposure, and that they respond by upregulating metallothionein (NIT) and heat shock protein (HSP70, HSP72) expression. Indirect support for the hypothesis that coelomocytes are capable of trafficking metals was also obtained. Coelomocytes were expelled from adult individuals of Eisenia fetida after 3-day exposure either to metal ions (Zn, Cu, Pb, Cd) or to distilled water (controls) via filter papers. The number of coelomocytes was significantly decreased after Cu, Pb, or Cd treatment. Cytospin preparations of coelomocytes were subjected to immunoperoxidase staining with monoclonal antibodies against human beat shock proteins (HSP70 or HSP72), or rabbit polyclonal antibodies raised against. metallothionein 2 (w-MT2) of Lumbricus rubellus. Applied antibodies detected the respective proteins of E. fetida and revealed that the expression of HSP70, HSP72 and w-MT2 proteins was either induced or significantly enhanced in coelomocytes from metal-exposed animals. In conclusion, stress protein expression in earthworm coelomocytes may be used as sensitive biomarkers of metal contaminations. Further experimentation is needed for quantitative analysis of kinetics of metal-induced stress protein expression in earthworm coelomocyte

Photosensitized oxidative stress to ARPE-19 cells decreases protein receptors that mediate photoreceptor outer segment phagocytosis

PURPOSE. To determine whether previously shown photodynamic (PD)-induced inhibition of specific photoreceptor outer segment (POS) phagocytosis by ARPE-19 cells is associated with reductions in receptor proteins mediating POS phagocytosis, and if PD treatment with merocyanine-540 (MC-540) produces additional effects leading to its inhibition of nonspecific phagocytosis. METHODS. ARPE-19 cells preloaded with MC-540 or rose bengal (RB) were sublethally irradiated with green light. Phagocytosis of POS was measured by flow cytometry and POS receptor proteins (Mer tyrosine kinase receptor [MerTK] and integrin subunits αv and β5) and β-actin were quantified by Western blotting at 0.5 and 24 hours after irradiation, with comparison to samples from nonsensitized control cultures. The intact integrin heterodimer αvβ5 was quantified by immunoprecipitation followed by blotting. The distribution of N-cadherin, ZO-1, and F-actin was visualized by fluorescence microscopy. RESULTS. Mild PD stress mediated by both photosensitizers that elicits no significant morphologic changes produces transient and recoverable reductions in MerTK. The individual αv and β5 integrin subunits are also reduced but only partially recover. However, there is sufficient recovery to support full recovery of the functional heterodimer. Light stress mediated by MC-540 also reduced levels of actin, which is known to participate in the internalization of particles regardless of type. CONCLUSIONS. After PD treatment POS receptor protein abundance and phagocytosis show a coincident in time reduction then recovery suggesting that diminution in receptor proteins contributes to the phagocytic defect. The additional inhibition of nonspecific phagocytosis by MC-540–mediated stress may result from more widespread effects on cytosolic proteins. The data imply that phagocytosis receptors in RPE cells are sensitive to oxidative modification, raising the possibility that chronic oxidative stress in situ may reduce the efficiency of the RPE's role in photoreceptor turnover, thereby contributing to retinal degenerations

Zeaxanthin and alpha-tocopherol reduce the inhibitory effects of photodynamic stress on phagocytosis by ARPE-19 cells

Zeaxanthin and α-tocopherol have been previously shown to efficiently protect liposomal membrane lipids against photosensitized peroxidation, and to protect cultured RPE cells against photodynamic killing. Here the protective action of combined zeaxanthin and α-tocopherol was analyzed in ARPE-19 cells subjected to photodynamic (PD) stress mediated by rose Bengal (RB) or merocyanine-540 (MC-540) at sub-lethal levels. Stress-induced cytotoxicity was analyzed by the MTT assay. The peroxidation of membrane lipids was determined by HPLC-EC(Hg) measurements of cholesterol hydroperoxides using cholesterol as a mechanistic reporter molecule. The specific phagocytosis of FITC-labeled photoreceptor outer segments (POS) isolated from bovine retinas was measured by flow cytometry, and the levels of phagocytosis receptor proteins αv integrin subunit, β5 integrin subunit and MerTK were quantified by Western blot analysis. Cytotoxicity measures confirmed that PD stress levels used for phagocytosis analysis were sub-lethal and that antioxidant supplementation protected against higher, lethal PD doses. Sub-lethal PD stress mediated by both photosensitizers induced the accumulation of 5α-OOH and 7α/β-OOH cholesterol hydroperoxides and the addition of the antioxidants substantially inhibited their accumulation. Antioxidant delivery prior to PD stress also reduced the inhibitory effect of stress on POS phagocytosis and partially reduced the stress-induced diminution of phagocytosis receptor proteins. The use of a novel model system where oxidative stress was induced at sub-lethal levels enable observations that would not be detectable using lethal stress models. Moreover, novel observations about the protective effects of zeaxanthin and α-tocopherol on photodynamic damage to ARPE-19 cell membranes and against reductions in the abundance of receptor proteins involved in POS phagocytosis, a process essential for photoreceptor survival, supports the importance of the antioxidants in protecting of the retina against photooxidative injury