Analysis of substances issuing from Zajdela ascitic hepatoma cells after exposure to UV radiation of different wave-lengths. I. Relationship to dose]

The release of substances from the Zaidela ascitic hepatoma cells after irradiation with physiological doses of short-wave (254 nm) and long-wave (300-380 nm) UV light (far and near UV light) has been studied spectrophotometrically. Within the range of 200-520 nm, the absorption spectra of releasing substances show maxima at 215 and 260 nm and are identical to spectra of non-irradiated cells. The amount of substances increases with dose making up, at the maximal alteration, 180-220%, of the amount releasing from non-irradiated cells. Irradiation with far UV light exceeds by one order that with near UV light. The effect of minimum doses is opposite to the action of high doses: the release of substances from irradiated cells is much less.     

Analysis of substances issuing from Zajdela ascitic hepatoma cells following exposure to UV radiation of different wavelength. II. Proton release]

The release of protein from the Zaidela ascitic hepatoma cells following irradiation with physiological doses of short-wave (254 nm) and long-wave (300--380 nm) UV light (far and near UV radiation) has been investigated. The amount of protein increases with dose making, upon the maximal radiation damage, 180 and 2 per cent of the protein against, resp., the protein amount releasing from non-treated cells and the total protein of the intact cell. The far UV light is by one order more efficient than the near UV light. Irradiation of cells with the former and the latter results in the release of high and low molecular proteins, resp. The near UV irradiation brings about heavier releasing of proteins than does the far UV light.     

Glycoprotein desorption from the surface of human peripheral blood lymphocytes after irradiation with short-wave UV rays

The vital quantitative method of the cell coat (outer perimembraneous layer-OPML) identification with alcian blue (AB), which was earlier developed for the rat hepatoma cells and human erythrocytes, has been adopted for human blood lymphocytes. AB is bind by glycoproteins, glycolipids and acid mucopolysaccharides of the cell surface. Under experimental condition to be used each lymphocyte adsorbed 1.1 X 10(-10) g of AB. Irradiation with non-lethal doses of UV light induced a decrease in AB sorption by 8-13%. At the same time, the release of substances took place, some properties of which are similar to those of glycoproteins. A conclusion is made that the lymphocyte OPML was destroyed by UV rays and its components released into the extracellular space. The role of this phenomenon is discussed in terms of the therapeutic effect of UV light.     

Destructive changes in the outer perimembrane layers (glycocalyx) of Zajdela ascites hepatoma cells under the action of UV radiation]

Immediately after far (254) nm and near (300--380 nm) UV light in small and moderate doses alcian blue sorption by glycocalix of Zaidela ascitic hepatoma cells decrease, which is indicative of destruction and solubilization. The effect of UV light on the cell surface is compared with the action of trypsin. Contribution of the damage of outer perimembrane layers to the lethal effect of UV light is discussed.     

Control of cytoplasmic calcium with photolabile tetracarboxylate 2-nitrobenzhydrol chelators.

This paper introduces nitr-2, a new Ca2+ chelator designed to release Ca2+ upon illumination with near UV (300-400 nm) light. Before illumination nitr-2 has Ca2+ dissociation constants of 160 and 630 nM in 0.1 and 0.3 M ionic strength respectively; after photoconversion to a nitrosobenzophenone the values shift to 7 and 18 microM, high enough to liberate substantial amounts of Ca2+ under intracellular conditions. The speed of release is limited by a dark reaction with rate constant 5 s-1. Aplysia central neurons injected with nitr-2 and exposed to UV light exhibit two separate Ca2+-dependent membrane currents: one carried by potassium ions and one a nonspecific cation current. A quantitative estimate of the spatial distribution of intracellular [Ca2+] changes in large cells filled with a high concentration of nitr-2 and exposed to an intense UV flash is offered.     

Plasmid ColE3 specifies a lysis protein.

Tn5 insertion mutations in plasmid ColE3 were isolated and characterized. Several of the mutants synthesized normal amounts of active colicin E3 but, unlike wild-type colicinogenic cells, did not release measurable amounts of colicin into the culture medium. Cells bearing the mutant plasmids were immune to exogenous colicin E3 at about the same level as wild-type colicinogenic cells. All of these lysis mutants mapped near, but outside of, the structural genes for colicin E3 and immunity protein. Cells carrying the insertion mutations which did not release colicin E3 into the medium were not killed by UV exposure at levels that killed cells bearing wild-type plasmids. The protein specified by the lysis gene was identified in minicells and in mitomycin C-induced cells. A small protein, with a molecular weight between 6,000 and 7,000, was found in cells which released colicin into the medium, but not in mutant cells that did not release colicin. Two mutants with insertions within the structural gene for colicin E3 were also characterized. They produced no colicin activity, but both synthesized a peptide consistent with their map position near the middle of the colicin gene. These two insertion mutants were also phenotypically lysis mutants--they were not killed by UV doses lethal to wild-type colicinogenic cells and they did not synthesize the small putative lysis protein. Therefore, the lysis gene is probably in the same operon as the structural gene for colicin E3.     

Replication of adeno-associated virus in cells irradiated with UV light at 254 nm.

Irradiation of simian virus 40 (ori mutant)-transformed Chinese hamster embryo cells (OD4 line) with UV light induced a cellular capacity which supported a full cycle of helper-independent adeno-associated virus replication. Monochromatic UV light at 254 nm was about 1,000-fold more effective than UV light at 313 nm, indicating that cellular nucleic acid is the primary chromophore in the UV-induced process leading to permissiveness for adeno-associated virus replication. The UV irradiation and the infection could be separated for up to 12 h without substantial loss of permissiveness. During this time interval, the induction process was partly sensitive to cycloheximide, suggesting a requirement for de novo protein synthesis.     

Effect of UV radiation of varying wave length on the tumoringenicity of Zajdela ascitic hepatoma cells determined by transplantations into the anterior chamber of the eye in rats]

Potentialities for evaluating the viability of the Zaidela ascitic hepatoma cells by the number of tumours formed after transplantation of cells to the rat's eye anterior chamber have been considered. Dose effect of far (254 nm) and near (300--380 nm) UV light on the cell tumoregenecity has been studied.     

Effect of far and near ultraviolet radiation on the DNA content in the unicellular flagellate alga Dunaliella salina]

Effects of far (254 nm) and near (above 300 nm) ultraviolet radiation on DNA in Dunaliella salina were compared. DNA synthesis was delayed by both the kinds of UV radiation applied in isoeffective doses (ld0 and LD90), but the extent of inhibition of DNA synthesis was much stronger after action of far UV rays.     

Photoreactivation of cells and phages injuried by ultraviolet radiation in the ecological long-wave band]

Photoreactivation (PR) was measured after inactivation by far (254 nm), middle (300-315 nm) and near (315-400 nm) UV radiation of Paramecium caudatum and 8 strains of Escherichia coli differing in PR and dark repair capability. PR volume was high and practically the same after irradiation by far and middle UV, but PR was not observed in near UV-inactivated cells of all the strains. It is proposed that pyrimidine dimers are not significant in near UV lethal lesions in cells, as near UV-irradiated phages (T7 and lambdacI 857) are not photoreactivated in undamaged host bacterial cells.     

Fatty Acid Induced Leaking of Organic Compounds from Boletus variegatus

Addition of octanoic acid (2· 10^-3M) to the suspending medium (final pH 4.85) of Boletus variegatus mycelium induced a marked leaking of UV absorbing substances from the cells. The material had an absorption maximum at 260 nm, a minimum at 240 nm, and the absorption ratios 250: 260 and 280:260 were 0.81 and 0.49. The material released immediately after addition of the acid consisted mainly of low molecular weight substances. These substances, listed according to decreasing rates of leaking, were identified as pentoses, pentosephosphates, nucleosides, and mono- and di-nucleotides. Also, purine and pyrimidine bases were released at this early stage of treatment. After 90 minutes' treatment, an outflux of oligoribonucleotides was observed. The oligoribonucleotides did not occur as single substances, but were forming complexes with peptides. Minor amounts of ribonucleic acid were also leaking out from the cells. Deoxyribose containing substances were never observed in the filtrates. The compounds were subjected to enzymatic degradation after they had left the cells. This was shown by a marked increase with time of inorganic phosphorus, pentose/pentosephosphates, and nucleosides in the filtrate. The leaking of low molecular weight substances immediately after acid addition is correlated to seriously reduced growth. However, the growth was wholly restored after a three days' lag period. On the other hand, when considerable amounts of oligoribonucleotide peptides had been released from the cells, growth could not be re-established.     

The macrocyclic effect and vasodilation response based on the photoinduced nitric oxide release from trans-[RuCl(tetraazamacrocycle)NO](2+)

Irradiation of trans-[RuCl(cyclam)(NO)](2+), cyclam is 1,4,8,11-tetraazacyclotetradecane, at pHs 1-7.4, with near UV light results in the release of NO and formation of trans-[Ru(III)Cl(OH)(cyclam)](+) with pH dependent quantum yields (from approximately 0.01 to 0.16 mol Einstein(-1)) lower than that for trans-[RuCl([15]aneN(4))(NO)](2+), [15]aneN(4) is 1,4,8,12-tetaazacyclopentadecane, (0.61 mol Einstein(-1)). After irradiation with 355 nm light, the trans-[RuCl([15]aneN(4))(NO)](2+) induces relaxation of the aortic ring, whereas the trans-[RuCl(cyclam)(NO)](2+) complex does not. The relaxation observed with trans-[RuCl([15]aneN(4))(NO)](2+) is consistent with a larger quantum yield of release of NO from this complex.     

Leakage of cellular material from Thiobacillus ferrooxidans in the presence of organic acids.

Thiobacillus ferroodixans cells released varying amounts of iron, phosphate, sugar, ribonucleic acid, deoxyribonucleic acid, and substances that absorbed light at both 260 and 280 nm, when exposed to 10(-2) to 10(-1) M concentrations of these organic acids: propionic, butyric, valeric, hexanoic, and oxalacetic. These acids also retarded iron oxidation by the cells. Electron microscope observation of cells after exposure to the organic acids showed varying degrees of cell envelope disruption, suggesting that the mode of inhibition of autotrophic iron oxidation in the cell involves interference with the function of the cell envelope, possibly the cell membrane.     

Blue- and green-light signals for gamete release in the brown alga, Silvetia compressa

The intertidal brown alga Silvetia compressa releases gametes from receptacles (the reproductive tissue) rapidly upon a dark transfer (following a photosynthesis-dependent period in the light, termed potentiation). In this study, the wavelength-dependence of this process was investigated. During the potentiation period in white light (WL), gametes are not released. However, gametes were released during potentiation in blue light (BL), or in low red light/blue light (RL/BL) ratios, but not in RL alone, high RL/BL ratios, or in broadband blue-green light (B-GL) (presence of BL, but absence of RL). RL was as effective as WL for potentiation, i.e., both lead to gamete release following transfer to darkness. Rates of linear photosynthetic electron transport were similar in RL and BL. Gamete release in BL was inhibited by equal amounts of additional narrow-waveband light between the green and red regions of the spectrum, with light-induced gamete release restricted between <491 nm and 509 nm. Very little light-induced gamete release occurred between 530 nm and 650 nm. It is proposed that a BL-responsive photoreceptor is responsible for light-induced gamete release. Transfer of WL-potentiated receptacles to GL near 530 nm resulted in significant de-potentiation and reduced gamete release during a subsequent dark transfer. This effect was not seen at 509 nm or 560 nm and revealed the presence of a second photoreceptor system repressing or counteracting potentiation in the light. We propose that the restriction of gamete release to periods when irradiance is blue-shifted may constitute a depth-sensing mechanism for this intertidal alga, allowing controlled release of gametes at high tide and/or less turbid periods, thus minimizing gamete dilution, and promoting fertilization success.     

Differential scattering of circularly polarized light in protonated DNA in the presence of spermine

The protonation of the spermine containing calf thymus DNA (molecular mass 15 and 5 MDa) solutions has been studied by means of circular dichroism method. It has been shown that the acid-induced transition from the low-protonated B(+)-form to the double-stranded structure with presumably Hoogsteen complementation of syn-G.C-base-pairs (S-form) in case of high-molecular partially condensed DNA is accompanied by differential scattering of circularly polarized light (DSCPL). The comparative study of protonation of partially and completely condensed low-molecular DNA enabled to obtain a family of DSCPL spectra. It has been demonstrated that the B+----S-transition in partially condensed high-molecular DNA is associated with formation of large intermolecular aggregates (with dimensions about 200 divided by 400 nm) which are destructed by acid-induced DNA denaturation.     

Blue Light Inhibits Mitosis in Tissue Culture Cells

Irradiation of the mitotic (prophase and prometaphase) tissue culture PK (pig kidney embryo) cells using mercury arc lamp and band-pass filters postponed or inhibited anaphase onset. The biological responses observed after irradiation were: (i) normal cell division, (ii) delay in metaphase and then normal anaphase and incomplete cytokinesis, (iii) exit into interphase without separation of chromosomes, (iv) complete mitotic blockage. Cell sensitivity to the light at wavelengths from 423 and 488 nm was nearly the same; to the near UV light (wavelength 360 nm) it was 5–10 times more; to the green light (wavelength >500 nm) it was at least 10 times less. To elucidate the possible mechanism of the action of blue light we measured cell adsorption and examined cell autofluorescence. Autofluorescence of cytoplasmic granules was exited at wavelengths of 450–490 nm, but not at >500 nm. In mitotic cells fluorescent granules accumulated around the spindle. We suppose blue light irradiation induces formation of the free radicals and/or peroxide, and thus perturb the checkpoint system responsible for anaphase onset.     

Role of the systems of DNA dark repair in determining bacterial and phage sensitivity to ultraviolet radiation in an ecological long-wave range]

A comparison has been made of sensitivity to far (254 nm), middle (300--315 nm) and near (315--400 nm) UV radiation of 12 strains of E. coli and 2 strains of B. subtilis differing in DNA dark repair (DR) capability. The mechanisms controlled by uvrA, uvrB, polA, recA, lon, and lexA genes are very effective in cells, irradiated by far and middle UV, but by 15--70% less effective in those irradiated by near UV. As the unirradiated bacteria poorly repair the near UV damaged phages (T7, lambda, SPPI), the low bacterial DR level after UV irradiation seems to be due to the unrepairable photoproduct formation in DNA.     

The contribution of fungal enzymes to the digestion of leaves by Gammarus fossarum Koch (Amphipoda)

Summary Gut extracts of Gammarus fossarum liberated reducing substances (at pH values 7) and amino acids (pH7) from freshly shed oak leaves only after removal of soluble leaf phenols. When carboxymethylcellulose was used at a concentration equal to that of leaf cellulose, release of reducing substances was considerably higher. Fungal enzymes extracted from decomposing leaves were active against structural carbohydrates but showed no proteolytic activity. At low pH values, they retained their full activity in the presence of gut enzymes of G. fossarum, at higher pH values they were inhibited. Conditioned leaves released larger amounts of reducing substances and amino acids when exposed to gut enzymes. The improvement varies with the fungal species used for conditioning and with the length of the conditioning period. The digestibility of leaf carbohydrates and proteins reached separate peaks and then declined. Fungal carbohydrases ingested by G. fossarum retained some activity for up to 4h.     

Characterization of the laser-induced blue, green and red fluorescence signatures of leaves of wheat and soybean grown under different irradiance

The blue, green and red fluorescence emission of green wheat ( Triticum aestivum L. var. Rector) and soybean leaves ( Glycine max L. var. Maple Arrow) as induced by UV light (nitrogen laser: 337 nm) was determined in a phytochamber and in plants grown in the field. The fluorescence emission spectra show a blue maximum near 450 nm, a green shoulder near 530 nm and the two red chlorophyll fluorescence maxima near 690 and 735 nm. The ratio of blue to red fluorescence, F450/F690, exhibited a clear correlation to the irradiance applied during the growth of the plants. In contrast, the chlorophyll fluorescence ratio, F690/F735, and the ratio of blue to green fluorescence, F450/F530, seem not to be or are only slightly influenced by the irradiance applied during plant growth. The blue fluorescence F450 only slightly decreased, whereas the red chlorophyll fluorescence decreased with increasing irradiance applied during growth of the plants. This, in turn, resulted in greatly increased values of the ratio, F450/F690, from 0.5 – 1.5 to 6.4 – 8.0. The decrease in the chlorophyll fluorescence with increasing irradiance seems to be caused by the accumulation of UV light absorbing substances in the epidermal layer which considerably reduces the UV laser light which passes through the epidermis and excites the chlorophyll fluorescence of the chloroplasts in the subepidermal mesophyll cells.     

Complex carbohydrates in the secretory epithelium of the goat prostate

Summary The complex carbohydrates in the epithelium of the goat prostate were studied with a battery of light microscopical histochemical methods that included peroxidase-labelled lectin-diaminobenzidine (PO-LT-DAB) procedures. The secretory epithelial cells were found to be of three types: (a) mucus-producing cells, (b) protein-secreting cells, and (c) cells intermediate between (a) and (b). All these cell types contained variable amounts of neutral and acidic complex carbohydrates. The histophysiological significances of these substances in the glandular cells are discussed with special reference to the function of the accessory sex glands.