Cytokine profiles in mice tissues after irradiation of the thymus projection area with femtosecond laser

The effects of pulsed femtosecond laser irradiation in the near ultraviolet region on the levels of cytokines in the thymus, blood, and skin of irradiated mice have been studied. Irradiation of the thymus projection area with low-intensity laser radiation in the near UV region of the spectrum showed significant changes in cytokine levels in the skin and thymus and, to a lesser extent, in the blood of irradiated mice. Laser irradiation with a power density of 20 mW/cm² affects the cytokine profile in the thymus: IFN-γ, IL-3, IL-4, IL-5, eotaxin, GM-CSF, and chemokine KC–factors that can affect differentiation and proliferation of the cells of the immune system in the gland. Conclusions: It is assumed that changes in the expression of cytokines in the thymus after laser irradiation are explained by the rearrangement of biochemical processes possibly associated with the maturation of cells in the gland.     

Radiation-induced molecular changes inEscherichia coli B and its S-30 and DNA-plus-membrane fractions

Oxygenated aqueous suspensions ofEscherichia coli B cells in the resting state were irradiated with 0.8-MeV electrons. Dried films of whole cells, the S-30 fraction, and the DNA-plus-membrane fraction were studied by using infrared spectroscopy in conjunction with the technique of attenuated total reflectance (ATR) in the range from 4000 cm^?1 to 800 cm^?1. Cells irradiated in the oxygenated or the anoxic state yield the same kind of molecular damage, the main difference being the lower doses (by a factor 4 or 5) required in well oxygenated systems. Results show that some bonds are more sensitive to radiation than others. Decreases in the PO₂ bands (1225 and 1084 cm^?1) indicate radiation-induced degradation of the DNA-RNA backbone. The increase in absorption between 1700 cm^?1 and 1750 cm^?1 indicates formation of C=O bonds upon exposure to ionizing radiation. Most of the radiation damage occurs in cells that have undergone lysis during irradiation, but the process of cell lysis, by itself, does not cause appreciable molecular bond damage as measured by ATR. Doses ranged from 0.1 Mrad to 1.1. Mrad.     

Investigation of DNA structural changes by infrared spectroscopy

DNA-oriented samples of various origins were studied under different conditions of humidiity and sodium chloride content by means of infrared spectroscopy. (1) Oriented DNA (M. Lysodeikticus, E. coli, calf thymus and salmon sperm) films at 3–4% sodium chloride yield polarized spectra which show drastic changes at relative humidities (r.h.) between 94% and 0% indicative of conformational changes: B form → a form → disordered form The measurements of the infrared dichroism at frequencies of about 1230 cm^?1 and at about 1090 cm^?1 allow one to determine the orientation of the phosphate group, whereas the measurements at 1710 cm^?1 characterize the base orientation. At humidities higher than 90% r.h. (B form) the bisector of OPO forms an angle of 70° relative to the helix axis, whereas at lower humidities, between 75% and 50% r.h. (A form) a rotation to about 45° is observed. Simultaneously, the 0—0 line of phosphate group changes its orientation from 55° to 65° to the helix when B → A transition takes place. The results are in general agreement with that of X-ray diffraction and allow one to determine the orientation of the phosphate group with greater precision. (2) The B–A conformational change is not observed for satellite DNA, isolated from Cancer pagurus, of which the guanine + cytosine content is below 5%. As a function of decreasing humidities, one observes the transition: B form → disordered form A diagram of conformational changes of DNA's as a function of base composition and of r.h., suggests that B–A transition will occur for DNA of relatively higher G + C content, whereas for high (A + T) content, base sequence may be of importance. The B–A transition is prevented in DNA at a relatively high or very low sodium chloride content.     

GC–MS explores the health care components in the extract of Pterocarpus pedatus Pierre

Pterocarpus a high-end, expensive furniture materials collectively. Pterocarpus and Pterocarpus products have a certain human health function. Therefore, this paper to Pterocarpus pedatus Pierre as an example, to study its extract on human health beneficial health care ingredients. FT-IR analysis showed that the infrared transmittance of Pterocarpus pedatus Pierre powder after ethanol/benzene extraction was the highest in the infrared spectrum of 400 cm⁻¹–800 cm⁻¹, 2750 cm⁻¹–3200 cm⁻¹ wave number. In the 1750 cm⁻¹–2400 cm⁻¹ wave segment, methanol, ethyl acetate and ethanol/benzene after the extraction of Pterocarpus pedatus Pierre powder infrared transmittance increased values are basically the same. GC–MS analysis, the health care ingredients in the Pterocarpus pedatus Pierre have cough and phlegm, heat detoxification, enhance human immunity, analgesic and anti-inflammatory and so on. Among them, Homopterocarpin is excellent in inhibiting and killing cancer cell activity; Cryptomeridiol is a natural product with anti-Alzheimer's disease and antispasmodic nature, and its medicinal value is remarkable. Scoparone has a wide range of pharmacological values.     

Interaction of palladium (II) with DNA

The nature of interaction of palladium (II) with calf thymus DNA was studied using viscometry, ultraviolet, visible and infrared spectrophotometry and optical rotatory disperison and circular dichroism measurements. The results indicate that Pd(II) interacts with both the phosphate and bases of DNA. The ORD/CD data indicate that the binding of Pd(II) to DNA brings about considerable conformational changes in DNA.     

Infrared spectrum of a DNA-RNA hybrid

The infrared absorption spectrum in the 4000–400 cm^? region of an oriented film of a DNA–RNA hybrid in its undeuterated and deuterated states was observed with the polarized radiation. Most of the stronger bands found in the double-helical DNA's and double-helical RNA's are identified in the spectrum of the hybrid. The absorption band at 1225 cm^?1 shows a perpendicular dichroism and that at 1085 cm^?1 shows almost no dichroism. These facts indicate that the orientation of the group with respect to the helix axis in the hybrid structure is not entirely the same as that in the double-helical Na DNA at, 75% RH., although the x-ray diffraction pattern of the hybrid is quite similar to that of the DNA A form. The PO₂^? orientation is not the same as that in the double-helical RNA either. The observed dichroism is explained, however, by considering that the PO₂^? group in the RNA moiety takes nearly the same orientation as that in the double-helical RNA, and the PO₂^? group in the DNA moiety takes nearly the same as that in the double-helical DNA.     

Isopycnic centrifugation of sheared L-cell chromatin in metrizamide gradients

Isopycnic gradient centrifugation of L-929 cell chromatin in a 38% (W/V) metrizamide solution yields two distinct fractions. The fraction banding at a density of 1.24 gm/cm² (H chromatin) contains about 10% of the DNA present in the fraction banding at a density of 1.18 gm/cm² (L chromatin). Both fractions contain the same proportions of satellite to main band DNA''s. Some differences can be seen in the DNA: protein ratios and types of proteins present in the H and L chromatin fractions.     

Buoyant densities of natural and synthetic DNA's in CsCl and Cs2SO4 considered as sequence-dependent properties

The buoyant densities of natural and synthetic DNA's can be accurately interrelated if second-neighbor influences are taken into account. We derive the following expressions, based partly on the buoyant densities of six synthetic DNA's, for the buoyant densities ρ (g/cm³) of DNA's having random sequences. In CsCl, and in Cs₂SO₄, . In these equations, H_G is the mole fraction of G : C base pairs in the DNA and the buoyant densities are calculated relative to densities for E. coli DNA of 1.703 and 1.426 (g/cm³) in CsCl and Cs₂SO₄, respectively.     

Pressure dependence of the helix-coil transition temperature for polynucleic acid helices

Thermal denaturation of DNA's and the corresponding helix–coil transformation of artificial polyribonucleic and polydeoxyribonucleic acids have been studied extensively both theoretically^1–13 and experimentally. ^14–30 Much less work has been carried out on the properties of these polynucleic acids at high pressure, and in particular, on the presure dependence of the helix–coil transition temperature.^31–33 Light-scattering techniques have been used in this study to measure the pressure dependence of the helix–coil transition temperature of the two- and three-stranded helices of polyriboadenylic and polyribouridilic acids and of calf thymus DNA. From the slopes of the transition temperature vs. pressure curves and heats of transition obtained from the literature,^20,34 the following volume changes from these helix–coil transitions have been obtained: (a) ?0.96 cc/mole of nucleotide base pairs for the poly (A + U) transition, (b) +0.35 cc/mole of nucleotide base trios for the poly (A + 2U) transition, and (c) +2.7 cc/mole of nucleotide base pairs for the DNA transition. The relative magnitudes and signs of these volume changes which show that poly (A + U) is destabilized by increased pressure, whereas poly (A + 2U) and calf thymus DNA are stabilized by increased pressure, indicates that further development of the helix–coil transition theory for polynucleotides is needed.     

The effect of low ionic strength on the radiation chemistry and physical properties of calf thymus DNA

Studies of ultraviolet and circular dichroism spectra of aqueous solutions of calf thymus (CT) DNA confirm the tendency of DNA to change conformation at low ionic strength. The qualitative shape and transition width of 260 nm melting curves below 1 mM NaCl differed significantly from those previously published for DNA solutions containing 1 mM NaCl and above. Neutral aqueous solutions of CT DNA at low ionic strengths (0.1 mM-10 mM NaCl) were irradiated with low doses of gamma-rays. The melting temperature, Tm, of irradiated DNA samples increased below 1 mM NaCl suggesting interstrand crosslinking of the denatured DNA or formation of regions of more thermally stable DNA conformation. The magnitudes of these radiation responses were found to be a function of the time elapsed between salt concentration changes and irradiation as well as time after irradiation. These results are consistent with the hypothesis that the purine and pyrimidine base chromophores in double stranded DNA are sheltered from radical attack by the sugar phosphate backbone. Low dose radiation studies (0.8-8.0 Gy) of CT DNA in 1 mM NaCl and below showed a split dose and dose rate dependence for the sample melting curves.     

Ultraviolet reactivation of lambda phage: assay of infectivity of DNA molecules by spheroplast transfection

Prior irradiation of non-lysogenic bacteria by ultraviolet light leads to an increase in the viability of infecting irradiated λ phage (ultraviolet reactivation). Similarly, u.v. irradiation of wild type or uvrD bacteria lysogenic for λcIind^? increased the fraction of closed circular duplex phage DNA molecules formed after infection with u.v.-irradiated λ phage. The closed circular molecules isolated from the irradiated lysogens were shown to be free from u.v. damage by a spheroplast transfection assay. The increase of closed circular molecules is sufficient to explain the ultraviolet reactivation observed by the increase of viability of irradiated phage.In ultraviolet reactivation, damage must be erased on irradiated DNA molecules and the repair is independent of total replication of phage genomes, exchange of sister chromatids or recombination between phage genomes. Protein synthesis is necessary to increase the level of closed circular molecules of irradiated λ phage after irradiation of bacteria.     

Differential labelling of DNA in higher plants

Under our experimental conditions labelling of DNA in higher plants with ³²P phosphate, ¹⁴C uridine and ¹⁴C thymidine shows two distinct species of labelled DNA: bulk-DNA and a satellite DNA. The bulk DNA (? = 1.696 g.cm⁻³) does not incorporate ³²P phosphate, but ¹⁴C thymidine. On the other hand the satellite-DNA (? = 1.720 g.cm⁻³) does not incorporate ¹⁴C thymidine, but ³²P phosphate. Both have ¹⁴C-Uridine as a precursor. An attempt has been made to establish the cellular localisation of these two DNA's.     

Purification and characterization of an endonuclease from calf thymus acting on irradiated DNA.

An endonuclease acting on DNA exposed to ultraviolet light or gamma-rays has been extensively purified from calf thymus. The enzyme has a pH optimum at pH 7.0-7.5, acts with equal efficiency in the presence of EDTA or divalent cations (Mg-2+ or Ca-2+), is inhibited by NaCl and tRNA and is inactivated by incubation at 50 degrees C. Its molecular weight, determined by Sephadex chromatography or sodium dodecylsulfate gel electrophoresis, is approx. 30 000. The enzyme catalyzes the formation of breaks with 5'-phosphate termini in double-stranded DNA irradiated with ultraviolet or gamma-rays. It does not act on unirradiated DNA or denatured DNA. Since in all these properties the enzymatic activity on ultraviolet- and gamma-irradiated DNA behaved similarly and since the two activities cochromatographed in all systems used during purification, we conclude that they are associated with the same protein. The site of action of the enzyme in ultraviolet-irradiated DNA is a photoproduct other than pyrimidine dimers. Such a photoproduct can also be induced by irradiation of the DNA in vivo, i.e. within the cells.     

Purification and immunological characterization of DNA polymerase-alpha from human acute lymphoblastic leukemia cells

DNA polymerase-alpha was purified from the cytosol of blast cells of a patient with acute lymphoblastic leukemia by ammonium sulfate fractionation and successive column chromatographies. The purified enzyme had a specific activity of 2943 units/mg protein with activated calf thymus DNA as a template. The enzyme sediments under high-salt conditions as a homogeneous band at 7.2 S and free from other DNA polymerases (beta, gamma) and terminal deoxynucleotidyl transferase activity. The native molecular weight of the enzyme from gel filtration and glycerol gradient centrifugation was found to be 175 000. The values of Stokes radius (53 A), diffusion coefficient (4.05 x 10(-7) cm2/s) and frictional ratio (1.42) determined by gel filtration suggest that the native enzyme is compact and globular. Antibodies to DNA polymerase-alpha were raised in rabbits. These antibodies, partially purified by 50% ammonium sulfate saturation and Sephadex G-200 chromatography, gave one precipitin band on immunodiffusion and inactivate DNA polymerase-alpha-. This antibody preparation also inhibited, in vitro, the activity of DNA polymerase-alpha from calf thymus, phytohemagglutinin-stimulated normal human lymphocytes, as well as that from other leukemic cells. Thus, DNA polymerase-alpha from calf thymus and human leukemic cells resemble each other in antibody specificity.     

Analysis of circular dichroism spectra of homopurine: homopyrimidine DNA's for sequence information and comparison of results with heterobase DNA

Procedures are developed which make a first neighbor frequency analysis possible from a CD spectrum of homopyrimidine: homopurine DNA's. The contribution to the CD spectrum from the various first neighbor frequencies present in homopyrimidine: homopurine-type DNA's has been determined, and hence the CD spectrum for any DNA of this type with known first neighbor frequencies can easily be calculated. An identical analysis is presented for the determination of extinction coefficients. It is further shown that unlike the more usual heteropurine–pyrimidine DNA's a random sequence does not lead to a simplified formalism. Finally, it is concluded that the homopyrimidine: homopurine DNA's have a structure that is different from that of the more usual heterobase DNA's. A procedure capable of determining first neighbor frequencies from a CD spectrum for heterobase and/or homopurine: homopyrimidine DNA's is described. This procedure is used to determine that there is only minimal interference between these two types of DNA in the first neighbor analysis.     

Isolation and some properties of a mammalian ribosomal DNA

The DNA coding for 28 S and 18 S ribosomal RNA, including the spacer regions, has been isolated from calf (Bos taurus) thymus gland. The method used included shearing of the total DNA to a highly homogeneous size population, selective heat denaturation and S 1 nuclease treatment to remove single stranded DNA. Repeated centrifugation on density gradients yields a 140-fold purified rDNA fraction with a GC content of 61.2%. Eco RI nuclease cleaves this DNA into two fragments of 16.4 and 4.9×10⁶ daltons. Hybridization of these fragments with 28 S and 18 S rRNA shows that the 28 S coding sequence is located mostly on the 4.9×10⁶ dalton fragment, while both the 16.4 and 4.9×10⁶ dalton fragments contain the 18 S sequence. The data indicate that the ribosomal RNA gene has a repeat unit of 21.3×10⁶ daltons which includes a nontranscribed spacer of about 12.5×10⁶ daltons.     

Effect of low-energy electron irradiation on DNA damage by Fe3+ ion

We investigated the combined effects of low-energy electron irradiation and Fe(3+) ion on DNA damage. We used lyophilized pBR322 plasmid DNA films with various concentrations (0 ~ 7 mM) of Fe(3+) ions and irradiation with monochromatic, low-energy 3 or 5 eV electrons for these studies. DNA-Fe(3+) films were recovered and analyzed by agarose gel electrophoresis to identify and compare the effects of Fe(3+) ions and/or low-energy electrons alone or in combination on DNA damage. In nonirradiated DNA-Fe(3+) films, there was little DNA damage observed (less than 10% of the total DNA loaded on the gel appeared damaged) for Fe(3+) ion up to 7 mM concentration. In irradiated DNA films without Fe(3+) ions, there was also very little DNA damage observed (less than 3% of the total DNA loaded on the gel appeared damaged). However, when DNA-Fe(3+) films, were irradiated with low-energy electrons, DNA damage was significantly increased compared to the sum of the damage caused both by either Fe(3+) ion or low-energy electrons irradiation alone. We proposed that both DEA and/or electron transfer processes might play a role in the enhanced DNA damage when DNA-Fe(3+) films were irradiated by low-energy electrons.     

Preparation and Antioxidant Activity of Acetylated Mung Bean Peel Polysaccharides

Mung bean peel polysaccharides are one of the main active components in mung bean peel. Acetylated mung bean peel polysaccharides were prepared by extracting and acetylating them, and characterized by infrared and ultraviolet methods to preliminarily understand the structural characteristics and activity of acetylated mung bean peel polysaccharides. Acetylation modification can improve the structure of polysaccharides, thereby causing changes in their properties. The product obtained after acetylation modification exhibited new characteristic absorption peaks at 1732 cm⁻¹, and the scavenging ability of hydroxyl radicals was improved. Therefore, acetylation modification of mung bean peel polysaccharides could enhance the activity by improving the structure, which provided an experimental basis for the application of mung bean peel polysaccharides.     

Changes in Raman vibrational bands of calf thymus DNA during the B-to-A transition

The B -to-A conformational transition of calf thymus DNA fibers was followed employing Raman spectroscopy. The transition was induced by soaking DNA fibers in water/ethanol mixtures increasing from 60 to 85% ethanol (v/v). Intensity changes of 17 Raman vibrational bands were quantified in the region from 400 to 860 cm^?1. Two bands at 500 and 784 cm^?1 were employed as internal standards. These bands do not appear to change in intensity with ethanol concentration. Large intensity changes relative to these two bands are observed between 70 and 74% ethanol for backbone vibrations at 708, 808, and 835 cm^?1, and base vibrations at 682, 730, and 750 cm^?1. These results indicate that a highly cooperative conformational change takes place between different portions of DNA in the B -to-A transition. Relative intensity changes preceding the onset of the major transition are observed in only two bands; at 835 cm^?1, assigned to a ribose–phosphate vibration, and at 750 cm^?1, assigned to thymine. The implications of these pretransition changes are discussed.