Interactions of a biocompatible water-soluble anthracenyl polymer derivative with double-stranded DNA

A water soluble cationic porphyrin derivative TMPipEOPP can preferentially photocleave G-quadruplex in the presence of double-stranded DNA, thus might be used as a human telomere-targeted photosensitizer for tumor photodynamic therapy.

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Guanine-specific DNA oxidation photosensitized by the tetraphenylporphyrin P(V) complex

Journal of Porphyrins and Phthalocyanines ◽

10.1142/s108842460600065x ◽

2006 ◽

Vol 10 (11) ◽

pp. 1285-1292 ◽

Cited By ~ 4

Author(s):

Kazutaka Hirakawa ◽

Shosuke Kawanishi ◽

Hiroshi Segawa ◽

Toru Hirano

Keyword(s):

Dna Damage ◽

Electron Transfer ◽

High Performance ◽

Near Infrared ◽

Infrared Emission ◽

Water Soluble ◽

Electron Transfer Mechanism ◽

Double Stranded Dna ◽

Dna Photodamage ◽

Water Soluble Porphyrin

Porphyrins have been studied as photosensitizers in photodynamic therapy. DNA is one of the most important targets of the sensitizer. In the present study, we have examined the photosensitized DNA damage caused by dihydroxo P ( V ) tetraphenylporphyrin ( P ( V ) TPP ), a cationic water-soluble porphyrin. P ( V ) TPP photosensitized guanine-specific damage to the DNA fragment. P ( V ) TPP induced severe photodamage to single-stranded rather than to double-stranded DNA. High performance liquid chromatography measurements confirmed the formation of 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxo-G), an oxidized product of 2'-deoxyguanosine, and showed that the content of 8-oxo-G in single-stranded DNA is larger than that in double-stranded DNA. The effects of reactive oxygen scavengers on DNA damage suggested the involvement of singlet oxygen (1 O 2). Photosensitized 1 O 2 formation was confirmed by near-infrared emission measurements. The results showed that 1 O 2 formation mainly contributes to the mechanism of DNA photodamage by P ( V ) TPP . Absorption spectrum measurements showed the interaction between P ( V ) TPP and DNA. This interaction is expected to enhance the 1 O 2-mediated DNA damage since the lifetime of 1 O 2 in a cell is very short. On the other hand, P ( V ) TPP induced DNA damage at the consecutive guanines in double-stranded DNA. Because the consecutive guanines act as a hole trap, this DNA-damaging pattern suggests the partial involvement of photo-induced electron transfer. The fluorescence of P ( V ) TPP was quenched by DNA, supporting the electron transfer mechanism. However, DNA damage by electron transfer was not a main mechanism possibly due to reverse electron transfer. In conclusion, P ( V ) TPP binds to DNA and induces guanine-specific, photo-oxidation mainly via 1 O 2 generation.

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A Resorcinol-Formaldehyde Resin as an Embedding Material for Electron Microscopy of Membranes

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010006355x ◽

1969 ◽

Vol 27 ◽

pp. 328-329 ◽

Cited By ~ 1

Author(s):

J. G. Robertson ◽

D. F. Parsons

Keyword(s):

Electron Microscopy ◽

Osmium Tetroxide ◽

Neutral Lipids ◽

Lipid Extraction ◽

Water Soluble ◽

Formaldehyde Resin ◽

Electron Microscope Autoradiography ◽

Resorcinol Formaldehyde ◽

Major Disadvantage ◽

Cell Organization

The extraction of lipids from tissues during fixation and embedding for electron microscopy is widely recognized as a source of possible artifact, especially at the membrane level of cell organization. Lipid extraction is also a major disadvantage in electron microscope autoradiography of radioactive lipids, as in studies of the uptake of radioactive fatty acids by intestinal slices. Retention of lipids by fixation with osmium tetroxide is generally limited to glycolipids, phospholipids and highly unsaturated neutral lipids. Saturated neutral lipids and sterols tend to be easily extracted by organic dehydrating reagents prior to embedding. Retention of the more saturated lipids in embedded tissue might be achieved by developing new cross-linking reagents, by the use of highly water soluble embedding materials or by working at very low temperatures.

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Ferritin Labeled Antibody Staining of Thin Sections

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100064013 ◽

1969 ◽

Vol 27 ◽

pp. 420-421

Author(s):

J. D. McLean ◽

S. J. Singer

Keyword(s):

Biological Material ◽

Water Soluble ◽

Thin Sections ◽

Antibody Staining ◽

Thin Sectioning ◽

Ultrathin Sections

The successful application of ferritin labeled antibodies (F-A) to ultrathin sections of biological material has been hampered by two main difficulties. Firstly the normally used procedures for the preparation of material for thin sectioning often result in a loss of antigenicity. Secondly the polymers employed for embedding may non-specifically absorb the F-A. Our earlier use of cross-linked polyampholytes as embedding media partially overcame these problems. However the water-soluble monomers used for this method still extract many lipids from the material.

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Discrimination of monomer from multimer DNA disk-shaped toruses by freezeetch TEM

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100111161 ◽

1984 ◽

Vol 42 ◽

pp. 210-211

Author(s):

George C. Ruben ◽

Kenneth A. Marx

Keyword(s):

Biochemical Data ◽

Dna Structures ◽

Ice Surface ◽

Double Stranded Dna ◽

Data Support ◽

Dna Organization ◽

Trivalent Cations

In vitro collapse of DNA by trivalent cations like spermidine produces torus (donut) shaped DNA structures thought to have a DNA organization similar to certain double stranded DNA bacteriophage and viruses. This has prompted our studies of these structures using freeze-etch low Pt-C metal (9Å) replica TEM. With a variety of DNAs the TEM and biochemical data support a circumferential DNA winding model for hydrated DNA torus organization. Since toruses are almost invariably oriented nearly horizontal to the ice surface one of the most accessible parameters of a torus population is annulus (ring) thickness. We have tabulated this parameter for populations of both nicked, circular (Fig. 1: n=63) and linear (n=40: data not shown) ϕX-174 DNA toruses. In both cases, as can be noted in Fig. 1, there appears to be a compact grouping of toruses possessing smaller dimensions separated from a dispersed population possessing considerably larger dimensions.

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The Effect of Benzene on Bluegill Olfactory Lamellae

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010011965x ◽

1985 ◽

Vol 43 ◽

pp. 574-575

Author(s):

D.R. Mattie ◽

J.W. Fisher

Keyword(s):

Surface Morphology ◽

Soluble Fraction ◽

Lepomis Macrochirus ◽

Sublethal Concentration ◽

Water Soluble ◽

Major Constituent ◽

Jet Fuels ◽

Aquatic Biota ◽

Normal Surface ◽

Cacodylate Buffer

Jet fuels such as JP-4 can be introduced into the environment and come in contact with aquatic biota in several ways. Studies in this laboratory have demonstrated JP-4 toxicity to fish. Benzene is the major constituent of the water soluble fraction of JP-4. The normal surface morphology of bluegill olfactory lamellae was examined in conjunction with electrophysiology experiments. There was no information regarding the ultrastructural and physiological responses of the olfactory epithelium of bluegills to acute benzene exposure.The purpose of this investigation was to determine the effects of benzene on the surface morphology of the nasal rosettes of the bluegill sunfish (Lepomis macrochirus). Bluegills were exposed to a sublethal concentration of 7.7±0.2ppm (+S.E.M.) benzene for five, ten or fourteen days. Nasal rosettes were fixed in 2.5% glutaraldehyde and 2.0% paraformaldehyde in 0.1M cacodylate buffer (pH 7.4) containing 1.25mM calcium chloride. Specimens were processed for scanning electron microscopy.

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Synchrony of Digestion of SV40 DNA by Exonuclease III Determined by EM

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100116991 ◽

1975 ◽

Vol 33 ◽

pp. 674-675

Author(s):

Ray Wu ◽

G. Ruben ◽

B. Siegel ◽

P. Spielman ◽

E. Jay

Keyword(s):

Dark Field ◽

Uranyl Acetate ◽

Enzyme Digestion ◽

Dna Molecules ◽

Exonuclease Iii ◽

Aluminum Films ◽

Double Stranded Dna ◽

Before And After ◽

Exo Iii ◽

Sv40 Dna

A method for determining long nucleotide sequences of double-stranded DNA is being developed. It involves (a) the synchronous digestion of the DNA from the 3' ends with EL coli exonuclease III (Exo III) followed by (b) resynthesis with labeled nucleotides and DNA polymerase. A crucial factor in the success of this method is the degree to which the enzyme digestion proceeds synchronously under proper conditions of incubation (step a). Dark field EM is used to obtain accurate measurements on the lengths and distribution of the DNA molecules before and after digestion with Exo III, while gel electrophoresis is used in parallel to obtain a mean length for these molecules. It is the measurements on a large enough sample of individual molecules by EM that provides the information on how synchronously the digestion proceeds. For length measurements, the DNA molecules were picked up on 20-30 Å thick carbon-aluminum films, using the aqueous Kleinschmidt technique and stained with 7.5 x 10-5M uranyl acetate in 90% ethanol for 3 minutes.

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