Antioxidant and anticataractogenic effects of topical captopril in diquat-induced cataract in rabbits.

1-[(2s)-3-Mercapto-2-methylpropionyl]-L-proline (captopril), an antihypertensive and free radical scavenger, protected the rabbit lens from peroxidative and oxidative damage induced by 1 mM diquat in vitro. To evaluate the anticataract efficacy of captopril, an experimental group of five rabbits was treated with topical captopril (1% in 0.15 M NaCl, w/v), and 50 microliters was instilled onto both eyes four times a day for a total of 8 weeks. Following the same procedure, the eyes of five rabbits were treated with topical 0.15 M NaCl as a control for captopril treatment. At the end of the first week of treatment, a single intravitreal dose of 120 nmole diquat in 30 microliters of 0.15 M NaCl was injected into the right eye of each rabbit of both the groups. As a control for intravitreal diquat injection, the left eye of all the rabbits were injected with the diluent, 30 microliters per eye. The intravitreal diquat or its diluent injection was only for one time. From slit-lamp biomicroscopic observation of the diquat-injected right eyes, the anticataract effect of captopril in the treatment group was indicated by the finding that in four of five rabbits the cataract did not advance; whereas in four of five rabbits treated with the diluent the cataract progressed to grade 3. The lenses in the diluent-injected control left eyes of the rabbits treated with the captopril or diluent were normal. However, since the number of animals used for the in vivo studies was few, further confirmation of the anticataract effect of captopril is necessary. In diquat-injected right eyes of animals treated with captopril, the integrated rate of O2- production was about 50% less (p less than .001) in the aqueous humor, vitreous humor, and lens, compared with O2-, 33.49 +/- 2.26 microM (mean +/- SEM) in the aqueous humor, 17.12 +/- 0.75 microM in the vitreous humor, and 31.44 +/- 1.29 nmole/g wet weight in the lens of the diquat-injected right eyes treated with the diluent. Similar significant (p less than .01) differences in the production of .OH and H2O2 in eye tissues were also observed.(ABSTRACT TRUNCATED AT 400 WORDS)     

Water-soluble cationic gallium(III) and indium(III) phthalocyanines for photodynamic therapy

The new tetra-non-peripheral and peripheral 2-mercaptopyridine substituted gallium(III) and indium(III) phthalocyanine complexes (np-GaPc, p-GaPc, np-InPc and p-InPc) and their quaternized derivatives (Qnp-GaPc, Qp-GaPc, Qnp-InPc and Qp-InPc) have been synthesized and characterized. The quaternized complexes show excellent solubility in water, which makes them potential photosensitizer for use in photodynamic therapy (PDT) of cancer. Photophysical and photochemical properties of these phthalocyanines were investigated. General trends are described for quantum yields of photodegradation, fluorescence and fluorescence lifetimes as well as singlet oxygen quantum yields of these compounds. In this study, the effects of the position of the substituents, the nature of the metal ion and quaternization of the substituents on the photophysical and photochemical parameters of the gallium(III) and indium(III) phthalocyanines are also reported. This study also presented the ionic gallium(III) and indium(III) phthalocyanines strongly bind to bovine serum albumin (BSA).     

Chromium(III) interactions with nucleotides. III

Some new derivatives of Cr(III) with 5′AMP, 5′ATP, 5′CMP, 5′GMP, 5′IMP and 5′UMP have been obtained by reaction of the starting complexes cis and trans-[Cr(en)₂Cl₂]Cl with the above nucleotides.The complexes were characterized by elemental analysis, conductivity, infrared and electronic spectroscopy, and EPR for the 5′UMP derivative.In all cases, chlorine has been substituted and one ethylenediamine eliminated. The interaction of Cr(III) with the nucleotide seems to occur through the phosphate group and additional interaction through the heterocyclic ring especially for the 5′GMP and 5′IMP derivatives.The 5′UMP complex seems to be a dimer and the other complexes are polymer.     

Bromodeoxyuridine (BrdU) immunocytochemistry by exonuclease III (Exo III) digestion.

A new procedure is described to generate single-stranded DNA by exonuclease III (Exo III) digestion for bromodeoxyuridine (BrdU) immunocytochemistry on tissue sections. We compared this procedure with the most widely used procedure of DNA denaturation with 2 N HCl. In vivo and in vitro pulse and continuous labelling of tissues and cells were used. The specimens were fixed in formalin, ethanol, glutaraldehyde, Carnoy's, Bouin's or Zamboni's fixative and embedded in paraffin or used unfixed as cryostat sections or cytospin preparations. After Exo III digestion, BrdU substituted DNA was detected irrespective of the fixation procedure applied. The optimal protocol for nuclease digestion appeared to be simultaneous incubation, of 10 Units Exo III per ml EcoRI buffer and anti-BrdU monoclonal antibody at 37 degrees C. The advantages of Exo III digestion for BrdU immunocytochemistry compared to acid denaturation were: less non-specific nuclear background reactivity, no DNA renaturation, less DNA loss, optimal nuclear morphology, increase in antibody efficiency and the possibility for simultaneous detection of acid-sensitive tissue constituents. Disadvantages of the Exo III digestion are decreased sensitivity and the need for more rigorous pepsin pretreatment. We conclude that Exo III digestion of DNA is an appropriate alternative for acid denaturation for BrdU immunocytochemistry on sections of pulse-labelled specimens.     

Kinetic studies on the interaction of gold (III) with nucleic acids. IV. RNA-Au (III) system.

The kinetics of the interaction of Au(III) with whole yeast RNA has been studied using UV-spectrophotometry. The reaction is second order with respect to the nucleotide unit of RNA and first order with respect to Au(III) in the respective stoichiometry of 2 : 1. The effects of initial composition, temperature, ionic strength, pH and chloride ion on the kinetics have been studied. Activation energy is found to be 11.5 kcal/mol. Effect of ionic strength indicates that both the positively charged and neutral species of Au(III) take part in the rate limiting step, the former being dominant at low ionic strength. A plausible mechanism has been proposed which involves the interaction of two nucleotide units of RNA with one species of Au(III) in the rate limiting step.     

Molecular differentiation of the rabbit ovum. III. Fertilization-autonomous polypeptide synthesis

Autoradiographic patterns of [1-³⁵S]methionine-labeled polypeptides separated by two-dimensional polyacrylamide gel electrophoresis were obtained from (1) rabbit oocytes that had undergone meiotic maturation to metaphase II in vivo or in vitro, (2) in vitro matured oocytes cultured for an additional 36 hr, or recovered from the reproductive tract at 36 hr after ovulation, (3) newly fertilized eggs, and (4) embryos developed in vivo or in vitro from the 1-cell stage to the 12- to 16-cell stage. The findings indicate that the detectable synthesis of a set of stage-specific (cleavage) polypeptides is autonomous of fertilization and appears to follow a timed, translational schedule initiated with the breakdown of the oocyte nucleus during the resumption of arrested meiosis.     

Photophysical properties of Pr(III) and Er(III) complexes of poly(pyrazolyl)borates.

The complexes [M(L(1))(2)(NO(3))] and [M(L(2))(NO(3))(2)](M = Pr, Er; L(1)= the tetradentate ligand dihydrobis-[3-(2-pyridyl)pyrazolyl]borate; L(2)= the hexadentate ligand hydrotris-[3-(2-pyridyl)pyrazolyl]borate) were prepared and their structural and photophysical properties studied. All complexes are 10-coordinate. Crystallographic analysis of [M(L(1))(2)(NO(3))](M = Pr, Er) showed that for the smaller Er(iii) ions steric congestion at the metal centre results in two of the Er-N(pyridyl) distances being particularly long, which does not occur with the larger Pr(iii) ion that is better able to accommodate 10-fold coordination. On UV irradiation, both Pr(iii) complexes show, in the visible region of their luminescence spectra, transitions originating from both the (3)P(0) level (at ca. 21,000 cm(-1)) and the (1)D(2) level (at ca. 17,000 cm(-1)), a consequence of the fact that the lowest triplet state of the coordinated pyrazolylborate ligands lies at ca. 24,000 cm(-1) in each case so is high enough in energy to populate both levels. This contrasts with Pr(iii) complexes based on diketonate ligands in which the lower triplet energies of the ligands result in emission from the (1)D(2) level only. At longer wavelengths, near-infrared luminescence arising from the (1)D(2) emissive level is observed with lifetimes (in both the solid state and solution) being in the range 50-110 ns. For both Er(iii) complexes, luminescence at 1530 nm occurs following UV excitation of ligand-centred transitions. In CH(2)Cl(2) both complexes gave dual-exponential luminescence, with the major component having a lifetime characteristic of an intact Er(iii) complex (approximately 1.5 micros) and the minor component being much shorter lived (0.2-0.5 micros), suggestive of a species in which a ligand is partially detached and the metal is solvated, with the two forms interconverting slowly. This behaviour is consistent with the steric congestion and long M-N(pyridyl) bonds that were observed in [Er(L(1))(2)(NO(3))]. In the solid state both Er(iii) complexes gave very weak luminescence, which could be fitted to a single exponential decay with a lifetime similar to the longer-lived of the solution components.     

Pulmonary metabolism of prostacyclin (PGI2) in the rabbit.

Metabolism of prostacyclin, [9-³H]PGI₂, was examined in the isolated perfused rabbit lung and the post-microsomal supernate of rabbit lung homogenate. Two major metabolites of [9-³H]PGI₂ from the lung perfusate were separated by thin-layer chromatography and radiometric gas-chromatography. These two products were identified as 6 keto-PGF_1α and 6,15 diketo-13,14 dihydro PGF_1α by mass-spectrometry; they represented 65% and 14% of the total radioactivity. When [9-³H]PGI₂ was incubated with the lung homogenate in the presence of either NAD⁺ or NADP⁺, more than 36% and 25%, respectively, was converted to the 6,15 diketo-13,14 dihydro metabolite.     

Aminopolycarboxylates of rare earths. 13. The apparent molal volumes of the lanthanide(III), cobalt(III) and chromium(III) ethylenediamine tetraacetate complexes

The apparent molal volumes of the complexes KLnedta (Ln = La, Ce, Pr, Nd, Sm, Eu, Gd, Dy, Er and Yb), KCoedta and KCredta were determined from density measurements. The apparent molal volumes of the complexes KLnedta show a non-monotonous change with increasing atomic number; there is a significant increase in the values between Nd and Gd. To explain the observed trend, a structural change is postulated to take place gradually on progressing from Gd towards La. With increasing ionic size, there is an increase in the metal-ligand bond distances and in the mobility of the functional groups, resulting in a jump in the number of coordinated water molecules by one between Gd and Nd. There is a further increase in the hydration of the central ions, connected with the gradual de-coordination of a carboxylate group. As a result of the gradual structural change there is one free carboxylate group on average in Laedta^? and about four water molecules are coordinated in the inner sphere, while in the complexes of the elements heavier than Gd the ligand is hexadentate and only two water molecules are in the inner sphere. The apparent molal volumes of KCoedta and KCredta are practically equal and are higher than those of the lanthanide complexes; this has been explained by the stronger hydration and the more significant water-ordering effect of the Ln³⁺ ions coordinated in the complexes Lnedta^?.The postulated structural changes correlate with the trend of the protonation constant values, as well as with the results of ¹H NMR studies on the complexes Lnedta^?.     

Vogeleier aus Kansu. (III)

Ohne ZusammenfassungI: J. f. O. 1929, p. 35–40. — II: Orn. Monatsber. 1929, p. 172–175.     

兔的一种新病毒：...

In the spring 1986, an acute infectious disease occurred in Wuhan Second Producing Medical Manufactory, and the rabbit almost died. We tested the mortal symptom and confirmed rabbit Hemorrhagic Disease (RHD) as same as Huang Yinyao report. Hubei Traditional Chinese Medicine Institute appear this RHD also. After we purified virus of above two source by low speed, high speed and sucrose density gradient centrifugation, they can react with antiserum of RHDV from Nanjing Agricultural University in agar gel immunodiffusion tests. These results proved that they belong to the same serotype. Data indicate RHDV have difference morphological superstructure, viral polypeptides and especially RHDV can't react with antiserum of standard Parvovirus of rabbit and so on, so we suggest RHDV is a new virus.     

Effects of paraquat on the green alga Dunaliella salina: protection by the mimic of superoxide dismutase, Desferal-Mn(IV)

Paraquat caused a time-, dose-, and light-dependent bleaching of the halophilic green alga Dunaliella salina. Sublethal levels of paraquat elicited increases in cell content of both superoxide dismutase and catalase, with changes in the pattern of electromorphs of these enzymes. Desferal-Mn(IV), which catalyzes the dismutation of O2- in vitro, protected against the toxic effect of paraquat. Desferal (desferoxamine mesylate) alone was toxic to D. salina, and the salts of Mn(II), Mn(III), and Mn(IV), in the absence of Desferal, did not protect. Desferal-Mn(IV) is green, but its absorbance was 15% or less than the peak absorbances due to the chlorophyll in D. salina under the conditions of exposure; hence, masking of incident light could not have been the basis of the protective effect of the complex. Incubation of the cells with Desferal-Mn(IV), for up to 8 h prior to the addition of paraquat, did not increase its protective action, and brief washing, following 30 min incubation with the complex, eliminated its protective effect. Neither catalase nor superoxide dismutase, added to the medium, provided protection against paraquat. These results support the view that Desferal-Mn(IV) gains entry into D. salina and protects against the lethal effect of paraquat by there catalyzing the dismutation of O2- into H2O2 + O2.     

Synthesis of rhodium(III), iridium(III) and osmium(III) complexes with tris(2-aminoethanethiolato)cobalt(III)

Polynuclear sulfur bridged complexes where the neutral complex tris(2-aminoethanethiolato)cobalt(III) acts as a tridentate ligand to rhodium(III), iridium(III) and osmium(III) have been prepared. These complexes have been characterized by electronic spectroscopy, vibrational spectroscopy and nuclear magnetic resonance spectroscopy. Along with the previously prepared complexes of iron(III), ruthenium(III) and cobalt(III), these complexes form two series of complexes with the group 8 and group 9 elements from all three transition series.     

Europium(III) luminescence and tyrosine to terbium(III) energy-transfer studies of invertebrate (octopus) calmodulin.

The effects of minor differences in the amino acid sequences between a vertebrate (bovine testes) and an invertebrate (octopus) calmodulin on metal ion binding were investigated via laser-induced Eu3+ and Tb3+ luminescence. Amino acid substitutions at residues which are coordinated to the metal ion do not produce any detectable changes in the 7F0----5D0 excitation spectrum of the Eu3+ ion bound to octopus calmodulin relative to bovine testes calmodulin; only minor differences in the excited-state lifetime values in D2O solution are observed. The dissociation constants for Eu3+ (1.0 +/- 0.2 microM) and Tb3+ (5 +/- 1 microM) from the weak lanthanide binding sites (III and IV, numbered from the amino terminus) of octopus calmodulin were measured using luminescence techniques. Both values agree well with those reported previously for bovine testes calmodulin [Mulqueen, P. M., Tingey, J. M., & Horrocks, W. D., Jr. (1985) Biochemistry 24, 6639-6645]. The measured dissociation constant of Eu3+ bound in the tight lanthanide binding sites (I and II) is 6 +/- 2 nM for octopus calmodulin and 12 +/- 2 nM for bovine testes calmodulin. The distances between sites I and II (12.4 +/- 0.5 A) and sites III and IV (11.7 +/- 0.8 A) were determined from F?rster-type energy transfer in D2O solutions of octopus calmodulin containing bound Eu3+ donor and Nd3+ acceptor ions. F?rster theory parameters for nonradiative energy transfer between Tyr138 and Tb3+ ions bound at sites III and IV of octopus calmodulin were comprehensively evaluated, including a dynamics simulation of the orientation factor kappa 2. This theory is found to account quantitatively for the observed energy-transfer efficiency as evaluated from the observed sensitized Tb3+ emission.     

Intensity of cooperative vibronic transitions in the Eu(III), Gd(III) and Tb(III) formates spectra

The luminescence and absorption spectra of the lanthanide ions in solids and coordination compounds are characterized by sharp pure electronic lines, which are accompanied by much weaker lines of vibronic transitions. The vibronic spectroscopy is a good probing tool for investigations of the properties of surrounding ion ligands. The lanthanides formates are efficient luminescent crystals and can be viewed as the elementary type in the whole class of the oxygen-containing lanthanide coordination compounds. The intensity of vibronic transitions in spectra of luminescence and excitation europium (⁵D₀→⁷F₂, ⁷F₀→⁵D₂), terbium (⁷F₆→⁵D₄), gadolinium (⁶P_7/2→⁸S_7/2) in anhydrous formates of the type Ln(HCOO)₃ (Ln = Eu, Tb, Gd) and Y(HCOO)₃.2H₂O doped with Eu³⁺ and Tb³⁺ (C ~1 mol%) are reported. Also, the infrared and Raman spectra were obtained for the same compounds. Related integral intensity vibronic sidebands depend on the type of electronic transition of the same ion and varies for the same electronic transitions in different crystals. The obtained experimental data referring to the rate constants of vibronic transitions and intensity distribution in vibronic spectra on normal vibrations of the formate groups are in agreement with the predictions based on the Stavola–Dexter theory of cooperative vibronic transitions.     

Fluorescence of phosphotyrosine--terbium(III) complexes.

Phosphotyrosine, a biologically important protein residue, was investigated for the ability to enhance terbium (Tb3+) fluorescence. Spectroscopic analysis of the Tb3+: phosphotyrosine interaction indicated the development of a new excitation peak at 275 nm and strong Tb+ fluorescence enhancement at 488 and 540 nm that was linear over a range from 0.5 to 100 microM amino acid. Subsequent experiments comparing the ability of phosphotyrosine, phosphothreonine, phosphoserine and 20 other common non-phosphorylated amino acids showed that only phosphotyrosine produced significant Tb3+ fluorescence enhancement. Analysis of various phospho-sugars and nucleotides showed (with the expected exception of GMP) that they produced little or no significant fluorescence enhancement, indicating a further selectiveness for the phosphotyrosine: Tb3+ fluorescence enhancement event. These results establish a basis for the future use of Tb3+ fluorescence enhancement as a unique probe for the investigation of phosphotyrosine residues.