Superoxide Dismutases and Anti-Oxidants Protected Mice from no-Reflow and Necrotic Damage Induced by Ischemia

A simple method in mice was established to screen anti-ischemic compounds. Thirteen times binding of rubber ring (1 × 1 mm, d = 42 mm) for 4.5 hrs, swelled the paws of 60% mice applied and 14 times binding swelled only of 5% mice. Critically reversible limit lay between these conditions. “All or none” rule dominated the paw swelling perhap due to different endogenous anti-oxidants' levels of individual mice. Determination of paw reversibility at 90 min of recirculation, was proved to be suitable. Swollen paws at this time returned normal and the paws with no-reflow dropped out by muscle necrosis after several days. Intravenous (i.v.) bovine Cu, Zn-SOD and bacterial Mn-SOD (3 - 10 × 10⁴ U/kg) or liposomal Cu, Zn-SOD (0.3 - 3 × 10⁴ U/kg) were protective (35-50%) by 14 times binding. Allopurinol (10-100 mg/kg) and D-mannitol (3-30mg/kg) was effective (25-55%). Catalase (i.v., up to 10⁵ U/kg) showed little protection, but local injection of 100 U/kg resulted in 50% protection. Glutathione (30 mg/kg) was suppressive only by local injection suggesting the importance of administration route. Desferal, heparin and nitric oxide synthesis inhibitor showed some protection, but indomethacin, mepyramine, ascorbate, vitamin E and dexamethasone were without effect. Excess dosing of all anti-oxidants tested, dramatically decreased their effects demanding caution for therapeutic trials.     

Binding and the nature of Cu(II) ion interaction with nucleosomes

The energetics of Cu (II) ion binding to mononucleosomes from C3HA mice liver and ascitic hepatoma 22A cells was determined from their binding isotherms by equilibrium dialysis and pulse high frequency inductively coupled plasma atomic emission spectroscopy. Anticooperative binding of copper ions with normal and tumor mononucleosomes were observed under various NaCl concentrations (0.002; 0.02; 0.2 M). The binding constants of Cu(II) ion with normal mononucleosomes in 0.002, 0.02, 0.2 M NaCl are 6.10×10⁴,5.22×10⁴,4.31×10⁴ respectively. The binding constants of Cu(II) ion with tumor mononucleosomes in 0.002, 0.02, 0.2 M NaCl are 6.68×10⁴,6.12×10⁴,4.82×10⁴ respectively.     

Heparin, A Potent Releasing Agent of Extracellular Superoxide Dismutase (EC-SOD C), Suppresses Ischaemic Paw Oedema in Mice

Heparin (2,000 U/kg, i.v.) increases the plasma superoxide dismutase (SOD) activity by 2-3 times after 5min. followed by a gradual decrease. A high dose of heparin (4 × 10³ and 10 × 10³U/kg) exhibits a lower increase in the release of SOD. Ischaemic paw oedema in mice was suppressed by various types of SOD and heparin also suppresses this oedema. The dose-dependent curve of heparin of oedema suppression corresponds well with the increased plasma level of SOD. Inducibility with heparin, slow clearance from the bloodstream and blocking of oedema suppression by the copper chelator, diethyldithiocarbamate (DDC), suggest that the oedema suppressing SOD was the extracellular (EC)-SOD C. Other anticoagulants such as citrate and EDTA had no effect. Chondroitin sulphate A and C or carrageenan exhibited weak suppression. A complex of EC-SOD C and heparin appears not to bind to the endothelium in contrast to the injected free EC-SOD C. When heparin is re-injected, more than 1 week was required to get the same degree of oedema suppression. This indicates the necessity of newly synthesized enzyme. A biological role for heparin-induced release of plasma SOD is demonstrated for the first time in this investigation.     

Biological Control and Use of Adjuvants against Multiple Seeded Cocklebur (Xanthium strumarium) in Comparison with Several Other Cocklebur Types

Common cocklebur has several biotypes including multiple seeded cocklebur (MSC), NCC-TX, and NCC-MS. Alternaria helianthi applied at 2.5×10⁴ conidia mL^-1 in a 50% micro-emulsion of unrefined corn oil (MESUCO) or 0.2% Silwet L 77 caused 60-75% mortality on NCC-TX and MSC. Increasing the conidial concentration to 5×10⁴ mL^-1 increased mortality to 100% on MSC and NCC-TX, and 75% on NCC-MS. At 10×10⁴ conidia mL^-1, A. helianthi caused 100% mortality in all three biotypes. No mortality occurred in any biotype at inoculation rates of 2.5 and 5×10⁴ conidia mL^-1 when applied in water. Increasing the dew period from 0 to 12 h increased mortality from 0 to 100% on all three biotypes at a rate of 2.5×10⁴ conidia mL^-1 in Silwet and MESUCO. MSC appears to be the most sensitive biotype.     

Free radical scavenging actions of metallothionein isoforms I and II

By employing electron spin resonance spectroscopy, we examined the free radicals scavenging effects of hepatic metallothionein (MT) isoforms I and II (MTs-I and II) on four types of free radicals. Solutions of 0.15mM of MT-I and 0.3mM of MT-II were found to scavenge the 1,1-diphenyl-2-picrylhydrazyl radicals (1.30 × 10¹⁵ spins/ml) completely. In addition, both isoforms exhibited total scavenging action against the hydroxyl radicals (1.75 × 10¹⁵ spins/ml) generated in a Fenton reaction. Similarly, 0.3mM of MT-I scavenged almost 90% of the superoxide (2.22 × 10¹⁵ spins/ml) generated by the hypoxanthine and xanthine oxidase system, while a 0.3mM MT-II solution could only scavenge 40% of it. By using 2,2,6,6-tetramethyl-4-piperidone as a “spin-trap” for the reactive oxygen species (containing singlet oxygen, superoxide and hydroxyl radicals) generated by photosensitized oxidation of riboflavin and measuring the relative signal intensities of the resulting stable nitroxide adduct, 2,2,6,6-tetramethyl-4-piperidine-1-oxyl, we observed that MT-II (0.3 mM) could scavenge 92%, while MT-I at 0.15 mM μl/ml concentrations could completely scavenge all the reactive species (2.15 × 10¹⁵ spins/ml) generated.

The results of these studies suggest that although both isoforms of MT are able to scavenge free radicals, the MT-I appears to be a superior scavenger of superoxide and 1,1 diphenyl-2-picrylhydrazyl radicals.     

Influence of Agricultural By-products in Liquid Culture on Chlamydospore Production by the Potential Mycoherbicide Fusarium oxysporum Foxy 2

Economically feasible inoculum mass production methods are required for successful application of Fusarium oxysporum Foxy 2 as a potential mycoherbicide. Therefore, different substrates (agricultural by-products) and the factors that influence the production of spores, especially chlamydospores, of Foxy 2 were investigated in liquid cultures. The substrates tested were cotton seed cake, maize stover, wheat and triticale stillage. The presence of plant fibers in the medium of unfiltered cotton seed cake (2.5%, w/v) significantly enhanced chlamydospore, micro- and macroconidia production by 150, 185, and 300%, respectively, compared to the filtered (fiber-free) medium. Regardless of the type of substrate tested, Foxy 2 was able to produce abundant chlamydospores (0.14-2.7×10⁷ mL^-1) in all growth media. Generally, increasing the concentrations of cotton seed cake and maize stover in the medium significantly increased chlamydospore formation; however, this was not the case for wheat-based stillage. To optimize conditions required for chlamydospore production of Foxy 2, the effect of near ultra-violet (NUV) light, substrate combinations (synergism), level of agitation, incubation time and their interaction were studied. A liquid culture of 2.5% (w/v) filtered cotton seed cake, exposed to continuous NUV for 15 days, doubled the yield of chlamydospores (4.7×10⁶ mL^-1) and macroconidia (5×10⁵ mL^-1), and increased microconidial production by one-third (1×10⁸ mL^-1) compared to natural light. An apparent synergistic effect of substrate combination was observed, since the addition of 20% (v/v) of either wheat or triticale-based stillage to maize stover medium (1%, w/v) increased the number of chlamydospores produced by 16 or 12 times, respectively, compared to maize stover alone (1.4×10⁶ mL^-1), and 2 times more than either of the stillages. A significantly positive effect between a high level of agitation tested during incubation and chlamydospore production of Foxy 2 was recorded. All in all, substantial chlamydospore production of Foxy 2 (4.3×10⁷ mL^-1) was successfully achieved within 12 days of incubation in a single-step liquid fermentation through the combination of 0.5% (w/v) maize stover plus 20% (v/v) wheat-based stillage and a high level of agitation (200 rpm).     

DNA-biding of IQ, Me-IQ and Me-IQx, strong mutagens found in broiled foods

Non-covalent DNA-binding has been studied of 2-amino-3-methylimidazo[4,5-f]quinoline (IQ), 2-amino-3,4-dimethylimidazo[4,5-f]quinoline (Me-IQ) and 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (Me-IQx), strong mutagens found in broiled foods. These mutagens are intercalated into DNA, as found by ultraviolet absorption gel electrophoresis. The binding of IQ is stronger with GC pairs than AT pairs in DNA. The binding constants with calf thymus DNA are 1.6 × 10⁶ (Me-IQ), 0.9 × 10⁶ (IQ) and 0.7 × 10⁶ M⁻¹ (Me-IQx) at pH 6.0. This order of DNA affinity agrees with the order of mutagenicity towards Salmonella typhimurium TA98.     

On the Specificity of Allopurinol and Oxypurinol as Inhibitors of Xanthine Oxidase. A Pulse Radiolysis Determination of Rate Constants for Reaction of Allopurinol and Oxypurinol with Hydroxyl Radicals

Allopurinol has been employed as a “specific” inhihitor of xanthine oxidase in studies of hypoxic/ reoxygenation injury. Pulse radiolysis was used to establish rate constants for the reactions of allopurinol and its major metabolite oxypurinol with hydroxyl radicals: values were (1.45 ± 0.241 × 10⁹ M^-1 s^-1 for allopurinol and (4.95 ± 0.84) × 10⁹ M^-1 s^-1 for oxypurinol. These rate constants show that, in view of the amounts of allopurinol that have been used in animal studies. hydroxyl radical scavenging by this molecule could contribute to its biological actions. especially if animals are pre-treated with allopurinol. so allowing oxypurinol to form. The ability of allopurinol to protect tissues not containing xanthine oxidase against reoxygenation injury may be related to radical scavenging by allopurinol and oxypurinol.     

Effects of environmental factors, age and genotype on sperm production and semen quality in Bos indicus and Bos taurus AI bulls in Brazil

The effects of ambient temperature and humidity, month, age and genotype on sperm production and semen quality in AI bulls in Brazil were evaluated. Data from two consecutive years were analyzed separately. Seven Bos indicus and 11 Bos taurus bulls from one artificial insemination (AI) center were evaluated in Year 1 and 24 B. indicus and 16 B. taurus bulls from three AI centers were evaluated in Year 2. Ambient temperature and humidity did not significantly affect sperm production and semen quality, probably because there was little variation in these variables. Month accounted for less than 2% of the variation in sperm production and semen quality. Increased bull age was associated with decreased sperm motility (P<0.10) and increased minor sperm defects (P<0.001) in Year 1. B. indicus bulls had greater (P<0.005) sperm concentration than B. taurus bulls in both years (1.7×10⁹/ml versus 1.2×10⁹/ml in Year 1 and 1.6×10⁹/ml versus 1.2×10⁹/ml in Year 2, respectively). Ejaculate volume was not significantly affected by genotype in Year 1 (6.6 ml versus 6.9 ml in B. indicus and B. taurus bulls, respectively), but B. indicus bulls had greater (P<0.05) total (11.4×10⁹ versus 8.2×10⁹) and viable (6.7×10⁹ versus 4.9×10⁹) numbers of spermatozoa in the ejaculate than B. taurus bulls. In Year 2, B. taurus bulls had greater (P<0.05) ejaculate volume than B. indicus bulls (8.2 ml versus 6.7 ml, respectively) and total and viable number of spermatozoa in the ejaculate were not significantly different between genotypes (10.3×10⁹ versus 9.1×10⁹ and 6.1×10⁹ versus 5.4×10⁹ in B. indicus and B. taurus bulls, respectively). Sperm motility was not significantly affected by genotype (mean, 59%). In Year 1, B. indicus bulls tended (P<0.10) to have more major sperm defects and had more (P<0.05) total sperm defects than B. taurus bulls (11.8% versus 8.7% and 13.6% versus 10.0%, respectively). In Year 2, B. indicus bulls tended (P<0.10) to have more total sperm defects than B. taurus bulls (16.2% versus 13.3%, respectively). In conclusion, neither ambient temperature and humidity nor month (season) significantly affected sperm production and semen quality. B. indicus bulls had significantly greater sperm concentration and B. taurus bulls had significantly fewer morphologically defective spermatozoa.     

Effect of reaction temperature and reaction time on the preparation of low-molecular-weight chitosan using phosphoric acid

Low-molecular-weight chitosan were prepared using 85% phosphoric acid at different reaction temperatures and reaction time. At room temperature, the viscosity average-molecular weights (M_v) of chitosan decreased to 7.1×10⁴ from 21.4×10⁴ after 35 days treatment. The degradation rate decreased with increasing hydrolysis time. The yields of chitosan also continuously decreased from 68.4 to 40.2% after 35 days. At 40, 60 and 80 °C, the molecular weight decreased to 3.70×10⁴, 3.50×10⁴ and 2.00×10⁴ on 8 h hydrolysis, respectively. The yields of chitosan remain at a high level compared with that at room temperature and were 86.5, 71.4 and 61.3% at 40, 60 and 80 °C treatment, respectively. The different reaction time gave chitosan with different molecular weights. At 60 °C, the molecular weight of products decreased to 7.40×10⁴ from 21.4×10⁴ within 4 h, then decreased slowly to 1.90×10⁴ in 15 h. It was also found that the water-solubility of chitosan increased as the molecular weight decreased. Results show the changes in yields and molecular weight of chitooligomers were strongly dependent on the reaction temperature and reaction time.     

Thymus-dependency of the immune response of mice to a primary infection with the nematode Trichuris muris

Adult mice which had been thymectomized, irradiated and given stem-cell protection were incapable of making a self-cure response to a primary infection with the nematode Trichuris muris. The capacity to mount a self-cure response was restored by the injection of 2·5 × 10⁶ mesenteric lymph node lymphocytes or by 2·5 × 10⁸, but not 2·5 × 10⁷, thymocytes. Restoration of the ability to respond to sheep red blood cells was achieved with all three cell injections. Suppression of the immune response was also achieved by injection of ALS during the second week of infection and at intervals thereafter. The results of thymectomy and ALS treatment show that immunity to T. muris is dependent upon the presence of an intact thymus and thymus-dependent cell population.     

Interaction of sialosyl cholesterol with the cell surface of rat astrocytes and its biological activities

The interaction between sialosyl cholesterol (- or neuraminyl cholesterol, - or β-SC) and the plasma membrane of astrocytes was investigated by the use of ¹⁴C-labeled - or β-SC. Both - and β-SC were dose-dependently and time-dependently bound to rat astrocytes. The Scatchard plot analyses showed that rat astrocytes bound apparently 9.69 × 10⁹ molecules of both -SC/cell (apparent K_d = 2.29 × 10⁻⁵ M) and β-SC/cell (apparent K_d = 5.39 × 10⁻⁵ M) at 37°C. Both the binding of -SC to astrocytes and the subsequent inhibition of DNA synthesis were decreased at the low temperature (4°C), and also suppressed by serum proteins including albumin. One molecule of bovine serum albumin (BSA) bound 2.3 molecules of -SC with the slightly lower K_d-value (8.03 × 10⁻⁶ M) than that for the binding site on astrocytes. BSA not only suppressed the -SC-binding to astrocytes but also increased its release from the cells to the culture media. Gangliosides such as GM1 and GM3 unaffected the -SC-binding, promoted the small release of -SC from the cell surface, and inhibited the morphological changes of astrocytes induced by -SC. The mechanism of -SC-binding to cultured astrocytes with reference to the effects of serum or gangliosides is discussed.     

Human fibroblast culture on a crosslinked dermal porcine collagen matrix

The use of a novel porcine-derived collagen biomaterial as a dermal tissue engineering matrix was examined. The matrix is derived from porcine dermis, and is processed to retain the native collagen (Type 1) and elastin structure. Human primary fibroblasts were cultured on the matrix to examine its potential for creating a dermal replacement. Attachment of fibroblasts on the collagen was compared to tissue culture plastic and PET membranes. Cell proliferation was assessed using the MTT assay and DAPI staining. For seeding densities of 5×10⁴ and 1×10⁵ cells cm⁻², PET and plastic demonstrated >95% attachment of seeded numbers after 3 h. The collagen matrix reached levels >80% after 3–4 h with no influence of the seeding density. Matrix samples with perforating pores of 40 μm diameter were also studied. After 216 h culture in static culture, with media replacement every 3 days, the final cell numbers reached 2.1×10⁵ (perforated) and 2.0×10⁵ cells cm⁻² (unperforated). In comparison fibroblast culture in a perfusion bioreactor, with continuous media replacement, reached 2.3×10⁵ (unperforated) and 2.5×10⁵ cells cm⁻² (perforated) after 216 h.     

Binding of divalent copper and calcium ions to poly I

The effect ot Cu²⁺ and Ca²⁺ ions, on the ultraviolet differential (UVD) spectra of single-stranded poly I was studied and the coordination (Δε_b) and conformation (Δε_c) conponents of the spectra calculated The comparison of Δε_b and the UVD spectrum of protonated IMP leads to the conclusion that N(7) ot inosine-5'-monophosphate (IMP) is a coordinating site tor Ca²⁺ and Cu²⁺ ions on the polymer bases. The binding ot Ca²⁺ and Cu²⁺ ions causes differently directed displacements of the four absorption bands of poly I, which are observed in the wavenumber range (50-34) × 10³ cm⁻¹ The calculation of concentration dependencies tor the association constants (K“) ot Ca²⁺ and Cu²⁺ ions binding to poly I bases shows that the binding is cooperative The K“ values for the poly I + Ca²⁺ complex are two orders of magnitude lower than those for the poly 1 + Cu²⁺ complex At low ion concentrations, binding to the poly I phosphates predominates and increases the degree of the polynucleotide helicity. At higher concentrations the spectra are mainly affected by the ion binding to bases, which results in melting of the helical parts of poly I At Ca²⁺ concentrations exceeding 10⁻³ M light-scattering aggregates are formed. The degree of monomer order in them is close to that observed in multistranded helices of poly I     

Development of a Microbial Consortium from a Contaminated Soil That Degrades Pentachlorophenol and Wood-Preserving Oil

An indigenous microbial consortium capable of degrading pentachlorophenol (PCP) and petroleum hydrocarbons (C₁₀-C₅₀) was produced from a soil contaminated with wood-preserving oil. Two 10-L stainless steel soil slurry (10% w/v) bioreactors were operated in fed-batch mode. To verify the growth and efficiency of PCP degraders in the presence of other contaminants, one bioreactor was fed with a PCP-based wood-preserving mixture (WPM) and a second reactor was fed with technical-grade NaPCP. During the 90-day period of activation, PCP, C₁₀-C₅₀, Cl^-, pH, and dissolved oxygen levels were monitored. The microbial community was monitored using specific most probably number (MPN) bacterial counts and mineralization tests. PCP degradation rates increased similarly in both reactors, from 19 to 132 mg/L-d in the NaPCP reactor, and from 41 to 112 mg/L-d in the WPM reactor. Contaminant loss calculations showed that 99.5% of PCP and 92.5% of C₁₀-C₅₀ added to the WPM reactor were biodegraded. It also revealed that 83% of polychlorinated dioxins and furans were removed. PCP-degrading bacteria increased from 7×10² to 1.6×10⁶ bacteria/mL in both reactors, and petroleum hydrocarbon degraders increased from 1.7×102 to 3.4×108 bacteria/mL in the WPM reactor, indicating that the activity of PCP degraders was not inhibited by the presence of microorganisms growing on petroleum hydrocarbons.     

Kinetics of laccase-catalysed TEMPO oxidation

The oxidation of TEMPO (2,2,6,6-tetramethyl-piperidine-1-oxyl radical) has been studied in the presence of recombinant laccases (benzenediol:oxygen oxidoreductase, EC 1.10.3.2) from Polyporus pinsitus (rPpL), Myceliophthora thermophila (rMtL), Coprinus cinereus (rCcL) and Rhizoctonia solani (rRsL) in buffer solution pH 4.5–7.3 and at 25 °C. At pH 5.5 the oxidation constant calculated from the initial rate of TEMPO oxidation was 1.7 × 10⁴, 1.4 × 10³, 7.8 × 10² and 5.2 × 10² M⁻¹ s⁻¹ for rPpL, rRsL, rCcL and rMtL, respectively. The maximal activity of rPpL-catalysed TEMPO oxidation was at pH 5.0. The pK_a obtained in neutral pH range was 6.2. The reactivity of laccases is in a good agreement with laccases copper type I redox potential.

TEMPO oxidation rate increased 541 times in the presence of 10-(3-propylsulfonate) phenoxazine (PSPX). The model of synergistic TEMPO and PSPX oxidation was proposed. Experimentally obtained rate constants for rPpL-catalysed PSPX oxidation were in a good agreement with those calculated from the synergistic model, therefore confirming the feasibility of the model. The acceleration of TEMPO oxidation with high reactive laccase substrates opens new possibilities for TEMPO application as a mediator.     

The effect of Ca on the sulfhydryl reactivity of troponin: Evidence for a Ca-induced conformational charge

Troponin prepared from rabbit skeletal muscle in the presence of dithiothreitol (SH-troponin) was found to have a sulfhydryl content of about 4 moles/1 × 10⁵ g in the presence of a Ca²⁺-chelating agent. The addition of physiological concentrations of Ca²⁺ reduced the reactive sulfhydryl content to 2.0–2.5 moles/1 × 10⁵ g. These sulfhydryl groups are evidently not direct participants in the inhibition of actomyosin superprecipitation, since treatment with N-ethylmaleimide had no effect on the Ca²⁺-sensitizing activity of SH-troponin.

Troponin prepared in the absence of dithiothreitol (S-S-troponin) showed a significant reduction in Ca²⁺-sensitizing activity, relative to SH-troponin. The sulfhydryl groups of S-S-troponin, approx. 2 moles/1 × 10⁵ g were not appreciably affected by Ca²⁺.

It is postulated that the Ca²⁺-sensitive sulfhydryl groups exist at a site which is essential for the regulatory function of troponin and which undergoes a conformational change upon the binding of Ca²⁺.     

ORG 33201: A new highly selective orally active aromatase inhibitor

Org 33201 has been selected as a very potent aromatase inhibitor. The compound is an enantiomer of a SC₂H₅ substituted imidazoylethylphenalene. Org 33201 inhibited human aromatase activity for 50% at a concentration of 2.2 × 10⁹ mol/l. More than 200-fold higher concentrations were needed for the inhibition of other cytochrome P-450 enzymes. In vivo the compound was active in rats (ED₅₀ = 0.035 mg/kg) and dogs (1 mg/kg gave 70% inhibition) after oral administration. It can be concluded that Org 33201 is a potent and highly selective orally active aromatase inhibitor.     

Inhibitory effects of carrier-immobilized synthetic histo-blood group A-, B-, H-, and SiaLe-oligosaccharides on H2O2 generation by human polymorphonuclear leukocytes

Carbohydrate-bearing polymers of biologically inert design are versatile tools to delineate functional aspects of oligosaccharides. Binding of synthetic N-substituted polyacrylamide (PAA) conjugates of histo-blood group (A_di, A_tri, B_di, B_tri, H_di, SiaLe^a, and SiaLe^x) to human polymorphonuclear leukocytes (PMNs), and effects on H₂O₂ generation elicited by different agonists such as digitonin, N-formyl-Met-Leu-Phe (FMLP) and the galactoside-specific lectin from Viscum album L. (VAA) were assessed. PMNs expressed binding sites for blood group-related neoglycoconjugates in the range of N10⁶–10⁷/cell with K_D-values in the μM range. Treatment of PMNs (2×10⁶ cells/ml) with PAA-probes (50 μg/ml) for 5 min did not activate the “respiratory burst”. However, it led to suppression (range 20–70%) of H₂O₂ generation of cells in the presence of elicitors. In detail, the FMLP-induced response was significantly decreased by A_di, A_tri, B_tri, H_di, SiaLe^a, and SiaLe^x conjugates, whereas for digitonin one only by A_di, A_tri, B_tri. All the seven tested PAA-probes were found to inhibit significantly VAA-mediated release of H₂O₂ from PMNs. In this case, interference can take place already, at the stage of initial binding, especially for B- and H-epitopes, but less prominently for A- and SiaLe-epitopes. These results support the notion that PAA-immobilized histo-blood group oligosaccharides can serve as effector molecules with the ability to reduce the H₂O₂-generation of PMNs, warranting further studies on the involved reaction pathway.     

Interaction of anthracycline antibiotics with biopolymers: 7. Equilibrium binding studies on the interaction of iremycin and DNA

We report spectrophotometric equilibrium studies of both the self-association of the new antibiotic iremycin and of its binding to calf thymus DNA in solution (ionic strength 0.2 M; pH 6.0). Iremycin forms dimers in this solution with a dimerization constant K₄=(1.19 ± 0.10) × 10³ M⁻¹. This equilibrium is taken into account in the evaluation of the interaction of iremycin with DNA. The binding behaviour can be completely described by a single binding mechanism of monomeric iremycin to DNA with allowance both for neighbour exclusion and for cooperativity of interaction. The three intrinsic binding parameters for the homogeneous model were determined simultaneously by a least squares fit of the original titration data: equilibrium constant of cooperative binding K = (2.72 ± 0.66) × 10⁵ M⁻¹ cooperativity parameter σ=0.38±3.27 ± 0.32. The binding parameters of iremycin and adriamycin and their microbial activities are compared.