Evolutionary stability of antibiotic protection in a defensive symbiosis.

The increasing resistance of human pathogens severely limits the efficacy of antibiotics in medicine, yet many animals, including solitary beewolf wasps, successfully engage in defensive alliances with antibiotic-producing bacteria for millions of years. Here, we report on the in situ production of 49 derivatives belonging to three antibiotic compound classes (45 piericidin derivatives, 3 streptochlorin derivatives, and nigericin) by the symbionts of 25 beewolf host species and subspecies, spanning 68 million years of evolution. Despite a high degree of qualitative stability in the antibiotic mixture, we found consistent quantitative differences between species and across geographic localities, presumably reflecting adaptations to combat local pathogen communities. Antimicrobial bioassays with the three main components and in silico predictions based on the structure and specificity in polyketide synthase domains of the piericidin biosynthesis gene cluster yield insights into the mechanistic basis and ecoevolutionary implications of producing a complex mixture of antimicrobial compounds in a natural setting.

Actinomadura roseirufa sp. nov., producer of semduramicin, a polyether ionophore.

The taxonomic position of 'Actinomadura roseorufa' LMG 30035T, a semduramicin-producing mutant of strain ATCC 53666P, which was isolated from a soil sample collected in Yamae Village, Kamamoto, Japan, was clarified in the present study using a polyphasic approach. This Gram-positive, aerobic actinomycete formed a well-developed, extensively branched, non-fragmenting substrate and aerial mycelia which differentiated into single, smooth-appearing spores. Based on analysis of nearly complete 16S rRNA gene sequence, strain LMG 30035T was found to be closely related to the type strains of Actinomadura fibrosa ATCC 49459T (98.88 %) and Actinomadura formosensis JCM 7474T (98.82 %) (pairwise similarity values in parentheses). Digital DNA-DNA hybridisation experiments revealed unambiguously that strain LMG 30035T represents a novel Actinomadura species (OrthoANIu values less than 83.1 %; dDDH values less than 27.2 % with type strains of validly named Actinomadura species). Analysis of the cell wall revealed the presence of meso-diaminopimelic acid in the peptidoglycan. The whole-cell sugars were glucose, madurose, galactose, ribose and rhamnose. The major polar lipids included phosphatidylinositol and diphosphatidylglycerol. The predominant menaquinones were MK-9(H6), MK-9(H8), MK-9(H4) and MK-9(H2). The major fatty acids were C16 : 00, 10-methyl C18 : 0, C18 : 1 ω9c and C18 : 00. The DNA G+C content of its genome was 72.5 mol%. In summary, these characteristics distinguish strain LMG 30035T from validly named species of the genus Actinomadura, and therefore, we propose to classify this strain formally as the novel species Actinomadura roseirufa sp. nov. with LMG 30035T (=CECT 9808T,=ATCC 53664T) as the type strain.

Seihai-to (TJ-90)-Induced Activation of Airway Ciliary Beatings of Mice: Ca2+ Modulation of cAMP-Stimulated Ciliary Beatings via PDE1.

Sei-hai-to (TJ-90, Qing Fei Tang), a Chinese traditional medicine, increases ciliary beat frequency (CBF) and ciliary bend angle (CBA) mediated via cAMP (3',5'-cyclic adenosine monophosphate) accumulation modulated by Ca2+-activated phosphodiesterase 1 (PDE1A). A high concentration of TJ-90 (≥40 µg/mL) induced two types of CBF increases, a transient increase (an initial increase, followed by a decrease) and a sustained increase without any decline, while it only sustained the CBA increase. Upon inhibiting increases in intracellular Ca2+ concentration ([Ca2+]i) by 10 µM BAPTA-AM (Ca2+-chelator, 1,2-Bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid tetrakis(acetoxymethyl ester) or Ca2+/calmodulin-dependent PDE1 by 8MmIBMX (a selective PDE1 inhibitor), TJ-90 (400 µg/mL) induced only the sustained CBF increase without any transient CBF increase. The two types of the CBF increase (the transient increase and the sustained increase) induced by TJ-90 (≥40 µg/mL) were mimicked by the stimulation with both procaterol (100 pM) and ionomycin (500 nM). Thus, TJ-90 stimulates small increases in the intracellular cAMP concentration ([cAMP]i) and [Ca2+]i in airway ciliary cells of mice. These small increases in [cAMP]i and [Ca2+]i cause inducing a transient CBF increase or a sustained CBF increase in an airway ciliary cells, depending on the dominant signal, Ca2+-signal, or cAMP-signal.

Interaction between NFATc2 and the transcription factor Sp1 in pancreatic carcinoma cells PaTu 8988t.

BACKGROUND: Nuclear factors of activated T-cells (NFATs) have been mainly characterized in the context of immune response regulation because, as transcription factors, they have the ability to induce gene transcription. NFAT proteins are found in several types of tumors, for instance, pancreatic carcinoma. The role of NFATs in carcinogenesis is regulating central genes in cell differentiation and cell growth. NFAT proteins are primarily located in cytoplasm and only transported to the cell nucleus after activation. Here, they interact with other transcription factors cooperating with NFAT proteins, thus influencing the selection and regulation of NFAT-controlled genes. To identify and characterize possible interaction partners of the transcription factor NFATc2 in pancreatic carcinoma cells PaTu 8988t. METHODS: NFATc2 expression and the mode of action of Ionomycin in the pancreatic tumor cell lines PaTu 8988t were shown with Western blotting and immunofluorescence tests. Potential partner proteins were verified by means of immunoprecipitation and binding partners, their physical interactions with DNA pull-down assays, siRNA technologies, and GST pull-down assays. Functional evidence was complemented by reporter-promoter analyses. RESULTS: NFATc2 and Sp1 are co-localized in cell nuclei and physically interact at the NFAT target sequence termed NFAT-responsive promotor construct. Sp1 increases the functional activity of its binding partner NFATc2. This interaction is facilitated by Ionomycin in the early stimulation phase (up to 60 min). CONCLUSIONS: Oncological therapy concepts are becoming more and more specific, aiming at the efficient modulation of specific signal and transcription pathways. The oncogenic transcription partner Sp1 is important for the transcriptional and functional activity of NFATc2 in pancreatic carcinoma. The binding partners interact in cells. Further studies are necessary to identify the underlying mechanisms and establish future therapeutic options for treating this aggressive type of tumor.

Release of relaxin-like gonad-stimulating substance from starfish radial nerves by lonomycin.

In starfish, the peptide hormone gonad-stimulating substance (GSS) secreted from nervous tissue stimulates oocyte maturation to induce 1-methyladenine (1-MeAde) production by ovarian follicle cells. Recently, GSS was purified from radial nerves of the starfish Asterina pectinifera and identified as a relaxin-like peptide. This study examines the mechanism of GSS secretion from radial nerves. When radial nerves isolated from A. pectinifera were incubated in artificial seawater containing ionomycin as a calcium ionophore, GSS release increased in a dose-dependent manner; 50% activity of GSS release was obtained with approximately 10 µM ionomycin. Another calcium ionophore, A23187, also stimulated GSS release from radial nerves. In contrast, membrane permeable cyclic AMP and cyclic GMP analogs failed to induce GSS release. These results suggest that GSS secretion is induced by intracellular Ca(2+) as a second messenger.

Development and validation of a multiresidue liquid chromatography/tandem mass spectrometry method for 11 coccidiostats in feed.

A confirmatory method for the determination of 11 regulated coccidiostats including the ionophore antibiotics lasalocid, maduramicin, monensin, narasin, salinomycin, and semduramicin and the chemical coccidiostats decoquinate, diclazuril, halofuginone, nicarbazin, and robenidine in animal feed was developed and validated. The procedure was intended for the identification and quantification of the coccidiostats at concentrations relating both to the unintentional carryover as stated in Regulation 574/2011 and to the authorized levels in target feed. The analytes were determined by LC/MS/MS in the positive or negative electrospray ionization mode. The method performance characteristics were estimated in the relevant application field from 0.003 to 200 mg/kg. Validation criteria of linearity, specificity, trueness, precision, LOD, and LOQ along with measurement uncertainty were estimated for all analytes. Absolute and relative matrix effects were also studied. The results proved that the method performance was satisfactory, and it was successfully applied to carryover control by analyzing 165 feed samples collected within regulatory monitoring plans. Finally, since the carryover phenomenon in feed may result in the presence of residues in food products of animal origin, a survey has been carried out on the occurrence of coccidiostats in 167 eggs and animal muscles.

Interlaboratory comparison of an analytical method for the determination of semduramicin in poultry feed at the authorized level using high-performance liquid chromatography coupled to postcolumn derivatization and spectrophotometric detection.

The performance characteristics of a method based on HPLC with postcolumn derivatization and spectrophotometric detection for the quantification of semduramicin in poultry feedingstuffs have been determined via a collaborative study. Semduramicin is a feed additive that is authorized for fattening chickens within the European Union at a minimum and maximum content of 20 and 25 mg/kg in feedingstuffs, respectively. The target concentration of semduramicin in the test samples ranged from 11.5 to 45.0 mg/kg. The study has been conducted with two different types of test material, namely, feedingstuff samples that have been previously ground in our laboratory and pelleted feedingstuffs. In the latter case, the laboratories participating in the study had to grind the samples prior to analysis. The obtained RSD for repeatability (RSD(r)) ranged from 2 to 10% for the ground materials, and from 2 and 7% for the pelleted materials. The RSD for reproducibility (RSDR) varied between 11 and 16% for the ground materials, and between 12 and 15% for the pelleted materials. These data indicated that grinding as an additional step in the analytical procedure did not influence the precision profile of the method. In addition, the HorRat values for all test materials were below or equal to 1.5, thus demonstrating that the obtained precision data were acceptable for the purpose of the method. Furthermore, an estimation of trueness based on statistical treatment of the results reported from the laboratories for spiked samples revealed acceptable mean recovery values of 88 +/- 4%. Based on the obtained performance profile, the method can be considered fully validated and transferable to control laboratories to be used within the framework of official control.

Nigericin and grisorixin methyl ester from the Algerian soil-living Streptomyces youssoufiensis SF10 strain: a computational study on their epimeric structures and evaluation of glioblastoma stem cells growth inhibition.

The present work describes the metabolites produced by a strain identified as Streptomyces youssoufiensis, whose secondary metabolites profile has not been studied so far. The crude ethyl acetate extract was analyzed by high performance liquid chromatography-electrospray ionization mass spectrometry, leading to the detection of the ionophoric polyethers nigericin, epinigericin, abierixin and the newly isolated grisorixin methyl ester. The presence of epimeric forms of nigericin/epinigericin and grisorixin/epigrisorixin has spurred density functional theory computational calculations. This analysis was able to provide the relative stability of the most favored epimers, setting the basis for general structural considerations applicable to several other polyethers. Both nigericin sodium salt and grisorixin methyl ester showed to affect glioblastoma stem cells proliferation in a dose-dependent manner, with a higher activity for the more lipophilic grisorixin methyl ester (GI50 values of 3.85 and 3.05 µM for VIPI and COMI human glioblastoma stem cells, respectively).

Differential radio-adaptive responses in BALB/c and C57BL/6 mice: pivotal role of calcium and nitric oxide signalling.

Purpose: Our earlier studies demonstrated that transient radio-adaptive responses (RAR) in BALB/c mice were due to MAPK hyperactivation. The objective of this study was to determine the time duration of this low dose induced MAPK activation in BALB/c mice and to find out if similar adaptive responses are observed in C57BL/6 mice. Materials and methods: Mice were irradiated with 0.1 Gy priming dose (PD), 2 Gy challenge dose (CD) with an interval of 4 h (P + CD) and radiation induced immunosuppression in splenic lymphocytes was monitored as the endpoint for RAR. Results: Time kinetics following 0.1 Gy demonstrated persistence of MAPK hyperactivation till 48 h. Similar experiments in C57BL/6 mice indicated absence of RAR at 24 h following CD, in spite of MAPK activation which was also confirmed by time kinetics. Therefore, upstream activators of MAPK, viz., reactive oxygen and nitrogen species (ROS, RNS) and calcium levels were estimated. There was increased intracellular calcium (Ca2+) and nitric oxide (NO) in BALB/c and an increase in intracellular ROS in C57BL/6 mice 24 h after PD. Inhibition of NO and calcium chelation abrogated RAR in BALB/c mice. In vitro treatment of spleen cells with combination of NO donor and Ca2+ ionophore mimicked the effect of PD and induced adaptive response after 2 Gy not only in BALB/c but also in C57BL/6 mice confirming their crucial role in RAR. Conclusions: These results suggest that low dose induced differential induction of Ca2+ and NO signaling along with MAPK was responsible for contrasting RAR with respect to immune system of BALB/c and C57BL/6 mice. Abbreviations [3H]-TdR: 3H-methyl-thymidine; BAPTA: 1,2-bis(o-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid; CD: Challenge Dose; CFSE: Carboxy Fluorescein Succinamidyl Ester; on A: Concanavalin A; DAF-FM: 4-amino-5-methylamino-2',7'-difluorescein; DCF-DA: 2',7'-dichlorofluorescein diacetate; DSB: Double Strand Break; ELISA: Enzyme Linked ImmunoSorbent Assay; ERK: Extracellular signal-Regulated protein Kinase; FBS: Fetal Bovine Serum; HIF-1A: Hypoxia-Inducible Factor 1-alpha; LDR: Low Dose Radiation; MAPK: Mitogen Activated Protein Kinase; MAPKK/MKK: MAPK Kinase; MAPKKK: MAPK Kinase Kinase; NO: Nitric Oxide; NOS: Nitric Oxide Synthase; P + CD: Priming + Challenge dose; PBS: Phosphate Buffered Saline; PBST: Phosphate Buffered Saline-Tween 20; PD: Priming Dose; PI3K: Phosphatidyl Inositol 3-Kinase; PKC: Protein Kinase C; RAR: Radio Adaptive Response; RNS: Reactive Nitrogen Species; ROS: Reactive Oxygen Species; RPMI-1640: Roswell Park Memorial Institute-1640 medium; SAPK/JNK: Stress-Activated Protein Kinase/ c-Jun NH2-terminal Kinase; SEM: Standard Error of Mean; SNAP: S-nitro amino penicillamine; TP53: Tumor Protein 53; γ-H2AX: Gamma- H2A histone family member X; Th1: Type 1 helper T cell responses; Th2: Type 2 helper T cell responses.

Calcium triggers calcineurin-dependent synaptic vesicle recycling in mammalian nerve terminals.

BACKGROUND: Following exocytosis at the synapse, synaptic vesicle components are recovered by endocytosis. Morphological analysis has suggested that this occurs by a clathrin-mediated pathway, and the GTPase dynamin is thought to be involved in 'pinching off' endocytosing vesicles. The finding that the calcium-dependent phosphatase calcineurin can dephosphorylate dynamin and two other proteins implicated in endocytosis (amphiphysin and synaptojanin) has suggested a potential role for calcium and dephosphorylation in regulating synaptic vesicle endocytosis. RESULTS: We tested this hypothesis with an endocytosis assay in isolated nerve terminals (synaptosomes) that relies on the use of the fluorescent dye FM2-10. In synaptosomes, vesicle recycling occurs predominantly via a pathway dependent on both dynamin and amphiphysin. We found that endocytosis could be stimulated maximally at calcium concentrations that yielded only low levels of exocytosis, suggesting that the two processes had different calcium sensitivities cyclosporin A and Fk506, we identified calcineurin as a calcium sensor for endocytosis and showed that its activity is essential for synaptic vesicle endocytosis in synaptosomes. CONCLUSIONS: Our results suggest that dynamin-dependent synaptic vesicle endocytosis is triggered by calcium influx occurring upon nerve-terminal depolarisation. An essential mediator of calcium's effect is calcineurin, the activation of which leads to dephosphorylation of at least four proteins implicated in endocytosis-dynamin, amphiphysin 1, amphiphysin 2 and synaptojanin. Our findings also imply that endocytosis and exocytosis may occur in tandem in vivo simply because they share a responsiveness to calcium influx, rather than because they are mechanistically coupled.

Tiamulin and semduramicin: effects of simultaneous administration on performance and health of growing broiler chickens.

The pleuromutilin antibiotic tiamulin (TIA) is known to produce a variety of negative interactive effects when it is administered in combination with several anticoccidial ionophores. A 35-d growth study was performed in cages to evaluate the compatibility of TIA when it was administered concurrently with the poly-ether ionophore anticoccidial semduramicin (SEM). Tiamulin and SEM, both alone and in combination, were administered to 10 replicates of female broilers arranged in a completely randomized block design. Tiamulin was administered in drinking water (250 mg of TIA/kg of water) from d 15 through 19 of the study, whereas SEM was incorporated in feed (25 mg/kg) from placement to the conclusion of the test. Water consumption was determined during the period of concurrent administration of the drugs and weekly measurements of feed intake and bird performance were recorded. In addition, hematocrit, blood cell counts, serum protein, albumin, glucose, uric acid, electrolytes, and activities of several enzymes were determined from blood samples taken at d 35. Results indicated that simultaneous administration of TIA and SEM during the third week of the trial reduced water and feed intake resulting in a temporary growth depression. Feed efficiency was transiently affected during the period of coadministration. However, during the fourth week of the test, negative effects in body weight were not observed for any treatment and feed conversion improved for birds concurrently receiving TIA + SEM. By the termination of the experiment, no adverse effects were observed in final performance for any treatment. Histopathological and hematological parameters were unaffected by treatment at d 35 of the test. These results demonstrated that simultaneous administration of TIA and SEM produced only temporary impairments of water and feed consumption that transiently influenced performance. Neither mortality nor long-term effects on performance variables occurred in broilers.

The influence of pH on the conductance of lipid bimolecular membranes in relation to the alkaline ion transport induced by carboxylic carriers grisorixin, alborixin and monensin.

The influence of the pH on the stability and stoichiometry of the complexes formed by carboxylic-antibiotics such as grisorixin, alborixin and monensin with alkaline and alkaline earth cations has been investigated. The maximum values of bimolecular lipid membrane conductance are obtained with grisorixin and potassium ion. The conductance-pH curves show a very pronounced maximum in the neutral pH range. The results are analysed on the basis of a dimeric form of the ionophore in the complex and the possibility of having several charged complexes resulting from an heterogeneous reaction, the number of each complexed form depending on the pH of the bulk solutions.

Study of a delta-hydroxy ketone-hemiketal equilibrium by 2D-NMR exchange spectroscopy for the antibiotic grisorixin: involvement in cationic transport through membranes.

Study of a delta-hydroxyketone-hemiketal equilibrium in the polyether antibiotic grisorixin was performed with 2D-NMR spectroscopy. The efficiency of 13C chemical exchange spectroscopy for the assignment of 1H and 13C resonances, in the 2 forms, was shown, making possible a conformational investigation of both forms. This equilibrium was observed for grisorixin in solvents of varying polarity, such as CD2Cl2, CDCl3, CD3CN, or CD3OD, but not in C6D12 or C6D6. Other related antibiotics with the same terminal heterocycle were described only in the closed hemiacetalic structure. The low ionic fluxes measured in a bulk chloroformic membrane for grisorixin were explained by this equilibrium, which competed unfavorably with the cation capture process at the water-chloroform interface. This equilibrium would not be present in a phospholipidic bilayer membrane containing the ionophore, published experimental results are taken into account. The peculiar tautomeric equilibrium observed for grisorixin could be linked to the specific axial stereochemistry of the C7-C8 bond, which creates tension in the globular conformation.

Effect of phosphate and ionophores on (14C)-NEM incorporation in mitochondrial membranes and relationships with phosphate carrier system.

Phosphate transport in mitochondria was investigated with respect to its inhibition by NEM. The reactivity of the Pi carrier SH groups was influenced by phosphate or ionophores during preincubation before the addition of NEM. Furthermore in order to obtain some mitochondrial protein fractions where the typical effects of phosphate and ionophores on [14C]-NEM fixations were observed, mitochondria were submitted to hypotonic treatment and sonication. The following results were obtained: 1. -- Phosphate and grisorixin (a new ionophore of the nigericin group) decreased the inhibition of phosphate transport by NEM. The same effect was observed for [14C]-NEM incorporation. 2. -- Valinomycin increased [14C]-NEM incorporation. The valinomycin effect was abolished by phosphate. ClCCP alone affected [14C]-NEM incorporation slightly. Valinomycin plus ClCCP decreased NEM inhibition of phosphate transport and [14C]-NEM incorporation like grisorixin. 3. -- The variability of SH group reactivity can be interpreted by a control of SH group accessibility by transmembrane delta pH as previously suggested. 4. -- Typical effects of phosphate or ionophores were observed in whole pig heart and rat liver mitochondria. These effects were enhanced in the same supernatant protein fraction resulting from sonication in pig heart mitochondria : phosphate decreased [14C]-NEM incorporation by 1,50 nmoles/mg protein, grisorixin by 0.95 nmoles, whereas valinomycin increased it by 0.75 nmoles. For rat liver mitochondria the phosphate effect and the valinomycin increased it by 0.75 nmoles. For rat liver mitochondria the phosphate effect valinomycin effect on [14C]-NEM incorporation were observed in the subparticular fraction obtained after sonification.

Secondary metabolites by chemical screening. 17. Nigericinol derivatives: synthesis, biological activities, and modeling studies.

The synthesis and the biological activity of C-1-reduced nigericin derivatives (nigericinols) are described and discussed. The dichloronigericinol 7 impressively demonstrated that the C-1 carboxylic acid moiety was not required for a distinct activity against bacteria and viruses. Based on the correlation between K+/H+ antiport activities and antibacterial activities it was deduced that the mode of action of the described nigericinols are related to their ionophoric properties. Molecular modeling studies showed that the efficiency of the nigericinols as ionophores correlates, qualitatively, with the probability of forming a cyclic structure, with the exception of 7.

Assessment of drugs by functional imaging of thallium-201 distribution in the dog.

The effect of drugs on thallium-201 biodistribution in the body has been studied in dogs using a new functional imaging method. The calculation is based upon the comparison of the activities of the two scintigraphic images obtained after two successive injections of thallium-201 separated by an interval of at least 10 min, during which some drug of interest can be administered. This imaging technique was applied in control dogs (n = 6) and in animals treated with dipyridamole (n = 7) or grisorixin (n = 7). As expected, these two vasoactive drugs increased mainly the myocardial uptake, whereas smaller variations were noted in the other organs studied: liver, kidney, lungs, and skeletal muscles. Thus this method should allow a rapid and reliable noninvasive assessment of cardiovascular drugs with thallium-201.

Myocardial imaging in dogs with thallium-201 and the ionophore grisorixin.

The effect of grisorixin, a monocarboxylic ionophore, upon the myocardium-to-background ratio in thallium-201 scintigrams has been studied in 20 dogs. Three sequences of injection and two doses of the ionophore have been used. A significant improvement was obtained when grisorixin was injected 10 min before thallium-201 and at a submaximal dose; the myocardium-to-background ratio was at least twice that of the controls during the first 20 min following the tracer injection.

Effects of grisorixin on the distribution of thallium-201 and on the oxidative metabolism in cultured myocardial cells.

Previous experiments in the dog and guinea-pig have shown that grisorixin, a monocarboxylic ionophore, can significantly increase the coronary blood flow and the myocardial uptake of 201Tl, as well as have a stimulant effect on the heart. In this study, cultures of myocardial cells were used in order to isolate the cells from the vascular and extracardiac influences so that any ionophorous effect on 201Tl could be evidenced. The effects of grisorixin on the oxidative metabolism were simultaneously studied. The technique described by Harary was used to prepare the cultures. The activity of the 14CCO2 produced by oxidation of [14C]glucose and [14C]octanoate added to the medium of culture and the intra/extracellular ratio of 201Tl concentrations (Tl i/e) were measured. In the controls (n = 8), the Tl i/e was 40 +/- 10 while it was 17 +/- 6 (p less than 0.05) in the cells that received 201Tl and grisorixin at the same time (n = 4), and 19 +/- 5 (p less than 0.05) in the flasks where 201Tl was injected after grisorixin (n = 7). A significant decrease of the [14C]octanoate oxidation was found in the flasks treated with grisorixin (n = 4, -50 +/- 16%, p less than 0.01) while the [14C]glucose oxidation was not significantly lowered (n = 3; -11 +/- 12%). The conclusion is that grisorixin decreases both the intracellular concentration of 201Tl and the fatty-acids oxidation in cultured myocardial cells. The beneficial effects previously observed in vivo were probably the consequence of the strong coronary dilatation and of an indirect stimulation.

Myocardial imaging in dogs treated with grisorixin: relationship between thallium-201 uptake and coronary blood flow.

Thallium-201 myocardial imaging was performed in dogs after pretreatment with grisorixin, which appeared to increase the myocardial uptake of Tl-201. This effect of grisorixin was found to be dose dependent, with an optimal dose of 60 microgram/kg. The myocardial-to-background ratio, which was 1.92 in the control dogs, rose to 4.45. The increase in the absolute myocardial uptake was demonstrated in guinea pigs that received Tl-201 after similar pretreatment with grisorixin. In the animals treated with 500 microgram/kg, the uptake of Tl-201 by the heart was 35% over the control value. With 60 microgram/kg grisorixin, the coronary blood flow increased from 40 to 176 ml/min 5 min after the injection. This dose, optimal for imaging, induced the maximum vasodilator effect with only a very slight concomitant increase in the left-ventricular pressure and myocardial contractility. Above 60 microgram/kg, grisorixin appeared to be a potent inotropic agent, whereas below this dose it showed only coronary vasodilator properties. Some evidence for an ionophore effect of this compound was found in dogs pretreated with 60 microgram/kg. In these the radionuclide was injected when the coronary vasodilatation had become insignificant, but a significant improvement of the M/B ratio was still evident.

Potentiation of ionophorous anticoccidials with dihydroquinolines: compatibility of lasalocid and semduramicin with duokvin.

Three battery tests were conducted to reveal whether or not there is an interaction between the new dihydroquinoline antioxidant, duokvin and lasalocid or the new anticoccidial, semduramicin, similar to that observed with some other ionophorous anticoccidials. In terms of body weight gain, no significant difference due to toxic interaction between duokvin and any dose of lasalocid or semduramicin was detected in chickens experimentally infected with oocysts of Eimeria tenella and E. mitis. Anticoccidial efficacy at reduced doses of both lasalocid and semduramicin in combination with duokvin showed numerical improvement; however, this again proved to be insignificant. The lack of incompatibility of this antioxidant with lasalocid or semduramicin allows their simultaneous administration on the one hand, but it fails to enable a substantial reduction of the chemoprophylactic concentration of anticoccidials in the broiler ration on the other.