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PubMed1999

Arginine biosynthesis in Campylobacter jejuni TGH9011: determination of the argCOBD cluster.

Arginine biosynthetic genes from Campylobacter jejuni TGH9011 were cloned by functional complementation of the respective Escherichia coli arginine biosynthetic mutants. Complementation of argA, argB, argC, argD, argE, argF, and argH auxotrophs was accomplished using a pBR322-based C. jejuni TGH9011 plasmid library. By cross-complementation analyses, the first four steps of arginine biosynthesis were shown to be closely linked on the genome. Two additional clones complementing the first (ArgA) and fifth (ArgE) steps in arginine biosynthesis were obtained. Neither recombinant showed linkage to the arg cluster, to each other, nor to other arginine biosynthetic functions by cross-complementation. Genes argF and argH were not linked to other arginine biosynthetic genes by cross-complementation analysis. Restriction enzyme patterns of recombinant plasmids fell into five groups. Group I contained the arg(ABCD) complementing locus. Group II and Group III were the two genetic loci corresponding to the argA and argE complementing genes. Group II contains the hipO gene encoding N-benzoylglycine-amino-acid amidohydrolase, also known as hippurate hydrolase. Group III contains the hipO homolog of C. jejuni. Group IV represents the argF gene. Group V is the argH gene. Functional complementation of mutations in the first four steps of the arginine biosynthetic pathway was obtained on recombinant plasmid pARGC2. The predicted order of gene complementation was argCargA(argBargD). The sequence of the insert in plasmid pARGC2 revealed direct homologs for argC, argB, and argD. However, sequence analysis of the gene complementing ArgA function in two separate E. coli argA mutants determined that the C. jejuni gene was not a canonical argA gene. The gene complementing the argA defect, which we call argO, showed limited homology to the streptothricin acetyltransferase gene (sat) of Escherichia coli. The flanking open reading frames in pARGC2 showed no homologies to arginine biosynthetic genes. The structure of the argCOBD gene arrangement is discussed with reference to the presence and location of other arginine biosynthetic genes on the genome of C. jejuni and other bacterial organisms.

Amino Acid Sequence↗

PubMed2001

The CNS midline cells control the spitz class and Egfr signaling genes to establish the proper cell fate of the Drosophila ventral neuroectoderm.

The spitz class genes, pointed (pnt), rhomboid frho), single-minded (sim), spitz (spi)and Star (S), as well as the Drosophila epidermal growth factor receptor (Egfr) signaling genes, argos (aos), Egfr, orthodenticle (otd) and vein (vn), are required for the proper establishment of ventral neuroectodermal cell fate. The roles of the CNS midline cells, spitz class and Egfr signaling genes in cell fate determination of the ventral neuroectoderm were determined by analyzing the spatial and temporal expression patterns of each individual gene in spitz class and Egfr signaling mutants. This analysis showed that the expression of all the spitz class and Egfrsignaling genes is affected by the sim gene, which indicates that sim acts upstream of all the spitz class and Egfr signaling genes. It was shown that overexpression of sim in midline cells fails to induce the ectodermal fate in the spi and Egfr mutants. On the other hand, overexpression of spi and Draf causes ectopic expression of the neuroectodermal markers in the sim mutant. Ectopic expression of sim in the en-positive cells induces the expression of downstream genes such as otd, pnt, rho, and vn, which clearly demonstrates that the sim gene activates the EGFR signaling pathway and that CNS midline cells, specified by sim, provide sufficient positional information for the establishment of ventral neuroectodermal fate. These results reveal that the CNS midline cells are one of the key regulators for the proper patterning of the ventral neuroectoderm by controlling EGFR activity through the regulation of the expression of spitz class genes and Egfr signaling genes.

Animals↗

PubMedDate not supplied

[Analysis and recognition of promoters for erythroid-specific genes based on degenerative oligonucleotide motifs].

We developed a method to search for degenerate oligonucleotide motifs specific for certain regions in eukaryotic gene promoters. A procedure of promoter recognition based on these motifs is presented. The methods are integrated within program package ARGO available for the Internet users (http://wwwmgs.bionet.nsc.ru/mgs/programs/argo). This method was applied to study erythroid-specific gene promoters. High efficiency of their recognition is demonstrated.

Base Sequence↗

PubMed2002

[Blood culture update].

BACKGROUND: Blood culture is one of the most important bacteriological examinations with important clinical and therapeutic consequences. Blood cultures should be ordered in all patients with signs suggesting septicemia, endocarditis or severe infection (pneumococcal pneumonia, bacterial meningitis with bloodstream dissemination). Blood culture methods have evolved considerably over the last twenty years. After using manual methods for many years, read by non-standardized visual methods, the development of media with defined compositions and supplemented to allow growth of bacteria difficult to culture has been associated with the development of automatic blood culture devices. AUTOMATIC DEVICES: These devices have undergone rapid improvement. Semi-automatic devices (Bactec NR-660) were rapidly followed by completely automatic techniques, including four devices currently available: since 1989 Bio-Argos (Rio-Rad) and Bact/Alert (Organon-Teknika) and in 1993, Bactec 9240 (Becton-Dickinson) and Vital (BioMérieux). All these devices allow automatic detection of CO2 produced during bacterial growth. Automatic reading systems provide continuous output avoiding the need for invasive methods and thus the risk of contamination in addition to saving time. Potential application to achieve quantitative blood cultures for intensive care units is in the development stage. CONSEQUENCES: The reliability of these devices is well recognized and their contribution to severe bacterial infection is undeniable. There are certain limitations however related to material cost and the non-identification of the pathogen involved. Molecular biology techniques open new perspectives in this field. The evolution of techniques, definitions, and pathogenic approach to septicemia must be revisited as new infectious situations have been identified at the same time as new investigation tools resulting from considerable technological progress. New methods of blood culture have largely contributed to this progress.

Bacteriological Techniques↗

PubMed2003

[Physical methods of dissection and coagulation in surgery].

The paper presents 20-year experience in using various devices for tissue dissection and coagulation of such as electrosurgical ones EN-57M, Efa-0201 (Russia), Karl Storz-endoscope (Germany), Force-300, Force-FX, LigaSure ("Valley-lab", USA), Erbotom ICC 350 ("Erbe"), Argo-3000 ("Soring", Germany), a Surgitrone radioknife ("Ellman", USA), ultrasonic Auto Sonix devices ("Auto Suture", USA), a harmonic Ultracision scalpel ("Ethicon Endo-Surgery", USA), laser Scalpel-1 and Romashka-1 devices (Russia), plasma Fakel-01 and Prometey devices (Russia). The results of 424 experimental studies and clinical application of different energies in 1683 patients are analyzed. The influence of different types of energy on tissue and reparative processes were revealed, morphological changes were studied. Advantages and shortcomings of each type of energy are demonstrated, recommendations for their optimum application are formulated.

Dissection↗

PubMed2003

The hierarchical relationship among the spitz/Egfr signaling genes in cell fate determination in the Drosophila ventral neuroectoderm.

The spitz class and Egfr signaling (spi/Egfr) genes are required for the proper establishment of cell fate in the Drosophila ventral neuroectoderm. We investigated the role of the central nervous system (CNS) midline cells, and the hierarchical relationship among the spi/Egfr genes, in this process by analyzing the spatial and temporal expression of several of the genes in selected spi/Egfr mutants. Our analysis showed that expression of all the spi/Egfr genes is severely reduced in the single-minded (sim) mutant, and ectopically induced in en-Gal4/UAS-sim embryos. This result indicates that sim acts upstream of all the other spi/Egfr genes. The CNS midline cells regulate rhomboid (rho) expression in the ventral neuroectoderm and activate the EGFR signaling pathway. We also found that argos (aos) and orthodenticle (otd) act downstream of pointed (pnt), and that aos represses expression of otd in the lateral neuroectoderm to establish differential cell fates in the ventral neuroectoderm. Our findings suggest the following hierarchical relationship among the spi/Egfr genes: [see text].

Animals↗

PubMed2004

In vivo caries formation in enamel following argon laser irradiation and combined fluoride and argon laser treatment: a clinical pilot study.

OBJECTIVE: This in vivo pilot study investigated the role of argon laser irradiation and combined fluoride and argon laser treatment in accelerated natural caries development in sound enamel surfaces beneath plaque-retentive orthodontic bands. METHOD AND MATERIALS: Five patients (3 female, 2 male, ages 19 to 28 years) requiring tooth extraction prior to orthodontic treatment, participated in the study. Buccal surfaces were treated with either: (1) argon laser (250 mW for 10 seconds, ARGO-MOD); (2) topical fluoride (0.5% fluoride ion, Thera-Flur-N) followed by argon lasing; or (3) no treatment (control). Orthodontic bands with plaque-retentive slots on buccal surfaces were placed on the teeth slated for extraction (n = 14). Following a minimum of 5 weeks of intraoral exposure, the teeth were extracted for laboratory analysis. The teeth underwent serial longitudinal sectioning (12 sections per tooth). The sections were imbibed in water, and lesion depths were determined with each section, using polarized light microscopy. Comparisons were made among treatment groups (analysis of variance, Duncan's multiple range test for paired samples). RESULTS: Mean lesion depths were: 261 +/- 24 microm for the no treatment control group (n = 84 sections); 147 +/- 18 microm for the argon laser group (n = 24 sections); and 99 +/- 12 microm for the fluoride and argon laser group (n = 60 sections). Both the argon laser (44%) and the fluoride and argon laser groups (62%) had significant lesion depth reductions compared to controls. The addition of fluoride treatment prior to argon lasing resulted in a 32% reduction in lesion depth compared to argon laser treatment alone. CONCLUSIONS: Within this clinical pilot study, in vivo natural caries formation was affected significantly by a single exposure to low fluence argon laser irradiation. Topical fluoride treatment in combination with argon lasing provided an even greater degree of resistance against in vivo enamel caries development. A simple technique for reducing the caries susceptibility of enamel may be a clinical reality.

Adult↗

PubMed2001

[A comparative study of craniofacial morphologic differences between groups with shallow and deep antegonial notch depth].

OBJECTIVE: To evaluate the correlation between craniofacial morphology and antegonial notch depth of the mandible. METHODS: 42 patients of ear ly permanent dentition from Nanchang city, 21 with shallow and 21 with deep antegonial notch were selected. Lateral cephalometric film before treatment were taken. Hard tissue angular and linear measurement were performed. The correlation between antegonial notch depth and craniofacial morphology were analysed statistically. RESULTS: The angular measurements of SN-GoGn, S-ArGo, FH-GoMe,and ANS-Xi-Pm were significantly greater in the deep antegonial notch group. Other angles as SNB, SNP-BaN-CCGn, FH-NPg and MD-AA were significantly greater in the shallow antegonial notch group. For the linear measurements, those items as N-Me, ANS-Me,Convexity and LM-GoGn were significantly greater in the deep antegonial notch group. CONCLUSION: The deep antegonial notch group showed steeper mandibular plane, smaller chin, greater anterior facial height and gonial angle, and has a mandible with more backward and downward growth.

English Abstract↗

PubMed2004

Calibration of a vector-MEG helmet system.

The MEG system Argos 500, recently installed at the University of Ulm, is designed for clinical application and routine use, to allow investigation of a large number of patients per day. To reach this goal, the system design meets the requirements of reliability, high field sensitivity, minimal set-up overhead before each measurement and an easy-to-handle user interface. The sensor system consists of a 163 vector-magnetometer array oriented and located in a suitable way to cover the whole head of the patient. Four additional triplets are available as references to build software gradiometers. To use this system at a high performance level, it must be properly calibrated, with these goals: to determine the actual geometry of the sensors array, which can deviate from the design specifications, and to determine the actual sensitivity of each sensor. The calibrating source consists of 31 coils placed at the corners of a head-size dodecahedron. Various details of the calibration system and process are presented here.

Calibration↗

PubMed1991

An investigation on the quinoprotein nature of some fungal and plant oxidoreductases.

The presence of pyrroloquinoline quinone (PQQ) as the organic cofactor of Dactylium dendroides galactose oxidase and lentil (Lens culinaris) seedling amine oxidase, purported PQQ-containing oxidoreductases (Van der Meer, R. A., Jongejan, J. A., and Duine, J. A. (1989) J. Biol. Chem. 264, 7792-7794; Citro, G., Verdina, A., Galati, R., Floris, G., Sabatini, S., and Finazzi-Argo', A. (1989) FEBS Lett. 247, 201-204), was reinvestigated using the nitro blue tetrazolium redoxcycling method (Paz, M. A., Gallop, P. M., Torrelio, B. M., and Flückiger, R. (1988) Biochem. Biophys. Res. Commun. 154, 1330-1337; Paz, M. A., Flückiger, R., Boak, A., Kagan, H. M., and Gallop, P. M. (1991) J. Biol. Chem. 266, 689-692) and the enzyme-linked immunosorbent assay with polyclonal antibodies against PQQ. The possible quinoprotein nature of the laccases from Polyporus versicolor and Rhus vernicifera was also investigated because of the similarities in spectroscopic and kinetic features of these enzymes and the laccase from Phlebia radiata, reported to be a PQQ protein (Karhunen, E., Niku-Paavola, M.-L., Viikari, L., Haltia, T., Van der Meer, R. A., and Duine, J. A. (1990) FEBS Lett. 267, 6-8). The presence of a quinonoid cofactor in lentil seedling amine oxidase is confirmed, whereas galactose oxidase and both laccases do not display any quinoprotein nature.

Amine Oxidase (Copper-Containing)↗

PubMedDate not supplied

[Export of metabolites by the proteins of the DMT and RhtB families and its possible role in intercellular communication].

The earlier published and new experimental data are summarized on the properties of the genes encoding the membrane proteins of the DMT family (RhtA (YbiF), EamA (YdeD), YijE, YddG, YedA, PecM, eukaryotic nucleoside phosphate sugar and hexose phosphate transporters), the RhtB/LysE family (RhtB, RhtC, LeuE, YahN, EamB (YfiK), ArgO (YggA), CmaU), as well as some other families (YicM, YdhC, YdeAB, YdhE (NorE)). These proteins are involved in the export of amino acids, purines, and other metabolites from the cell. The expression of most of the genes encoding these proteins is not induced by the substrates they transport but is controlled by the global regulation systems, such as the Lrp protein, and activated by the signal compounds involved in the intracellular communication. The level of expression, assessed in experiments on translational fusion of the corresponding bacterial genes with the beta-galactosidase gene, depends on the growth phase of the bacterial culture, composition of the medium, and some stress factors, such as pH osmolarity or decreased aeration. The efflux of normal cell metabolites is assumed to be the natural function of these proteins. This function may play a role in density-dependent behavior of cell populations (quorum sensing). It may have been enhanced in the course of evolution via specialization of these proteins in the efflux of compounds derived from metabolic intermediates and adjusted to the role of transmitters.

Amino Acids↗

PubMed1991

[Comparison between alpha-amylase from B. amyloliquefaciens and B. licheniformis].

Two sequence-homologous alpha-amylases from B. amyloliquefaciens and B. licheniformis were studied with respect to their stability against heat denaturation and were compared with respect to common structure-stabilizing principles. The investigated alpha-amylases were isolated from culture broth of B. amyloliquefaciens and B. licheniformis. The molecular parameters (molecular weight and isoelectric point) are similar. The thermostability was determined by changes of the protein structure (changes of the fluorescence emission spectra). At pH 5.0 the thermostable alpha-amylase from B. licheniformis showed a rate of denaturation which was achieved by the thermolabile alpha-amylase from B. amyloliquefaciens at a temperature 15 degrees lower. The alpha-amylase from B. licheniformis exhibits a marked stability also at the alkaline pH-range in contrast to the alpha-amylase from B. amyloliquefaciens. From measurements in the presence of EDTA and Ca2+ follows that both enzymes are stabilized by binding of calcium ions. An analysis of preferred amino acid exchanges between the two sequence-homologous alpha-amylases showed correspondences and differences to the well-known diagram of ARGOS. Possibly an increased thermic stability can already be achieved by special amino acid exchanges without significant changes in the protein structure.

Bacillus↗

PubMed1990

A review of the biochemical roles, toxicity and interactions of zinc, copper and iron: III. Iron.

Because of its essentiality, iron occurs in tissues of all living matter especially aerobes. Iron compounds have been utilized in various aspects of argo-industry. Iron-binding proteins and their vital role in intestinal uptake, distribution and storage of iron are concisely examined. Ceruloplasmin (cuproenzyme) is imperative for iron mobilization from storage sites for hemoglobin synthesis. There is evidence of relationship between enhance susceptibility to infection and iron deficiency. The extent to which behavioral abnormalities and iron depletion are related remains to be established. The efficacy of chelating agents such as desferroxamine, bicarbonates and ethylene diamine tetraacetic acid (EDTA) against iron overload has been tested.

Anemia, Hypochromic↗

PubMed1990

Ca2(+)-induced conformational change and aggregation of chromogranin A.

Chromogranin A, the most abundant protein in bovine adrenal chromaffin granules, bound calmodulin in a Ca2(+)-dependent manner, and the calmodulin-binding property was utilized to purify chromogranin A. Chromogranin A has been described in the past as a "random-coil polypeptide" with little alpha-helix or beta-sheet conformation. However, circular dichroism measurements with pure, native chromogranin A revealed relatively high alpha-helical contents (40% at the intravesicular pH of 5.5). Fluorescence studies confirmed previous observations that chromogranin A binds Ca2+ with low affinity. Considering the high concentration of Ca2+ in the secretory vesicle, the effect of Ca2+ on the secondary structure and self-association of chromogranin A was examined. Ca2+ induced a decrease of alpha-helicity of chromogranin A from 40 to 30% at pH 5.5. In contrast, at pH 7.5 the same amount of Ca2+ increased alpha-helicity of the protein from 25 to 40%. Boiling of the adrenal extract, a commonly used purification procedure for chromogranin A, resulted in the isolation of conformationally distinct chromogranin A molecule. Unlike secretory protein-I of the parathyroid gland (Gorr, S.-V., Dean, W. L., Radley, T. L., and Cohn, D. V. (1988) Bone Mineral 4, 17-25), chromogranin A aggregated rapidly in the presence of Ca2+. The extent and rate of aggregation were highly dependent on Ca2+ concentration. Although both the rate and extent of aggregation at pH 7.5 were much lower than those at pH 5.5, aggregation of chromogranin A proceeded at both pH's. In this respect, chromogranin A differs from human chromogranin C which was shown by Gerdes et al. (Gerdes, H.-H., Rosa, P., Phillips, E., Baeuerle, P. A., Frank, R., Argos, P., and Huttner, W. B. (1989) J. Biol. Chem. 264, 12009-12015) to aggregate at pH 5.2 but not at pH 7.4.

Adrenal Medulla↗

PubMed1988

The relationship between the structures of four beta-lactamases obtained from Bacillus cereus.

Bacillus cereus has proved to be one of the most interesting microorganisms in the study of beta-lactamases. It secrets these enzymes very efficiently and, frequently, in multiple forms. Three different forms are produced by strain 569/H; mutant 5/B of the same microorganism is constitutive for the secretion of beta-lactamases I and II. The present study, based on secondary structure prediction by two independent methods, states the relationship among the structures of beta-lactamases I, II and III produced by B. cereus 569/H and beta-lactamase I from the strain 5/B of this microorganism. A strong similarity is also established for the enzyme type III of B. cereus and the enzyme type I produced by B. licheniformis which could have an evolutionary explanation. A structural analysis of the leader peptide regions of these enzymes by the method of Mohana and Argos is also reported.

Bacillus cereus↗

PubMed1983

Cloning of the Bacillus subtilis glutamine phosphoribosylpyrophosphate amidotransferase gene in Escherichia coli. Nucleotide sequence determination and properties of the plasmid-encoded enzyme.

The Bacillus subtilis gene encoding glutamine phosphoribosylpyrophosphate amidotransferase (amidophosphoribosyltransferase) was cloned in pBR322. This gene is designated purF by analogy with the corresponding gene in Escherichia coli. B. subtilis purF was expressed in E. coli from a plasmid promoter. The plasmid-encoded enzyme was functional in vivo and complemented an E. coli purF mutant strain. The nucleotide sequence of a 1651-base pair B. subtilis DNA fragment was determined, thus localizing the 1428-base pair structural gene. A primary translation product of 476 amino acid residues was deduced from the DNA sequence. Comparison with the previously determined NH2-terminal amino acid sequence indicates that 11 residues are proteolytically removed from the NH2 terminus, leaving a protein chain of 465 residues having an NH2-terminal active site cysteine residue. Plasmid-encoded B. subtilis amidophosphoribosyltransferase was purified from E. coli cells and compared to the enzymes from B. subtilis and E. coli. The plasmid-encoded enzyme was similar in properties to amidophosphoribosyltransferase obtained from B. subtilis. Enzyme specific activity, immunological reactivity, in vitro lability to O2, Fe-S content, and NH2-terminal processing were virtually identical with amidophosphoribosyltransferase purified from B. subtilis. Thus E. coli correctly processed the NH2 terminus and assembled [4Fe-4S] centers in B. subtilis amidophosphoribosyltransferase although it does not perform these maturation steps on its own enzyme. Amino acid sequence comparison indicates that the B. subtilis and E. coli enzymes are homologous. Catalytic and regulatory domains were tentatively identified based on comparison with E. coli amidophosphoribosyltransferase and other phosphoribosyltransferase (Argos, P., Hanei, M., Wilson, J., and Kelley, W. (1983) J. Biol. Chem. 258, 6450-6457).

Amidophosphoribosyltransferase↗

PubMed1981

Protein thermostability. Correlations between calculated macroscopic parameters and growth temperature for closely related thermophilic and mesophilic bacilli.

The amino acid composition of more than 20 enzymes and protein from various closely related mesophilic and thermophilic micro-organisms (esp. Bacillus) have been used to calculate a variety of macroscopic parameters. These included the hydrophobic index (H phi ), the ratio of polar to non-polar volumes (rho), the ratios of Arg/(Arg + Lys), and (Arg + Lys) or (Glx + Asx) to total amino acids, % H-bonding amino acids, % alpha-helix- or beta-sheet-forming amino acids, the theoretical melting temperature (TCalcm), the total volume to total amino acid ratio (VR), and the % non-polar residues (NPS). In contrast to previous similar comparisons with proteins from divergent sources, it was found that thermophilic vs mesophilic proteins from the same genus show correlations between thermostability and increased H phi, decreased rho, and increased Arg/(Arg + Lys), as well as increased alpha-index and beta-index. Weaker correlations were seen for VR, TCalcm, aliphatic index, and NPS, all derived from, or related to, H phi. No correlations existed for the other calculated parameters. These results are consistent with recent results of Argos et al. (1979) [Biochemistry 18, 5698-5703] on sequence analyses of glyceraldehyde-3-P dehydrogenases, where thermophilic proteins showed multiple amino acid replacements that caused increased internal hydrophobicity and increased external polarity. No trends were observed in any of the parameters calculated from amino acid compositions for crude cytoplasmic protein extracts from mesophilic vs thermophilic Bacilli.

Amino Acids↗

PubMed1995

Bradykinin acting on B2 receptors contracts colon circular muscle cells by IP3 generation and adenylate cyclase inhibition.

Receptor subtypes and intracellular signaling events involved in bradykinin-evoked contraction of colonic circular muscle are unknown. We studied the roles of inositol trisphosphate (IP3) and cyclic AMP generation and the selectivity for B1 and B2 receptors in guinea pig colon. Bradykinin induced concentration-dependent contraction of circular muscle strips with an EC50 of 2 x 10(-8) M that was inhibited by the B2 antagonist D-Argo-(Hyp3,Thi5,8,D-Phe7)-bradykinin but not the B1 antagonist des-Arg9-[Leu8]bradykinin. The B1 agonist des-Arg9-bradykinin did not evoke contraction or relaxation. Bradykinin induced concentration-dependent shortening of isolated myocytes from circular muscle with an EC50 of 2 x 10(-11) M that was inhibited by the B2 but not the B1 antagonist, confirming the myogenic nature of the bradykinin receptors. Persistence of myocyte contraction in a calcium-free medium with EGTA confirmed the lack of dependence on extracellular calcium. In colon muscle tissue, bradykinin evoked concentration-dependent IP3 generation with an EC50 of 10(-7) M and a maximal level of 58 +/- 17 pmol/mg of protein at 10(-4) M that was inhibited by the B2 but not the B1 antagonist. Bradykinin, acting on B2 receptors, inhibited cyclic AMP formation after forskolin (10(-5) M) with an EC50 of 3 x 10(-8) M and maximal inhibition of 48% at 10(-5) M. In conclusion, bradykinin induces colon muscle contraction via myogenic non-B1 receptors, which are likely of the B2 subtype, with phosphoinositide turnover activation and adenylate cyclase inhibition.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenylyl Cyclase Inhibitors↗