Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 20 de 20
Filter
Add more filters










Publication year range
1.
SAR QSAR Environ Res ; 35(5): 343-366, 2024 May.
Article in English | MEDLINE | ID: mdl-38776241

ABSTRACT

Most of pharmaceutical agents display a number of biological activities. It is obvious that testing even one compound for thousands of biological activities is not practically possible. A computer-aided prediction is therefore the method of choice in this case to select the most promising bioassays for particular compounds. Using the PASS Online software, we determined the probable anti-inflammatory action of the 12 new hybrid dithioloquinolinethiones derivatives. Chemical similarity search in the World-Wide Approved Drugs (WWAD) and DrugBank databases did not reveal close structural analogues with the anti-inflammatory action. Experimental testing of anti-inflammatory activity of the synthesized compounds in the carrageenan-induced inflammation mouse model confirmed the computational predictions. The anti-inflammatory activity of the studied compounds (2a, 3a-3k except for 3j) varied between 52.97% and 68.74%, being higher than the reference drug indomethacin (47%). The most active compounds appeared to be 3h (68.74%), 3k (66.91%) and 3b (63.74%) followed by 3e (61.50%). Thus, based on the in silico predictions a novel class of anti-inflammatory agents was discovered.


Subject(s)
Anti-Inflammatory Agents , Carrageenan , Quantitative Structure-Activity Relationship , Quinolines , Animals , Mice , Anti-Inflammatory Agents/chemistry , Anti-Inflammatory Agents/pharmacology , Quinolines/chemistry , Quinolines/pharmacology , Inflammation/drug therapy , Inflammation/chemically induced , Thiones/chemistry , Thiones/pharmacology , Male , Edema/drug therapy , Edema/chemically induced
2.
SAR QSAR Environ Res ; 33(6): 485-497, 2022 Jun.
Article in English | MEDLINE | ID: mdl-35703013

ABSTRACT

Despite the greatest achievement in the development of anti-inflammatory agents in the last two decades, the current clinical drugs suffer from a variety of complications in community settings and hospital. There is still an urgent need to design novel molecules with better safety profile and with different molecular targets from those in current clinical use. The aim of this research was to discover a series of benzothiazole-based thiazolidinones with lipoxygenase (LOX) inhibitory activity as a mechanism of anti-inflammatory action. Carrageenan-induced mouse foot paw oedema assay was carried out to determine the anti-inflammatory activity, while LOX inhibition was examined through the conversion of sodium linoleate to 13-hydroperoxylinoleic acid. Molecular docking studies were performed using AutoDock 4.2 software. The anti-inflammatory activity of the title compounds was determined in a range of 18.4%-69.57%, where compound #3 was found to be the most potent (69.57%) and also to be more active than the reference drug indomethacin (47%). Moreover, compound #3 showed the highest LOX inhibitory activity with IC50 of 13 µM being less potent to that of the reference NDGA (IC50 = 1.3 µM). Compound #3 has been identified as lead compound for further modification in an attempt to improve anti-inflammatory and LOX inhibitory activities.


Subject(s)
Lipoxygenase Inhibitors , Lipoxygenase , Animals , Anti-Inflammatory Agents/pharmacology , Anti-Inflammatory Agents, Non-Steroidal/pharmacology , Benzothiazoles/pharmacology , Edema/chemically induced , Edema/drug therapy , Lipoxygenase/therapeutic use , Lipoxygenase Inhibitors/pharmacology , Mice , Molecular Docking Simulation , Quantitative Structure-Activity Relationship , Glycine max , Structure-Activity Relationship
3.
Horm Metab Res ; 53(3): 204-206, 2021 Mar.
Article in English | MEDLINE | ID: mdl-33652492

ABSTRACT

Currently, we are experiencing a true pandemic of a communicable disease by the virus SARS-CoV-2 holding the whole world firmly in its grasp. Amazingly and unfortunately, this virus uses a metabolic and endocrine pathway via ACE2 to enter our cells causing damage and disease. Our international research training programme funded by the German Research Foundation has a clear mission to train the best students wherever they may come from to learn to tackle the enormous challenges of diabetes and its complications for our society. A modern training programme in diabetes and metabolism does not only involve a thorough understanding of classical physiology, biology and clinical diabetology but has to bring together an interdisciplinary team. With the arrival of the coronavirus pandemic, this prestigious and unique metabolic training programme is facing new challenges but also new opportunities. The consortium of the training programme has recognized early on the need for a guidance and for practical recommendations to cope with the COVID-19 pandemic for the community of patients with metabolic disease, obesity and diabetes. This involves the optimal management from surgical obesity programmes to medications and insulin replacement. We also established a global registry analyzing the dimension and role of metabolic disease including new onset diabetes potentially triggered by the virus. We have involved experts of infectious disease and virology to our faculty with this metabolic training programme to offer the full breadth and scope of expertise needed to meet these scientific challenges. We have all learned that this pandemic does not respect or heed any national borders and that we have to work together as a global community. We believe that this transCampus metabolic training programme provides a prime example how an international team of established experts in the field of metabolism can work together with students from all over the world to address a new pandemic.


Subject(s)
COVID-19 , Diabetes Mellitus , Education, Medical, Continuing , Obesity , Pandemics , SARS-CoV-2 , COVID-19/epidemiology , COVID-19/therapy , Diabetes Mellitus/epidemiology , Diabetes Mellitus/therapy , Humans , Obesity/epidemiology , Obesity/therapy
4.
Bioorg Med Chem ; 21(2): 532-9, 2013 Jan 15.
Article in English | MEDLINE | ID: mdl-23219856

ABSTRACT

As a part of our ongoing studies in developing new derivatives as dual antimicrobial/anti-inflammatory agents we describe the synthesis of novel 5-arylidene-2-(1,3-thiazol-2-ylimino)-1,3-thiazolidin-4-ones. All newly synthesized compounds were tested for their anti-inflammatory activity using carrageenan mouse paw edema bioassay. Their COX-1/LOX inhibitory activities were also determined. Moreover, all compounds were evaluated for their antimicrobial and antifungal activities against a panel of Gram positive, Gram negative bacteria and moulds. All tested compounds exhibited better antimicrobial activity than commercial drugs, bifonazole, ketoconazole, ampicillin and streptomycin.


Subject(s)
Anti-Infective Agents/chemical synthesis , Anti-Inflammatory Agents/chemical synthesis , Thiazoles/chemistry , Animals , Anti-Infective Agents/chemistry , Anti-Infective Agents/pharmacology , Anti-Inflammatory Agents/chemistry , Anti-Inflammatory Agents/therapeutic use , Cyclooxygenase 1/chemistry , Cyclooxygenase 1/metabolism , Cyclooxygenase 2/chemistry , Cyclooxygenase 2/metabolism , Cyclooxygenase Inhibitors/chemistry , Cyclooxygenase Inhibitors/pharmacology , Cyclooxygenase Inhibitors/therapeutic use , Drug Evaluation, Preclinical , Edema/chemically induced , Edema/drug therapy , Gram-Negative Bacteria/drug effects , Gram-Positive Bacteria/drug effects , Mice , Microbial Sensitivity Tests , Thiazoles/pharmacology , Thiazoles/therapeutic use
5.
Genomics ; 92(5): 301-8, 2008 Nov.
Article in English | MEDLINE | ID: mdl-18616996

ABSTRACT

WISP1 and CTGF are members of the CCN family of growth factors encoding extracellular matrix proteins participating in several developmental and tumorigenic processes. Both are induced by the WNT signaling pathway, and microarray data suggest that expression of WISP1 and CTGF is repressed by Neurogenin3 (Ngn3 (NEUROG3)), a transcription factor directing specification of the endocrine pancreas. Single-cell reverse transcription polymerase chain reaction analysis suggested that this was a cell autonomous effect. To identify possible common regulatory networks involved in WISP1 and CTGF gene expression, their genomic regions were searched for common transcription factor motifs using a combination of in silico approaches and documented knowledge concerning pancreas development. This analysis revealed the presence of a conserved enhancer in both CTGF and WISP1 regulatory regions in 10 species covering a wide evolutionary distance. This enhancer contains binding sites for Ngn1/3 (NEUROG1/3) and transcription factors that are critically involved in pancreas development. Furthermore, it contained binding sites for three additional transcription factor families, which may indicate novel players are involved in this process.


Subject(s)
Connective Tissue Growth Factor/genetics , Enhancer Elements, Genetic/genetics , Gene Expression Regulation, Developmental , Oncogene Proteins/genetics , Pancreas/growth & development , Phylogeny , Animals , Basic Helix-Loop-Helix Transcription Factors/genetics , Basic Helix-Loop-Helix Transcription Factors/metabolism , CCN Intercellular Signaling Proteins , Computational Biology , Connective Tissue Growth Factor/metabolism , Conserved Sequence , Humans , Mice , Nerve Tissue Proteins/genetics , Nerve Tissue Proteins/metabolism , Oligonucleotide Array Sequence Analysis , Oncogene Proteins/metabolism , Pancreas/metabolism , Proto-Oncogene Proteins , Regulatory Sequences, Nucleic Acid , Reverse Transcriptase Polymerase Chain Reaction
6.
Cardiovasc Hematol Agents Med Chem ; 5(4): 249-64, 2007 Oct.
Article in English | MEDLINE | ID: mdl-17979687

ABSTRACT

Cardiovascular disease remains the leading cause of death worldwide. The renin-angiotensin-aldosterone system (RAAS) plays a key role in the regulation of blood pressure, through the actions of angiotensin (Ang) II. Excessive RAAS activity may lead to hypertension and associated target organ damage. Indeed, RAAS blockade with angiotensin converting enzyme inhibitors (ACEIs) and/or angiotensin receptor AT (1) blockers (ARBs) has proved to be successful treatment for arterial hypertension, heart failure and diabetes. Accumulating evidence suggests that arterial stiffness is an important and independent predictor of cardiovascular risk. More recently, a role for advanced glycation end-products (AGEs) in the development of arterial stiffening has been suggested. Advanced glycation end-products form by a nonenzymatic reaction between reducing sugars and biological proteins. Mechanisms underlying these alterations include AGE cross-linking of collagen and AGE interactions with circulating proteins and AGE receptors. New pharmacologic agents that prevent AGE formation, break cross-links, or block AGE receptors reduce vascular and myocardial stiffness, inhibit atherosclerotic plaque formation, and improve endothelial function. These agents promise to reduce the risk of isolated systolic hypertension, diastolic dysfunction, diabetes and thus, heart failure.


Subject(s)
Antihypertensive Agents/chemistry , Glycation End Products, Advanced/metabolism , Renin-Angiotensin System/drug effects , Angiotensin-Converting Enzyme Inhibitors/chemistry , Antihypertensive Agents/therapeutic use , Calcium Channel Blockers/chemistry , Diabetes Complications/metabolism , Endothelin Receptor Antagonists , Heart Failure/prevention & control , Humans , Hypertension/complications , Hypertension/drug therapy
7.
Development ; 128(15): 3017-27, 2001 Aug.
Article in English | MEDLINE | ID: mdl-11532923

ABSTRACT

Hoxa1 and Hoxb1 have overlapping synergistic roles in patterning the hindbrain and cranial neural crest cells. The combination of an ectoderm-specific regulatory mutation in the Hoxb1 locus and the Hoxa1 mutant genetic background results in an ectoderm-specific double mutation, leaving the other germ layers impaired only in Hoxa1 function. This has allowed us to examine neural crest and arch patterning defects that originate exclusively from the neuroepithelium as a result of the simultaneous loss of Hoxa1 and Hoxb1 in this tissue. Using molecular and lineage analysis in this double mutant background we demonstrate that presumptive rhombomere 4, the major site of origin of the second pharyngeal arch neural crest, is reduced in size and has lost the ability to generate neural crest cells. Grafting experiments using wild-type cells in cultured normal or double mutant mouse embryos demonstrate that this is a cell-autonomous defect, suggesting that the formation or generation of cranial neural crest has been uncoupled from segmental identity in these mutants. Furthermore, we show that loss of the second arch neural crest population does not have any adverse consequences on early patterning of the second arch. Signalling molecules are expressed correctly and pharyngeal pouch and epibranchial placode formation are unaffected. There are no signs of excessive cell death or loss of proliferation in the epithelium of the second arch, suggesting that the neural crest cells are not the source of any indispensable mitogenic or survival signals. These results illustrate that Hox genes are not only necessary for proper axial specification of the neural crest but that they also play a vital role in the generation of this population itself. Furthermore, they demonstrate that early patterning of the separate components of the pharyngeal arches can proceed independently of neural crest cell migration.


Subject(s)
Body Patterning , Branchial Region/embryology , Homeodomain Proteins/physiology , Neural Crest/embryology , Transcription Factors/physiology , Animals , Cell Lineage , Cell Movement , Cell Survival , Culture Techniques , Homeodomain Proteins/genetics , Mice , Mutation , Neural Crest/cytology , Transcription Factors/genetics
8.
J Biol Chem ; 276(23): 20506-15, 2001 Jun 08.
Article in English | MEDLINE | ID: mdl-11278854

ABSTRACT

Regionally restricted expression patterns of Hox genes in developing embryos rely on auto-, cross-, and para-regulatory transcriptional elements. One example is the Hoxb1 auto-regulatory element (b1-ARE), which drives expression of Hoxb1 in the fourth rhombomere of the hindbrain. We previously showed that HOXB1 and PBX1 activate transcription from the b1-ARE by binding to sequences required for the expression of a reporter gene in rhombomere 4 in vivo. We now report that in embryonal carcinoma cells, which retain characteristics of primitive neuroectodermal cells, the b1-ARE displays higher basal and HOX/PBX-induced activities than in other cell backgrounds. We have identified a bipartite-binding site for SOX/OCT heterodimers within the b1-ARE that accounts for its cell context-specific activity and is required for maximal transcriptional activity of HOX/PBX complexes in embryonal carcinoma cells. Furthermore, we found that in an embryonal carcinoma cell background, HOXB1 has a significantly higher transcriptional activity than its paralog HOXA1. We map the determinants for this differential activity within the HOXB1 N-terminal transcriptional activation domain. By using analysis in transgenic and HOXA1 mutant mice, we extended these findings on the differential activities of HOXA1 and HOXB1 in vivo, and we demonstrated that they are important for regulating aspects of HOXB1 expression in the hindbrain. We found that mutation of the SOX/OCT site and targeted inactivation of Hoxa1 both impair the response of the b1-ARE to retinoic acid in transgenic mice. Our results show that Hoxa1 is the primary mediator of the response of b1-ARE to retinoic acid in vivo and that this function is dependent on the binding of SOX/OCT heterodimers to the b1-ARE. These results uncover novel functional differences between Hox paralogs and their modulators.


Subject(s)
Enhancer Elements, Genetic , Gene Expression Regulation , Homeodomain Proteins/metabolism , Transcription Factors/metabolism , Animals , Base Sequence , Binding Sites , Cell Line , DNA Probes , Homeodomain Proteins/genetics , Homeodomain Proteins/physiology , Humans , Transcription Factors/physiology , Transcriptional Activation/physiology , Tretinoin/pharmacology
9.
Curr Opin Genet Dev ; 10(4): 380-6, 2000 Aug.
Article in English | MEDLINE | ID: mdl-10889064

ABSTRACT

Retinoid signalling has been implicated in regulating a wide variety of processes in vertebrate development. Recent advances from analyses on the synthesis, degradation and distribution of retinoids in combination with functional analysis of signalling components have provided important insights into the regulation of patterning the nervous system and the hindbrain in particular.


Subject(s)
Body Patterning/physiology , Rhombencephalon/embryology , Signal Transduction/physiology , Tretinoin/metabolism , Animals , Humans , Morphogenesis , Retinoids/metabolism
10.
J Pharm Pharmacol ; 50(6): 583-91, 1998 Jun.
Article in English | MEDLINE | ID: mdl-9680067

ABSTRACT

The synthesis of aroylaminoalcohols and 3-amino-substituted 1-phenylpropanols is described. These novel basic compounds have potent anti-inflammatory activity, significantly inhibiting rat paw oedema induced by a variety of phlogistic agents as a result of the release of inflammatory mediators such as histamine, 5-hydroxytryptamine, kinins, prostaglandins or leukotrienes. The biological activity of a selected, representative number of these compounds was examined on adjuvant-induced arthritis, a good animal model for rheumatoid arthritis in man. The results show that 3-(1-hydroxymethylpropylamino)-1-phenylpropan-1-one hydrochloride (2) and 3-(3-hydroxypiperidin-1-yl)-1-phenylpropan-1-one hydrochloride (4) effectively suppress the secondary lesions of adjuvant arthritis. 2-(3-Hydroxy-3-phenylpropylamino)-butan-1-ol hydrochloride (6) had no preventive activity in this animal model. In addition, several of the compounds suppressed the mitogenic responses of T-lymphocytes to concanavalin A, suggesting direct or indirect action on lymphocytes and therefore possible immunosuppressive properties. Finally, we investigated the in-vitro effects of compounds 2 and 4 on production of interleukins 1 and 2 (IL-1 and IL-2). We found that compound 4 suppressed the in-vitro production or action of IL-2 and IL-1. In contrast, compound 2 significantly increased the IL-1 activity of arthritic macrophages while reducing the ability of normal and arthritic splenocytes to produce or release IL-2. Our data suggest that compounds 2 and 4 have immunomodulatory properties that influence T lymphocyte functions.


Subject(s)
Adjuvants, Immunologic/chemical synthesis , Adjuvants, Immunologic/pharmacology , Alcohols/chemical synthesis , Alcohols/pharmacology , Anti-Inflammatory Agents/chemical synthesis , Anti-Inflammatory Agents/pharmacology , Arthritis, Rheumatoid/immunology , Arthritis, Rheumatoid/metabolism , Propanols/chemical synthesis , Propanols/pharmacology , Propiophenones/chemical synthesis , Propiophenones/pharmacology , Adjuvants, Immunologic/chemistry , Alcohols/chemistry , Animals , Anti-Inflammatory Agents/chemistry , Propanols/chemistry , Propiophenones/chemistry , Rats , Rats, Inbred F344
11.
Int J Dev Biol ; 42(3): 393-401, 1998.
Article in English | MEDLINE | ID: mdl-9654024

ABSTRACT

Hox genes are segmentally expressed in the developing vertebrate hindbrain, neural crest cells and pharyngeal arches suggesting an important role in patterning these structures. Here we discuss the cellular and molecular mechanisms controlling segmentation and specification in the branchial region of the head. In addition, based on the recent phenotypical and molecular analysis of loss-of-function mutants in the mouse, we speculate that Hox genes may act like Drosophila selector genes in this system.


Subject(s)
Body Patterning/genetics , Genes, Homeobox/physiology , Genes, Insect/physiology , Rhombencephalon/embryology , Animals , Drosophila/embryology , Drosophila/genetics , Gene Expression Regulation, Developmental , Homeodomain Proteins/genetics , Homeodomain Proteins/physiology , Mice , Rhombencephalon/cytology
12.
Development ; 125(6): 1025-36, 1998 Mar.
Article in English | MEDLINE | ID: mdl-9463349

ABSTRACT

In the developing vertebrate hindbrain Hoxa1 and Hoxb1 play important roles in patterning segmental units (rhombomeres). In this study, genetic analysis of double mutants demonstrates that both Hoxa1 and Hoxb1 participate in the establishment and maintenance of Hoxb1 expression in rhombomere 4 through auto- and para-regulatory interactions. The generation of a targeted mutation in a Hoxb1 3' retinoic acid response element (RARE) shows that it is required for establishing early high levels of Hoxb1 expression in neural ectoderm. Double mutant analysis with this Hoxb1(3'RARE) allele and other targeted loss-of-function alleles from both Hoxa1 and Hoxb1 reveals synergy between these genes. In the absence of both genes, a territory appears in the region of r4, but the earliest r4 marker, the Eph tyrosine kinase receptor EphA2, fails to be activated. This suggests a failure to initiate rather than maintain the specification of r4 identity and defines new roles for both Hoxb1 and Hoxa1 in early patterning events in r4. Our genetic analysis shows that individual members of the vertebrate labial-related genes have multiple roles in different steps governing segmental processes in the developing hindbrain.


Subject(s)
Homeodomain Proteins/genetics , Rhombencephalon/embryology , Rhombencephalon/metabolism , Transcription Factors/genetics , Animals , Base Sequence , DNA Primers/genetics , Female , Gene Expression Regulation, Developmental , Gene Targeting , Genes, Homeobox , Genes, Reporter , Heterozygote , Homozygote , Lac Operon , Mice , Mice, Inbred C57BL , Mice, Mutant Strains , Phenotype , Polymerase Chain Reaction , Pregnancy
13.
Development ; 125(6): 1123-36, 1998 Mar.
Article in English | MEDLINE | ID: mdl-9463359

ABSTRACT

The analysis of Hoxa1 and Hoxb1 null mutants suggested that these genes are involved in distinct aspects of hindbrain segmentation and specification. Here we investigate the possible functional synergy of the two genes. The generation of Hoxa1(3'RARE)/Hoxb1(3'RARE) compound mutants resulted in mild facial motor nerve defects reminiscent of those present in the Hoxb1 null mutants. Strong genetic interactions between Hoxa1 and Hoxb1 were uncovered by introducing the Hoxb1(3'RARE) and Hoxb1 null mutations into the Hoxa1 null genetic background. Hoxa1(null)/Hoxb1(3'RARE) and Hoxa1(null)/Hoxb1(null )double homozygous embryos showed additional patterning defects in the r4-r6 region but maintained a molecularly distinct r4-like territory. Neurofilament staining and retrograde labelling of motor neurons indicated that Hoxa1 and Hoxb1 synergise in patterning the VIIth through XIth cranial nerves. The second arch expression of neural crest cell markers was abolished or dramatically reduced, suggesting a defect in this cell population. Strikingly, the second arch of the double mutant embryos involuted by 10.5 dpc and this resulted in loss of all second arch-derived elements and complete disruption of external and middle ear development. Additional defects, most notably the lack of tympanic ring, were found in first arch-derived elements, suggesting that interactions between first and second arch take place during development. Taken together, our results unveil an extensive functional synergy between Hoxa1 and Hoxb1 that was not anticipated from the phenotypes of the simple null mutants.


Subject(s)
Branchial Region/embryology , Cranial Nerves/embryology , Homeodomain Proteins/genetics , Rhombencephalon/embryology , Transcription Factors/genetics , Animals , Branchial Region/abnormalities , Cell Movement/genetics , Cranial Nerves/abnormalities , Ear/embryology , Facial Nerve/abnormalities , Facial Nerve/drug effects , Facial Nerve/embryology , Female , Gene Expression Regulation, Developmental , Homozygote , Mice , Mutation , Neural Crest/cytology , Phenotype , Pregnancy , Retinoids/pharmacology , Rhombencephalon/abnormalities
14.
Development ; 124(19): 3693-702, 1997 Oct.
Article in English | MEDLINE | ID: mdl-9367425

ABSTRACT

Segmentation plays an important role in neuronal diversification and organisation in the developing hindbrain. For instance, cranial nerve branchiomotor nuclei are organised segmentally within the basal plates of successive pairs of rhombomeres. To reach their targets, motor axons follow highly stereotyped pathways exiting the hindbrain only via specific exit points in the even-numbered rhombomeres. Hox genes are good candidates for controlling this pathfinding, since they are segmentally expressed and involved in rhombomeric patterning. Here we report that in Hoxa-2(-/-) embryos, the segmental identities of rhombomere (r) 2 and r3 are molecularly as well as anatomically altered. Cellular analysis by retrograde dye labelling reveals that r2 and r3 trigeminal motor axons turn caudally and exit the hindbrain from the r4 facial nerve exit point and not from their normal exit point in r2. Furthermore, dorsal r2-r3 patterning is affected, with loss of cochlear nuclei and enlargement of the lateral part of the cerebellum. These results point to a novel role for Hoxa-2 in the control of r2-r3 motor axon guidance, and also suggest that its absence may lead to homeotic changes in the alar plates of these rhombomeres.


Subject(s)
Axons/physiology , Body Patterning , Gene Expression Regulation, Developmental , Genes, Homeobox , Motor Neurons/physiology , Rhombencephalon/embryology , Animals , Axons/ultrastructure , Cerebellum/abnormalities , Cerebellum/embryology , Cranial Nerves/abnormalities , Cranial Nerves/embryology , Embryonic and Fetal Development , Mice , Mice, Knockout , Motor Neurons/cytology , Polymerase Chain Reaction
15.
J Biol Chem ; 270(5): 2403-10, 1995 Feb 03.
Article in English | MEDLINE | ID: mdl-7836476

ABSTRACT

GPAT and AIRC encode two enzymes that catalyze steps 1 and 6 plus 7, respectively, of the de novo purine biosynthetic pathway. The chicken genes are closely linked and divergently transcribed from an approximately 230-base pair intergenic region. The promoter was scanned by deletion mutagenesis in a bireporter vector that allowed assay of transcriptional activity in both directions in transfected HepG2 and chicken LMH cells. Three classes of deletions were obtained: those affecting bidirectional transcription, those predominantly affecting GPAT transcription, and those predominantly affecting AIRC transcription. Defects in bidirectional transcription resulted from removal of an initiator-like element overlapping the AIRC transcription start site, as well as deletions removing a series of GC and CCAAT boxes from the AIRC proximal half of the promoter and a CCAAT-containing segment from the GPAT side. Several regions in the GPAT proximal half of the promoter, including an octamer-like motif downstream from the transcription start site, were required predominantly for GPAT expression. Evidence for interaction of HeLa nuclear proteins with some of these sites was obtained by gel retardation, DNase I, and methylation interference assays. Overall, the results showed that the intergenic region is an integrated bidirectional promoter and that a novel initiator-like element plays a central role in coordinating expression of the divergently transcribed AIRC and GPAT genes.


Subject(s)
Amidophosphoribosyltransferase/genetics , Carboxy-Lyases/genetics , Promoter Regions, Genetic , Purine Nucleotides/biosynthesis , Animals , Base Sequence , Chickens , DNA-Binding Proteins/metabolism , Gene Expression Regulation, Enzymologic , Molecular Sequence Data , Protein Binding , RNA, Messenger/genetics , Sequence Deletion , Sp1 Transcription Factor/metabolism , Structure-Activity Relationship , Transcription, Genetic
16.
J Biol Chem ; 269(7): 5313-21, 1994 Feb 18.
Article in English | MEDLINE | ID: mdl-8106516

ABSTRACT

A cDNA encoding human glutamine phosphoribosylpyrophosphate amidotransferase for step one in de novo purine nucleotide synthesis was cloned, sequenced, and expressed in Chinese hamster ovary cells to yield functional enzyme. Enzyme function was dependent upon removal of an 11-amino-acid propeptide. A mutant enzyme having three propeptide amino acid replacements was not processed and was not active. The human genes GPAT, encoding the amidotransferase, and AIRC, encoding a bifunctional enzyme for steps six and seven in the pathway, were cloned and characterized. GPAT and AIRC are closely linked and divergently transcribed from an intergenic region of approximately 625 base pairs. Expression of a luciferase reporter from the GPAT promoter was approximately 3-4-fold higher than from the AIRC promoter. The GPAT gene was mapped to the q12 region of chromosome 4.


Subject(s)
Amidophosphoribosyltransferase/genetics , Carboxy-Lyases/genetics , Chromosomes, Human, Pair 4 , Purine Nucleotides/biosynthesis , Transcription, Genetic , Amidophosphoribosyltransferase/isolation & purification , Amidophosphoribosyltransferase/metabolism , Amino Acid Sequence , Animals , Bacillus subtilis/enzymology , Base Sequence , Blotting, Southern , CHO Cells , Carboxy-Lyases/isolation & purification , Carboxy-Lyases/metabolism , Chickens , Chromosome Mapping , Cloning, Molecular , Cricetinae , DNA/analysis , Escherichia coli/enzymology , Genes , Genetic Linkage , HeLa Cells , Humans , In Situ Hybridization, Fluorescence , Molecular Sequence Data , Promoter Regions, Genetic , Pseudogenes , Rats , Restriction Mapping , Saccharomyces cerevisiae/enzymology , Sequence Homology, Amino Acid , Sequence Homology, Nucleic Acid , Transfection
17.
Int J Neurosci ; 74(1-4): 259-64, 1994.
Article in English | MEDLINE | ID: mdl-7928110

ABSTRACT

The anti-inflammatory activity of two novel opioids PM and PO as well as of pethidine was studied. The mouse paw edema, induced by various phlogistic agents, was significantly inhibited after the administration of opioids, fact that was independent of their antioxidant properties. The anti-inflammatory action of the above opioids was not reversed by naloxone. These results suggest that a variety of complex regulatory activities may be performed by opioid agonists via naloxone-sensitive or naloxone insensitive receptors on inflammatory cells, directly or indirectly by the inhibition of cytokines and mediators involved in inflammation.


Subject(s)
Anti-Inflammatory Agents/pharmacology , Meperidine/pharmacology , Naloxone/pharmacology , Narcotics/agonists , Narcotics/pharmacology , Animals , Anti-Inflammatory Agents/immunology , Cells, Cultured , Edema/physiopathology , Extremities/physiopathology , Lipid Peroxidation/drug effects , Meperidine/immunology , Mice , Naloxone/immunology , Narcotics/immunology , Spleen
18.
Mol Cell Biol ; 13(8): 4784-92, 1993 Aug.
Article in English | MEDLINE | ID: mdl-8336716

ABSTRACT

Two avian genes encoding essential steps in the purine nucleotide biosynthetic pathway are transcribed divergently from a bidirectional promoter element. The bidirectional promoter, embedded in a CpG island, directs coexpression of GPAT and AIRC genes from distinct transcriptional start sites 229 bp apart. The bidirectional promoter can be divided in half, with each half retaining partial activity towards the cognate gene. GPAT and AIRC genes encode the enzymes that catalyze step 1 and steps 6 plus 7, respectively, in the de novo purine biosynthetic pathway. This is the first report of genes coding for structurally unrelated enzymes of the same pathway that are tightly linked and transcribed divergently from a bidirectional promoter. This arrangement has the potential to provide for regulated coexpression comparable to that in a prokaryotic operon.


Subject(s)
Amidophosphoribosyltransferase/genetics , Carbon-Nitrogen Ligases , Chickens/genetics , Gene Expression Regulation, Enzymologic , Ligases/genetics , Promoter Regions, Genetic , Purine Nucleotides/chemistry , Animals , Base Sequence , Genes , Genetic Linkage , Molecular Sequence Data , RNA, Messenger/genetics , Restriction Mapping , Transcription, Genetic
19.
Arzneimittelforschung ; 41(5): 537-41, 1991 May.
Article in English | MEDLINE | ID: mdl-1898425

ABSTRACT

The effect of the novel agent gamma-(2-aminoethylamino)-2-butyrothienone (gamma-ABT) on local and systemic changes of rats with adjuvant induced disease (AID) was investigated, gamma-ABT showed potent inhibitory effect on adjuvant primary inflammation and almost totally inhibited the secondary lesions. gamma-ABT improved the changes in lymphoid organ weight except of the thymus. gamma-ABT improved the change in albumin/globulin ratio, which is a parameter of systemic inflammatory reaction but did not improve the body weight gain in AID and normal rats. In addition gamma-ABT did not affect directly T or B lymphocytes as shown by the lymphocyte responses to mitogen (Con A) and a T cell dependent antigen (SRBC) but rather inhibited the suppressor cells found in AID. Although structurally gamma-ABT is completely different from known non-steroidal anti-inflammatories, immunosuppressive drugs behave to a great extent like them.


Subject(s)
Adjuvants, Immunologic/pharmacology , Anti-Inflammatory Agents, Non-Steroidal/pharmacology , Arthritis, Experimental/drug therapy , Thiophenes/pharmacology , Animals , Antibody Formation/drug effects , Antibody-Producing Cells/drug effects , Arthritis, Experimental/pathology , B-Lymphocytes/drug effects , Body Weight/drug effects , Concanavalin A/pharmacology , Female , Foot/pathology , Liver Glycogen/metabolism , Male , Mitogens , Mycobacterium , Rats , Rats, Inbred F344 , T-Lymphocytes/drug effects
20.
Arzneimittelforschung ; 41(4): 423-6, 1991 Apr.
Article in English | MEDLINE | ID: mdl-1859517

ABSTRACT

A series of 4-hydroxy-(or-amino)-ethyl-amino butyrophenones or butyrothienones were synthesized. For detail studies on antiinflammatory effects, gamma-(2-aminoethylamino)-2-butyrothienone (gamma-ABT) was chosen as representative of these new non-steroidal anti-inflammatories (NSAID). The effect on mouse paw edema induced by various phlogistic agents was first investigated. The inhibitory effect of gamma-ABT on carrageenin-induced edema was remarkable and nearly equal to that of indometacin. Similarly to indometacin, gamma-ABT inhibited the early and late stage of yeast-induced edema in contrast to concanavalin A (Con A) induced edema which was only inhibited by gamma-ABT. Both the above induced edema are supposed to be unrelated to prostaglandins in the rat system. gamma-ABT displayed an inhibitory effect on nystatin-induced edema similar to indometacin suggesting that gamma-ABT has significant membrane stabilizing action and a strong blocking action on synthesis of prostaglandins. gamma-ABT inhibited as well the sustained edema induced by mustard. In conclusion gamma-ABT is an effective agent not only on acute but also on subacute and chronic inflammation and its mode of action appears similar to other NSAID. gamma-ABT posses in addition the advantage of antioxidant activity which is not shared by selective cyclooxygenase inhibitors and thus should be potentially effective in autoimmune diseases.


Subject(s)
Anti-Inflammatory Agents, Non-Steroidal/pharmacology , Edema/drug therapy , Thiophenes/pharmacology , Animals , Carrageenan , Concanavalin A , Edema/chemically induced , Edema/pathology , Female , Foot/pathology , Indomethacin/pharmacology , Male , Mice , Mice, Inbred C57BL , Mice, Inbred Strains , Mustard Plant , Nystatin , Plant Lectins , Plants, Medicinal , Yeasts
SELECTION OF CITATIONS
SEARCH DETAIL
...