acid-phosphatase and Staphylococcal-Infections

The identity of Staphylococcus aureus virulence factors involved in chronic osteomyelitis remains unresolved. SapS is a class C non-specific acid phosphatase and a well-known virulence factor that has been identified in S. aureus strain 154 but in protein extracts from rotting vegetables.. To identify the SapS gene and characterize the activity of SapS from S. aureus strains: 12 isolates from bone infected samples of patients treated for chronic osteomyelitis and 49 from a database with in silico analysis of complete bacterial genomes.. The SapS gene was isolated and sequenced from 12 S. aureus clinical isolates and two reference strains; 49 S. aureus strains and 11 coagulase-negative staphylococci were tested using in silico PCR. Culture media semi-purified protein extracts from the clinical strains were assayed for phosphatase activity with p-nitro-phenylphosphate, O-phospho-L-tyrosine, O-phospho-L-serine, and OphosphoL-threonine in conjunction with various phosphatase inhibitors.. SapS was detected in the clinical and in-silico S. aureus strains, but not in the in silico coagulase-negative staphylococci strains. Sec-type I lipoprotein-type N-terminal signal peptide sequences; secreted proteins, and aspartate bipartite catalytic domains coding sequences were found in the SapS nucleotide and amino acid sequence analysis. SapS dephosphorylated with p-nitro-phenyl-phosphate and ophosphoLtyrosine were selectively resistant to tartrate and fluoride, but sensitive to vanadate and molybdate.. SapS gene was found in the genome of the clinical isolates and the in silico Staphylococcus aureus strains. SapS shares biochemical similarities with known virulent bacterial, such as protein tyrosine phosphatases, suggesting it may be a virulence factor in chronic osteomyelitis.. Se desconoce la identidad de los factores de virulencia de Staphylococcus aureus implicados en la osteomielitis crónica. Sin embargo, SapS, una fosfatasa ácida no específica de clase C, es un factor de virulencia reconocido y ya fue identificada en la cepa 154 de S. aureus, pero en extractos proteicos de vegetales podridos.. Detectar el gen SapS y caracterizar la actividad de la fosfatasa SapS en cepas de S. aureus aisladas de pacientes con osteomielitis crónica y en las reportadas en una base de datos de análisis in silico de genomas bacterianos completos.. Se aisló y secuenció el gen SapS en los 12 aislamientos clínicos de S. aureus y en dos cepas de referencia; estas secuencias se analizaron junto con las secuencias de las cepas reportadas en la base de datos de genomas bacterianos: 49 cepas de S. aureus y 11 cepas de estafilococos negativos para coagulasa. Se evalúo la actividad de la fosfatasa SapS, presente en los extractos de los sobrenadantes de los cultivos de las cepas clínicas, mediante la hidrólisis de fosfato p-nitrofenil, O-fosfo-Ltirosina, O-fosfo-L serina y O-fosfo-L treonina junto con varios inhibidores de fosfatasas.. Se detectó el gen SapS en el genoma de las cepas clínicas y en las 49 cepas de S. aureus analizadas in silico, pero no en las 11 cepas de estafilococos negativos para coagulasa. La secuenciación de SapS reveló un péptido señal presente en el extremo N-terminal de proteínas extracelulares y los dominios bipartitos de aspartato (DDDD) en su sitio catalítico. SapS hidroliza selectivamente el fosfato p-nitrofenil y la O-fosfo-L-tirosina, pero es sensible a vanadato y molibdato.. Se encontró SapS en el genoma de S. aureus de las cepas clínicas y de las cepas de simulación computacional. La SapS con actividad específica para la hidrólisis de la O-fosfo-L-tirosina comparte similitudes bioquímicas con las fosfatasas-tirosina bacterianas, por lo que puede formar parte de la red de factores de virulencia de la osteomielitis crónica.

Topics: Acid Phosphatase; Coagulase; Humans; Staphylococcal Infections; Staphylococcus; Staphylococcus aureus

Topics: Acid Phosphatase; Animals; Bacterial Proteins; Mice; Staphylococcal Infections; Staphylococcus aureus; Virulence; Virulence Factors; Zebrafish

Tartrate-resistant acid phosphatase (TRAP) is a lysosomal di-iron protein of mononuclear phagocytes and osteoclasts. Hitherto, no role for the enzyme in immunity has been identified; however, knockout mice lacking TRAP have a skeletal phenotype caused by an intrinsic osteoclast defect. To investigate a putative function for TRAP in macrophages (Mphi), we investigated proinflammatory responses and systemic microbial clearance in knockout mice compared with age- and gender-matched congenic wild-type mice. Phorbol 12-myristate 13-acetate (PMA)-stimulated and interferon-gamma (IFN-gamma)-induced superoxide formation was enhanced in peritoneal Mphi lacking TRAP; nitrite production in response to stimulation with lipopolysaccharide (LPS) and IFN-gamma was also increased. In addition, secretion of the proinflammatory cytokines, tumour necrosis factor-alpha (TNF-alpha), interleukin (IL)-1beta and IL-12, was significantly greater in TRAP-deficient Mphi when stimulated with LPS, with or without addition of either TNF-alpha or IFN-gamma. The activity of tartrate-sensitive (lysosomal) acid phosphatase was increased in Mphi from the knockout mice but activities of the lysosomal hydrolases N-acetyl beta-glucosaminidase and acid beta-glucuronidase were unchanged, indicating selective activation of compensatory acid phosphatase activity. Evidence of impaired Mphi function in vivo was obtained in TRAP knockout mice, which showed delayed clearance of the microbial pathogen, Staphylococcus aureus, after sublethal intraperitoneal inoculation. After microbial challenge, peritoneal exudates obtained from TRAP knockout mice had a reduced population of Mphi. As peritoneal Mphi and neutrophils lacking TRAP were able to phagocytose and kill S. aureus normally in vitro, TRAP may directly or indirectly influence recruitment of Mphi to sites of microbial invasion. Our study shows that TRAP participates in the inflammatory response of the Mphi and influences effector signalling pathways in innate immunity.

Topics: Acid Phosphatase; Animals; Bone Marrow; Cytokines; Female; Free Radicals; Immunophenotyping; Inflammation; Isoenzymes; Lysosomes; Macrophages; Macrophages, Peritoneal; Male; Mice; Mice, Knockout; Phagocytosis; Staphylococcal Infections; Staphylococcus aureus; Superoxides; Tartrate-Resistant Acid Phosphatase

Topics: Acid Phosphatase; Animals; Chronic Disease; Electric Stimulation Therapy; Fallopian Tubes; Female; Glucuronidase; Rabbits; Salpingitis; Staphylococcal Infections

The study of the phosphatase activity of chick fibroblasts in the process of staphylococcal infection by the electron-histochemical method has revealed the presence of correlation between the degree of cytoplasmic destruction in fibroblasts and the level of acid phosphatase activity. Changes in the phosphatase activity of fibroblasts in the process of bacterial infection, depending on the presence of homologous bacteriophages in the system used as a model, have also been studied.

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Cells, Cultured; Chick Embryo; Fibroblasts; Histocytochemistry; Microscopy, Electron; Phosphoric Monoester Hydrolases; Staphylococcal Infections

Lysosomes form an integral part of the degradative mechanisms of the phagocytic cells. Mice were injected with suramin, a lysosomotrophic drug, to investigate the effects of lysosomal pathology on the cell biology and in situ bactericidal activity of the pulmonary macrophage. Treatment with suramin resulted in marked alterations in the cell biology of the macrophage: (i) increased vacuolization and protein content, (ii) suppressed intracellular phagosome-lysosome fusion, (iii) decreased activity of the lysosomal enzymes beta-glucuronidase and N-acetyl-glucosaminidase, and (iv) enhanced exocytosis of acid phosphatase during phagocytosis. Addition of suramin, in vitro, to cell lysates resulted in a reduction in the catalytic activities of acid phosphatase, beta-glucuronidase, and N-acetyl-glucosaminidase; thereby suggesting that selective interaction, in vivo, between suramin and lysosomes containing beta-glucuronidase and N-acetyl-glucosaminidase may have occurred. Plasma membrane 5'-nucleotide phosphodiesterase activity was increased in macrophages recovered from suramin-treated animals. Although the "resting-state" reduction of nitroblue tetrazolium (NBT) was lower in these macrophages, cells stimulated by a phagocytic challenge demonstrated normal increases in NBT reduction. Phagocytosis, in vitro, and pulmonary bactericidal activity were not altered. These data demonstrate that suramin altered numerous aspects of the phagocyte's lysosomal system. Despite these changes in the cell biology of the pulmonary macrophage, the cell's defense functions were not reduced.

Topics: Acetylglucosaminidase; Acid Phosphatase; Animals; Glucuronidase; Lung; Lung Diseases; Lysosomes; Macrophages; Male; Mice; Phagocytosis; Proteus Infections; Receptors, Fc; Staphylococcal Infections; Suramin

Topics: Acid Phosphatase; Animals; Glucuronidase; Injections, Intraperitoneal; Kinetics; Male; Mannans; Mice; Muramidase; Polysaccharides; Saccharomyces cerevisiae; Staphylococcal Infections

The role of lysosomal enzymes in the inactivation of inhaled bacteria by alveolar macrophages was studied in rats infected with aerosols of Staphylococcus aureus and then exposed for 5 hr to 2.5 ppm of ozone to determine whether pollutant-induced defects in phagocytic killing were associated with reduction in enzyme activity. Rates of bacterial ingestion and the activities of cellular acid phosphatase and beta-glucuronidase were measured simultaneously in in situ perfused right lungs by sequential staining of frozen sections for enzyme and bacteria. Quantitative measurements of enzyme activity within macrophages without ingested bacteria were made with a computer-controlled cytospectrophotometry system. Exposure to ozone resulted in diminished rates of bacterial clearance and ingestion, large increases in numbers of intra- and extracellular staphylococcal microcolonies, and an absence of enzyme activity for macrophages containing bacterial microcolonies. Enzyme activity was unimpaired in macrophages without ingested bacteria. These results, in which absence of enzyme activity occurred only in macrophages subjected to the dual insults of ozone exposure and ingested bacteria, prove a relationship between impairment in bactericidal capacity and cellular activities of lysosomal enzymes.

Topics: Acid Phosphatase; Aerosols; Animals; Glucuronidase; Lung; Lysosomes; Macrophages; Ozone; Phagocytosis; Rats; Staining and Labeling; Staphylococcal Infections; Staphylococcus aureus

The activity of redox and hydrolytic enzymes was studied in the blood lymphocytes and neutrophils of rabbits infected with hemolitic Staphylococcus albus. The changes in the enzymatic activity of blood cells in generalized and localized forms of the inflammatory process depended on the form, course and duration of the disease. At early periods the indices of the acid phosphatase activity of lymphocytes had the greatest informative value for the diagnosis of the inflammatory process. The diagnostic significance of the activity of the alkaline phosphatase of neutrophils consisted in redistribution of cells by the degree of the enzyme activity.

Topics: Abscess; Acid Phosphatase; Alkaline Phosphatase; Animals; Clinical Enzyme Tests; Histocytochemistry; Lymphocytes; Neutrophils; Rabbits; Sepsis; Staphylococcal Infections; Succinate Dehydrogenase

Topics: Acid Phosphatase; Alkaline Phosphatase; Dysentery, Bacillary; Hepatitis A; Humans; Meningitis, Meningococcal; Neutrophils; Staphylococcal Infections

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Escherichia coli Infections; Lymphocyte Activation; Lymphocytes; Macrophages; Mice; Phagocytosis; Staphylococcal Infections; Streptococcal Infections; Uracil

Topics: Acid Phosphatase; Adult; Agranulocytosis; Alkaline Phosphatase; Bacterial Infections; Chediak-Higashi Syndrome; Chemotaxis; Female; Glucuronidase; Humans; Leukocytes; Lysosomes; Male; Muramidase; Peroxidases; Skin Window Technique; Staphylococcal Infections; Streptococcal Infections

Topics: Acid Phosphatase; Agglutination Tests; Antibodies; Antigen-Antibody Complex; Arthus Reaction; Complement Fixation Tests; Complement System Proteins; Humans; Immune Adherence Reaction; Neutrophils; Peptide Hydrolases; Pharynx; Respiratory Tract Infections; Staphylococcal Infections; Streptococcal Infections; Streptococcus pyogenes; Vascular Diseases

Topics: Acid Phosphatase; Aminosalicylic Acids; Ampicillin; Animals; Anti-Bacterial Agents; Benzimidazoles; Cephaloridine; Dihydrostreptomycin Sulfate; Drug Synergism; Escherichia coli Infections; Kanamycin; Liver Glycogen; Macrophages; Mice; Neomycin; Oxacillin; Penicillin G; Penicillin Resistance; Staphylococcal Infections; Tetracycline

Topics: Acid Phosphatase; Aerosols; Air Pollution; Alkaline Phosphatase; Animals; Environmental Exposure; Germ-Free Life; Histocytochemistry; Lung; Mice; Nitrogen Dioxide; Orthomyxoviridae Infections; Oxygen Consumption; Staphylococcal Infections

Topics: Acid Phosphatase; Animals; Bacterial Proteins; Bacteriophage Typing; Culture Media; Drug Resistance, Microbial; Hemolysis; Lipids; Mice; Oxidoreductases; Oxygen Consumption; Pigments, Biological; RNA; Staphylococcal Infections; Staphylococcus; Succinate Dehydrogenase; Virulence

Topics: Acid Phosphatase; Alkaline Phosphatase; Animals; Dihydrolipoamide Dehydrogenase; Gastric Mucosa; Gastroenteritis; Glucosephosphate Dehydrogenase; Haplorhini; Ileum; In Vitro Techniques; Jejunum; Staphylococcal Infections; Succinate Dehydrogenase; Toxins, Biological

Topics: Acid Phosphatase; Carrier State; Catalase; Coagulase; Humans; Oxidoreductases; Staphylococcal Infections; Staphylococcus; Urease; Virulence

Topics: Acid Phosphatase; Bacteriophage Typing; Biomedical Research; Carrier State; Child; Diarrhea; Diarrhea, Infantile; Glutamates; Humans; Ketones; Mice; Pleurisy; Pneumonia; Research; Staphylococcal Infections; Staphylococcus; Suppuration

Topics: Acid Phosphatase; Mercury; Penicillin Resistance; Penicillins; Staphylococcal Infections; Staphylococcus; Staphylococcus aureus

Cannon, Frances D. (The Mary Imogene Bassett Hospital, Cooperstown, N.Y.), and Clinton V. Z. Hawn. Phosphatase activity of Staphylococcus aureus: correlation of enzyme production with resistance to penicillin and phage pattern. J. Bacteriol. 86:1052-1056. 1963.-A simplified method for the determination of the phosphatase activity in different strains of staphylococci is described. The relationship between phosphatase production and penicillin resistance was studied and found inconstant. A close relationship between phosphatase production and phage group and type was observed.

Topics: Acid Phosphatase; Bacteriophages; Metabolism; Penicillin Resistance; Penicillins; Research; Staphylococcal Infections; Staphylococcus; Staphylococcus aureus

Pan, Yue-Liang (University of Michigan Medical School, Ann Arbor), and Harold J. Blumenthal. Correlation between acid phosphatase and coagulase production or phage type of Staphylococcus aureus. J. Bacteriol. 82:124-129. 1961.-A quantitative assay of acid phosphatase in 26 coagulase-negative and 73 coagulase-positive strains of Staphylococcus aureus was performed using an assay procedure modified to estimate extracellular, as well as the cellular, phosphatase. Some of the factors affecting the formation of acid phosphatase were studied. On the average, coagulase-negative strains produced 4.0 units and coagulase-positive strains 19.6 units although some negative strains produced more phosphatase than positive strains. More phosphatase was produced by strains of phage-group I than by strains of any other group.

Topics: Acid Phosphatase; Bacteriophages; Coagulase; Phosphoric Monoester Hydrolases; Staphylococcal Infections; Staphylococcus; Staphylococcus aureus