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1.
Colorectal Dis ; 21(11): 1259-1269, 2019 Nov.
Article in English | MEDLINE | ID: mdl-31282600

ABSTRACT

AIM: Volatile organic compounds (VOCs) are potential biomarkers for diagnosing colorectal cancer (CRC). We characterized urinary VOCs from CRC patients, their spouses/cohabitors (spouses) and first-degree relatives (relatives) to determine any differences. Correlation with stool-derived microbiomes was also undertaken. METHODS: Urine from 56 CRC patients, 45 spouses and 37 relatives was assayed using liquid chromatography, field asymmetric ion mobility spectrometry (FAIMS), mass spectrometer technology. Analysis was performed using five-fold cross-validation and a random forest classifier. Faecal microbiome 16S rRNA was sequenced using Illumina MiSeq protocols and analysed using UPARSE and QIIME pipelines. VOC and microbiome profiles were also compared before and after cancer treatment. RESULTS: Urinary VOC profiles of CRC patients were indistinguishable from either spouses or relatives. When spouses and relatives were grouped together to form a larger non-cancer control group (n = 82), their VOC profiles became distinguishable from those of CRC patients (n = 56) with 69% sensitivity and specificity, area under the curve 0.72 (P < 0.001). Microbiome analysis identified > 1300 operational taxonomic units across all groups. The analysis of similarity R value was 0.067 (P < 0.001), with significantly different bacterial abundances in 82 operational taxonomic units (6.2%) by Kruskal-Wallis testing. CRC patients' VOC or stool microbiome profiles were unchanged after treatment. CONCLUSION: Although CRC patients' urinary VOC profiles cannot be differentiated from those of spouses or relatives they can be differentiated from a larger non-cancer control group. Comparison of the groups' microbiomes confirmed differences in bacterial species abundance. The current FAIMS-based assay can detect a unique, but modest, signal in CRC patients' urinary VOCs, which remains unaltered after treatment.


Subject(s)
Colorectal Neoplasms/microbiology , Colorectal Neoplasms/urine , Feces/microbiology , Gastrointestinal Microbiome/genetics , Ion Mobility Spectrometry/statistics & numerical data , Volatile Organic Compounds/urine , Aged , Area Under Curve , Biomarkers, Tumor/urine , Case-Control Studies , Family , Female , Humans , Male , Middle Aged , RNA, Ribosomal, 16S/analysis , Reproducibility of Results , Sensitivity and Specificity , Spouses/statistics & numerical data
2.
Genome Announc ; 3(6)2015 Nov 12.
Article in English | MEDLINE | ID: mdl-26564037

ABSTRACT

Pandoraea species, in particular Pandoraea apista, are opportunistic, multidrug-resistant pathogens in persons with cystic fibrosis (CF). To aid in understanding the role of P. apista in CF lung disease, we used Illumina MiSeq and nanopore MinION technology to sequence the whole genome of the P. apista LMG 16407(T).

3.
Br J Dermatol ; 154(6): 1175-9, 2006 Jun.
Article in English | MEDLINE | ID: mdl-16704651

ABSTRACT

BACKGROUND: Melanocytes represent a static component of the epidermis, and the role of apoptosis in basal melanocyte function and melanocytic tumour formation has not been fully elucidated. OBJECTIVES: The aim of this study was to investigate the expression of Bcl-2 anti-apoptotic and Bax apoptotic proteins in congenital naevi in correlation with p-27 protein and Ki-67 proliferative index. METHODS: Our material comprised 30 congenital naevi (eight giant) excised from children aged from 15 days to 14 years old. The immunohistochemical streptavidin-biotin method was performed on paraffin sections for the detection of Bcl-2 (cl100/D5), Bax (cl2D2) , Ki-67 (MIB-1) and p-27 (1B4) proteins with monoclonal antibodies. RESULTS: Bcl-2 protein was detected in all cases showing a strong diffuse cytoplasmic expression in >70% of the naevocytes and was preserved in the deeper parts of the naevi. On the other hand, Bax was detected in 13 of the cases, showing a fainter cytoplasmic expression in 40-50% of the naevocytes without any particular topographic distribution. Ki-67 was detected in all cases showing a limited expression in 1-2% of the nuclei mainly in the junctional and upper dermal components. p-27 protein showed a broad diffuse nuclear expression (>70% of the nuclei) in all cases with a particular increase in the deeper parts of the naevi. Bcl-2 expression showed a parallel correlation with p-27 protein. CONCLUSIONS: Broad Bcl-2 expression in congenital naevi suggests that suppression of apoptosis may play an important role in the maintenance of naevocytes despite the low proliferative activity.


Subject(s)
Nevus/metabolism , Proto-Oncogene Proteins c-bcl-2/metabolism , Skin Neoplasms/metabolism , bcl-2-Associated X Protein/metabolism , Adolescent , Apoptosis , Cell Proliferation , Child , Child, Preschool , Cyclin-Dependent Kinase Inhibitor p27/metabolism , Female , Humans , Immunoenzyme Techniques , Infant , Infant, Newborn , Ki-67 Antigen/metabolism , Male , Neoplasm Proteins/metabolism , Nevus/congenital , Nevus/pathology , Skin Neoplasms/congenital , Skin Neoplasms/pathology
4.
Biochem Soc Trans ; 34(Pt 1): 104-7, 2006 Feb.
Article in English | MEDLINE | ID: mdl-16417494

ABSTRACT

RNA was isolated from cultures of Escherichia coli strain MG1655 and derivatives defective in fnr, narXL, or narXL with narP, during aerobic growth, or anaerobic growth in the presence or absence of nitrate or nitrite, in non-repressing media in which both strain MG1655 and an fnr deletion mutant grew at similar rates. Glycerol was used as the non-repressing carbon source and both trimethylamine-N-oxide and fumarate were added as terminal electron acceptors. Microarray data supplemented with bioinformatic data revealed that the FNR (fumarate and nitrate reductase regulator) regulon includes at least 104, and possibly as many as 115, operons, 68 of which are activated and 36 are repressed during anaerobic growth. A total of 51 operons were directly or indirectly activated by NarL in response to nitrate; a further 41 operons were repressed. Four subgroups of genes implicated in management of reactive nitrogen compounds, NO and products of NO metabolism, were identified; they included proteins of previously unknown function. Global repression by the nitrate- and nitrite-responsive two-component system, NarQ-NarP, was shown for the first time. In contrast with the frdABCD, aspA and ansB operons that are repressed only by NarL, the dcuB-fumB operon was among 37 operons that are repressed by NarP.


Subject(s)
DNA-Binding Proteins/metabolism , Escherichia coli Proteins/metabolism , Escherichia coli/physiology , Iron-Sulfur Proteins/metabolism , Nitrates/metabolism , Nitrites/metabolism , Oxygen/metabolism , Anaerobiosis , DNA-Binding Proteins/genetics , Escherichia coli/genetics , Escherichia coli Proteins/genetics , Gene Expression Profiling , Gene Expression Regulation, Bacterial , Iron-Sulfur Proteins/genetics , Oligonucleotide Array Sequence Analysis , Operon , Regulon
5.
J Bacteriol ; 183(9): 2937-42, 2001 May.
Article in English | MEDLINE | ID: mdl-11292815

ABSTRACT

Three potential regulators of flagellar expression present in the genome sequence of Campylobacter jejuni NCTC 11168, the genes rpoN, flgR, and fliA, which encode the alternative sigma factor sigma(54), the sigma(54)-associated transcriptional activator FlgR, and the flagellar sigma factor sigma(28), respectively, were investigated for their role in global regulation of flagellar expression. The three genes were insertionally inactivated in C. jejuni strains NCTC 11168 and NCTC 11828. Electron microscopic studies of the wild-type and mutant strains showed that the rpoN and flgR mutants were nonflagellate and that the fliA mutant had truncated flagella. Immunoblotting experiments with the three mutants confirmed the roles of rpoN, flgR, and fliA in the expression of flagellin.


Subject(s)
Bacterial Proteins/genetics , Campylobacter jejuni/genetics , DNA-Binding Proteins/genetics , DNA-Directed RNA Polymerases/genetics , Flagella/genetics , Genes, Bacterial , Sigma Factor/genetics , Trans-Activators , Transcription Factors/genetics , Amino Acid Sequence , Campylobacter jejuni/cytology , Cloning, Molecular , Flagella/metabolism , Molecular Sequence Data , Mutagenesis, Insertional , PII Nitrogen Regulatory Proteins , RNA Polymerase Sigma 54 , Sequence Alignment
6.
J Oral Pathol Med ; 30(3): 148-53, 2001 Mar.
Article in English | MEDLINE | ID: mdl-11271629

ABSTRACT

The oral manifestations of chronic graft-versus-host disease (cGVHD) in eight allogeneic bone marrow transplant (BMT) paediatric recipients were studied clinically, and lip biopsies were performed in seven of them. A prominent lichenoid reaction was observed in four patients, two with accompanying ulceration. Superficial mucoceles were present in three children. Clinically obvious xerostomia was seen in seven patients. Lip biopsies were positive and correlated with the clinical manifestations. Both clinical and histological findings confirmed the diagnosis of cGVHD. In three additional children, with systemic manifestations indicating cGVHD, the oral mucosa was clinically and histologically normal, and the systemic manifestations were, thus, attributed to drug reactions. The above findings indicate the high value of oral examination in diagnosing or confirming paediatric cGVHD. Superficial mucoceles, reported for the first time in paediatric recipients, seem to be important in the early diagnosis of cGVHD.


Subject(s)
Bone Marrow Transplantation/adverse effects , Graft vs Host Disease/complications , Mouth Diseases/etiology , Mouth Mucosa/pathology , Adolescent , Biopsy , Child , Chronic Disease , Female , Graft vs Host Disease/diagnosis , Graft vs Host Disease/etiology , Humans , Infant , Lichen Planus, Oral/etiology , Lip/pathology , Male , Mucocele/etiology , Xerostomia/etiology
7.
J Periodontol ; 72(1): 107-12, 2001 Jan.
Article in English | MEDLINE | ID: mdl-11210066

ABSTRACT

BACKGROUND: The purpose of this paper is to present the first case of gingival overgrowth, premature root resorption, and alveolar bone loss, which preceded the diagnosis of a stage IVB Hodgkin's lymphoma (HL) in a 9-year-old boy. METHODS: The child presented complaining of gingival pain which first appeared 3 months prior. Clinical examination revealed inflamed, hyperplastic gingivae, while x-ray showed premature root resorption and alveolar bone loss. Medical work-up was significant for cervical lymphadenopathy. Gingival biopsy, followed by lymph node resection, was performed twice. RESULTS: Histological examination of both gingival biopsies disclosed a mixed inflammatory infiltrate, while classical Hodgkin's lymphoma of the nodular sclerosis type was diagnosed from the second lymph node biopsy. Chemotherapy was instituted with mustard-vincristine-procarbazine-prednizone and adriamycine-bleomycine-vinblastine-dacarbazine. Remission of the lymphoma was observed with concomitant regression of the gingival overgrowth. CONCLUSIONS: The inflammatory gingival overgrowth, premature root resorption of deciduous teeth, and alveolar bone loss in this case, in conjunction with the regression of gingival overgrowth which followed the completion of chemotherapy, are strongly indicative of a paraneoplastic manifestation of HL. The postulated mechanism for the development of the manifestation is the constitutive activation of the transcription factor NF-kB. The gingival inflammatory reaction was probably further aggravated by the bacterial-stimulated cytokine secretion released by monocytes.


Subject(s)
Gingival Overgrowth/etiology , Hodgkin Disease/complications , Paraneoplastic Syndromes/etiology , Alveolar Bone Loss/etiology , Antineoplastic Combined Chemotherapy Protocols/administration & dosage , Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Biopsy , Bleomycin/administration & dosage , Child , Dacarbazine/administration & dosage , Doxorubicin/administration & dosage , Gingival Hyperplasia/etiology , Humans , Lymph Nodes/pathology , Male , Mechlorethamine/administration & dosage , Neoplasm Staging , Prednisone/administration & dosage , Procarbazine/administration & dosage , Remission Induction , Root Resorption/etiology , Tooth, Deciduous/pathology , Vinblastine/administration & dosage , Vincristine/administration & dosage
8.
J Bacteriol ; 181(16): 4798-804, 1999 Aug.
Article in English | MEDLINE | ID: mdl-10438747

ABSTRACT

Microaerophiles like Campylobacter jejuni must resist oxidative stresses during transmission or infection. Growth of C. jejuni 81116 under iron limitation greatly increased the expression of two polypeptides of 26 and 55 kDa. The identification of these proteins by N-terminal amino acid sequencing showed both to be involved in the defense against oxidative stress. The 55-kDa polypeptide was identical to C. jejuni catalase (KatA), whereas the N terminus of the 26-kDa polypeptide was homologous to a 26-kDa Helicobacter pylori protein. The gene encoding the C. jejuni 26-kDa protein was cloned, and the encoded protein showed significant homology to the small subunit of alkyl hydroperoxide reductase (AhpC). The upstream region of ahpC encoded a divergent ferredoxin (fdxA) homolog, whereas downstream sequences contained flhB and motB homologs, which are involved in flagellar motility. There was no evidence for an adjacent homolog of ahpF, encoding the large subunit of alkyl hydroperoxide reductase. Reporter gene studies showed that iron regulation of ahpC and katA is achieved at the transcriptional level. Insertional mutagenesis of the ahpC gene resulted in an increased sensitivity to oxidative stresses caused by cumene hydroperoxide and exposure to atmospheric oxygen, while resistance to hydrogen peroxide was not affected. The C. jejuni AhpC protein is an important determinant of the ability of this microaerophilic pathogen to survive oxidative and aerobic stress.


Subject(s)
Campylobacter jejuni/enzymology , Gene Expression Regulation, Bacterial/drug effects , Iron/pharmacokinetics , Oxidative Stress/physiology , Oxidoreductases/metabolism , Peroxidases , Aerobiosis , Bacterial Proteins/genetics , Base Sequence , Campylobacter jejuni/genetics , Campylobacter jejuni/growth & development , DNA Mutational Analysis , Gene Expression Regulation, Enzymologic/drug effects , Genes, Reporter , Molecular Sequence Data , Peptides/genetics , Peroxiredoxins , Sequence Homology, Amino Acid , Transcription, Genetic/drug effects
9.
Pediatr Hematol Oncol ; 16(4): 341-6, 1999.
Article in English | MEDLINE | ID: mdl-10407871

ABSTRACT

Non-Hodgkin's lymphomas (NHL) were often erroneously diagnosed as other malignancies and treated accordingly. In this study cisplatin combined with vincristine, cyclophosphamide, and Adriamycin was used incidentally as a front-line treatment in seven children with NHL, because the initial histologic diagnosis was that of a sarcoma. After reevaluation three patients had Ki-1 anaplastic large cell lymphoma of T-cell origin, two abdominal B-cell diffuse high-grade NHL, one mediastinal diffuse large B-cell lymphoma, and one B-cell lymphoma in the stomach. They received at least two courses of cisplatin combined regimen and continued with other protocols for NHL. All patients showed an extremely good response from the first course of therapy and the masses vanished completely. They were followed up for a mean time of 29.5 months and are all in complete remission. The data indicate that cisplatin is active against NHL and might be a promising alternative front-line therapy.


Subject(s)
Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Cisplatin/administration & dosage , Lymphoma, Non-Hodgkin/drug therapy , Adolescent , Bone Marrow/drug effects , Child , Child, Preschool , Cisplatin/adverse effects , Female , Humans , Male
10.
Res Microbiol ; 149(2): 95-107, 1998 Feb.
Article in English | MEDLINE | ID: mdl-9766213

ABSTRACT

In order to study a 19-kDa protein (p19) of Campylobacter jejuni, we purified this protein to homogeneity from C. jejuni strain 81,176 by anion exchange chromatography. The molecular weight of the native protein is 19,000 daltons. P19 was found to be acidic with an isoelectric point of 4.8 and was located in the periplasmic space of the bacteria. The 20 N-terminal amino acids were sequenced and no significant similarities with known proteins were shown. A monoclonal antibody showed that p19 is conserved in the 2 species C. jejuni and C. coli. Analysis of sera from 23 patients with a Campylobacter-related infection indicated that p19 is not immunogenic during natural infection in man. The gene encoding p19 was cloned and no strong homologies with known sequences were identified. The preparation of a knockout mutant in p19 will enable the investigation of the function of this cell wall component of Campylobacter.


Subject(s)
Bacterial Proteins/isolation & purification , Campylobacter Infections/immunology , Campylobacter coli/chemistry , Campylobacter jejuni/chemistry , Membrane Proteins/isolation & purification , Periplasm/chemistry , Periplasmic Proteins , Amino Acid Sequence , Animals , Antibodies, Monoclonal , Bacterial Proteins/chemistry , Bacterial Proteins/genetics , Base Sequence , Blotting, Western , Campylobacter coli/genetics , Campylobacter jejuni/genetics , Child , Chromatography, High Pressure Liquid , Cloning, Molecular , DNA, Bacterial/chemistry , Electrophoresis, Gel, Two-Dimensional , Electrophoresis, Polyacrylamide Gel , Female , Humans , Isoelectric Point , Membrane Proteins/chemistry , Membrane Proteins/genetics , Mice , Mice, Inbred BALB C , Molecular Sequence Data , Molecular Weight , Sequence Analysis, DNA , Sequence Homology, Amino Acid
11.
Pediatr Hematol Oncol ; 15(3): 265-9, 1998.
Article in English | MEDLINE | ID: mdl-9615326

ABSTRACT

Anaplastic large cell lymphoma is a rare malignancy in childhood. We describe the case of a 6-year-old boy with Ki-1 lymphoma of the thymus who presented with an endocardiac mass. The first histologic analysis suggested a high-grade undifferentiated sarcoma, but reevaluation and immunohistochemistry confirmed the CD30+ lymphoid derivation of the process. The patient was given chemotherapy and 24 months later he remains in complete remission. It is noted that echocardiography was repeated many times to detect heart lesion.


Subject(s)
Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Heart Neoplasms/drug therapy , Heart Neoplasms/physiopathology , Heart Neoplasms/surgery , Lymphoma, Large-Cell, Anaplastic/drug therapy , Lymphoma, Large-Cell, Anaplastic/physiopathology , Lymphoma, Large-Cell, Anaplastic/surgery , Child , Humans , Male , Remission Induction
12.
FEMS Microbiol Lett ; 152(2): 205-11, 1997 Jul 15.
Article in English | MEDLINE | ID: mdl-9231413

ABSTRACT

Although flagellar motility is essential for the colonisation of the stomach by Helicobacter pylori, little is known about the regulation of flagellar biosynthesis in this organism. We have identified a gene in H. pylori, designated fliI, whose deduced amino acid sequence revealed extensive homology with the FliI/LcrB/InvC family of proteins which energise the export of flagellar and other virulence factors in several bacterial species. An isogenic mutant of fliI was non-motile and synthesised reduced amounts of flagellin and hook protein subunits. The majority (> 99%) of mutant cells were completely aflagellate. These results suggest that FliI is a novel ATPase involved in flagellar export in H. pylori.


Subject(s)
Adenosine Triphosphatases/physiology , Bacterial Proteins/physiology , Flagella/metabolism , Helicobacter pylori/enzymology , Proteins/physiology , Proton-Translocating ATPases , Adenosine Triphosphatases/genetics , Amino Acid Sequence , Bacterial Proteins/analysis , Bacterial Proteins/genetics , Flagellin/analysis , Genes, Bacterial/genetics , Helicobacter pylori/genetics , Helicobacter pylori/ultrastructure , Molecular Sequence Data , Mutation , Proteins/genetics , Sequence Analysis, DNA , Sequence Homology, Amino Acid
13.
Microb Pathog ; 20(2): 87-100, 1996 Feb.
Article in English | MEDLINE | ID: mdl-8722097

ABSTRACT

In previous work, a factor which enhances the ability of cytidine 5'-monophospho-N-acetyl neuraminic acid (CMP-NANA) to sialylate gonococcal lipopolysaccharide (LPS) was liberated at 4 degrees C in diffusates from high M(r) fractions of blood cell sonicates. The diffusates also contained CMP-NANA and converted serum susceptible gonococci to resistance. The enhancer has now been separated from CMP-NANA and material absorbing at 260 nm by HPLC on mu Bondapak-10 NH2. Resistance inducing activity was found only in fractions containing CMP-NANA and recovery was poor (about 25%). However, addition of enhancer fractions to CMP-NANA substantially increased its resistance inducing activity. Blood cell sonicates dialysed at 18-20 degrees C released enhancer in diffusates. These were ultrafiltered (nominal cut off 3000 Da) and fractionated on Biogel P2 which removed saccharides and most material absorbing at 260 nm. Over 90% of a fraction which was enhancer-active in nanogram quantities was identified by nuclear magnetic resonance (NMR) spectroscopy and gas chromatography/mass spectometry (GC/MS) as lactic acid. A fraction with similar properties was obtained from a different batch of diffusate by fractionation on Dowex 1. Authentic lithium L-lactate in nanogram quantities enhanced LPS sialyation by CMP-NANA and increased its serum resistance inducing activity. These results have important implications for gonococcal pathogenicity.


Subject(s)
Blood Cells/chemistry , Cytidine Monophosphate N-Acetylneuraminic Acid/chemistry , Lactic Acid/isolation & purification , Lipopolysaccharides/metabolism , Chromatography, High Pressure Liquid , Cytidine Monophosphate N-Acetylneuraminic Acid/metabolism , Immunity, Innate , Lactic Acid/chemistry , Lactic Acid/metabolism , Neisseria gonorrhoeae/chemistry
14.
Microb Pathog ; 18(3): 187-95, 1995 Mar.
Article in English | MEDLINE | ID: mdl-7565013

ABSTRACT

A stable, sialyltransferase-deficient mutant of Neisseria gonorrhoeae strain F62 totally defective in CMP-NANA-dependent lipopolysaccharide (LPS) sialylation was isolated by insertion mutagenesis with transposon Tn 1545-delta 3 and screened for unlabelled colonies following incubation with CMP-14C-NANA. In contrast to the parental strain which became serum resistant on incubation with CMP-NANA or blood cell extracts, the mutant, JB1, remained serum sensitive. French press extracts of strain F62 catalysed LPS sialylation, but corresponding extracts of the mutant were inactive. Five LPS components were detected by SDS-PAGE in the parental strain. Five components of the same Mr were also found in the mutant. Three identical components were detected by Western blotting using MAb 3F11, which recognizes the Gal beta 1-4GlcNAc groups in the conserved LPS components of F62 which can be sialylated. The mutant, JB1, is therefore deficient in the sialyltransferase that is essential for both LPS sialylation and conversion of serum-sensitive gonococci to serum resistance by either CMP-NANA or blood cell extracts. No evidence was obtained for an LPS sialylation pathway by blood cell extracts that is independent of CMP-NANA.


Subject(s)
Blood Bactericidal Activity , Neisseria gonorrhoeae/genetics , Antibodies, Monoclonal , Carbohydrate Sequence , Cytidine Monophosphate N-Acetylneuraminic Acid/pharmacology , DNA Transposable Elements , Humans , In Vitro Techniques , Lipopolysaccharides/chemistry , Molecular Sequence Data , Mutagenesis, Insertional , Neisseria gonorrhoeae/drug effects , Neisseria gonorrhoeae/enzymology , Sialic Acids/chemistry , Sialyltransferases/genetics
15.
Microb Pathog ; 16(6): 413-21, 1994 Jun.
Article in English | MEDLINE | ID: mdl-7830528

ABSTRACT

Serum resistance of gonococci in most patients is due to sialylation of a Gal beta 1-4GlcNAc group on a conserved 4.5 kDa lipopolysaccharide (LPS) component by host cytidine 5'-monophospho-N-acetyl neuraminic acid (CMP-NANA) catalysed by a gonococcal sialyl transferase. This sialylation is enhanced by a low M(r) factor(s) which, like CMP-NANA, is released in diffusates from high M(r) fractions obtained from sonicates dialysed at 4 degrees C. Also, as shown here, this factor(s) is released when the sonicates are dialysed at 18-20 degrees C. The enhancement of sialylation, first demonstrated using enzymes in gonococcal extracts, has been shown to occur in live gonococci and hence probably to have a role in pathogenicity. Gonococci, emerging from lag phase and incubated for 2 h with CMP-14CNANA fixed up to 90% more radiolabel than controls when the second factor(s) was present; their LPS separated by SDS-PAGE contained more radiolabel than control samples and label was not detected in any other component. Fractions with enhancing activity absorbed maximally at about 260 nm but a mixture of UDP-galactose (UDP-Gal), UDP-N-Acetyl galactosamine (UDP-GalNAc), UDP-glucose (UDP-Glc) and UDP-N-Acetyl glucosamine (UDP-GlcNAc) showed no significant enhancing activity. The enhancing action of the low M(r) fractions was unaffected by incubation with beta-galactosidase.


Subject(s)
Cytidine Monophosphate N-Acetylneuraminic Acid/metabolism , Lipopolysaccharides/metabolism , Neisseria gonorrhoeae/metabolism , Blood Bactericidal Activity , Carbohydrate Sequence , Gonorrhea/blood , Gonorrhea/microbiology , Humans , In Vitro Techniques , Lipopolysaccharides/chemistry , Lipopolysaccharides/immunology , Molecular Sequence Data , Neisseria gonorrhoeae/drug effects , Neisseria gonorrhoeae/immunology , Sialic Acids/chemistry , Sialic Acids/metabolism , Uridine Diphosphate Sugars/pharmacology
16.
Microb Pathog ; 14(4): 329-35, 1993 Apr.
Article in English | MEDLINE | ID: mdl-8326856

ABSTRACT

In previous work, a low M(r) component from human blood which converts serum-sensitive gonococci to resistance was shown to be indistinguishable from cytidine 5'-monophospho-N-acetyl neuraminic acid (CMP-NANA) by seven criteria. However, the presence of CMP-NANA was not proved by physicochemical methods. Purified, high M(r) fractions from human blood cells, which confer serum resistance on gonococci and enhance the transfer of sialyl groups from CMP-NANA to lipopolysaccharide (LPS) by a sialyltransferase in gonococcal extracts, were rechromatographed on DEAE Sepharose CL-6B. Both activities co-eluted from the column but on dialysis were found in the diffusate. After desalting the diffusate with Sephadex G10, the presence of CMP-NANA was proved by mass spectrometry. This confirmed previous work and is the first unequivocal demonstration of CMP-NANA in constituents of human blood cells.


Subject(s)
Blood Cells/immunology , Cytidine Monophosphate N-Acetylneuraminic Acid/blood , Cell Fractionation , Cytidine Monophosphate N-Acetylneuraminic Acid/immunology , Diffusion , Humans , Mass Spectrometry , Molecular Weight , Neisseria gonorrhoeae/immunology
17.
Microb Pathog ; 12(6): 421-32, 1992 Jun.
Article in English | MEDLINE | ID: mdl-1522797

ABSTRACT

A high relative molecular mass (M(r)) component which confers serum resistance on gonococci has been purified about 300-fold from a dialysed sonicate of human blood cells. Serum resistance conferred by the high M(r) factor (RIF), like that induced by cytidine-5' monophospho-N acetyl neuraminic acid (CMP-NANA), decreased when gonococci were incubated with neuraminidase. Also, the resistance-inducing activities of both high M(r) RIF and CMP-NANA were inhibited by CMP and inactivated at pH 4.0. These activities were not additive but synergistic. Neuraminidase decreased the activity of high M(r) RIF but not CMP-NANA. In tests with 14C CMP-NANA and gonococcal lipopolysaccharide, no sialyltransferase activity was detected, even in highly active samples of high M(r) RIF under conditions in which low activities of rat liver sialyltransferase were readily detected. Conversely, rat liver sialyltransferase was neither active in the RIF assay nor able to enhance the RIF activity of CMP-NANA. Nevertheless, high M(r) RIF greatly enhanced the sialyltransferase activity of a gonococcal extract; this enhancement suggests an explanation for the synergism between CMP-NANA and high M(r) RIF in inducing serum resistance in gonococci.


Subject(s)
Blood Cells/immunology , Cytidine Monophosphate N-Acetylneuraminic Acid/immunology , Neisseria gonorrhoeae/immunology , Cytidine Monophosphate N-Acetylneuraminic Acid/metabolism , Humans , Immunity, Innate/immunology , Lipopolysaccharides/metabolism , Molecular Weight , Neuraminidase , Sialyltransferases/metabolism
18.
Pediatr Radiol ; 22(6): 465-6, 1992.
Article in English | MEDLINE | ID: mdl-1437378

ABSTRACT

Myelolipoma of the adrenal gland is a rare benign tumour which seldom produces symptoms unless it attains considerable size or hemorrhages into itself. Histologically the tumor is composed of varying proportions of fat and bone marrow elements. We present a case of a male child, with homozygous beta thalassemia and asymptomatic myelolipoma.


Subject(s)
Adrenal Gland Neoplasms/diagnosis , Lipoma/diagnosis , Adolescent , Humans , Infant, Newborn , Male
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