Biomedical subjects

J McFadden

Publications and source records attributed to J McFadden.

At least 19 recordsLinked to original sources

PubMed1995

Gene replacement by homologous recombination in Mycobacterium bovis BCG.

Gene replacement by homologous recombination is a powerful tool for fundamental studies of gene function, as well as allowing specific attenuation of pathogens, but has proved difficult to achieve for Mycobacterium tuberculosis. We have used a plasmid-based test system to demonstrate the occurrence of homologous recombination in the tuberculosis vaccine strain Mycobacterium bovis BCG, and we have successfully replaced a target gene in BCG by homologous recombination, using a shuttle plasmid. Specific inactivation of selected genes will facilitate study of virulence factors and drug resistance as well as allowing rational attenuation of M. tuberculosis for the production of new vaccines.

BCG Vaccine↗

PubMed1995

Mycobacterial granulomatous inflammation in the ulcerated caecum of indomethacin-treated rats.

We report mycobacterial granulomatous inflammation in the ulcerated caecum of rats that received indomethacin. Two groups of male rats were treated with dietary indomethacin 3 mg/kg/day or untreated diet for 3 weeks. Six out of 8 indomethacin treated rats showed both ulceration and inflammation of the caecal mucosa. Two of the rats showing caecal ulceration also showed distinct granulomatous inflammation of the caecal mucosa and acid-fast bacilli were identified within granulomata. None of the other indomethacin treated or control rats contained acid-fast bacilli within caecal tissue sections but they were present, in the same sections, within the lumen of most rats in both groups. Polymerase chain reaction analysis identified the mycobacterial 65 kDa GroEL gene within control and granulomatous caecal tissue. In a repeat of indomethacin administration to a third group of rats, culture of both non-granulomatous caecal tissue (containing histologically identified luminal acid-fast bacilli) and faecal samples for mycobacteria was negative.

Animals↗

PubMed1994

Varicella-associated acute supraglottitis.

Acute bacterial supraglottitis is a well-recognized pediatric entity. There are reports in the literature of numerous supraglottic infections not limited to the usual organism, Haemophilus influenzae type b. We present a case of acute supraglottitis that is associated with varicella infection.

Airway Obstruction↗

PubMed1994

Characterization of the virulence of Mycobacterium avium complex (MAC) isolates in mice.

The virulence of different isolates of MAC was studied in naturally susceptible BALB/c mice. In preliminary experiments, MAC bacteria forming smooth transparent colonies on solid media (SmT variants) were found to be virulent for BALB/c mice, causing progressive infection; smooth opaque (SmOp) were generally avirulent, being slowly eliminated from the infected organs; and rough (Rg) variants were either avirulent or as virulent as SmT variants. We chose to compare the virulence of different isolates of MAC of different origins, studying only the SmT morphotype. Strains of MAC isolated from naturally infected animals were those that most consistently caused progressive infections. AIDS patients-derived isolates were of intermediate virulence or devoid of virulence in mice. The environmental strains were eliminated from mice or did not proliferate. Strains of MAC isolated from individuals who were not infected by HIV varied in virulence from completely avirulent to highly virulent. There was no close correlation between virulence and restriction fragment length polymorphism (RFLP) type, although all highly virulent strains were of the A/I type. There was also no correlation between virulence analysed in vivo and the ability to grow in cultured macrophages.

Animals↗

PubMed1993

Construction and use of integrative vectors to express foreign genes in mycobacteria.

We have constructed a mycobacterial integrative vector by placing two copies of the insertion sequence IS900 flanking a kanamycin-resistance gene into a 'suicide' vector unable to replicate in mycobacteria. The Mycobacterium leprae gene encoding the M. leprae 18 kDa protein was cloned between the two copies of IS900 to provide expression signals. Constructs were introduced into Mycobacterium species smegmatis, vaccae and bovis BCG by electroporation and selection for kanamycin resistance. The expression of the 18 kDa gene was analysed by Western blotting. Integration of the vector into the M. smegmatis chromosome was analysed by Southern blotting. One to five copies of the vector were detected in each transformant. The SIV gag p27 gene and the foot-and-mouth disease virus VP1 140-160 epitope were successfully cloned into the 18 kDa gene and expression in M. smegmatis was obtained.

Amino Acid Sequence↗

PubMed1992

Polymerase chain reaction for diagnosis of meningococcal meningitis.

Meningococcal disease is normally suspected on clinical grounds but confirmed by isolation of Neisseria meningitidis from blood or cerebrospinal fluid (CSF), or by detection of gram-negative diplococci in CSF. After parenteral antibiotics are started the isolation rate of meningococci from blood cultures drops from 50% to less than 5% and the chances of CSF being positive by culture or microscopy are also reduced. We used the polymerase chain reaction (PCR) in a blinded study to detect meningococcal DNA in 54 CSF samples from patients with meningococcal disease or from controls. The PCR primers were specific for the meningococcal insertion sequence IS1106. The sensitivity and specificity of this PCR for diagnosis of meningococcal meningitis were both 91%. Sensitivity was not affected by prior antibiotic treatment. The IS1106 PCR is a rapid and sensitive test for confirmation of the diagnosis of meningococcal meningitis.

DNA, Bacterial↗

PubMed1992

Detection of mycobacterial DNA in sarcoidosis and tuberculosis with polymerase chain reaction.

The cause of sarcoidosis is unknown. However, the histological similarity between the disorder and tuberculosis suggests that mycobacteria might contribute to the pathogenesis of sarcoidosis. We have used the polymerase chain reaction (PCR) to detect mycobacterial DNA in clinical samples from patients with sarcoidosis. 104 patients were included in the study (62 referred for possible tuberculosis and 20 for possible sarcoidosis, and 22 control patients who had undergone bronchoscopy for other reasons). Bronchoalveolar lavage samples, bronchial washings, and tissue specimens (1 from each patient) underwent assay by PCR as well as bacteriological, histological, and cytological examination. We used two PCR reactions: in the first the complex-specific insertion sequence IS986/IS6110 was used to specifically detect DNA from Mycobacterium tuberculosis complex bacteria; in the second, conserved sequences of the mycobacterial groEL gene were used to detect DNA from mycobacteria other than M tuberculosis. The PCR was more sensitive than culture for diagnosis of tuberculosis. However, the false-positive PCR rate for M tuberculosis was 9%. M tuberculosis DNA was found in half the sarcoidosis patients, and non-tuberculosis mycobacterial DNA in a further 20%. The findings that a significant proportion of the sarcoidosis patients in this study have mycobacteria in their lungs and that most of these mycobacteria belong to M tuberculosis complex suggest an aetiological role for mycobacteria in sarcoidosis.

Base Sequence↗

PubMed1992

First International Conference on the Pathogenesis of Mycobacterial Infections: a summary.

Two hundred scientists representing 25 countries assembled for the First International Conference on the Pathogenesis of Mycobacterial Infections, which was held in Stockholm on 27-29 June 1990. A total of 40 speeches and 80 poster presentations covered nearly all of the recent progress made in the field of mycobacterial pathogenicity. The present report is intended as a brief summary of the research results presented during the conference and focuses on findings of broad scientific interest. However, the omission of other presented findings from this report does not constitute any judgment of their scientific quality.

Antigens, Bacterial↗

PubMed1992

Topically applied verapamil hydrochloride inhibits tuberculin-induced delayed-type hypersensitivity reactions in human skin.

Calcium channel antagonists have been reported to possess inhibitory effects on lymphocyte migration and activation in vitro, and on cell-mediated immune reactions in the skin of experimental animals. We have therefore studied the effects of topically applied 8% (w/v) verapamil hydrochloride in propylene glycol on tuberculin-induced delayed-type hypersensitivity reactions in the skin of normal human volunteers. There was significant inhibition of the tuberculin reactions by the verapamil preparation compared to vehicle controls, as determined by forearm skin fold thickness measurement and assessment of the density of mononuclear cell infiltrates in skin biopsies. The precise mechanism of action of verapamil hydrochloride remains unclear, but could include effects on T-cell migration and activation, on antigen-presenting cells, and/or on other cells. The potential for the use of topical calcium channel antagonist preparations in inflammatory skin diseases warrants further study.

Administration, Topical↗

PubMed1992

Unilateral subungual hyperkeratosis following a cerebrovascular incident in a patient with psoriasis.

The prevalence of nail abnormalities in psoriasis is reported to be between 50 and 90% of affected adults. These include pitting, nail discoloration, onycholysis and subungual hyperkeratosis. Although nail changes may be associated with neurological disease, there are no previous reports of psoriatic nail changes following cerebrovascular events. We report the first case to our knowledge of unilateral subungual hyperkeratosis in a psoriatic patient which developed following a cerebrovascular event.

Cerebrovascular Disorders↗

PubMed1992

Mycobacteria in Crohn's disease: DNA probes identify the wood pigeon strain of Mycobacterium avium and Mycobacterium paratuberculosis from human tissue.

Mycobacterium paratuberculosis is known to cause Johne's disease, a granulomatous ileitis in ruminants, and may be involved in some cases of Crohn's disease. Like M. paratuberculosis, the wood pigeon strain of Mycobacterium avium may also show mycobactin dependence on primary isolation that is attenuated on further subculturing. A wood pigeon strain, M. avium restriction fragment length polymorphism (RFLP) type A/I, is also capable of causing granulomatous ileitis in experimental animal models but is not known to cause disease in humans. M. avium RFLP type A is associated with disease in immunocompromised hosts. Three DNA probes, pMB22 and the two subclones pMB22/S4 and pMB/S12, were found to be capable of distinguishing among M. paratuberculosis, M. avium type A, and M. avium type A/I (wood pigeon strain) on the basis of RFLPs. These DNA probes were used to identify two mycobacterial isolates (M. paratuberculosis and M. avium type A/I, wood pigeon strain) derived from the intestinal tissues of two patients with Crohn's disease. In addition, the wood pigeon strain of M. avium was identified from a patient with ulcerative colitis, and M. avium RFLP type A was identified from a patient with colonic carcinoma. This is the first time that M. avium A/I (wood pigeon strain) is known to have been isolated from human tissue. There are too few isolates to speculate about the etiological significance of mycobacteria and inflammatory bowel disease, but it is reasonable to conjecture that M. paratuberculosis may be responsible for some cases of Crohn's disease and that the wood pigeon strain of M. avium may also be an inflammatory bowel disease pathogen in humans.

Crohn Disease↗

PubMed1991

Origins of AIDS.

Explore the source record for details and available documents.

Acquired Immunodeficiency Syndrome↗

PubMed1991

Cross-reactivity between streptococcal M surface antigen and human skin.

Psoriasis can be triggered by haemolytic streptococcal infections. As M protein is a major pathogenic surface antigen in these streptococci, the cross-reactivity between streptococcal M protein surface antigens and human epidermis was investigated. The conserved component common to the few M proteins investigated consists of an alpha-helical 'coiled-coil' configuration, similar to sub-units of human keratin. The amino acid sequence of protein M6, one of the M proteins that has been fully sequenced, was compared with that of 4721 ubiquitous peptides, by computer-assisted analysis using a protein-sequence data bank. Of all human proteins in the data bank 50-kDa keratin type 1 showed the closest homology with protein M6. Further evaluation revealed that this homology mainly involved the heptapeptide repeat patterns, which form the alpha-helical 'coiled-coil' structure, in both M6 and 50-kDa keratin. Cryostat sections of normal, involved and uninvolved psoriatic skin were studied for cross-reactivity with rabbit antisera raised against 10 different M proteins. All these antisera reacted with the stratum corneum of normal and psoriatic epidermis to a variable extent. Staining of keratinocyte cytoplasm was also observed, but this tended to be more prominent in lesional than in uninvolved and normal skin. Some of the M antisera also stained dendritic cells in the upper dermis as well as endothelium and smooth muscle. These cross-reactivities might be relevant to the pathogenesis of post-streptococcal psoriasis.

Amino Acid Sequence↗

PubMed1991

IS900-promoted stable integration of a foreign gene into mycobacteria.

An artificial mycobacterial transposon was constructed by placing two copies of the insertion sequence IS900 flanking a kanamycin resistance gene into a non-(mycobacterial) replicating vector. Constructs were introduced into mycobacteria by electroporation and transposition events conferring kanamycin resistance were selected. Integration of IS900 into several genomic sites was analysed by Southern blotting and shown to involve both simple insertions and cointegrate formation, suggesting that IS900 can transpose by a replicative mechanism. Kanamycin resistance of IS900-integrated transformants was shown to be stable in the absence of selection.

Blotting, Southern↗

PubMed1990

Identification of a UK outbreak strain of Neisseria meningitidis with a DNA probe.

To investigate the epidemiology of meningococcal disease, a specific DNA probe pUS210 (carrying insert DNA which is repeated in the meningococcal genome) was isolated. The ability of this probe to hydridise with multiple polymorphic fragments in Southern blots was exploited to examine genetic relations within strains. Two geographically distinct foci of prolonged meningococcal disease (Gloucester and Plymouth, UK) are due to a clonal population of virulent strains that are distinct from those found elsewhere in the UK.

DNA Probes↗

PubMed1990

Renal function after long-term low-dose cyclosporin for psoriasis.

Two groups of patients receiving cyclosporin A (CyA) for psoriasis had their renal function assessed by measurement of glomerular filtration rate (GFR) and effective renal plasma flow (ERPF). The first group comprised 13 patients who had taken low dose (average 3 mg/kg per day) CyA for an average period of 2.5 years. Seven of the 13 had a normal GFR of 108 (77-121) ml/min (median; range). In the other six patients the GFR was low at 63 (50-77) ml/min and CyA was discontinued for periods ranging from 3 to 17 weeks. The GFR rose in all six patients, to 79 (60-91) ml/min; this change was significant (P less than 0.05). The six patients restarted CyA because their psoriasis recurred and after a mean interval of 15 weeks the GFR had fallen in all six to 63 (46-80) ml/min (P less than 0.05) and the ERPF decreased from 339 (231-414) ml/min to 244 (177-321) ml/min (P less than 0.05). In the second group of 11 patients measurements were made prior to starting CyA and after taking CyA for a mean of 9 weeks. The GFR fell in eight out of 11 subjects, the GFR for the 11 patients being 117 (72-128) ml/min before taking CyA and 97 (51-122) ml/min after CyA (P less than 0.02). The ERPF was measured in nine of the 11 patients and fell in seven of the nine. The ERPF for the nine patients before CyA was 490 (296-642) ml/min and for the 11 patients after CyA was 410 (195-543) ml/min (P less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

PubMed1990

Nasal glioma. Presenting as neonatal respiratory distress. Definition of the tumor mass by MRI.

Nasal gliomas are benign congenital midline tumors with the potential for intracranial extension. They are most commonly seen in neonates and children but rarely in adults. The treatment of choice is surgical excision. Inadequate primary excision results in a 4 to 10 percent recurrence. Hence, a thorough preoperative evaluation is essential to delineate the exact site and extension of the tumor and to plan the appropriate surgical approach. Computerized tomographic (CT) scans are useful in visualizing bony defects, but are not well suited for soft tissue imaging. Magnetic resonance imaging (MRI) offers superior soft tissue contrast, without ionizing radiation. This is a report of a neonate with unexplained early respiratory distress. On day 5, a soft nasal mass became apparent. CT scans were inconclusive, so MRI scan was used to demonstrate intracranial extension. MRI is superior for imaging brain tissue, so it should be used preferentially to delineate intracranial extension and to help guide the surgical approach.

Diagnosis, Differential↗

PubMed1989

Long-term cyclosporin for psoriasis.

Thirteen patients with severe persistent psoriasis, intolerant of, or unresponsive to, other current treatments have been treated with cyclosporin (Cys) for periods varying from 12-25 (mean 18) months. The dose ranged from 1-4 mg/kg/day (mean 2.8 mg). There was a 72% reduction in the mean PASI score at 4 weeks, and at the end of the study, an 81% reduction. Adjuvant therapy with topical steroids was used in 11 of the 13 patients after the first 3 months of Cys treatment to persistent patches on an intermittent basis with beneficial effect. Six patients developed mild to moderate hypertension, in three this was controlled by a reduction in the dose of Cys, and in the other three by hypotensive agents. The mean serum creatinine rose from 72 to 90 microM/l during the study. Hypertrichosis occurred in seven of the 13 patients. Low dosage Cys is an effective treatment for clearing psoriasis and maintaining improvement on a long-term basis.

Adult↗