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Biomedical subjects

R J Fallon

Publications and source records attributed to R J Fallon.

At least 55 records · Page 3Linked to original sources
PubMed1991

Seasonal variation of Legionnaires' disease in Scotland.

A summer and autumn peak in incidence is a characteristic epidemiological feature of Legionnaires' disease, a fact which may help in diagnosis. Previous reports from Scotland have reported a winter peak. This study examined the seasonal variation in Legionnaires' disease in Scotland in each year and by category of diagnosis. A characteristic summer/autumn peak was seen for travel-related infection. For non-travel infection, there was an autumnal peak between 1978 and 1982 but an early winter peak between 1983 and 1986. Overall, the pattern could be described as a summer/autumn plateau reaching a peak in early winter. This pattern, seen in men and women, was clearest in the Greater Glasgow area. Nosocomial cases were clustered in the months of October, November and December. In non-travel cases the classical seasonal pattern was not observed and clinicians cannot rely upon this epidemiological feature for diagnosis. However, the relative infrequency of the disease between January and May was a consistent and potentially useful feature. Most reports on seasonal variation are based on aggregated data on travel and non-travel cases. The present observations question the hypotheses developed to explain the seasonal pattern and call for further studies.

Cross Infection↗
PubMed1991

Variation in time and space of non-outbreak Legionnaires' disease in Scotland.

The main aim of this study was to measure and explain geographic variations in the incidence of Legionnaires' disease in Scotland, particularly to help understand the source of non-outbreak infection. Between 1978 and 1986 the overall mean annual incidence rate was 7.9 per million (range 3.1-20.2), and for non-outbreak, non-travel cases it was 5.6. There were geographical variations by health board, by city and within cities, e.g. the mean annual incidence rate per million for non-travel, non-outbreak disease was 1.2 in Tayside Health Board, 3.7 in Lanarkshire, 5.6 in Lothian and 14.4 in Greater Glasgow. In Greater Glasgow Health Board non-travel cases lived in and around the city centre and in some postcode sectors there, the mean annual incidence rate exceeded 100. Travel-related cases lived in peripheral areas. These variations could not be explained by differences in access to and use of diagnostic services, surveillance, or host susceptibility (as reflected by socioeconomic status and frequency of other respiratory disease). The explanation probably lay in environmental factors, though differences in agent virulence were not excluded. The two main conclusions are, that non-outbreak cases were not truly sporadic, and that the space-time variations in incidence support the hypothesis that cooling towers were an important source of infection for non-travel, non-outbreak cases. If so such infection is potentially preventable.

Adolescent↗
PubMed1990

Endocytosis and degradation of alpha 1-antitrypsin-protease complexes is mediated by the serpin-enzyme complex (SEC) receptor.

Alpha 1-Antitrypsin (alpha 1-AT) is similar to other members of the serine protease inhibitor (serpin) supergene family in that it undergoes structural rearrangement during the formation of a covalently stabilized inhibitory complex with its cognate enzyme, neutrophil elastase. We have recently demonstrated an abundant, high-affinity cell surface receptor on human hepatoma cells and human mononuclear phagocytes which recognizes a conformation-specific domain of the alpha 1-AT-elastase complex as well as of other serpin-enzyme complexes (Perlmutter, D. H., Glover, G. I., Rivetna, M., Schasteen, C. S., and Fallon, R. J. (1990) Proc. Natl. Acad. Sci. U.S.A. 87, 3753-3757). Binding to this serpin-enzyme complex (SEC) receptor activates a signal transduction pathway for increased expression of the alpha 1-AT gene and may be responsible for clearance of serpin-enzyme complexes. In this study, we show that there is time-dependent and saturable internalization of alpha 1-AT-elastase and alpha 1-AT-trypsin complexes in human hepatoma HepG2 cells. Internalization is mediated by the SEC receptor as defined by inhibition by synthetic peptides corresponding to residues 359-374 of alpha 1-AT. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of intracellular radioactivity demonstrated that intact 75- and 66-kDa alpha 1-AT-trypsin complexes were internalized. Kinetic analysis of internalization at 37 degrees C showed that a single cohort of 125I-alpha 1-AT-trypsin complexes, prebound to cells at 4 degrees C, disappeared from the cell surface and accumulated intracellularly within 5-15 min at 37 degrees C. The intracellular concentration of radiolabeled complexes then decreased rapidly coincident with appearance of acid-soluble degradation products in the extracellular culture fluid. Intracellular degradation was inhibited by internalization at 18 degrees C or by internalization at 37 degrees C in the presence of weak bases ammonium chloride, primaquine, and chloroquine, indicating that degradation is lysosomal. These results indicate that in addition to its role in signal transduction the SEC receptor participates in internalization and delivery of alpha 1-AT-protease complexes to lysosome for degradation.

Carcinoma, Squamous Cell↗
PubMed1990

Staurosporine inhibits a tyrosine protein kinase in human hepatoma cell membranes.

Membranes from the human hepatoma cell line HepG2 mediate the phosphorylation on tyrosine of the asialoglycoprotein receptor. Manganese was the preferred divalent for phosphorylation although magnesium was effective at an 8-fold higher concentration. Calcium was ineffective at promoting phosphorylation and zinc was inhibitory. The protein kinase inhibitor staurosporine blocked asialoglycoprotein receptor phosphorylation on tyrosine in nanomolar concentrations (IC50 = 70 nM). In contrast another protein kinase C inhibitor, H7, was not inhibitory, suggesting that the effect of staurosporine was not mediated by protein kinase C inhibition. Concentrations of staurosporine that inhibit receptor phosphorylation by greater than 90% did not inhibit the phosphorylation of other protein substrates identified on SDS-polyacrylamide gels. These data suggest that staurosporine selectively and directly inhibits a membrane-associated tyrosine protein kinase.

1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine↗
PubMed1990

Tyrosine phosphorylation of the asialoglycoprotein receptor.

The asialoglycoprotein (ASGP) receptor undergoes constitutive endocytosis through the coated pit/coated vesicle pathway in hepatocytes. Studies on HepG2 cells have shown that the receptor is phosphorylated at serine under control conditions and following protein kinase C stimulation. This study examined whether the ASGP receptor could also serve as a substrate for a tyrosine kinase in HepG2 cells. 32P labeling was performed in membrane preparations, in permeabilized cells at 4 degrees C, and in intact cells at 37 degrees C. The phosphorylated ASGP receptor was isolated by immunoprecipitation, hydrolyzed in 6 N HCl at 110 degrees C, and analyzed by two-dimensional high voltage electrophoresis. The receptor isolated from a membrane preparation incubated in vitro with [gamma-32P]ATP incorporated radiolabel predominantly (greater than 90%) into phosphotyrosine. ASGP receptor phosphorylation at both tyrosine and serine was detected in intact cells incubated with phosphatase inhibitors for 60 min at 37 degrees C. The presence of both phenylarsine oxide (20 microM) and sodium orthovanadate (200 microM) was required for tyrosine phosphorylation. Use of these inhibitors together resulted in a 16.4-fold increase in phosphorylation of the immunoprecipitated ASGP receptor, whereas phosphorylation of total HepG2 membrane proteins was not significantly augmented by this procedure. Selective proteolytic digestion of ASGP receptors in isolated vesicles demonstrated that the phosphorylation site identified in these studies is located at tyrosine 5 in the cytoplasmic tail.

Asialoglycoprotein Receptor↗
PubMed1990

A cumulative review of studies on travellers, their experience of illness and the implications of these findings.

A cumulative review of illness experienced by 13,816 travellers returning to Scotland since 1977, shows an overall attack rate of 36%. Alimentary complaints predominated; 18% of travellers had these alone and a further 10% had other symptoms as well as their gastro-intestinal disorder. Higher attack rates were noted in those taking package holidays. Inexperience of travel, smoking, more southerly travel and younger age (particularly those between 20- and 29-years-old) were other contributing factors. A similar pattern emerged from a I year study of hospital in-patients with travel related admissions. Serological studies of 470 travellers showed that 20% had incomplete immunity to poliomyelitis; 25% of those tested (312 travellers) had serological evidence of typhoid immunisation, I.9% (of 760 travellers) had antibodies to Legionella pneumophila, 64% (5II travellers tested) had antibodies to hepatitis A, 87% (288 tested) had adequate levels of tetanus antitoxin but only 40% of the 225 travellers tested had adequate levels of diphtheria antitoxin. Amongst a subgroup of 645 travellers the travel agent was the most frequently consulted source of pre-travel health advice. This carries particular significance for the dissemination of relevant advice in view of the inadequacies found from study of the health information in travel brochures. These findings, viewed against the perspective of the continuing growth in international travel, means that travellers, the medical profession, the travel trade, health educators, global health agencies and health authorities in those countries accepting and encouraging tourists, will be required to recognise the health implications of further tourism development if this problem of illness associated with travel is to be brought under control.

Adolescent↗
PubMed1990

Secretor status, smoking and carriage of Neisseria meningitidis.

A survey of ABO blood groups, secretor status and smoking habits among 389 students and staff of a school in which there was an outbreak of meningococcal disease found no difference in the distribution of the ABO blood groups but a significantly higher proportion of non-secretors (37.6%) in the population examined compared with that reported for previous surveys of the neighbouring population in Glasgow (26.2%) (P less than 0.0005). There was also a significantly higher proportion of non-secretors among carriers of meningococci (47%) compared with non-carriers (32%). Increased carriage of meningococci among non-secretors might contribute to the increased susceptibility of individuals with this genetic characteristic to meningococcal disease observed in previous studies. Although passive exposure to cigarette smoke has been associated with meningococcal disease, there was no association between passive smoking and carriage. There was, however, a significant association between active smoking and carriage.

ABO Blood-Group System↗
PubMed1990

Identification of a serpin-enzyme complex receptor on human hepatoma cells and human monocytes.

Formation of the covalently stabilized complex of alpha 1-antitrypsin (alpha 1-AT) with neutrophil elastase, the archetype of serine proteinase inhibitor serpin-enzyme complexes, is associated with structural rearrangement of the alpha 1-AT molecule and hydrolysis of a reactive-site peptide bond. An approximately 4-kDa carboxyl-terminal cleavage fragment is generated. alpha 1-AT-elastase complexes are biologically active, possessing chemotactic activity and mediating increases in expression of the alpha 1-AT gene in human monocytes and macrophages. This suggested that structural rearrangement of the alpha 1-AT molecule, during formation of a complex with elastase, exposes a domain that is recognized by a specific cell surface receptor or receptors. To test this hypothesis, the known three-dimensional structure of alpha 1-AT and comparisons of the primary structures of the serpins were used to select a potentially exteriorly exposed and highly conserved region in the complexed form of alpha 1-AT as a candidate ligand (carboxyl-terminal fragment, amino acids 359-374). We show here that synthetic peptides based on the sequence of this region bind specifically and saturably to human hepatoma cells and human monocytes (Kd = 4.0 X 10(-8) M, 4.5 X 10(5) plasma membrane receptors per cell) and mediate increases in synthesis of alpha 1-AT. Binding of peptide 105Y (Ser-Ile-Pro-Pro-Glu-Val-Lys-Phe-Asn-Lys-Pro-Phe-Val-Tyr-Leu-Ile) is blocked by alpha 1-AT-elastase complexes, antithrombin III (AT III)-thrombin complexes, alpha 1-antichymotrypsin (alpha 1-ACT)-cathepsin G complexes, and, to a lesser extent, complement component C1 inhibitor-C1s complexes, but not by the corresponding native proteins. Binding of peptide 105Y is also blocked by peptides with sequence corresponding to carboxy-terminal fragments of the serpins AT III and alpha 1-ACT, but not by peptides having the sequence of the extreme amino terminus of alpha 1-AT. The results also show that peptide 105Y inhibits binding of 125I-labeled alpha 1-AT-elastase complexes. Thus, these studies demonstrate an abundant, relatively high-affinity cell surface receptor which recognizes serpin-enzyme complexes (SEC receptor). This receptor is capable of modulating the production of at least one of the serpins, alpha 1-AT. Since the ligand specificity is similar to that previously described for in vivo clearance of serpin-enzyme complexes, the SEC receptor may also be involved in the clearance of certain serpin-enzyme complexes.

Amino Acid Sequence↗
PubMed1990

Microbiological investigations into an outbreak of Pontiac fever due to Legionella micdadei associated with use of a whirlpool.

In the investigation of a large outbreak of non-pneumonic legionellosis at a leisure complex in Lochgoilhead, Scotland all direct cultures of environmental samples were initially negative for legionellae. Legionellae readily infect appropriate protozoa under suitable conditions, and following immunofluorescence to select specimens for special study, Legionella micdadei was isolated from whirlpool water via co-cultivation with Acanthamoeba polyphaga. L micdadei was also isolated, along with host amoebae, from the whirlpool filter. The use of amoebae in the isolation of legionellae from environmental (and other) sources can be of great value, especially if specimens shown by indirect immunofluorescence to contain legionellae fail to yield legionellae on routine culture.

Acanthamoeba↗
PubMed1989

A cytokine-selective defect in interleukin-1 beta-mediated acute phase gene expression in a subclone of the human hepatoma cell line (HEPG2).

Several well-differentiated human hepatoma cell lines (HepG2, Hep3B) have been used to identify factors which regulate hepatic gene expression during the host response to inflammation/tissue injury (acute phase response). Studies in these cell lines, as well as in primary cultures of rat, rabbit, and mouse hepatocytes, have demonstrated that interleukin-1 beta (IL-1 beta), tumor necrosis factor (TNF-alpha), and interferon-beta 2 (IFN-beta 2) each mediate changes in expression of several hepatic acute phase genes. In this study we identify a subclone of the HepG2 cell line in which there is a selective defect in IL-1 beta-mediated acute phase gene expression. Recombinant human IL-1 beta mediates an increase in synthesis of the positive acute phase complement protein factor B and a decrease in synthesis of negative acute phase protein albumin in the parent uncloned HepG2 cell line (HG2Y), but not in the subclone HG2N. Recombinant human IFN-beta 2 and TNF-alpha, however, regulate acute phase protein synthesis in the subclone HG2N; i.e. IFN-beta 2 and TNF-alpha increase synthesis of factor B and decrease synthesis of albumin in both HG2Y and HG2N cells. Equilibrium binding analysis with 125I-rIL-1 beta at 4 degrees C showed that both HG2N and HG2Y cells bind IL-1 beta specifically and saturably. HG2N and HG2Y possess 3.8 and 4.0 x 10(3) plasma membrane receptors/cell with affinities of 0.96 and 1.07 x 10(-9) M, respectively. Thus, the defect in this subclone of the HepG2 cell line is likely to involve the signal transduction pathway for the biological activity of IL-1 beta and will be useful in elucidation of this signal transduction pathway.

Acute-Phase Proteins↗
PubMed1989

Lochgoilhead fever: outbreak of non-pneumonic legionellosis due to Legionella micdadei.

Analysis of case histories from 187 people who had visited a hotel and leisure complex in Lochgoilhead, a village on the west coast of Scotland, indicated that 170 had had an acute illness characterised by headache, fatigue, arthralgia, myalgia, cough, and breathlessness. These symptoms were consistent with Pontiac fever-like illness. Legionella micdadei was isolated from the leisure complex whirlpool spa at the time that 60 of 72 individuals with symptoms seroconverted to L micdadei antigen. This outbreak is thought to be the first of a Pontiac fever-like illness ascribed to L micdadei and the first large-scale outbreak of its kind to have occurred outside North America. Whirlpool spas can be a major reservoir of legionella organisms; they must therefore be properly maintained and operated to prevent outbreaks of infection.

Acute Disease↗
PubMed1988

Asialoglycoprotein receptor phosphorylation and receptor-mediated endocytosis in hepatoma cells. Effect of phorbol esters.

The asialoglycoprotein (ASGP) receptor on Hep G2 cells undergoes constitutive recycling and ligand endocytosis in the presence of phorbol dibutyrate, at a 50% reduced rate relative to control cells (Fallon, R. J., and Schwartz, A. L. (1986) J. Biol. Chem. 261, 15081-15089). The relevance of receptor phosphorylation to these events was investigated by selective immunoprecipitation of surface receptors with polyclonal anti-human ASGP antiserum and pulse-chase labeling with [32P]orthophosphate to identify subcellular locations of initial receptor phosphorylation events as well as the eventual fate of phosphorylated receptor during recycling. The surface immunoprecipitation method recovers greater than 95% of surface ASGP receptors and only 5% or less of intracellular (brief[35S]methionine pulse-labeled) receptors. With this assay we detected low levels of ASGP receptor phosphorylation at the cell surface in control cells (0.1 mol of P/mol of R) which were rapidly (less than 1 min) stimulated 20-fold by 400 nM phorbol dibutyrate addition (1.7 mol of P/mol of R). Staurosporine, a protein kinase C inhibitor, blocks this stimulation by phorbol. Receptor phosphorylation at early time points in the presence of phorbol esters was restricted to the plasma membrane. Subsequent chase in the presence of excess unlabeled phosphate and phorbol esters lowered [32P] ATPi specific activity by 68% at 1 h. Surface immunoprecipitation during this chase period showed the phosphorylated ASGP receptors were rapidly lost from the cell surface (t1/2 = 20 min). In contrast, examination of intracellular receptor during the pulse-chase experiment in phorbol dibutyrate-treated cells showed the presence of phosphorylated pool(s) of ASGP receptors which were detectable for 6 h of chase. Since no labeled receptor can be detected at the cell surface at this time, the described intracellular phosphorylated receptors are in a non-recycling pool.

Adenosine Triphosphate↗