Novel protein pathways in development and progression of pulmonary sarcoidosis.

Pulmonary involvement occurs in up to 95% of sarcoidosis cases. In this pilot study, we examine lung compartment-specific protein expression to identify pathways linked to development and progression of pulmonary sarcoidosis. We characterized bronchoalveolar lavage (BAL) cells and fluid (BALF) proteins in recently diagnosed sarcoidosis cases. We identified 4,306 proteins in BAL cells, of which 272 proteins were differentially expressed in sarcoidosis compared to controls. These proteins map to novel pathways such as integrin-linked kinase and IL-8 signaling and previously implicated pathways in sarcoidosis, including phagosome maturation, clathrin-mediated endocytic signaling and redox balance. In the BALF, the differentially expressed proteins map to several pathways identified in the BAL cells. The differentially expressed BALF proteins also map to aryl hydrocarbon signaling, communication between innate and adaptive immune response, integrin, PTEN and phospholipase C signaling, serotonin and tryptophan metabolism, autophagy, and B cell receptor signaling. Additional pathways that were different between progressive and non-progressive sarcoidosis in the BALF included CD28 signaling and PFKFB4 signaling. Our studies demonstrate the power of contemporary proteomics to reveal novel mechanisms operational in sarcoidosis. Application of our workflows in well-phenotyped large cohorts maybe beneficial to identify biomarkers for diagnosis and prognosis and therapeutically tenable molecular mechanisms.

Adalimumab induced pulmonary sarcoid reaction.

Sarcoidosis is a multisystem granulomatous inflammatory disease of unknown etiology. There is evidence that Tumor Necrosis Factor alpha (TNF-α) antagonists are useful in the treatment of advanced or refractory disease. However, sarcoidosis-like reaction has been reported with TNF-α blockade in other inflammatory conditions. Here we report a case of sarcoid-like reaction in a patient with psoriatic arthritis shortly after initiation of adalimumab therapy. Stopping adalimumab and systemic anti-inflammatory therapy with corticosteroids resulted in resolution of pulmonary symptoms and chest radiographic findings. Though TNF-α plays a critical role in pathogenesis of sarcoidosis, the development of sarcoid reaction with TNF-α blockade is paradoxical and the mechanism of this response remains unknown. TNF-α induced sarcoid-reaction could involve multiple organs. Its development with one agent does not preclude therapy with other TNF-α blockers.

Campylobacter immunity and quantitative excretion rates in Thai children.

Campylobacter species were isolated from 61 (15%) of 416 Thai children < 5 years old with diarrhea. Although the baseline levels of Campylobacter-specific antibody increased with age, 80.3% of Campylobacter-infected children seroconverted compared with 12.9% of 45 Shigella-infected patients used as controls. The response to acute infection was greatest in the 6- to 12-month-old group. Nonseroconverters had higher initial IgG levels than did seroconverters (P = .001). Quantitative cultures showed a range of 1-8 log10 Campylobacter cfu/g of stool (median, 6.0 log10), and the seroconversion rate was highest in those with the highest Campylobacter excretion. Fecal Campylobacter excretion was inversely related to age (chi 2 for trend, P = .03). These studies indicate that endemic Campylobacter exposure frequently induces seroconversion in young children, whether Campylobacter is isolated as a single pathogen or one of multiple pathogens, and that fecal excretion of the organism is inversely related to the age-related immune response to infection.

Serum antibody responses to bacterial enteropathogens in Swedish travelers to south-east Asia.

The possibility that serological analysis may be more sensitive than bacteriological examinations of stool samples to detect enteric infections was evaluated in 80 Swedish travellers to South-East Asia. Serum and faecal specimens were collected before, during and after their travel. Serological analyses of pre-travel and any later serum specimen identified infection with enterotoxinogenic Escherichia coli (ETEC), Salmonella or Campylobacter jejuni in 28% of the travellers. The seroconversion rate was 72% in travellers excreting the homologous pathogen in their stool; all symptomatic cases and half of those who had an asymptomatic infection seroconverted. Bacteriological examinations of stool samples collected repeatedly during travel identified an enteropathogen in 20% of the travellers. However, the isolation rate decreased to 11%, when only a single routine faecal specimen was examined. Our findings suggest that serological analyses of pre- and post-travel specimens are sufficiently specific and may be at least as sensitive as conventional bacteriology to identify infections with bacterial enteropathogens in travellers. However, reliable serodiagnosis requires collection of pre-travel sera and might therefore only be useful in prospective studies of travellers' diarrhoea.

Human disease associated with "Campylobacter upsaliensis" (catalase-negative or weakly positive Campylobacter species) in the United States.

Catalase-negative or weakly positive (CNW) thermotolerant campylobacteria, first isolated from dogs in 1983, were recently recognized as a new species, "Campylobacter upsaliensis," but their association with human illness has not been established. Twelve human isolates received at the Centers for Disease Control between 1980 and 1986 were identified as CNW campylobacteria by biochemical tests, cellular fatty acid composition, and antimicrobial susceptibility patterns. Eleven CNW Campylobacter strains tested by DNA-DNA hybridization (hydroxyapatite method) were all highly related and were related to two "C. upsaliensis" strains at the species level (86% under optimal conditions and 76% under stringent conditions). Clinical information was obtained for 11 human isolates from three stool and eight blood specimens. They were isolated from four female and seven male patients 6.5 months to 83 years of age residing in 10 different states. The patients had a wide spectrum of illnesses. The stool isolates were obtained from two previously healthy persons during episodes of acute gastroenteritis and from one immunocompromised patient with persistent diarrhea and fever. The blood isolates were obtained from two infants with fever and respiratory symptoms; a young woman with a ruptured ectopic pregnancy; three elderly men with underlying chronic diseases; and two immunocompromised adults. In a bactericidal assay to assess sensitivity to serum, seven of eight blood isolates showed some resistance to killing by pooled normal human serum. These observations suggest that "C. upsaliensis" is a potential human pathogen associated with both gastroenteritis and bacteremia in normal hosts and with opportunistic infection in immunocompromised individuals.

Persistent Campylobacter jejuni infections in patients infected with the human immunodeficiency virus (HIV).

We identified Campylobacter jejuni infections in four patients infected with the human immunodeficiency virus (HIV); three had persistent and severe C. jejuni infections. Multiple isolates obtained from each patient had the same biochemical and serotypic characteristics, indicating recurrent infection rather than reinfection with unrelated strains. Serum antibody responses to C. jejuni group antigens by enzyme-linked immunosorbent assay were markedly impaired in the three patients with persistent infection compared with forty-two immunocompetent C. jejuni-infected controls and with the HIV-infected patient who readily cleared the organism. One patient was bacteremic; his blood isolate was killed by normal serum but was resistant to his own serum, whereas a simultaneous stool isolate of a different serotype was sensitive. Failure of two patients to eradicate the organism and long-term administration of erythromycin therapy led to the in-vivo development of resistance to this antibiotic, which is most frequently used to treat C. jejuni infections.

Pneumobilia as the sole radiographic finding in emphysematous cholecystitis.

Emphysematous cholecystitis is an uncommon variant of acute cholecystitis. Pneumobilia with air in the biliary radicles is rarely seen in the disease and has been previously described in only four cases which the author could find in the English literature. Herein, we report a fifth case and a brief review of the literature.

The active site structure of Na+- and K+-stimulated ATPase. Location of a specific fluorescein isothiocyanate reactive site.

Fluorescein 5'-isothiocyanate (FITC) has been shown to specifically inactivate the Na+- and K+-stimulated adenosine triphosphatase ((Na,K)-ATPase) at low concentrations (Karlish, S. J. D. (1979) Na+,K+ATPase Structure and Kinetics 115-128). The site of modification of purified dog kidney (Na,K)-ATPase by FITC has been investigated by enzymatic cleavage and fluorescence resonance energy transfer. The binding of FITC, which occurs at a stoichiometry of approximately one site per ATP binding site, causes an ATP-protectable inactivation of ATPase activity suggesting that it is reacting at the ATP hydrolysis site. The FITC reaction site apparently is located near the center of the COOH-terminal 77,000-dalton peptide fragment obtained by chymotryptic cleavage of the alpha subunit. Addition of ouabain to the native enzyme in the presence of chymotrypsin enhances cleavage at this site and releases the fluorescein moiety from the membrane. It is further shown that the distance from the FITC reaction site to the ouabain binding site, as judged by fluorescence resonance energy transfer from anthroyl ouabain to FITC, is approximately 74 A. These results demonstrate that ouabain inhibits the (Na,K)-ATPase by causing a protein conformational change which extends an unusually large distance across the membrane.