Cell surface engineering by a modified Staudinger reaction.

Selective chemical reactions enacted within a cellular environment can be powerful tools for elucidating biological processes or engineering novel interactions. A chemical transformation that permits the selective formation of covalent adducts among richly functionalized biopolymers within a cellular context is presented. A ligation modeled after the Staudinger reaction forms an amide bond by coupling of an azide and a specifically engineered triarylphosphine. Both reactive partners are abiotic and chemically orthogonal to native cellular components. Azides installed within cell surface glycoconjugates by metabolism of a synthetic azidosugar were reacted with a biotinylated triarylphosphine to produce stable cell-surface adducts. The tremendous selectivity of the transformation should permit its execution within a cell's interior, offering new possibilities for probing intracellular interactions.

Treatment of chlamydial pneumonia of infancy.

Infants with untreated chlamydial pneumonia shed Chlamydia trachomatis and are symptomatic for may weeks. We used sulfisoxazole, 150 mg/kg/day, or erythromycin ethyl succinate, 40 mg/kg/day, for approximately 14 days to treat 32 patients with chlamydial pneumonia of infancy, and observed them for nasopharyngeal shedding of C trachomatis and changing clinical status. All infants stopped shedding chlamydiae soon after treatment was started. After treatment, three of the 25 infants tested again became culture positive (but did not have clinical relapse). All infants improved clinically. In 24 (83%) of 29 infants, where the onset of improvement could be times, improvement began within seven days of starting treatment. Progression to complete recovery was observed in 27 of 28 infants examined between two weeks and two months of treatment completion. Neither the existence of concomitant viral infection nor the duration of illness or hospitalization before starting treatment influenced the interval between initiation of treatment and onset of clinical improvement. While these observations do not prove, they are at least compatible with the hypotheses that C trachomatis plays a central etiologic role in this illness and that termination of chlamydial infection is beneficial clinically. Pending the availibility of data from controlled studies, we believe that either of the treatment programs outlined warrant consideration in the clinical management of patients with chlamydial pneumonia of infancy.

Clinical characteristics of the afebrile pneumonia associated with Chlamydia trachomatis infection in infants less than 6 months of age.

Respiratory tract colonization with Chlamydia trachomatis commonly occurs in natally acquired chlamydial infection and is sometimes associated with a chronic, afebrile pneumonia that has relatively distinctive clinical characteristics. To further define the frequency and clinical characteristics of lower respiratory tract disease associated with C trachomatis, we grouped 56 infants aged less than 6 months with afebrile pneumonia according to nasopharyngeal shedding of Chlamydia and viruses and compared their illnesses. Forty-one (73%) were positive for C trachomatis (23 had C trachomatis only, while 18 had C trachomatis plus a virus [cytomegalovirus, respiratory synctial virus, adenovirus, rhinovirus, or enterovirus]), and 15 were C trachomatis negative (nine had a virus only, and six had neither C trachomatis nor virus). The 41 infants with C trachomatis alone or C trachomatis plus a virus were similar clinically and differed significantly from other infants in several ways: (1) onset of symptoms before 8 weeks of age; (2) gradually worsening symptoms; (3) presentation for care at 4 to 11 weeks of age; (4) presence of conjunctivitis and ear abnormalities; (5) chest roentgenograms showing bilateral, symmetrical, interstitial infiltrates and hyperexpansion; (6) peripheral blood eosinophils greater than or equal to 300/cu mm; and (7) elevated values for serum immunoglobulins M, G, and A. Pediatrics 63:192--197, 1979, Chlamydia trachomatis, pneumonia, afebrile pneumonia, interstitial pneumonia.

A "traceless" Staudinger ligation for the chemoselective synthesis of amide bonds.

[reaction: see text] Here we report a novel modification of our previously reported "Staudinger ligation" that generates an amide bond from an azide and a specifically functionalized phosphine. This method for the selective formation of an amide bond, which does not require the orthogonal protection of distal functional groups, should find general utility in synthetic and biological chemistry.

Diagnosis of measles by fluorescent antibody and culture of nasopharyngeal secretions.

An indirect fluorescent antibody test (IFA) was evaluated using commercial mouse anti-measles monoclonal antibody and FITC-labeled goat anti-mouse immunoglobulin. For measles isolation, specimens were inoculated into Rhesus monkey kidney (RMK) cells and, when available, CV-1 cells. 381 specimens were tested by IFA and 408 specimens were cultured from patients suspected of having measles. For the 381 specimens tested by both methods, IFA and culture were positive for 31%, culture alone for 14%, IFA alone for 15%, and both negative for 40%. This study indicates that both IFA and culture are required for maximum measles virus detection. Of the positive specimens, 48% were detected either by IFA only (24%) or culture only (24%). IFA was positive in 69% of the culture-positive specimens and therefore, provided rapid diagnosis for many patients.

Diagnostic virology in clinical practice.

Viral illnesses are among the most common clinical problems encountered in medicine. Some have important epidemiologic implications and some are preventable by immunization, improved sanitation or other means. Recently, drugs have been developed which show promise for specific prophylaxis or treatment. It is thus increasingly important that physicians in practice be acquainted with methods of identification of common viral illnesses so they can intervene with appropriate counsel, preventive measures or treatment for their own patients. Physicians should also consider the larger epidemiologic implications of many viral illnesses and cooperate with local, state and national efforts to accurately determine the number of patients involved and the severity of their illnesses. It is only with this sort of information that an estimate of medical, social and economic impact of the common viral diseases can be made, so that attention will be given to development of better prevention and methods of control.

Deaths in Australian freshwater fishes associated with Chilodonella hexasticha infection.

The cause of the deaths of bony bream and other native fish in the Finke River near Alice Springs in winter 1984 was infection with the protozoan ciliate Chilodonella hexasticha. The parasites induced severe generalised epithelial hyperplasia in the gills, which would have compromised respiratory exchange and killed the fish through hypoxaemia. It is suggested that similar winter epizootics of ectoparasites have contributed to previous deaths of fish observed in the Finke River, in association with dry season conditions of low water temperatures and flow rates.

Respiratory-tract colonization and a distinctive pneumonia syndrome in infants infected with Chlamydia trachomatis.

To learn if Chlamydia trachomatis causes in young infants a distinctive penumonia characterized by chronic, afebrile course, diffuse lung involvement and elevated serum immunoglobulins G and M, 47 black infants four to 24 weeks of age were examined for nasopharyngeal shedding of C. trachomatis and serum immunofluorescent antibody to lymphogranuloma venereum Type I. Nasopharyngeal C. trachomatis was found in 18 of 20 with the pneumonia syndrome, two of 15 with various other illnesses and 10 of 12 with inclusion conjunctivitis but without lower respiratory illness. Chlamydial antibody titers of infants with the pneumonia syndrome were significantly elevated (geometric mean-1, pneumonia vs. conjunctivitis = 24,833 vs. 1024 P less than 0.001). No other commonly recognized respiratory pathogens were consistently associated with the pneumonia syndrome. We believe these findings demonstrate an association between the distinctive pneumonia syndrome and C. trachomatis. This, in turn, is a particular facet of a more general event consisting of frequent colonization of the respiratory tract by C. trachomatis in natally acquired infection.

Chemical and biological strategies for engineering cell surface glycosylation.

Oligosaccharides play a crucial role in many of the recognition, signaling, and adhesion events that take place at the surface of cells. Abnormalities in the synthesis or presentation of these carbohydrates can lead to misfolded and inactive proteins, as well as to several debilitating disease states. However, their diverse structures, which are the key to their function, have hampered studies by biologists and chemists alike. This review presents an overview of techniques for examining and manipulating cell surface oligosaccharides through genetic, enzymatic, and chemical strategies.

Chlamydia trachomatis and parainfluenza 2 virus: a shared antigenic determinant?

A tracheal aspirate from which parainfluenza 2 (PI-2) virus but not chlamydiae was isolated demonstrated positive immunofluorescence of elementary body- and reticulate body-like particles on direct examination with anti-chlamydia monoclonal antibodies (Syva Co.). In subsequent studies, we found this reagent to show specific staining of this strain of PI-2 as well as of 3 of 18 additional PI-2 strains that were evaluated.