An in vitro DNA hybridization assay was used to test 281 newborns for congenital infection with cytomegalovirus. The assay utilized an abbreviated method for DNA preparation and a dot blot assay that provided good sensitivity (100%) and specificity (98.9%) when compared with standard tissue culture, yet substantially reduced the total time of analysis. This assay would be a useful adjunct to tissue culture to diagnose newborns with congenital infection with cytomegalovirus.

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A technique based on polymerase chain reaction (PCR) amplification was developed to facilitate the study of the epidemiology of cytomegalovirus (CMV). Consensus oligonucleotide primers from repetitive DNA sequences were designed to amplify interspersed repetitive sequences in an area of heterogeneity within the L-S junction region of the CMV genome, and PCR products were detected by gel electrophoresis. Purified CMV DNAs from 25 CMV isolates, 13 from members of five families in which person-to-person transmission was documented, 9 random clinical isolates of CMV, and 3 laboratory reference strains of CMV (Towne, Davis, and AD169), were analyzed. The gel electrophoretic patterns of DNA bands, or PCR profiles, produced by amplification with the L-S primers were unique for epidemiologically unrelated strains and laboratory reference strains, yet similar patterns were observed for epidemiologically related strains isolated from members of the same family. This method of rapid fingerprinting of CMV DNA within the hypervariable L-S junction region by PCR to produce strain-specific, variably sized PCR products should simplify the molecular epidemiologic analysis of CMV.

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In any medical and social service setting, patient data must be readily shared among multiple providers for delivery of expeditious, quality care. This paper describes the development and implementation of a generalized social and medical services data model for an ambulatory population. The model, part of the Collaborative Social and Medical Services System Project, is based on the data needs of the Baylor College of Medicine Teen Health Clinics and follows the guidelines of the ANSI HISPP/MSDS JWG for a Common Data Model. Design details were determined by informal staff interviews, operational observations, and examination of clinic guidelines and forms. The social and medical services data model is implemented using object-oriented data modeling techniques and will be implemented in C++ using an Object-Oriented Database Management System.

This paper describes the design and implementation of a client application for the Baylor College of Medicine Teen Health Clinics. The application is the front end to the Collaborative Social and Medical Services System (CSMSS) under development by Baylor's Medical Informatics and Computing Research Program [8]. The application provides distributed access to an underlying object oriented database system. A process driven and patient centered design will provide staff members with a complete set of services, including forms for data entry and viewing, query, and access management to facilitate efficient and effective delivery of services. Role-specific interfaces will be supplied for clerks, nurses, nurse practitioners, physicians, and social workers. The client application is being designed using object oriented methodologies and technologies with the C++ programming language, and will operate within a Microsoft Windows operating environment utilizing Object Linking and Embedding for application interoperability.

This paper presents the development of the Collaborative Social and Medical Services System's (CSMSS) data security mechanism. This mechanism was synthesized from an analysis of the CSMSS problem domain, and from a study of the methods used by modern operating systems and database management systems. The resulting mechanism is more flexible and expressive than traditional access control methods and is generally applicable to the management of privacy and multi-provider access.

Investigations of polymorphonuclear leukocyte (PMN) function were performed in a 5-yr-old white female with delayed umbilical cord separation, impaired pus formation, and a severe defect of PMN chemotaxis. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis demonstrated an almost total deficiency of a high molecular weight glycoprotein(s) (GP138) in the granule and membrane fractions of the patient's cells, and NaB3H4-galactose oxidase labeling demonstrated the absence of a major glycoprotein complex on the surface of her PMNs. Monoclonal antibodies (MAb) were employed in flow cytometry experiments to demonstrate that two previously characterized glycoproteins (Mo1 and LFA1) were undetectable on the surface of the patient's PMNs and monocytes. Immunoprecipitation of 125I-labeled patient cells with subunit specific MAbs confirmed that the alpha-subunits of Mo1 (155 kD) and LFA1 (177 kD) and their common beta-subunit (94 kD) were totally deficient. Functional analyses of patient PMNs demonstrated severe impairment of adherence- and adhesion-dependent cell functions including spreading, aggregation, orientation in chemotactic gradients, antibody-dependent cellular cytotoxicity, and phagocytosis of particles (Oil-Red-0-paraffin, zymosan) selectively opsonized with C3-derived ligands. Patient PMNs demonstrated a normal capacity to rosette with IgG or C3b-coated sheep erythrocytes, but rosette formation with C3bi-coated erythrocytes was profoundly diminished. Adhesion-independent functions including shape change, N-formyl-methionyl-leucyl-3H-phenylalanine binding, and O-2 generation or secretion elicited by soluble stimuli were normal. Membrane fluidity, surface charge, and microtubule assembly were also normal. These findings provide new evidence that critical PMN surface glycoproteins are required to facilitate multiple adhesion-dependent cellular functions of the inflammatory response.

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Studies were performed to characterize chemotactic activity generated by Haemophilus influenzae type b (HiTb) in serum or elaborated independent of serum. Neutrophil aggregometry, Sephadex G-75 gel chromatography, and anti-C5 neutralization studies were used to demonstrate that the complement fragment C5a represented the major chemotactic moiety derived from HiTb-serum interactions. HiTb elaborated minimal chemotactic activity independently. Maximal C5a generation by HiTb as measured by neutrophil response in chemotaxis, shape change, and aggregation assays required specific antibody to the capsular polysaccharide, polyribosyl ribitol phosphate (PRP). Significantly more C5a was generated in pooled normal human serum containing high titers of anti-PRP (determined by an enzyme-linked immunosorbent assay) than in hypogammaglobulinemic serum. Furthermore, C5a generated in hypogammaglobulinemic serum reconstituted with purified high-titer immunoglobulin G, hyperimmune rabbit serum or heat-inactivated normal human serum was comparable to that generated in normal human serum. Absorption of antibody with PRP versus whole HiTb showed a contribution by non-PRP-directed antibody. As shown with the use of C4-deficient guinea pig serum, C5a generation occurred via the alternative complement pathway in nonimmune serum, and activation of the alternative complement pathway was facilitated by specific anti-PRP. C5a generation in test sera was proportional to its opsonic activity for HiTb as assessed by a luminol-chemiluminescence assay. Overall low levels of C5a activity were observed in 13 pediatric patient serum samples obtained during the acute phase of HiTb meningitis, and no pulmonary symptoms or radiographic abnormalities consistent with a leukocyte aggregation syndrome were observed in these patients.