Joseph V. Baublis

Isolation of measles virus from cell cultures of brain from a patient with subacute sclerosing panencephalitis.

Abstract Isolation of measles virus from the brain of a patient with subacute sclerosing panencephalitis was accomplished by in vitro propagation of the patients brain cells and cocultivation of the brain cells with a line of green-monkey-kidney cells. This observation offers direct evidence that measles virus has a a role in the etiology of the disease. Before the production of detectable, mature virus by the cell cultures, there was a prolonged period when they produced syncytia containing measles antigen, but they failed to hemadsorb.

POSSIBLE PATHOGENIC ROLE OF ENDOTOXIN IN REYE'S SYNDROME

Evidence of circulating endotoxin was sought in children with Reyes syndrome, on the thesis that severe hepatic failure is likely to result in loss of capacity to detoxify intestinal endotoxins entering the circulation. A modification of the Limulus assay was used to demonstrate high levels of endotoxin-like activity (E.L.A.) in nine comatose patients with Reyes syndrome and in one of the two non-comatose patients. The symptom-free sibling of one patient had raised liver enzymes and a negative Limulus test. Plasma E.L.A. correlated significantly with degree of electroencephalographic disturbance early in the course of the illness. E.L.A. was also found in both of two cerebrospinal fluids evaluated. Preliminary in-vitro characterisation of this substance indicated that it resembled endotoxin derived from anaerobic intestinal bacteria. Intestinally derived endotoxin could be one factor in the pathogenesis of encephalopathy and other features of Reyes syndrome.

Specific Response of the Immunoglobulins to Rubella Infection

The teratogenic consequences of rubella infection early in pregnancy are well known. However, experience with the rubella epidemic in 1964 revealed unexpected virologic and serologic consequences of maternal infection. Despite the fact that the rubella syndrome was caused by maternal infection prior to the immunologic maturity of the fetus, serologic immunity rather than tolerance was observed in older children with the syndrome (1, 2). Furthermore, rubella virus was found to persist in the affected infants for many months despite the presence of serum antibodies (3-5). Attempts to equate the presence of rubella antibodies in these infants with an active immune response by means of conventional serology was complicated by the presence of passively acquired maternal antibodies of the IgG variety which persist for several months. Since IgA and IgM do not cross the placental barrier readily, the association of antibody activity within the IgM and/or IgA class of globulins of the infants serum would indicate active immunity. The following studies were carried out in order to characterize the classes of immunoglobulin which responded specifically to rubella infection and to determine their chronology. Observations were also made regarding the classes of rubella antibodies in sera from “normal” newborns and infants with congenital rubella. Materials and Methods. Rubella antibodies were demonstrated by the indirect fluorescent antibody method as described by Brown et al. (6). Whereas conventional methods of serum fractionation and antibody titration were cumbersome, this technique provided a rapid and reliable method for identification of the antigenic class of antibodies (IgG, IgA, or IgM) in whole serum which had combined with viral antigen. Cover slip cultures of a chronically infected line of monkey kidney cells (7) fixed in acetone were employed as an antigen source. The conjugated antisera used in this study were purchased from Hyland Laboratories as fluorescein conjugated antisera prepared in goats against human IgG, IgA, and IgM globulins, respectively. Before use these sera were fractionated by DEAE chromatography as described by Riggs et al. (8).

Reservoirs of Pseudomonas in an intensive care unit for newborn infants: Mechanisms of control

Potential reservoirs of pseudomonas within a neonatal ICU were evaluated. Colonization of infants by the same pseudomonas pyocin types could be classified as a cluster colonization (occurring over three to ten days), or serial colonization (occurring over longer times). Hands of personnel, sink surfaces, and solutions used to rinse nasopharyngeal catheters were identified as the principle reservoirs. Utilization of a liquid iodophor agent for hand washing and of acetic acid for rinsing suction catheters was associated with a significant reduction in the histologic evidence of sepsis and of pneumonia observed among autopsied infants.

Reye Syndrome Associated with Vaccination with Live Virus Vaccines An Exploration of Possible Etiologic Relationships

To determine whether vaccination with live virus vaccines may be etiologi cally related to Reye syndrome, we examined 404 cases reported to the Center for Disease Control. Fifteen of 269 children with Reye syndrome had been inoculated with live virus vaccines within 30 days before onset of illness. Although this temporal relationship may have occurred by chance, seasonal distribution and clustering of incubation periods suggests that live virus vaccines may occasionally serve as cofactors in the etiology of Reye syndrome through undefined mechanisms.

Parainfluenza type 3 parotitis in two immunodeficient children

nous obstruction of the hepatic portion of the inferior vena cava: A clinical study of nine cases, Surgery 72:551, 1972. 5. Schaffner F, Gadboys HL, Safran AP, Baron MG, and Anfses AH Jr: Budd-Chiari syndrome caused by a web in the inferior vena cava, Am J Med 42:838, 1967. 6. Taneja A, Mitra SK, Moghe PD, Rao PN, Samanta N, and Kumar L: Budd-Chiari syndrome in childhood secondary to inferior vena caval obstruction, Pediatrics 63:808, 1979. 7. Kimura C, Shirotani H, Kuma T, Hirooka M, Hayashi K, Tsunekava K, and Matsuda S: Transcardiac membranotomy for obliteration of the inferior vena cava in the hepatic portion, J CardiovascSurg 3:393, 1962. 8. Yamamoto S, Yokoyama Y, Takeshige K, and Iwatsuki S: Budd-Chiari syndrome with obstruction of the inferior vena cava, Gastroenterology 54:1070, 1968. 9. Voorhees AB, Harris RC, Britton RC, Price JB, and Santulli TV: Portal hypertension in children, Surgery 58:540, 1965. 10. Raffensperger JG, Shkolnik AA, Boggis JD, and Swenson O: Portal hypertension in children, Arch Surg 105:249, 1972.

In Vitro Effect Of Metrizamide (amipaque) On Bacterial Growth

Previous workers have demonstrated that the ionic contrast materials sodium diatrizoate and sodium acetrizoate are bacteriocidal or bacteriostatic in vitro for certain strains of gram negative bacteria. If such an effect were present for the non-ionic contrast agent metrizamide, cultures of body fluids obtained subsequent to exposure to metrizamide might be negative when bacteria were present in the specimen. Five strains each of five bacterial species (Staphylococcus aureus, Staphylococcus epidermidis, Pseudomonas aeruginosa, Hemophilus influenza, and Klebsiella pneumoniae) were exposed in vitro to a metrizamide solution. Following isolation of each strain on appropriate agar, several colonies were used to inoculate a nutrient broth which was then incubated for one hour at 37° C. Aliquots of this broth were transferred to three 5 ml. tubes of nutrient broth to which one ml. of metrizamide solution had been added and to another three tubes of broth which served as controls. Quantitative cultures were then performed of the six culture tubes on an hourly basis from zero to five hours after inoculation. The nutrient agar plates were counted with the naked eye following incubation at 37° C for 24 to 48 hours. No significant inhibition of bacterial growth was detected when compared to controls. The presence of metrizamide in a body fluid would not be expected to interfere with bacterial cultures from subsequently obtained specimens.

Characteristics of Measles Virus Isolated from Patients with SSPE

We have isolated measles virus from brain cell cultures derived from two children less than one year following immunization with a live virus vaccine. Recovery of ‘mature’ measles virus was accomplished only after prolonged serial subcultivation of cells from the patients, or cells from the patient cultured together with BS-C-1 cells or primary human kidney cells. Cell cultures before the 12 th level of subcultivation showed the cytopathology and antigen associated with measles infection but produced no free virus. Isolation of measles virus from earlier passage levels of these cells was accomplished by intracerebral inoculation of newborn hamsters. Characterization studies completed to date have shown that the isolates are antigenically indistinguishable from other strains of measles virus. However, they have a distinctive CPE characterized by extensive syncytial formation and produce very large plaques under agar overlay. In continuous lines of monkey kidney cells they yield a low titer of virus, but grow poorly or not at ail in continuous human cell lines. They are distinguishable from other strains of measles by their resistance to thermal inactivation at 40 °C. Both strains produce fatal, acute encephalitis in baby hamsters. The lack of information regarding viral markers and their stability for vaccine and wild strains of measles, has made it impossible to determine the origin of the isolates on the basis of laboratory studies. This situation stresses the need for continuing emphasis on epidemiologic surveillance of SSPE, as well as laboratory research on measles virus and its markers.