A comparison of the effects of oral inoculation with Rotashield and pentavalent reassortant rotavirus vaccine (WC3-PV) on suckling CB17scid mice.

The effects of oral inoculation into infant CB17(scid) mice of two reassortant rotavirus vaccines were compared. The vaccines were Rotashield and WC3-PV, a mixture of five reassortants (G1, G2, G3, G4 and P1; pentavalent reassortant vaccine). Control mice were inoculated with a placebo.

Immune responses in rhesus rotavirus-challenged BALB/c mice treated with bifidobacteria and prebiotic supplements.

Bifidobacterium species (B. bifidum and B. infantis), with or without prebiotic compounds (arabino-galactan, short-chain fructo-oligosaccharide, iso-malto-dextrins), were orally fed to Balb/c pups (n = 192) to evaluate their potential synergistic effects on modulating the course of rhesus rotavirus (RRV) infection, as well as their ability to mediate the associated mucosal and humoral immune responses.

Epidemiology of group C rotavirus infection in Western New York women of childbearing age.

Umbilical cord serum samples (380), an average of 10 per month for 3 years (1990 to 1992), were tested by indirect immunofluorescence assay for group C rotavirus immunoglobulin G. Thirty percent were positive, suggesting that approximately one-third of women of childbearing age in western New York have experienced group C rotavirus infection.

Polymerase chain reaction assays for the detection of cytomegalovirus in organ and bone marrow transplant recipients.

Cytomegalovirus (CMV) infection is ubiquitous and results in a wide spectrum of clinical manifestations ranging from asymptomatic infection to severe life threatening disease. Infection in normal children and adults usually causes no symptoms but in the immunocompromised host, CMV may result in severe opportunistic infections with high morbidity and mortality. Historically, virus detection was dependent on culture of the virus or on a centrifugation culture system referred to as a shell vial assay.

Rapid, non-radioactive detection of virus infection by polymerase chain reaction.

Background: Polymerase chain reaction (PCR) diagnosis of infectious diseases, especially virus diseases, offers a very sensitive and specific technique for clinical diagnosis. However, detection systems for amplified DNA requiring radioactive probe hybridization or signal development using blot transfer or nucleotide capture require overnight incubation or specially labeled probe molecules for analysis of amplified DNA.

Objectives: To place this technology in the clinical laboratory, rapid and sensitive methods are needed for the detection of amplified DNA which are applicable to the assay of multiple specimens representing many different organisms and requiring a minimum of manipulation.

Detection of group C rotavirus in infants with extrahepatic biliary atresia.

The purpose of this retrospective study was to examine liver tissue from patients with cholestatic disease for the presence of group C rotavirus RNA. The reverse transcriptase-polymerase chain reaction (PCR) for genes 5 and 6 was used, and the PCR products were subjected to liquid hybridization with a 32P-labeled probe. A second amplification with nested primers was also used.

Reduction of virus shedding by B. bifidum in experimentally induced MRV infection. Statistical application for ELISA.

The protective effect of a human strain of Bifidobacterium bifidum (B. bifidum) against murine group A rotavirus (MRV) was examined in the intestines of BALB/c infected mice. In experiments designed to determine whether B.

Comparative analysis of VP8* sequences from rotaviruses possessing M37-like VP4 recovered from children with and without diarrhoea.

Rotavirus strains belonging to G types 1 to 4 and having a P3 genotype (M37-like VP4) were recovered from children with symptomatic and asymptomatic infections. Partial sequences of their VP4 genes were determined in an attempt to characterize these strains further. The genomic regions encoding VP8*, the connecting and putative fusion peptides and three other regions in VP5* were sequenced.

Effectiveness of Bifidobacterium bifidum in mediating the clinical course of murine rotavirus diarrhea.

Human Bifidobacterium sp strain bifidum (B. bifidum) was administered to BALB/c lactating mice (n = 58) and their litters (n = 327 pups) to evaluate the ingested strain's adherent properties and ability to inhibit murine rotavirus (MRV) infection. ELISA and anaerobic bacteriologic techniques were used to measure MRV shedding and colonization of Bifidobacterium in the small intestine.

VP4 genotyping of human rotavirus in the United States.

The VP4 (P) and VP7 (G) types of 171 rotavirus isolates obtained from children with diarrhea in the United States were characterized by PCR typing assays. Strains P1G1 predominated (71%); this was followed by strains P1G3 (20%) and P2G2 and P1G4 (2% each). Mixed types were identified in five (3%) specimens.

Effectiveness of Bifidobacterium bifidum in experimentally induced MRV infection: dietary implications in formulas for newborns.

The protective effect of a human strain of Bifidobacterium bifidum (B. bifidum) against murine Group A rotavirus (MRV) was examined in the intestines of BALB/c infected mice. In experiments designed to determine whether B.

Group A rotaviruses produce extrahepatic biliary obstruction in orally inoculated newborn mice.

Extrahepatic biliary atresia is a devastating disease occurring in 1 in 10,000 to 14,000 infants annually in the United States. We have recently described preliminary data suggesting an association of group C rotavirus with biliary atresia in two infants. However, a group C rotavirus animal model of biliary atresia is not presently available.

Comparison of rapid immunofluorescence assay to cell culture isolation for the detection of influenza A and B viruses in nasopharyngeal secretions from infants and children.

In the hospital setting it is often critical to isolate patients appropriately in order to prevent nosocomial infection. This is especially true with respiratory infection in infants and young children. At the present time a rapid immunofluorescence assay (IFA) for respiratory syncytial and parainfluenza viruses is routinely carried out in our laboratory.

Six-year retrospective surveillance of gastroenteritis viruses identified at ten electron microscopy centers in the United States and Canada.

To identify the prevalence, seasonality and demographic characteristics of patients with viral gastroenteritis, we reviewed 6 years of retrospective data on viral agents of gastroenteritis screened by electron microscopy at 10 centers in the United States and Canada. From 52,691 individual electron microscopic observations, a virus was detected in 16% of specimens, and the yearly positive detection rate among centers ranged from 8 to 34%. Rotavirus was the agent most commonly observed (26 to 83%), followed by adenoviruses (8 to 27%, respiratory and enteric combined), and small round viruses (SRVs) (0 to 40%) which were second most common at two of the centers.

Serotypes and electropherotypes of human rotavirus in the USA: 1987-1989.

The epidemiology of rotavirus gastroenteritis was investigated for two consecutive seasons (1987-1988 and 1988-1989) in seven locales in the continental USA. The 281 representative fecal samples obtained from children with diarrhea were electropherotyped and serotyped by an enzyme immunoassay with serotype-specific monoclonal antibodies and a new amplification typing technique (polymerase chain reaction typing). Serotype 1 was predominant in both years, particularly in the North and East; serotype 3 was second in frequency and found most often in the South; serotype 2 was detected only occasionally; serotypes 4, 8, and 9 were never found.

Effect of rotavirus infection and malnutrition on uptake of a dietary antigen in the intestine.

Intestinal absorption of ovalbumin (OVA), a dietary macromolecule, was studied in malnourished and normally nourished suckling mice after experimentally induced infection with rotavirus. All mice developed diarrhea within 24 to 48 h postinoculation. The malnourished animals exhibited more severe symptoms and an increased number of rotavirus-containing enterocytes in intestinal sections as compared to well-nourished mice when examined 3 d postinoculation, at the peak of diarrhea.

Extramucosal spread and development of hepatitis in immunodeficient and normal mice infected with rhesus rotavirus.

The pathogenic profiles of two heterologous animal rotaviruses, rhesus rotavirus strain MMU 18006 and bovine rotavirus strain WC3, were evaluated in mice with severe combined immunodeficiency (SCID mice) and normal BALB/c mice. Control animals were inoculated with homologous murine strain EDIM 5099 or a tissue culture-adapted murine rotavirus. Heterologous infection with rhesus rotavirus resulted in hepatitis in 84% of SCID and 21% of BALB/c mice, with mortality rates of 27 and 0%, respectively.

Vasoactive intestinal peptide as a laboratory supplement to clinical activity index in inflammatory bowel disease.

Circulating levels of vasoactive intestinal peptide (VIP) in plasma were measured in gauging activity in inflammatory bowel disease (IBD). One hundred-fifteen adult IBD patients were studied cross-sectionally and prospectively, 48 with ulcerative colitis (UC) and 67 with Crohn's disease (CD). Sequential samples of plasma were assayed for VIP by specific radioimmunoassay.

Effect of nutritional deprivation on mucosal viral infections.

Suckling BALB-c mice, subjected to nutritional deprivation in artificially expanded litters (18 to 20 pups), were compared to normally nourished pups (7-9 per litter) in a series of experiments designed to provide data on morphologic and functional alterations of the small intestine during malnutrition and infection. The effects of protein calorie malnutrition (PCM) on the viral replication pattern and severity of clinical disease were examined in suckling mice infected with murine rotavirus (MRV). The infection in nutritionally deprived animals was characterized by a significant decrease in the minimal infectious dose and in the incubation period for the onset of diarrhea as well as increased severity of disease when compared to well nourished controls.

Distribution of rotavirus antigen in intestinal lymphoid tissues: potential role in development of the mucosal immune response to rotavirus.

Groups of suckling BALB/c mice were inoculated with murine rotavirus (MRV) at 5 days of age and sequentially tested for the presence of MRV antigen (Ag) in intestinal villus epithelium (VE), Peyer's patch (PP), mesenteric lymph node (MLN), spleen, liver and lung, using indirect immunofluorescence techniques (IF). MRV Ag was first detected in VE 24 h after oral administration of the virus and remained in VE for as long as 10 days post-inoculation (pi). MRV Ag was observed in the epithelium overlying PP at 3-7 days pi and in subepithelial and interfollicular areas in the PP between 3 and 20 days pi.

Immunological aspects of interaction between rotavirus and the intestine in infancy.

Rotaviruses are important pathogens causing severe diarrhoea in human infants and young animals. Available information on the pathogenic mechanisms of and the immune response to rotaviruses is reviewed here. Studies in our laboratory using the suckling mouse model have focused on elucidating the nature of interaction between the virus and the gut, and on the importance of T and B cell mediated immunity in protection and recovery from the disease.

Persistent rotavirus infection in mice with severe combined immunodeficiency.

Rotaviruses are important pathogens of human infants and the infants of many animal species. The disease produced by these viruses can be described as an acute, self-limiting diarrheal disease, with virus replication localized to the differentiated epithelial enterocytes of the small intestine. Immunologically normal infants shed virus for approximately 5 to 12 days after the onset of infection.

Immune responses to herpes simplex virus in patients with recurrent herpes labialis: I. Development of cell-mediated cytotoxic responses.

Groups of subjects during acute (0-3 days) and convalescent (2-3 weeks) phase of recurrent herpes labialis (RHL), and other subjects seropositive or seronegative for herpes simplex virus type 1 (HSV-1) antibody without any history of RHL, were tested for the appearance of cell-mediated cytotoxic responses by stimulating peripheral blood leukocytes (PBL) in vitro with ultraviolet-inactivated HSV-1 antigen, using the release of radiolabelled chromium (51Cr) from HSV-1-infected autologous, or allogeneic lymphocytes and K562 erythroleukemia cell line as nonspecific targets. Development of HSV specific cytotoxic response using autologous targets was essentially limited to subjects with RHL and in HSV antibody seropositive control subjects. Peak activity was observed during the acute phase of the disease, compared to the activity in the convalescent phase in seropositive subjects with RHL, and was preceded by high lymphoproliferative response to HSV.