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Biomedical subjects

G L Barnes

Publications and source records attributed to G L Barnes.

At least 19 recordsLinked to original sources
PubMed1992

Severity of rotavirus infection in relation to serotype, monotype and electropherotype.

The aim of this study was to determine whether the severity of symptoms associated with rotavirus infection was related to the serotype of the infecting virus. Severity of clinical symptoms in 108 children admitted to hospital for treatment of rotavirus diarrhoea was retrospectively assessed using a scoring system for frequency and duration of vomiting and diarrhoea, degree of fever, acidosis and dehydration, and presence of electrolyte imbalance. Children were 6-30 months old and were fully weaned at onset of symptoms prior to admission to hospital. No other enteric pathogens were detected during the course of the illness. Serotypes and monotypes were identified using a panel of monoclonal antibodies. Gel electrophoresis of rotavirus RNA was performed to determine electropherotypes. Children surveyed were infected with serotype 1 (47), serotype 2 (15) or serotype 4 (46) rotaviruses. Comparisons of severity of clinical symptoms according to infecting serotype revealed no statistically significant differences between serotype 1, 2 or 4 infections. In addition, no differences were detected between different rotavirus strains within each serotype (as judged by electropherotype) including monotypes 1a or 1c. This study failed to reveal differences in virulence between rotavirus strains of different VP7 serotypes infecting young children.

Child, Preschool↗
PubMed1992

Normalization of vitamin B12 absorption after ileal resection in children.

Impaired Vitamin B12 absorption after significant ileal resection has been reported to be permanent, although partial recovery after ileal bypass can occur. Three children are presented in whom Vitamin B12 malabsorption returned to normal 6-8 years after ileal resection. This was due probably to adaptation of the remaining small bowel, although spontaneous resolution of bacterial overgrowth is a possible explanation. An abnormal Schilling test after ileal resection does not automatically imply the need for life-long Vitamin B12 injections.

Child↗
PubMed1992

Role of coproantibody in clinical protection of children during reinfection with rotavirus.

Rotavirus is the major cause of severe, dehydrating infantile gastroenteritis. Infection is limited to the gut, but the relative roles of serum and secretory copro-immunoglobulin A (IgA) in protection are unclear. Specific copro-IgA is predictive of duodenal antirotaviral IgA and correlates with virus-neutralizing coproantibody. Copro-IgA conversion is a more sensitive marker of rotavirus reinfection than seroconversion. We measured rotavirus reinfections by copro-IgA conversion prospectively in 35 children recruited at a time of severe rotavirus illness. The children were followed up longitudinally for 14 to 31 months to determine whether high coproantibody levels correlated with clinical protection against rotavirus disease. Ninety-four percent of the children experienced reinfection, and 38% developed persistent elevations in specific copro-IgA termed plateaus. Plateau children had a higher mean annual rate of rotavirus infection and a lower ratio of symptomatic to total number of rotavirus reinfections than did nonplateau children. The annual rates of rotavirus infection and disease were significantly higher outside the plateau than inside it in children experiencing antirotavirus copro-IgA plateaus. Frequent rotavirus infection of children appears to stimulate production of a specific copro-IgA plateau which correlates with protection against an excess of infection and symptomatic disease.

Age Factors↗
PubMed1991

Epidemiology of rotavirus serotypes in Melbourne, Australia, from 1973 to 1989.

Fecal rotavirus strains collected between 1973 and 1989 from 943 children admitted with acute diarrhea to one hospital in Melbourne, Australia, were serotyped by using an enzyme-linked immunosorbent assay. The assay incorporated neutralizing monoclonal antibodies specific for VP7 of the four major human serotypes (1 through 4). A serotype could be assigned to 690 of 943 specimens (73.2%). Typeable strains comprised serotype 1 (72.5%), serotype 2 (6.8%), serotype 3 (2.9%), or serotype 4 (15.4%). Monotypes 1a and 1c comprised 52 and 44%, respectively, of serotype 1 strains. All serotypes and monotypes exhibited polymorphic genomic RNAs. Specimens reacting as mixed serotypes were rare (3.2%) and included intertypic strains (0.7%) and mixed infections (1.0%). Nontypeable strains for which an electropherotype could be determined appeared to be identical with typeable strains present concurrently in the community. Serotypes exhibited various epidemiological patterns. Serotype 1 strains were dominant except during three successive winters when 60 to 90% of the disease was caused by serotype 2. Serotype 4 strains showed an episodic pattern of appearance, recurring at peak incidence approximately every 3 years. Fecal rotavirus strains collected from 145 newborn babies housed in Melbourne obstetric hospitals between 1974 and 1986 were also serotyped. All 135 typeable strains (93.1%) belonged to serotype 3. It is hypothesized that endemic infection with serotype 3 rotaviruses in nurseries for the newborn influenced the epidemiology of rotavirus serotypes responsible for severe clinical disease in young children in the same community.

Age Factors↗
PubMed1990

Comparison of rotavirus immunoglobulin A coproconversion with other indices of rotavirus infection in a longitudinal study in childhood.

In order to determine the sensitivity and reliability of antirotaviral fecal immunoglobulin A (IgA) as an indicator of rotavirus reinfection, the antibody responses to rotavirus of 44 infants with severe rotavirus gastroenteritis recruited on admission to a hospital were studied. Feces were collected daily during hospitalization and weekly thereafter, and sera were obtained every 4 months, for 6 to 32 months (median, 17 months). Antirotaviral IgG, IgA, and IgM were measured by enzyme immunoassay in all samples. Rotavirus antigen, rotavirus-neutralizing antibody, and total IgA were measured in feces. The results showed that use of an IgA index (ratio of specific IgA to total IgA) was unnecessary to identify copro-IgA conversion to rotavirus. The other markers of rotavirus infection tested showed a high level of predictive accuracy of coproconversion in rotavirus-neutralizing antibody. Copro-IgM, serum IgM, and virus in feces were insensitive measures of neutralizing antibody coproconversion. Seroconversion in IgG or IgA was detected in 46% of neutralizing coproconversions. The most sensitive marker, present in 92% of neutralizing coproconversions, was antirotaviral fecal IgA conversion. This correlation of fecal IgA with fecal neutralizing antibody suggests that coproconversions in IgA represent true elevations in antirotaviral IgA with neutralizing capacity. A coproconversion in IgA appears to indicate genuine rotavirus infection. Copro-IgA conversions in feces collected weekly are likely to be more sensitive markers of rotavirus reinfection than are seroconversion and virus detection combined in epidemiological studies of acute diarrhea in children and in rotavirus vaccine trials.

Antibodies, Viral↗
PubMed1989

Rotavirus serotypes causing acute diarrhoea in hospitalized children in Yogyakarta, Indonesia during 1978-1979.

Rotavirus strains in stool specimens from 111 children aged 3-24 months admitted to hospital in Yogyakarta, Indonesia for treatment of acute diarrhoea were serotyped using VP7 serotype specific monoclonal antibodies in a double sandwich enzyme immunoassay. A serotype could be assigned to 59 of 111 specimens (53%). Inability to assign a serotype to 47% of specimens was probably due to loss of the outer capsid during transport of specimens from Indonesia to Australia. All four major human rotavirus serotypes were detected during the 15 month survey from June 1978 to August 1979, including one serotype 1, 5 serotype 2, 31 serotype 3, and 21 serotype 4 strains. One additional strain reacted with serotype 3 and 4 Mabs. Serotype 3 strains showed intratypic variation. The relative frequency of serotypes 2, 3, and 4 varied during the 15 months and appeared to be influenced by climatic changes associated with dry and wet seasons. Vaccine strategies must take account of comparatively rapid changes of predominant serotypes in a community and are only likely to be successful if comprehensive immunity can be established simultaneously against the four major human serotypes.

Acute Disease↗
PubMed1989

Evaluation of end-point titration, single dilution and capture enzyme immunoassays for measurement of antirotaviral IgA and IgM in infantile secretions and serum.

In order to facilitate measurement of antirotaviral IgA in large collections of faeces and secretions, adaptations of enzyme immunoassay methods for estimating antirotaviral IgA and IgM in duodenal fluid, saliva, faeces and serum were studied. To quantitate specific IgA, a single dilution of each sample was assayed. Results were expressed as antirotaviral IgA units derived from a standard curve. Units were calculated by log-logit analysis on computer. There was strong correlation between antirotaviral IgA units and end-point titres in 257 faecal samples (correlation coefficient r = 0.92) and in 182 duodenal fluids and salivary samples (correlation coefficient r = 0.74). The assay was validated using acute and convalescent faeces from children with or without rotavirus infection. Immune conversions in IgA were detected in 33 (75%) of the children by units and 34 (77%) by titres. None of nine children with gastroenteritis due to other infectious agents showed immune conversions to rotavirus. A monoclonal capture IgM assay showed similar end-point titres and numbers of immune conversions when compared with a direct assay for antirotaviral IgM in serum and secretions. Use of the capture method eliminated false-positive reactions with the cell control. The assay for antirotaviral IgA units in secretions is simple, rapid, reproducible and reliable, and has proven of value in longitudinal epidemiological studies of rotavirus coproIgA profiles. Both the capture IgM technique and the single dilution IgA method permit analysis of large numbers of specimens and are appropriate for examination of immune responses to natural rotavirus infection or during vaccine trials.

Antibodies, Viral↗
PubMed1989

Infectious diarrhea in children undergoing bone-marrow transplantation.

Fecal flora of 12 children undergoing bone-marrow transplantation was monitored prospectively using comprehensive microbiological techniques. Diarrhea developed at least once in ten of the 12 children (83%), and a total of 24 episodes were recorded. Recognised gut pathogens were isolated from 11/21 (52%) diarrheal episodes where fecal specimens were obtained. Enteric pathogens identified included viral pathogens in 19% (rotaviruses, 'enteric' adenoviruses), parasites in 19% (cryptosporidium, Giardia lamblia) and cytotoxic C. difficile (14%). Excretion of clostridial species (including cytotoxin negative C. difficile, C. innocuum) occurred in 90% of diarrheal episodes when no enteric pathogen was identified. These results suggest that infection is often responsible for diarrhea associated with bone-marrow transplantation. Prophylaxis against enteric infection might reduce the morbidity and mortality associated with severe diarrhea in bone-marrow transplanted children.

Adolescent↗
PubMed1988

Sweat testing by capillary collection and osmometry: suitability of the Wescor Macroduct System for screening suspected cystic fibrosis patients.

A new method of collecting and analysing sweat (Wescor Macroduct) has advantages of simplicity of collection and direct reading of results by osmometry. Forty-seven children with cystic fibrosis and 47 normal children had sweat tests performed simultaneously by the Gibson and Cooke method and by the Wescor Macroduct method. The new method had a higher rate of inadequate collection (19% vs 6%) which was more marked in children under 5 years of age. This was due partly to the difficulty of fitting a 2.5 cm pilocarpine gel disc to small arms. When an adequate collection was obtained, results were reliable with no false negatives occurring during this study. The Wescor Macroduct sweat test is a reliable method for use in peripheral centres to screen patients suspected of having cystic fibrosis. All children with an inadequate collection or a positive result should be referred to a reference centre for confirmation of the diagnosis. However, the majority will be saved the expense and disruption of travel.

Adolescent↗
PubMed1988

Comparison of serum and mucosal antibody responses following severe acute rotavirus gastroenteritis in young children.

The development of mucosal immunity is presumed to be the most important marker of rotavirus infection. The practical difficulties of obtaining small-bowel secretions stimulated this study of the antibody response to acute rotavirus infection at other sites. Forty-four infants admitted to the hospital with rotavirus gastroenteritis had serum, saliva, and feces collected at the acute phase (median, 5.5 days), during convalescence (median, 33.5 days), and 4 months later (median, 12.2 weeks). A subgroup of 19 children also had duodenal juice collected in parallel. Rotavirus-specific immunoglobulin G (IgG), IgA, secretory immunoglobulin, and IgM were measured and compared in all samples. The results showed that the estimation of antirotavirus serum IgM, serum IgG, duodenal juice IgA, and duodenal juice IgM by an enzyme immunoassay indicated an immune response to severe primary rotavirus infection in all children. Four months later, the levels of serum IgG and IgA served as the most sensitive markers of the preceding rotavirus infection. The predictive accuracies of immune responses at different sites in relation to a positive IgA immune response in the duodenum were calculated. Fecal IgA predicted duodenal IgA rotavirus antibodies with accuracies of 86% at 1 month and 92% at 4 months. The high sensitivity of serum IgM and IgG in detecting rotavirus infection and the high predictive accuracy of fecal IgA as an indicator of duodenal IgA abrogates the need for duodenal intubation to detect (or monitor) an immune response to rotavirus infection. This finding has important practical implications for epidemiological studies of acute diarrhea in children and in rotavirus vaccine trials.

Antibodies, Viral↗
PubMed1988

'Normal' disaccharidase levels in children.

Results of disaccharidase assays in small bowel biopsies from 887 children over a 3 year period were analysed to establish normal values. Abnormal histology, the presence of giardia trophozoites or total absence of sucrase and isomaltase were found in 307 cases and these were excluded from further consideration. The results for maltase, sucrase and lactase from the remaining 580 children have been graphed as percentiles at various ages. They represent results which are as close to normal as it is possible ethically to obtain.

Age Factors↗
PubMed1987

Relative incidence of Crohn's disease and ulcerative colitis in six Melbourne hospitals.

Information on the relative incidence of Crohn's disease and ulcerative colitis was obtained by a prospective investigation at six Melbourne teaching hospitals. One hundred and eleven patients who presented with chronic inflammatory bowel diseases between 1980-1981 were admitted to the study. Forty (36%) patients were diagnosed as having Crohn's disease and 63 (57%) patients as having ulcerative colitis. The type of chronic inflammatory bowel disease could not be determined in eight (7%) patients. These findings suggest that the relative frequency of Crohn's disease and ulcerative colitis in Melbourne hospitals is within the range that is reported for northern Europe and the United States.

Adolescent↗
PubMed1987

Cultivation and characterization of rotavirus strains infecting newborn babies in Melbourne, Australia, from 1975 to 1979.

Twenty-three rotavirus strains obtained from the stools of 71 newborn babies were adapted to growth in MA-104 cells. Babies were housed in newborn nurseries of eight different obstetric hospitals in Melbourne between 1975 and 1979. All strains belonged to serotype 3 when reacted with serotype-specific neutralizing monoclonal antibodies in an enzyme immunoassay. Genome RNA of these 23 strains and of one stool virus not adapted to cell culture were compared by coelectrophoresis of mixtures of RNA. When strains were compared by coelectrophoresis of RNA for 4 h at 40 mA current, the majority appeared to be identical. Coelectrophoresis at 4 degrees C for 17 h at 10 mA current with 0.75-mm-thick polyacrylamide gels resulted in increased resolution of segments, revealing more genetic diversity than previously observed. Seventeen different electropherotypes showing slight variations in migration of one to seven segments were identified. Segments 5 and 7, 8, 9, 10, and 11 varied more frequently than segments 1, 2, 3, 4, and 6. Strains endemic in one hospital from 1975 to 1983 showed increased numbers of segmental changes over time. Differing patterns of reaction with two neutralizing monoclonal antibodies reacting with VP3 and VP7 were observed. Comparison of electropherotypes of three neonatal strains with a serotype 3 community strain showed marked differences in segment migration. The serotypic similarity, electropherotypic dissimilarity from community strains, and asymptomatic nature of most infections are additional evidence that these viruses infecting newborn babies form a unique group of rotaviruses.

Antibodies, Monoclonal↗
PubMed1986

Gastrointestinal hypersensitivity to cow's milk protein: the diagnostic value of gut function tests.

Thirty-six children with suspected gastrointestinal hypersensitivity to cow's milk protein were investigated before and after challenge with cow's milk protein by one or more of four tests of gut function: the appearance of small bowel mucosa, mucosal disaccharidase levels, a 1-h blood-xylose test, and a 50 g-lactose breath-hydrogen test. These tests were not always abnormal in children who had definite adverse reactions to milk. Conversely changes were seen in some with negative milk challenges. Although small bowel biopsy, assessment of disaccharidase activity and perhaps the breath-hydrogen test have an important place in the pre-challenge assessment to exclude other causes of gastrointestinal symptoms, these gut function tests and the 1-h xylose test done following milk provocation do not appear to have any advantage over careful clinical observation.

Biopsy↗
PubMed1986

Comparison between children treated at home and those requiring hospital admission for rotavirus and other enteric pathogens associated with acute diarrhea in Melbourne, Australia.

The etiology of acute diarrhea in children less than 42 months of age attending one pediatric hospital in Melbourne, Australia, was studied during a 7-month period encompassing the winter of 1984. Pathogens identified in 157 children treated as outpatients with mild disease were compared with those in 232 children hospitalized with severe disease. The pathogens (and frequencies among outpatients and inpatients, respectively) detected were rotaviruses (32.5 and 50.9%), enteric adenoviruses (8.9 and 7.4%), Campylobacter jejuni (7.2 and 1.3%), and Salmonella sp. (4.0 and 1.7%). Electropherotypes of rotavirus strains from outpatients and inpatients were compared. Two strains predominated during the 7 months of this study and were observed with equal frequency from outpatients and inpatients. Rotaviruses of the same electropherotype caused a wide spectrum of disease, with symptoms ranging from mild to severe, life-threatening diarrhea. The similarity of etiological agents identified from children with mild and severe forms of acute diarrhea suggests that the etiology of community enteric illness can be reasonably inferred from the etiology of inpatient disease in children in the same geographic area. During the winter epidemic period, the severity of symptoms associated with rotavirus infection in young children is likely to be determined by the inherent susceptibility of the host rather than by genetic differences in the strains of infecting rotaviruses.

Acute Disease↗
PubMed1986

Heterologous protection against rotavirus-induced disease in gnotobiotic piglets.

Administration per os of 2 X 10(6) fluorescent cell-forming units of a human serotype 3 rotavirus (RV-3) protected all of nine gnotobiotic piglets against severe diarrheal disease when they were challenged 10 to 14 days later with 8 X 10(3) fluorescent cell-forming units of virulent wild-type porcine rotavirus (AT/76). The porcine virus was similar antigenically to porcine prototype strain OSU, previously described as antigenically distinct from all four recognized human serotypes. Administration of RV-3 was associated with the development of serum-neutralizing antibody to both RV-3 and AT/76 in piglets that excreted RV-3. Neutralizing antibody levels to RV-3 and AT/76 increased rapidly postchallenge. Vaccinated piglets were not immune to infection with AT/76 but showed no or minimal gastrointestinal symptoms after challenge. Control nonvaccinated piglets that were fed AT/76 developed severe dehydrating diarrhea and low levels of neutralizing antibody to AT/76 alone. The apparent heterologous clinical protection observed in this study could have been predicted from results of in vitro assays. Neutralization tests with reduction of fluorescence focus indicated a one-way cross-reaction between RV-3 and AT/76 such that hyperimmune antiserum to RV-3 neutralized porcine virus to moderate titer, but not vice versa. The results emphasize the importance of neutralizing antibody in protection against disease and the need to determine reciprocal cross-neutralization titers, rather than serotype alone, in order to predict the ability of rotavirus strains to cross protect.

Animals↗