Induction of immune response to the 17 kDa OMPA Burkholderia cenocepacia polypeptide and protection against pulmonary infection in mice after nasal vaccination with an OMP nanoemulsion-based vaccine.

Burkholderia cepacia complex (Bcc) are opportunistic bacteria associated with life-threatening illness in persons with cystic fibrosis. Once Bcc colonization is established, these antimicrobial-resistant and biofilm-forming bacteria are difficult to eradicate and are associated with increased rates of morbidity and mortality. At present, no vaccines are available to prevent the Bcc infection. There is currently a paucity of published information regarding the development of vaccines designed to prevent Burkholderia colonization. This work expands on the recent studies published by Bertot et al. [Infect Immun 75(6):2740-2752, 2007], where successful protective immune responses were generated in mice using a B. multivorans OMP-based vaccine. Here, we evaluate an experimental mucosal vaccine against Bcc using a novel mucosal adjuvant (nanoemulsion) and a novel B. cenocepacia-based OMP antigen. The OMP antigen derived from B. cenocepacia was mixed with either nanoemulsion or with PBS and delivered intranasally to CD-1 mice. Serum analysis showed robust IgG and mucosal secretory IgA immune responses in vaccinated versus control mice. The antibodies had cross-neutralizing activity against both B. cenocepacia and B. multivorans species. We found that immunized mice were protected against pulmonary colonization with B. cenocepacia. We have also identified that a 17 kDa OmpA-like protein highly conserved between Burkholderia and Ralstonia species as a new immunodominant epitope in mucosal immunization.

Levels of BAFF in serum in primary biliary cirrhosis and autoimmune diabetes.

Levels of the B-cell activating cytokine BAFF are increased in serum in various autoimmune disease, and particularly Sjögren's syndrome in which there is evident B-lymphocyte proliferation. Studies in two autoimmune disease in which B-cell proliferation is less evident, primary biliary cirrhosis (PBC), and adult-onset Type 1 diabetes, showed serum levels of BAFF to be mostly in the normal range. A single raised level among eight sera tested in one patient studied with autoimmune hepatitis (AH) coincided with a relapse of the disease. Increased levels of BAFF in human sera, indexing a potent antigenic drive on B-cell production and survival in some autoimmune diseases, may mark only particular stages in the evolution of such diseases.

Molecular characterisation of mouse and human TSSC6: evidence that TSSC6 is a genuine member of the tetraspanin superfamily and is expressed specifically in haematopoietic organs.

Previous analyses of the murine and human TSSC6 (also known as Phemx) proteins were not carried out using the full length sequence. Using 5'-RACE and cDNA library screening, we identified an additional 5' sequence for both the murine Tssc6 cDNA and its human homologue TSSC6. This novel sequence encodes a 5' exon encoding an in frame, upstream start codon, an N-terminal cytoplasmic domain and a transmembrane domain. The deduced, and now full length, murine and human TSSC6 proteins contained four hydrophobic regions together with other features characteristic of the tetraspanin superfamily. Computational analyses of the full length sequences show that TSSC6 is a genuine, albeit relatively divergent member of this superfamily. Using RNA from a number of mouse tissues, we identified seven splice variants of Tssc6. Splice variants of the human gene were also detected. Tssc6 expression was detected early in embryogenesis in primitive blood cells and was confined to haematopoietic organs in the adult mouse. Tssc6 expression was detected in many haematopoietic cell lines and was highest in cell lines of the erythroid lineage.

Clonogenic analysis reveals reserve stem cells in postnatal mammals: I. Pluripotent mesenchymal stem cells.

Clonal populations of lineage-uncommitted pluripotent mesenchymal stem cells have been identified in prenatal avians and rodents. These cells reside in the connective tissue matrices of many organs and tissues. They demonstrate extended capabilities for self-renewal and the ability to differentiate into multiple separate tissues within the mesodermal germ line. This study was designed to determine whether such cells are present in the connective tissues of postnatal mammals. This report describes a cell clone derived by isolation from postnatal rat connective tissues, cryopreservation, extended propagation, and serial dilution clonogenic analysis. In the undifferentiated state, this clone demonstrates a high nuclear-to-cytoplasmic ratio and extended capacity for self-renewal. Subsequent morphological, histochemical, and immunochemical analysis after the induction of differentiation revealed phenotypic markers characteristic of multiple cell types of mesodermal origin, such as skeletal muscle, smooth muscle, fat cells, cartilage, and bone. These results indicate that this clone consists of pluripotent mesenchymal stem cells. This report demonstrates that clonal populations of reserve stem cells are present in mammals after birth. Potential roles for such cells in the maintenance, repair, and regeneration of mesodermal tissues are discussed.

BAFF mediates survival of peripheral immature B lymphocytes.

B cell maturation is a very selective process that requires finely tuned differentiation and survival signals. B cell activation factor from the TNF family (BAFF) is a TNF family member that binds to B cells and potentiates B cell receptor (BCR)-mediated proliferation. A role for BAFF in B cell survival was suggested by the observation of reduced peripheral B cell numbers in mice treated with reagents blocking BAFF, and high Bcl-2 levels detected in B cells from BAFF transgenic (Tg) mice. We tested in vitro the survival effect of BAFF on lymphocytes derived from primary and secondary lymphoid organs. BAFF induced survival of a subset of splenic immature B cells, referred to as transitional type 2 (T2) B cells. BAFF treatment allowed T2 B cells to survive and differentiate into mature B cells in response to signals through the BCR. The T2 and the marginal zone (MZ) B cell compartments were particularly enlarged in BAFF Tg mice. Immature transitional B cells are targets for negative selection, a feature thought to promote self-tolerance. These findings support a model in which excessive BAFF-mediated survival of peripheral immature B cells contributes to the emergence and maturation of autoreactive B cells, skewed towards the MZ compartment. This work provides new clues on mechanisms regulating B cell maturation and tolerance.

Relationship of small bowel motility to ileoanal reservoir function.

Some patients with an ileoanal reservoir have a high defecation frequency, despite a good anatomical result and the absence of pouchitis. This study aimed to determine whether variation in function is related to a difference in small bowel motility proximal to the reservoir and if small bowel motility is propagated into the reservoir. Ambulatory small bowel and reservoir motility was studied for 24 hours in five patients with good function (median bowel frequency 4 per day, range 3-6) and seven subjects with poor function (median bowel frequency 12 per day, range 10-20). Five solid state pressure sensors were positioned in the small bowel and one in the reservoir. During the fasting nocturnal period (2300-0800 h), patients with poor function had a median of 10 (range 5-13) migrating motor complexes (MMC), significantly greater (p = 0.03) than the corresponding median number of 3 (range 2-7) in patients with good function. A total of 120 MMCs were observed in the whole series of 12 patients. Of these only two were propagated from the small bowel into the reservoir. Discrete clustered contractions were not propagated into the reservoir, although prolonged propagated contractions did pass into the reservoir in one patient. Patients with poor function had similar 24 hour stool output and radiological reservoir size to those with good function, but the median maximum tolerated volume on reservoir distension was 290 ml (range 160-450) for patients with poor function compared with 475 ml (range 460-550) for patients with good function (p = 0.005). Small bowel motility proximal to the reservoir bears an important relationship to pouch function and defecation frequency. Propagation of coordinated proximal small intestinal motility into the reservoir is rare.

Pouch-vaginal fistula.

Between January 1976 and June 1991, 161 women underwent restorative proctocolectomy. Seventeen (10.6 per cent) developed pouch-vaginal fistula. There was no difference in the incidence of inflammatory (15 of 141; 10.6 per cent) and non-inflammatory (two of 20; 10 per cent) disease, the severity of colitis or the number of stages in the operation. In 15 patients the fistula involved the ileoanal anastomosis; in two it had possibly occurred at the dentate line. There was no significant difference in the proportion of hand-sewn (ten of 120; 8.3 per cent) and stapled (seven of 41; 17 per cent) ileoanal anastomosis. Of the 17 women, six had pelvic sepsis in the immediate postoperative period and five had an anastomotic complication. There was no case of Crohn's disease. Three patients developed a pouch-vaginal fistula before closure of the ileostomy. The remaining 14 fistulas occurred a median of 7 (range 1-144) months after closure. Five further patients were referred from elsewhere to give a total of 22 for analysis of treatment and outcome. The fistula developed before ileostomy closure in five patients (group 1) and after closure in 17 (group 2). Following treatment, six fistulas had healed with five patients still undergoing treatment. In group 1, three of the five fistulas healed, whereas in group 2 only three of the 17 healed. Eight patients required excision of the pouch. The prognosis appears to be worse when pouch-vaginal fistula occurs after ileostomy closure. The optimal management is not yet established.

Anti-endothelial cell antibodies in inflammatory bowel disease.

Antibodies to endothelial cells may be involved in the pathogenesis of vasculitic disorders. In view of recent evidence implicating intestinal vascular injury in the pathogenesis of inflammatory bowel disease, we have sought anti-endothelial cell antibodies in affected patients, examined their relationship to vascular injury, and tested their ability to mediate endothelial cell cytotoxicity in vitro. Anti-endothelial cell antibody levels were elevated in ulcerative colitis (P < 0.0001) and Crohn's disease (P < 0.05) compared with healthy controls. In ulcerative colitis, anti-endothelial cell antibody levels were related to disease activity and correlated with circulating levels of von Willebrand factor (r = 0.58, P < 0.01), a marker of vascular injury. Anti-endothelial cell antibodies, however, were not directly cytotoxic to endothelial cells in vitro. These data indicate, for the first time, an association between anti-endothelial cell antibody levels and vascular injury in vivo and suggest that they may be important in the pathogenesis of inflammatory bowel disease, particularly ulcerative colitis.

Ileoanal pouch compliance and motor function.

Measurement of intrapouch pressure during continuous distension with water (pouchmetrography) was performed in ten patients with good pouch function after restorative proctocolectomy and a 'normal' baseline pressure curve was constructed from the mean pressures at 50-ml intervals. Eight other patients with poor pouch function were studied and the two groups were compared. Patients with poor function had significantly lower maximum tolerated volumes (297.5 versus 565 ml, P less than 0.02) and volumes which caused urgency (135 versus 265 ml, P less than 0.02). Baseline pressure curves were above the upper limit of the normal range (mean plus two standard deviations) for a substantial proportion of the recording in six of the patients with poor function. Large, isolated contraction waves were recorded in six of ten patients with good function and in three of those with poor function. Rhythmic waves were frequently seen in both groups but were more prominent in patients with poor function. Pouchmetrography is a provocative test of pouch motor function which may unmask abnormal muscle activity resulting from reduced compliance or a primary motility disorder.

The correspondence between temperament and peer interactions for normally developing and mildly delayed preschool children.

The correspondence between temperament characteristics based on the Behavioural Style Questionnaire and the peer relations of normally developing and mildly developmentally delayed preschool-age children was examined in this study. Measures of the peer relations of 64 3- and 4-year-old boys were obtained through an analysis of children's social interactions in a series of specially designed playgroups. Maternal temperament ratings were found to be similar for the younger (3-year-olds) and older (4-year-olds) normally developing children as well as for the mildly delayed group (4-year-olds). Multiple regression analyses indicated that maternal-rated temperament dimensions of persistence and activity level were associated in a logical pattern with both the positive and negative aspects of children's peer relations. Issues related to the construct validity and situation-specificity of the temperament measure were discussed.

Pediatricians' perceptions of the effectiveness of early invention for at-risk and handicapped children.

To evaluate pediatricians' perceptions of the effectiveness of early intervention for handicapped and at-risk children, all board-certified pediatricians in the state of Ohio were surveyed. A detailed questionnaire was developed which included the use of brief but specific cases identifying eight different children with documented handicaps and two cases of children at developmental risk. Overall, the results indicated that pediatricians judged early intervention to produce at least a modest effect on children's general development and function. A more optimistic perspective was held for families in that involvement in early intervention was viewed as improving substantially the ability of families to cope with the problems of a handicapped or at-risk child and to provide a supportive and stimulating environment. However, perceptions of effectiveness did vary for specific disability and at-risk groups. The correspondence between perceived effectiveness and existing research findings also was discussed.

The peer relations of mildly delayed and nonhandicapped preschool children in mainstreamed playgroups.

Previously unacquainted groups of normally developing and mildly developmentally delayed preschool-age boys (N = 64) were brought together to form a series of 8 mainstreamed playgroups. Each playgroup consisted of 3 normally developing 3-year-olds, 3 normally developing 4-year-olds, and 2 mildly developmentally delayed 4-year-olds. The delayed children were matched with the normally developing older group for chronological age and with the normally developing younger group for developmental level. Each playgroup operated 5 days per week for 2 hours per day for a 4-week period. During that time, the peer-related social and play interactions of each child were videotaped, and peer sociometric ratings were obtained at the completion of each playgroup. Analyses of social participation and individual social behavior measures revealed that the analogue playgroup setting was appropriate for evaluating peer interactions, as expected developmental patterns emerged despite the presence of children heterogeneous with respect to chronological age and developmental status. The existence of a deficit in peer-related social interactions for mildly delayed children was supported in this investigation--a deficit that could not be attributed to reputational factors, the unavailability of responsive peers, inadequate matching procedures, unusual sample characteristics, or similar factors. Selected observational measures, peer preference patterns during free play, and peer sociometric ratings also indicated that the delayed children were perceived to be less competent and of lower social status. However, despite their relative isolation, important developmental opportunities were available for mildly delayed children in the mainstreamed playgroups. Possible processes responsible for these outcomes were discussed.

Dyadic peer interactions of mildly delayed and nonhandicapped preschool children.

Mildly developmentally delayed and nonhandicapped 3- and 4-year-old children were paired systematically in a series of dyadic play sessions to evaluate the effects of companion status on important aspects of peer-related social and play behavior. Mildly delayed children were paired with younger nonhandicapped children matched in terms of developmental level, with nonhandicapped children matched in terms of CA, and with other mildly delayed companions. Mixed-age and same-age pairings for the nonhandicapped children were also arranged. Results indicated that mildly delayed children's peer interactions improved substantially when paired with nonhandicapped older children in comparison to pairings with other mildly delayed children. Pairings with nonhandicapped younger children, although matched in terms of developmental level, had no influence on the peer interactions of mildly delayed children. Nonhandicapped children appeared to be able to maintain a consistent level of interaction irrespective of companion status. Explanations for these findings in terms of the directive role adopted by nonhandicapped older children and their developmental implications were discussed.

Correlates of peer-related social competence of developmentally delayed preschool children.

The peer-related social interactions of 33 developmentally delayed preschool children were examined. Measures of social participation and individual social behaviors were obtained during free-play periods and correlated with assessments of language development, MA, and teacher-rated social competence and behavior problems. Results suggested the existence of major deficits in peer-related social interactions for delayed children and the absence of specific individual social behaviors highly associated with peer-related social competence. Mental age was positively correlated with social play but unrelated to not playing at all. Although language comprehension was related to social interaction, expressive language did not correlate with any of the key measures. Teacher-rated behavior problems were associated with not playing, even when MA was controlled. Findings were discussed in terms of the congruence between the developmental processes of normally developing and delayed children as well as their clinical implications.

[Experimental infection of fattening swine with rotavirus: study of local immunity using coproantibodies]. / Infection expérimentale du porc à l'engrais par le rotavirus: étude de l'immunité locale par les coproanticorps.

Fattened pigs were experimentally infected with porcine rotavirus (OSU strain). Viral antigens found in the faeces were analysed over a period of two months (ELISA method). We have found no correlation between virus excretion and the clinical manifestation of disease. The local immunity has been analysed in the faeces by the coproantibodies antirotavirus. The different classes of antibody, G, M, A and immune complexes have been determined using the ELISA method. The level of IgA antibodies was higher (10 fold) than IgG and IgM, and the timing of the appearance of these antibodies antivirus was also different. No correlation was found between local immunity and virus excretion in the faeces.

Cardiac arrhythmias during outpatient dental anaesthesia: comparison of halothane with enflurane.

In 75 young female patients undergoing extraction of 3rd molar teeth during halothane or enflurane anaesthesia, the electrocardiogram was recorded on magnetic tape and analysed subsequently for arrhythmias, using a high-speed analyser. Enflurane induced a much lower frequency of arrhythmia during surgery than halothane, but there was otherwise little difference between the two drugs in the quality of anaesthesia or recovery. Many arrhythmias occurred before exposure of the patient to enflurane or halothane; the significance of this is discussed.

Hypotension following cimetidine administration during cardiopulmonary bypass.

Intravenous administration of cimetidine may occasionally cause profound hypotension. Cimetidine 200 mg was administered as a bolus injection to patients whilst on cardiopulmonary bypass and subsequent changes in systemic arterial pressure were recorded. A statistically significant fall in arterial pressure was observed (p less than 0.001), which was attributable to a fall in systemic vascular resistance.

Multidirectional in vivo strain analysis of the equine radius and tibia during dynamic loading with and without a cast.

Rosette strain gauges were applied to the equine radius and tibia. Three sites were examined on each bone on separate occasions (proximal metaphysis, middiaphysis, and distal metaphysis). At each site, 4 rosette gauges were applied around the bone (ie, cranial, caudal, medial, and lateral). Strain recordings were made while walking the horse with and without a full-limb plaster cast. The principal axis of tensile strain was on the craniolateral aspect of the radius. Distally, on the radius, the largest strains were torsional. The cast changed the principal axis of tensile strain on the radius from the craniolateral surface to the caudal surface. The principal tensile strain of the tibia was just to the lateral side of cranial in the proximal and diaphyseal regions. Distally, the principal axis of tensile strain was craniolateral; however, the largest strains measured here were torsional. The cast changed the principal axis of tensile strain on the distal metaphysis of the tibia, but it did not reduce the magnitude of the strains measured. Casts may not aid stabilization of radial or tibial fractures repaired with internal fixation and may in fact place additional stress on the fracture site.