Epidemiology of the Bacteroides fragilis group in the colonic flora in 10 patients with colonic cancer

We report the relative frequencies of members of the Bacteroides fragilis group in the faeces, in colon lavage fluid obtained pre-operatively, and in colonic tissue specimens obtained at operation from 10 patients with colonic cancer. B. vulgatus was the most and B. fragilis and B. ovatus were the least frequently isolated Bacteroides spp. in the faeces of the 10 subjects. B. uniformis and B. thetaiotaomicron ranked second and third in the faeces. The relative frequencies of all species except B. fragilis were lower in the lavage fluid and in cultures of mucosa. The relative frequency of B. fragilis increased from 4% in faeces to 39% in the final lavage fluid and to 42% in the colonic mucosa culture. Our results suggest that B. fragilis has a more intimate association with the gut mucosa than other members of the B. fragilis group, which might be one explanation for the high incidence of this species in gut-associated intra-abdominal infections.     

Human faecal immunoglobulins in healthy infants and children, and in some with diseases affecting the intestinal tract or the immune system.

IgA, IgG and IgM is faeces were quantified by single radial immunodiffusion using extracts of freeze-dried faeces. IgA in small specimens of faeces seemed to mirror the total amound of IgA secreted into the gut at the time of sampling. Presumptive normal values of faecal IgA concentrations in infants and children were established. Agglutinins to rabbit erythrocytes served as markers for the antibody activity. Infants and children just recovered from enteritis of probably infectious origin had higher concentrations of both IgA and agglutinins in faeces. Faeces from three out of five patients with ulcerative colitis in remission, contained IgG in markedly increased concentrations. Two patients with IgA deficiency had no detectable IgA in faeces, but had increased levels of faecal IgM which also agglutinated rabbit erythrocytes. Immunoglobulins were not demonstrated in faeces from three patients with agammaglobulinaemia. The findings indicate that faeces can be used for assay of immunoglobulins of the intestinal tract.     

Detection of antigen-specific antibodies on lung tissue in a patient with hypersensitivity pneumonitis

Summary A patient with hypersensitivity pneumonitis showed positive Ouchterlony's immunodiffusion tests against pigeon faecal extract, Cephalosporium and Pullularia antigens. Deposits of immunoglobulins - IgG and IgM antibodies - were detected in a subendothelial position in arterial and venous vessels and on alveolar macrophages in the lung tissue. The IgG desposits in blood vessels belonged to IgG₁, IgG₂ and IgG₃ subclasses and the absorbed IgG on alveolar macrophages to all IgG subclasses. The detection of allergen specific antibodies in lung tissue was made by indirect immunofluorescent staining with FITC conjugated antigen extracts from pigeon faeces and demonstrated the aetiology of this hypersensitivity pneumonitis.Abbreviation HP hypersensitivity - FITC fluorescein-isothiocyanate - PBS phosphate buffered saline     

An investigation of possible immunologic reactions to human serum albumin used as a stabilizer in allergy extracts

Two hundred fifteen patients who had received allergy immunotherapy for at least 1 year with an extract preparation containing 0.03% human serum albumin (HSA) were tested for evidence of an immunologic reaction to HSA by immediate skin testing. Sera from 39 of these subjects and from control subjects were studied in a microtiter enzyme-linked immunosorbent assay for anti-HSA antibodies. The 39 subjects' sera were examined by immunoelectrophoretic analysis for evidence of "tailing albumin" indicative of HSA bound to immunoglobulins. All studies were negative. We conclude that in these patients HSA-containing allergy extracts used in immunotherapy failed to induce the production of specific antibodies against HSA. This suggests that HSA was a safe extract stabilizing agent for allergy extracts in these representative allergic patients.     

Humoral immune responses within the lung of bone marrow transplant recipients studied by bronchoalveolar lavage.

We investigated 20 bone marrow transplant recipients with pneumonitis using bronchoalveolar lavage (BAL) to assess the humoral immune response in the lung. We measured the levels of total IgG, IgM and IgA in bronchoalveolar lavage fluid and serum, and albumin measurements were used to correct for simple diffusion of immunoglobulins from serum into the lung. We found evidence for the local production of immunoglobulins G, M and A in 15 patients. This was independent of the cause of the pneumonitis. We also found that, although seven patients who recovered from their pulmonary problem had evidence of considerable local production of immunoglobulin, eight patients who died were also producing immunoglobulins in the lung. Death due to pneumonitis in BMT recipients cannot, therefore, be ascribed to a failure of the local humoral immune response.     

Antibody responses and interferon titers in the respiratory tracts of mice after aerosolized exposure to influenza virus.

We studied the temporal appearance of immunoglobulins (immunoglobulins G1, G2, M, and A) and interferon in lung lavage fluids of mice after aerosol exposure to influenza virus in six animal groups in which mortality rates ranged from 0 to 24%. Immunoglobulin levels in the lung lavage fluids were markedly higher in mouse groups with higher mortality rates (16, 20, and 24%) than in those with low mortality rates (0, 2.5, and 7.5%). Analysis of serum albumin in the respiratory secretions as an index of edema indicated that increased immunoglobulin G levels during the early phase of infection were due to increased vascular permeability. The detection of virus-neutralizing antibodies and antibodies reactive with influenza virus antigens in the lavage fluids at 6 to 8 days postinfection suggested local immunoglobulin synthesis as a result of antigenic stimulation. Both systemic and local antibody productions contributed to immunoglobulin levels in the respiratory secretions after aerosolized influenza virus infection. Peak levels of interferon in the lavage fluids were reached before detection of significant levels of virus-neutralizing antibody in the serum or the lung lavage.     

Procollagen III peptide in bronchoalveolar lavage fluid. A potential marker of altered collagen synthesis reflecting pulmonary disease in sarcoidosis

Type III procollagen N-terminal peptide was not detectable in bronchoalveolar lavage fluid from healthy volunteers but was present in fluid from the majority of patients with pulmonary sarcoidosis (N = 110); the mean concentration was 0.6 micrograms/liter returned fluid, range less than or equal to 0.2 to 19.5 or, expressed in relation to the amount of albumin recovered, 9.5 mg/gm albumin (range less than or equal to 1 to 45). The serum concentrations in the patients with sarcoidosis were normal. Significant inverse correlations were found between lavage fluid procollagen peptide and vital capacity (p less than 0.001), forced expiratory volume (p less than 0.01), and diffusion capacity (p less than 0.01). Lavage fluid procollagen peptide was also related to pulmonary radiological findings (p less than 0.001) and serum levels of angiotensin converting enzyme (p less than 0.001). These findings support the hypothesis that procollagen peptide in lavage fluid is a potential marker of activated pulmonary fibroblasts or an expanded fibroblast mass associated with interstitial lung fibrosis.     

Tumor necrosis factor alpha-induced pulmonary vascular endothelial injury.

Tumor necrosis factor alpha (TNF-alpha) mediates components of the acute-phase response, stimulates granulocyte metabolism, and induces endothelial cell surface changes. We studied whether human recombinant TNF-alpha (rTNF-alpha) could increase pulmonary edema formation and pulmonary vascular permeability. Rabbits preinfused with 125I-albumin were administered rTNF-alpha or saline. Animals were sacrificed, and lung wet/dry weight ratios as well as bronchoalveolar lavage fluid and plasma 125I activities were determined. rTNF-alpha increased lung wet/dry weight ratios by 151% (P less than 0.02) and bronchoalveolar lavage fluid/plasma 125I activity ratios by 376% (P less than 0.01) compared with values for saline controls. Electron microscopy of lung sections demonstrated endothelial injury, perivascular edema, and extravasation of an ultrastructural permeability tracer. To demonstrate that rTNF-alpha could directly increase pulmonary vascular endothelial permeability in vitro, we studied albumin transfer across cultured porcine pulmonary artery endothelial cell monolayers. rTNF-alpha induced time-dependent dose-response increments in transendothelial albumin flux in the absence of granulocyte effector cells. These observations suggest that rTNF-alpha can provoke acute pulmonary vascular endothelial injury in vivo as well as in vitro.     

Quantitation of Sperm Bindable IgA and IgG in Seminal Fluid

ABSTRACT: Seminal fluid and serum from 95 infertile males were assayed for sperm bindable immunoglobulins using an indirect ELISA with whole target sperm. The ELISA method was compared to seminal fluid and serum immobilization and agglutination assays (functional assays). In this infertile group, the ELISA assay was positive in 22% of seminal fluids (>1.2 fg IgA/ sperm and >0.3 fg IgG/sperm). The seminal fluid antibodies were IgA and had an accompanying elevated IgG component in 78% of patients. There was a 96% correlation between negative seminal fluid functional assays and negative ELISA, and a 95% correlation between positive seminal fluid functional assays and positive ELISA. Positive serum sperm antibody tests were found in 71% of the infertile males with positive seminal fluid sperm antibodies, but 29% of the infertile males with strongly positive IgA seminal fluid sperm antibodies showed normal levels of serum sperm antibodies by either ELISA or functional assays. The ELISA method gives reproducible quantitation of sperm antibodies in seminal fluid and correlates well with accepted functional assays. Comparisons with serum sperm antibody assays suggests that seminal fluid sperm antibody analysis complements the serum analysis of sperm antibodies.     

Virus-specific immune response in the lungs of mice infected with influenza virus.

The time course of primary humoral immune response in NFS/N mice infected with the adapted influenza virus A/Aichi 2/68(H3N2) was followed by determination of the different class immunoglobulins in lungs, lung washings, and in blood serum. The quantity of antibody-producing cells (APC) was estimated by local haemolysis in gel. The presence of antibodies in serum and lavage fluid was tested by the methods of radial haemolysis and radial immunodiffusion. It was shown that the local immune response had developed earlier than systemic antibodies occurred in the serum.     

Immunoglobulin and antibody levels in bronchoalveolar lavage fluid from symptomatic and asymptomatic pigeon breeders.

Twenty-one symptomatic subjects with pigeon breeders' lung (PBL) and 10 asymptomatic pigeon breeders, with a similar exposure to pigeon antigens, underwent bronchoalveolar lavage. Total IgG, IgM and IgA in lavage fluid were determined as were specific antibody levels against antigens in pigeon serum and droppings. Results were converted to levels in epithelial lining fluid (ELF) using lavage and serum urea ratios. It was found that symptomatics represent a group that is hyperreactive to pigeon antigens compared with the asymptomatic group with significantly higher IgG, IgM, IgA levels as well as specific antibody levels against pigeon serum and droppings. Paired serum and ELF samples from 12 symptomatic subjects showed significantly elevated IgG, IgM and IgA levels in ELF compared with serum when values were expressed in terms of albumin. This strongly supports the concept of local production of immunoglobulins within the lung after inhaling immunogens as opposed to their diffusion from the vasculature. Results for IgA indicate that any putative protective role for this immunoglobulin is not valid in relation to the prevention of extrinsic allergic alveolitis. Analysis of smoking habits, lung immunoglobulins and response to inhalation challenge confirm the negative influence of smoking on total and functional lung immunoglobulins; however, levels in the ELF of ex-smokers suggest that the effect of smoking is not permanent. Smoking did not prevent responses to inhalation challenge.     

Use of Pseudomonas aeruginosa lipopolysaccharide immunoadsorbents to prepare high potency, mono-specific antibodies

Potent, mono-specific anti-Pseudomonas immunoglobulins were isolated from serum and lung lavage fluid from patients with cystic fibrosis using immunotype specific Pseudomonas aeruginosa lipopolysaccharide (LPS) substituted immunoadsorbent gel. Iodinated monovalent Pseudomonas LPS somatic antigens, Fisher immunotypes, were used as ligands for two different insoluble gel matrices. LPS iodination, using the water insoluble chloroglycoluril reagent, permitted quantitation of the percent LPS bound as a ligand. The coupling efficiencies of epoxy-activated and cyanogen bromide-activated Sepharose matrices for various Pseudomonas immunotype specific LPS preparations were compared. Although each of the 7 LPS somatic antigens produced an equivalent amount of coupling, higher percentages of coupling were found using the cyanogen bromide-activated gel when compared to the epoxy-activated gel. IgG fractions prepared from cystic fibrosis sera and lung lavage fluid were passed through the LPS affinity gels, and Pseudomonas LPS immunotype specific antibodies were eluted. The presence of specific antibody activity against individual Pseudomonas immunotypes was determined with a passive micro-hemagglutination assay. Bronchial lavage fluid seemed to be as effective as serum as a source of Pseudomonas specific antibody. Use of such a LPS substituted gel permits direct recovery of Pseudomonas monospecific antibodies suitable for physical-chemical analyses and for biologic functional assays.     

Salivary IgA from the sublingual compartment as a novel noninvasive proxy for intestinal immune induction

Whole-saliva IgA appears like an attractive noninvasive readout for intestinal immune induction after enteric infection or vaccination, but has failed to show consistent correlation with established invasive markers and IgA in feces or intestinal lavage. For reference, we measured antibodies in samples from 30 healthy volunteers who were orally infected with wild-type enterotoxigenic Escherichia coli. The response against these bacteria in serum, lavage, and lymphocyte supernatants (antibody-in-lymphocyte-supernatant, ALS) was compared with that in targeted parotid and sublingual/submandibular secretions. Strong correlation occurred between IgA antibody levels against the challenge bacteria in sublingual/submandibular secretions and in lavage (r=0.69, P<0.0001) and ALS (r=0.70, P<0.0001). In sublingual/submandibular secretions, 93% responded with more than a twofold increase in IgA antibodies against the challenge strain, whereas the corresponding response in parotid secretions was only 67% (P=0.039). With >twofold ALS as a reference, the sensitivity of a >twofold response for IgA in sublingual/submandibular secretion was 96%, whereas it was only 67% in the parotid fluid. To exclude that flow rate variations influenced the results, we used albumin as a marker. Our data suggested that IgA in sublingual/submandibular secretions, rather than whole saliva with its variable content of parotid fluid, is a preferential noninvasive proxy for intestinal immune induction.     

Selective recruitment of eosinophils by substance P after repeated allergen exposure in allergic rhinitis

We have investigated the nasal response to substance P after pollen exposure in seasonal allergic rhinitic patients. Seven patients with strictly seasonal allergic rhinitis were studied during the pollen season, 24 h after nasal challenge with pollen. They received increasing doses of nebulized substance P (0 to 80 nmol) in each nostril. Responses were assessed by measurement of nasal airway resistance by posterior rhinomanometry and quantification of albumin, histamine, and inflammatory cells in the nasal lavage fluid. Nasal airway resistance increased in a dose-dependent manner after substance P challenge. Protein and albumin in nasal lavage fluids increased after administration of substance P: from 2.6 ± 0.3 to 6.8 ± 1.1 mg for protein (P≤0.01) and from 0.2 ± 0.1 to 3.1 ± 0.6 mg for albumin (P≤0.02). Expressed as a percentage of total protein, albumin increased from 10.5 ± 3.6% to 39.9 ± 3.5% (P≤0.02), suggesting occurrence of plasma leakage. No histamine release was observed after challenge with substance P. Total cell counts significantly increased from 11.4 ± 2.4 to 41.8 ± 17.3 × 10³ cells/ml after substance P (P≤0.05). Eosinophils were already numerous before substance P challenge (2.1 ± 0.7 × 10³ cells/ml), and the number of eosinophils markedly increased in all patients after substance P (for the whole group, 25.8 ± 13.3 cells/ml, P≤0.05). In contrast, the number of neutrophils only slightly increased in five patients, and changes did not reach significance for the group as a whole. Our results show that substance P induces nasal obstruction and albumin extrusion in allergic rhinitic patients after repeated pollen exposure. These vascular phenomena are associated with recruitment of eosinophils. Since substance P is known to be released after nasal allergen challenge, our data suggest a role for substance P in the chronic eosinophilic inflammation of the nasal mucosa observed in symptomatic allergic rhinitis.     

Origin of immunoglobulins in respiratory tract secretion and saliva of sheep.

The origin of the immunoglobulins in the upper respiratory tract secretion of sheep was determined by measuring the distribution between plasma and secretion of radiolabelled purified immunoglobulins and albumin. By calculation of the ratio of specific activity for each immunoglobulin between plasma and secretion, it was estimated that about 81% of IgA in secretion was of local origin, whereas IgM, IgG1, IgG2 and albumin were wholly derived from plasma. Estimates of the selectivity of transport of IgA and IgM into both respiratory tract secretion and saliva were obtained by calculation of a selective index relative to IgG1 or IgG2, which do not bind secretory component (SC). This was based on radioactivity ratios after the simultaneous injection of immunoglobulin labelled with different isotopes (IgA or IgM injected with either IgG1 or IgG2). These calculations revealed that both IgA and IgM were selectively transported into respiratory tract secretion and saliva. This provides further support for the proposition that SC-binding immunoglobulins may be transported from serum into secretions at a variety of mucosal sites dependent on SC availability. Since the IgA in serum of sheep is predominantly of gut origin, this provides an opportunity, in addition to relocation of gut-derived plasma cell precursors, by which the gut may contribute to extraintestinal mucosal responses.     

Comparison of 7.2% hypertonic saline - 6% hydroxyethyl starch solution and 6% hydroxyethyl starch solution after the induction of anesthesia in patients undergoing elective neurosurgical procedures

OBJECTIVE:

The ideal solution for fluid management during neurosurgical procedures remains controversial. The aim of this study was to compare the effects of a 7.2% hypertonic saline - 6% hydroxyethyl starch (HS-HES) solution and a 6% hydroxyethyl starch (HES) solution on clinical, hemodynamic and laboratory variables during elective neurosurgical procedures.

METHODS:

Forty patients scheduled for elective neurosurgical procedures were randomly assigned to the HS-HES group or the HES group. After the induction of anesthesia, patients in the HS-HES group received 250 mL of HS-HES (500 mL/h), whereas the patients in the HES group received 1,000 mL of HES (1000 mL/h). The monitored variables included clinical, hemodynamic and laboratory parameters. Chictr.org: ChiCTR-TRC-12002357

RESULTS:

The patients who received the HS-HES solution had a significant decrease in the intraoperative total fluid input (p<0.01), the volume of Ringer''s solution required (p<0.05), the fluid balance (p<0.01) and their dural tension scores (p<0.05). The total urine output, blood loss, bleeding severity scores, operation duration and hemodynamic variables were similar in both groups (p>0.05). Moreover, compared with the HES group, the HS-HES group had significantly higher plasma concentrations of sodium and chloride, increasing the osmolality (p<0.01).

CONCLUSION:

Our results suggest that HS-HES reduced the volume of intraoperative fluid required to maintain the patients undergoing surgery and led to a decrease in the intraoperative fluid balance. Moreover, HS-HES improved the dural tension scores and provided satisfactory brain relaxation. Our results indicate that HS-HES may represent a new avenue for volume therapy during elective neurosurgical procedures.     

Immunoglobulins and Other Serum Proteins in Feces from Infants and Children

IgA in variable amounts was found in extracts of feces from all of 14 infants and children with no apparent diseases involving the gastrointestinal tract or the immune system, whereas IgG and IgM were only found in some. No age variation was evident's even one-month-old infants seemed fully able to secrete intestinal immunoglobulins. Freeze-drying of the feces facilitated the extraction, and the amounts of immunoglobulins were related to the dry weight of fecas. Most of the immunoglobulins were easily dissolved in saline. Of other serum proteins α₁ antitrypsin was constantly present; α₁ antichymotrypsin and α₂ macroglobulin were found in many. Only traces of albumin were sometimes demonstrated.     

A Staphylococcus aureus capsular polysaccharide (CP) vaccine and CP-specific antibodies protect mice against bacterial challenge.

The efficacy of capsular polysaccharide (CP)-specific antibodies elicited by active immunization with vaccines composed of Staphylococcus aureus types 5 and 8 CP linked to Pseudomonas aeruginosa exoprotein A or with immune immunoglobulin G (I-IgG) obtained from vaccinated plasma donors was tested in lethal and sublethal bacterial mouse challenge models. A dose of 2 x 10(5) CFU of S. aureus type 5 CP per mouse administered intraperitoneally (i.p.) with 5% hog mucin was found to cause 80 to 100% mortality in BALB/c mice within 2 to 5 days. Mice passively immunized i.p. 24 h earlier or subcutaneously 48 h earlier with 0.5 ml of I-IgG showed significantly higher average survival rates than animals receiving standard IgG or saline (P < 0.01) following the bacterial challenge. Animals actively immunized with the monovalent type 5 CP-P. aeruginosa exoprotein A conjugate showed a survival rate of 73% compared with 13% in phosphate-buffered saline-immunized animals. The prechallenge geometric mean titer of type 5 CP antibodies in animals that died was significantly (P < 0.05) lower than that of animals which survived the challenge (95.7 versus 223.6 micrograms/ml, respectively). The IgG was further evaluated in mice challenged i.p. with a sublethal dose of 5 x 10(4) CFU per mouse. Serial blood counts were performed on surviving animals at 6, 12, 24, and 48 h. Surviving animals were sacrificed at 72 h, and bacterial counts were performed on their kidneys, livers, and peritoneal lavage fluids. Animals receiving I-IgG had lower bacterial counts in blood samples and lower bacterial densities in kidneys, livers, and peritoneal lavage samples than mice immunized with standard IgG (P < 0.05). These data suggest that S. aureus type 5 CP antibodies induced by active immunization or administered by passive immunization confer protection against S. aureus infections.     

Occupational rhinitis and bronchial asthma due to morphine: evidence from inhalational and nasal challenges

A case of occupational bronchial asthma due to morphine in a nonatopic 46-year-old woman is presented. The following diagnostic tests were used: a workplace trial with bronchodilator and placebo, and single-blind, placebo-controlled nasal and bronchial challenge with 0.5% morphine HC1. For the nasal challenge, four asthmatic patients were selected as a control group. The nasal washings were done before and 30 min, 3 h, 24 h, and 48 h after all challenges. In the nasal lavage fluid, the total numbers of eosinophils, neutrophils, basophils, and mast cells were counted, and, after the nasal challenge, total protein and albumin levels were measured. During the workplace trial, the PEF variability ratio increased from 5% to 38%. After the challenges, a decrease in the spirometric parameters (VC and FEV) of about 30–40% was observed, with minimums at 24 and 48 h. An influx of granulocytes with an increase in the relative number of eosinophils and basophils from 3 h until 48 h after the challenge was observed in the nasal lavage fluid. The protein level in the nasal lavage fluid increased from 190 to 1275 μg/ml 24 h after the challenge with an increase of relative albumin level from 24% to 40% at 24 h. In the control group, no changes in relative number of basophils and eosinophils and albumin/total protein ratio in the nasal lavage fluid or in the spirometric parameters were observed after the challenge.     

Nasal secretory response to allergen provocation

Nasal provocation with ragweed pollen extract was performed on ragweed-sensitive and non-atopic subjects. Nasal lavage fluids were collected 15 min after saline and allergen challenges, and assayed for total protein, albumin, potassium, lysozyme activity and peroxidase activity. There was no statistically significant increase in any of these lavage fluid constituents in non-atopic subjects after allergen provocation, compared with after saline provocation. The lavage fluids of ragweed-sensitive subjects had significant rises in each of the constituents following allergen provocation. This method provides a simple mechanism for quantitating the nasal secretory response to allergen provocation.