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[From atopic eczema to respiratory allergy].

The prevalence of atopic eczema in childhood is increasing. This is probably related with a deviation of immune system to Th2 directed to allergen instead of Th2 directed to microorganisms. Atopic eczema is genetically linked, but its expression is principally determined by environmental factors, allergens and infections. In children with atopic eczema, the immune system is altered. The number of Langheran's cells is augmented. Th2 cells infiltrate the skin and produce cytokines which mediate both immediate and late allergic reaction. Most of eczematous children is allergic to food. A IgE-mediated reaction take place at the mucosal intestinal site. The food challenge test is the most reliable mean to identify offending foods. Soy formula can be used in the treatment of cow milk allergy. The tolerance of formulae based on cow milk hydrolysed protein should be tested under medical supervision in children with cow milk allergy. Prevention of atopic disease with dietary interventions has not been successful. To prevent asthma, the administration of immunotherapy in children with allergic rhinitis seems to be promising. In eczematous children with elevated total IgE and specific IgE to inhalant allergens, cetarizine seems to prevent onset of asthmatic symptoms.

Asthma↗

[Molecular and cellular bases of the virulence of Shigella flexneri].

Shigella flexneri, a Gram negative bacillus, causes bacillary dysentery, an ulcerative disease of the human colon, by invading intestinal epithelial cells. Entry into epithelial cells occurs via an induced phagocytic process which involves the actino-myosin complex. The host-cell receptor and the transmembrane signal which initiate reorganization of the cytoskeleton are under study. Binding to integrins has recently been demonstrated in related models such as the entry of Yersinia pseudotuberculosis and Bordetella pertussis into cells. Bacterial genes necessary to achieve entry are located on five contiguous loci covering 30 kb on a 220 kb virulence plasmid in S. flexneri. Locus 2 has been particularly studied. Six genes organized as an operon encode highly immunogenic proteins among which IpaB (62 kD) and IpaC (48 kD) are the invasins of this microorganism which subsequently grows very rapidly within infected cells due to its capacity to lyse the membrane bound phagocytic vacuole. Once free within the cytoplasm, bacteria interact again with the cell cytoskeleton. They first express Olm (organelle like movement), a phenotype reflecting intracellular movement along actin stress cables. They subsequently express lcs (intracellular spread), a phenotype by which intracellular bacteria induce nucleation and polymerization of actin followed by accumulation of this material at one end of the bacillus. This process causes rapid random movement leading to the formation of protrusions which allow passage to adjacent cells. A combination of these two movements achieves bacterial colonization of the epithelium.

Chromosome Mapping↗

Regression of rectal mucosa-associated lymphoid tissue lymphoma but persistence of Helicobacter pylori infection of gastric mucosa after administration of levofloxacin: report of a case.

PURPOSE: Several articles have reported regression of rectal lymphoma of mucosa-associated lymphoid tissue after anti- Helicobacter pylori therapy. It remains unclear, however, whether lymphoma of rectal mucosa-associated lymphoid tissue is related to Helicobacter pylori infection. METHODS: A 60-year-old woman visited our hospital with a complaint of hematochezia. On colonoscopy a pale, ulcerated protrusion approximately 3.5 cm in diameter was found in the rectum. Pathologic diagnosis of a biopsy specimen was low-grade mucosa-associated lymphoid tissue lymphoma. The patient received a ten-day course of levofloxacin, and the cycle was repeated three times over seven months. RESULTS: Although persistence of Helicobacter pylori infection was confirmed, the tumor regressed completely. CONCLUSION: Our findings in this case suggest that rectal lymphoma of mucosa-associated lymphoid tissue may be related to unknown microorganisms other than Helicobacter pylori. Levofloxacin may be effective for treatment of rectal lymphoma of mucosa-associated lymphoid tissue regardless of Helicobacter pylori infection.

Anti-Infective Agents↗

The influence of ofloxacin versus trimethoprim-sulfamethoxazole on the aerobic flora in granulocytopenic subjects.

Ofloxacin (300 mg twice a day) and trimethoprim-sulfamethoxazole (TMP-SMZ) (160 mg trimethoprim and 800 mg sulfamethoxazole twice a day) were given prophylactically to 19 adult patients with acute leukemia undergoing induction chemotherapy. The influence of the two regimens on the bacterial aerobic flora was evaluated. Both of the prophylactic regimens conditioned the aerobic microflora of the patients. Both groups acquired new microorganisms, prevalently gram-positive cocci, but also gram-negative bacteria with the TMP-SMZ regimen. Both treatment groups acquired yeasts.

Adult↗

Molecular cloning and sequencing of two phospho-beta-galactosidase I and II genes of Lactobacillus gasseri JCM1031 isolated from human intestine.

Lactobacillus (Lb.) gasseri JCM1031, which is classified into the B1 subgroup of the Lb. acidophilus group of lactic acid bacteria, characteristically produces two different phospho-beta-galactosidases (P-beta-gal) I and II in the same cytosol as reported in our previous papers [Biosci. Biotech. Biochem., 60, 139-141, 708-710 (1996)]. To clarify the functional and genetic properties of the two enzymes, the structural genes of P-beta-gal I and II were cloned and sequenced. The structural gene of P-beta-gal I had 1,446 bp, encoding a polypeptide of 482 amino acid residues. The structural gene of P-beta-gal II had 1,473 bp, encoding a polypeptide of 491 amino acid residues. The deduced relative molecular masses of 55,188 and 56,243 agreed well with the previous value obtained from the purified P-beta-gal I and II protein, respectively. Multiple alignment of the protein sequence of P-beta-gal I and II with those of P-beta-gals from 5 microorganisms had 30-35% identity on the amino acid level, but those with phospho-beta-glucosidases from 5 microorganisms had the relatively high identity of about 50%. Considering that this strain grows on lactose medium and shows no beta-galactosidase activity, and that purified P-beta-gal I and II can obviously hydrolyze o-nitrophenyl-beta-D-galactopyranoside 6-phosphate (substrate), and also the conservation of a cysteine residue in the molecule, the P-beta-gal I and II were each confirmed as a novel P-beta-gal enzyme.

Amino Acid Sequence↗

Probiotics: clinics and/or nutrition.

The primary claim of probiotics is their beneficial influence on the intestinal ecosystem, which, in turn, may provide protection against gastro-intestinal infections. The positive actions on human health include antagonistic activity against pathogens, anti-allergic effects and other effects on the immune system. In recent years, the concept of probiotics evolved from food with beneficial, even if not clearly established, effects on well-being, to the new area of the nutraceuticals or functional foods. This concept was developed further into the biotherapeutics, which are the clinical applications of probiotic microorganisms originally selected for food use. Whereas several of these health claims remain controversial, well-planned clinical trials increasingly support the claims for some carefully selected probiotic strains. Aim of this review is to provide a first attempt to evaluate these two areas of use of probiotics and to draw a borderline between them. Only a more in-depth knowledge of the mechanisms underlying the probiotic effects will allow a clear discrimination between the food use and the clinical use.

Bifidobacterium↗

Defensins: antimicrobial and cytotoxic peptides of mammalian cells.

Defensins are antimicrobial and cytotoxic peptides that contain 29-35 amino acid residues, including six invariant cysteines whose intramolecular disulfide bonds cyclize and stabilize them in a complexly folded, triple-stranded beta-sheet configuration. Generated by the proteolytic processing of 93-95 amino acid precursor peptides, the constitute > 5% of the total cellular protein in human and rabbit neutrophils (polymorphonucleated neutrophils--PMN) and are also produced by rabbit lung macrophages and by mouse and rabbit small intestinal Paneth cells. Despite their prominence in rat PMN, defensins are not found in murine PMN. The antimicrobial spectrum of defensins includes gram positive and gram negative bacteria, mycobacteria, T. pallidum, many fungi, and some enveloped viruses. Defensins exert nonspecific cytotoxic activity against a wide range of normal and malignant targets, including cells resistant to TNF-alpha and NK-cytolytic factor. They appear to kill mammalian target cells and microorganisms by a common mechanism, which involves initial electrostatic interactions with negatively charged target cell surface molecules (likely the head groups of polar membrane lipids), followed by insertion into the cell membranes which they permeabilize, forming voltage-regulated channels. In addition to their antimicrobial and cytotoxic properties, some defensins act as opsonins, while others inhibit protein kinase C, bind specifically to the ACTH receptor and block steroidogenesis or act as selective chemoattractants for monocytes. Defensins are a newly delineated family of effector molecules whose contribution to host defense, inflammation, and cytotoxicity may be considerable for humans, even though it is unlikely to be revealed by experimentation with mice.

Amino Acid Sequence↗

Microsporidia in the small intestine of HIV-infected patients. A new diagnostic technique and a new species.

OBJECTIVES: To determine whether microsporidian infections occur in Australian patients infected with human immunodeficiency virus (HIV), to assess the incidence, and to discuss microscopic detection methods. DESIGN AND PATIENTS: 180 consecutive HIV-infected patients (109 with chronic diarrhoea and 71 with other indications) underwent upper gastrointestinal tract endoscopy and pinch biopsies of the second part of the duodenum. The biopsies were handled by a protocol: four levels, with haematoxylin and eosin stain (H&E) at each level, periodic acid Schiff reagent after diastase (DiPAS) and auramine stain at the second level, and Warthin-Starry (WS) stain and cytomegalovirus early antigen immunoperoxidase study at the third level. Electron microscopy was carried out on samples from the first 95 patients, and thereafter from selected patients. SETTING: The patients came from the HIV Medicine Unit of a teaching hospital and from the practice of a gastroenterologist. MAIN OUTCOME MEASURES: Diagnosis of microsporidia was to based on the H&E stain, with electron microscopy as the definitive test because the microsporidia are often difficult to see with H&E. Empirically, the WS stain was found to stain the microorganisms and it replaced electron microscopy during the study as the screening diagnostic test. RESULTS: Microsporidia were present in 36 of the 109 patients with diarrhoea (33%) and one of 71 patients without diarrhoea. The WS stain in all cases showed developing spores in the enterocytes and in four cases in macrophages as well. The H&E stain showed non-specific duodenitis and was not diagnostic in some cases. Electron microscopy on samples from the first 95 consecutive patients showed 100% concordance with the WS stain. In 33 cases, electron microscopy showed the multinucleated plasmodia and the spores of Enterocytozoon bieneusi and in the four cases confirmed the spores in macrophages and showed a new Encephalitozoon-like species with a septate parasitophorous vacuole. Other causes of duodenal infection were cytomegalovirus (11 cases), mycobacteria (8), cryptosporidia (8) and Giardia lamblia (5). CONCLUSION: E. bieneusi was the commonest microorganism found in our series of 180 consecutive patients. The actual prevalence of the two microsporidia species within the HIV-positive population and general community awaits further study. The WS stain provides a sensitive diagnostic test for the presence of E. bieneusi and the new Encephalitozoon-like species, avoiding the cost and potential sampling error of electron microscopy. The detailed ultrastructure and taxonomy of the new species requires further study.

AIDS-Related Opportunistic Infections↗

Ecotoxicological profile of xylenes. Working Party on Ecotoxicological Profiles of Chemicals.

The xylenes (o, m, p), which are produced in large amounts and widely used, must be considered as environmental contaminants, as they have been identified in the atmosphere of many countries. Their acute and chronic toxicity in mammals is moderate. Mutagenic and teratogenic properties can be excluded. Carcinogenic potential has been at present inadequately studied. In man some disturbances are evident at the concentration of 45 ppm. Hepatic metabolism is a valid process of detoxification and its high rate limits the risk of accumulation of the xylenes in adipose tissue. Ecotoxicological data refer almost exclusively to water. No long-term tests have been made. However, the high volatility, elevated elimination, and the low factor of accumulation suggest the impact on aquatic environment should be limited. Xylenes are easily biodegraded by many microorganisms. Photoxydation is one of the main degradation processes, responsible for the disappearance of the xylenes from the atmosphere.

Animals↗

Changes in fecal flora and comparative multiple-dose pharmacokinetics of ceftibuten, cefpodoxime proxetil and amoxycillin/clavulanate.

OBJECTIVE: To investigate changes in fecal flora and multiple-dose pharmacokinetics with the oral antibiotics ceftibuten 400 mg daily and cefpodoxime proxetil (CPX) 200 mg every 12 h, compared to amoxycillin/clavulanate 500/125 mg every 8 h during and following 1 week of medication. METHODS: In an open randomized triple crossover design, 18 (nine female, nine male) healthy volunteers received each drug for 7 days, followed by a 'washout' period of 4 weeks. Serum and urine levels of the substances were determined by bioassay, and for ceftibuten isomers by high-pressure liquid chromatography. Statistical analysis of quantitative aerobic and anaerobic cultures of feces was performed, and beta-lactamase activity was determined. RESULTS: Ceftibuten showed a mean Cmax of 18.9 (SD 3.0) mg/L, a terminal half-life of 2.89 h, and an AUCtot of 100 (21.8) mg.h/L; protein binding was 63.7 (5.1)%, and accumulation was marginal. Cefpodoxime proxetil had a Cmax of 1.92 (0.61) mg/L, a terminal half-life of 1.97 (0.42) h and an AUCtot of 10.8 (3.3) mg.h/L; no accumulation was seen. Amoxycillin and clavulanate had Cmax values of 7.15 (2.16) mg/L and 3.39 (1.31) mg/L, terminal half-life values of 1.03 (0.15) h and 0.93 (0.17) h, AUCtot values of 20.0 (4.2) mg.h/L and 8.87 (3.10) mg.h/L, and there was no accumulation. Statistical analysis for ech microorganism in fecal samples showed significant differences between amoxycillin/clavulanate and the two third-generation cephalosporins, but virtually no differences between ceftibuten and cefpodoxime proxetil. Eleven of 12 volunteers reported loose stools (days 2-7, mean duration 4.4 (SD 2.7) days) with amoxycillin/clavulanate, but nobody during ceftibuten administration and one volunteer during cefpodoxime proxetil administration. CONCLUSIONS: Ceftibuten showed excellent and cefpodoxime favorable pharmacokinetic properties, with significantly less pronounced fecal flora changes and intestinal side effects compared to amoxycillin/clavulanate. The multiple crossover design allows powerful microbiological statistical analysis and pharmacokinetic parameter comparisons.

Journal Article↗

[Epithelial cells as sentinels in mucosal immune barrier].

The mucosal surface of the body is exposed to a vast array of exogenous antigens and microorganisms. Epithelial cells evoke minimal immune response to food ingredients and commensal bacteria, while they release an array of antimicrobial peptides and CXC chemokines in response to bacterial invasion or inflammatory stimuli. The mucosal antigens are transported from the gut lumen to organized lymphoid follicles by specialized epithelial M cells residing in follicle-associated epithelium (FAE). An alternative pathway of antigen uptake with neonatal Fc receptor (FcRn) is also reported. Furthermore, intestinal dendritic cells underneath epithelium directly take up luminal antigens, where epithelial fractalkine expression plays a critical role in the guidance of dendrite extrusion. Epithelial cells express polymeric Ig receptor (pIgR) that is essential for the luminal secretion of dimeric IgA produced in the lamina propria. Furthermore, soluble factors released by mucosal epithelial cells condition dendritic cells, which in turn promote Th2 response. These multiple lines of evidence clearly suggest the significant role of epithelial cells at the front line of mucosal immune defense.

Animals↗

The importance of Campylobacter jejuni to the meat industry: a review.

Campylobacter jejuni is a microorganism that only recently has been implicated in gastroenteritis in humans. As appropriate methods used for detection of the bacterium have been developed, the rates of illness caused by the pathogen were found to approach or surpass those attributed to Salmonella. Substantial evidence has been gathered to document that the route for human infection is through the ingestion of adulterated food and drink. Some slaughter animals harbor this potential pathogen among the intestinal flora and, consequently, transfer of the organism to carcasses and to the resulting meat products does occur. The most frequently implicated meat is poultry, with an incidence of recovery of C. jejuni from the store-bought poultry meat reported to be at least 50%. Red meat from slaughter animals have also yielded this bacterium from carcasses, but at lower incidence levels. Foodborne disease has been associated most frequently with the ingestion of raw milk, but poultry, hamburger, and other foods have all been implicated as potential sources. However, cause and effect relating the presence of C. jejuni in meat and human gastroenteritis has not been demonstrated. Additional research is needed to determine whether C. jejuni isolated from meat causes gastroenteritis and whether all strains of the organism are virulent. Recognition of C. jejuni as a potential meatborne pathogen by the meat industry is necessary, and appropriate sanitary practices to prevent passage of the organism through meat products should be implemented.

Adult↗

[The vaginal Bifidobacterium flora in women of reproductive age].

The composition of vaginal bifidoflora in 56 clinically healthy women of reproductive age was studied. The study revealed that four species of bifidobacteria, viz. Bifidobacterium bifidum, B. breve, B. adolescentis 2 and B. longum, dominated in the composition of this bifidobacterial population. Nine out of 11 isolated strains were found to be capable of inhibiting indicator microorganisms Staphylococcus aureus and Enterococcus faecalis when tested in vitro; in addition, strains B. adolescentis 2 F1, B. bifidum G1, B. breve P2 and B. longum Z4 inhibited Klebsiella ozaenae, Pseudomonas aeruginosa, Escherichia coli and were also active acid producers. Three of these 4 bifidobacterial strains were capable of adhesion to vaginal epitheliocytes, while B. bifidum G1 was practically incapable of adherence to these cells, similarly to B. bifidum strain 791 of intestinal origin. In addition, the spectra of antibiotic susceptibility varied from strain to strain, but all bifidobacterial strains were susceptible to benzylpenicillin and resistant to lomefloxacin, most of them being also resistant to cyprofloxacin and gentamicin. Thus the data presented in this work are indicative of the possibility and advantages of using bifidobacterial strains belonging to this ecological niche as probiotics for the correction of the microflora of the urogenital tract in females.

Adult↗

Clinical use of selective decontamination: the concept.

Infections can be classified according to: (1) the type of offending microorganism (virus, bacteria, fungi, parasites), (2) according to the clearance by the defence system (T cell dependent/independent) and (3) in case bacteria are the causative agents in Gram-positive and Gram-negative infections. The latter classification in Gram-positive and Gram-negative infections has appeared to have a practical consequence. Gram-negative bacteria, often involved in major infections and yeasts, appear to play practically no role in the intestinal ecological system. Consequently, it is nowadays increasingly attempted to eliminate Gram-negative bacteria and yeasts selectively from the digestive tract with antimicrobial agents. Selective suppression of Gram-positive bacteria may severely affect the ecosystem of the digestive tract. This selective suppression of Gram-negatives must be continued as long as patients are immunocompromised (locally or systemically) and is called selective decontamination of the digestive tract.

Anti-Bacterial Agents↗

Effects of the oral administration of the products derived from milk fermentation by kefir microflora on immune stimulation.

Nutritional status has a major impact on the immune system. Probiotic effects ascribed to fermented dairy products arise not only from whole microorganisms but also from metabolites (peptides, exopolysaccharides) produced during the fermentation. We recently demonstrated the immunomodulating capacity of kefir in a murine model. We now aimed at studying the immunomodulating capacity in vivo of the products derived from milk fermentation by kefir microflora (PMFKM) on the gut. BALB/c mice received the PMFKM for 2, 5 or 7 consecutive days. IgA+ and IgG+ cells were determined on histological slices of the small and large intestine. IL-4, IL-6, IL-10, IL-12, IFNgamma and TNFalpha were determined in the gut, intestinal fluid and blood serum. IL-6 was also determined in the supernatant of a primary culture of small intestine epithelial cells challenged with PMFKM. PMFKM up-regulated IL-6 secretion, necessary for B-cell terminal differentiation to IgA secreting cells in the gut lamina propria. There was an increase in the number of IgA+ cells in the small and large intestine. The increase in the number of IgA+ cells was accompanied by an increase in the number of IL-4+, IL-10+ and IL-6+ cells in the small intestine. Effects of PMFKM in the large intestine were less widely apparent than the ones observed at the small intestine lamina propria. All cytokines that increased in the small intestine lamina propria, also did so in blood serum, reflecting here the immunostimulation achieved in the gut mucosa. We observed that the PMFKM induced a mucosal response and it was able to up and down regulate it for protective immunity, maintaining the intestinal homeostasis, enhancing the IgA production at both the small and large intestine level. The opportunity exists then to manipulate the constituents of the lumen of the intestine through dietary means, thereby enhancing the health status of the host.

Administration, Oral↗

[Occurrence and characteristic of P-like adhesin among Klebsiella strains].

Colonisation and remaining of microorganism on mucus membrane of microorganism is tightly connected with adhesion mechanisms and determine the first step of physiological settlement of the organism or the first stage of clinically demonstrated infection. In Klebsiella rods there are known three types of fimbrial adhesins (type 1, 3 and KPF-28) and non-fimbrial adhesin CF29K. It is stated that Klebsiella strains adhesions are responsible for their adherence to the epithelial cells of both respiratory and urinary tracts and to intestine epithelium. The in vitro research affirmed Klebsiella rods adherence to protein matrix. The aim of our work was the establishment of character, receptor specificity and the appearance frequency of P-like called adhesin. The frequency of expression of P-like adhesin was estimated among 380 isolated from the patients strains on the basis of agglutinating methods. The amorphic character of P-like adhesin was proved using electron microscopy method. The isolation and purification of P-like protein with a help of affinity chromatography enabled to estimate the receptor specificity of the adhesin. The receptor specificity was established as similar to E.coli PapG adhesin.

Adhesins, Bacterial↗

[The presence of bacteriophages in human feces and their potential importance].

Bacteriophages are widely distributed throughout the environment as well as in the bodies of humans and animals (feces, urine, saliva, sputum). Higher presence of Escherichia coli phages compared with Bacteroides fragilis and Salmonella phages was noticed in the feces of healthy human individuals and patients, mainly those with gastro-intestinal tract diseases. A strict correlation exists between the number of bacteria and of phages in the feces of healthy individuals as well as of patients with different diseases. The presence of phages in human feces correlates with the character of the coexisting disease. The frequency of phages in the feces depends on the different indicator bacterial host strains and the numbers of indicator strains. The role of bacteriophages in protecting against pathogenic microorganisms and controlling bacterial flora in the human organism is of major significance.

Animals↗

Observations on the feeding mechanism, diet and digestive physiology of Histriobdella homari Van Beneden 1858: an aberrant polychaete symbiotic with North American and European lobsters.

1. The aberrant annelid Histriobdella homari (Polychaeta:Eunicida) lives in the branchial chambers of the marine lobsters Homarus americanus and H. vulgaris where it feeds on the rich microflora of bacteria, blue-green algae and related organisms which grow on the inner surface of the branchial chamber, the setae fringing the edges of the carapace, the gill filaments and, especially, the surfaces and setae of the epipodite plates between the gills. H. homari, therefore, is to be regarded as an epizoic microphagous cleaning symbiote of the lobsters. 2. The alimentary canal consists of mouth, buccal cavity, oesophagus, proventriculus, stomach, intestine and anus. The much-modified proboscis lies ventrally below the oesophagus and proventriculus, with its anterior portions protruding into the rear of the buccal cavity. 3. The proboscis consists of two fixed parallel mandibles, a transverse carrier which slides upon the mandibles and to which is attached, posteriorly, a median flexible dorsal rod and, anteriorly, four pairs of movable articulated maxillae, paired external and internal retractor muscles and various tensor, flexor and extensor muscles. 4. Contraction of the retractor muscles withdraws the carrier and maxillae posteriorly, causing bowing of the dorsal rod which is fixed at its posterior end. Relaxation of the muscles allows the rod to straighten and, thus, causes protraction of the carrier and protraction and lateral expansion of the maxillae. Contraction and relaxation of the relaxation of the retractor muscles are supplemented by appropriate changes in the other muscular components of the proboscis. 5. During feeding the serrated anterior ends of the mandibles are applied to the food, the maxillae are fully expanded and then dawn ventro-posteriorly toward the mid-line by contraction of the retractor muscles in the effective movement of the feeding mechanism. This draws the food organisms across the anterior ends of the mandibles, detaching them from the substratum and allowing their ingestion by ciliary action. The first pair of maxillae are also capable of independent action and can be used while the remainder of the proboscis apparatus is held in the protracted position. 6. Detached microorganisms are entangled in a sticky mucous secretion from the salivary glands; other salivary secretions provide a transport medium for the clumped particles and a third set contain C-esterases which initiate digestion. 7. Ingested food is held briefly in the proventriculus, then passed to the stomach where gland cells secrete A- and C-esterases which continue and extend the digestion initiated by the salivary C-esterases. 8. Some soluble products of gastric digestion are taken up by absorptive cells in the stomach wall and their digestion is completed intracellularly by enzymes which include beta-glucuronidase. Others pass into the intestine for absorption and completion of digestion by cells similar to the gastric absorptive cells but which lack beta-glucuronidase...

Animals↗