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PubMed1999

Probiotics and gut inflammation.

The intestine's mucosal surface provides a defense barrier against antigens encountered by the enteric route. In this system a balance is generated and maintained between host and microfloral bacteria. In intestinal inflammation, the integrity of the barrier is disrupted, a greater amount of antigens traverses the mucosal barrier, and the routes of transport are altered, possibly evoking aberrant immune responses and release of proinflammatory cytokines with further impairment of the barrier function. Nutritional therapy remains an attractive tool in the management of intestinal inflammation. The advances this past year are related to the ecologic system provided by specific strains of gut microflora, the concept of healthy microflora, and ways in which gut barrier function could be strengthened by consumption of mono-and mixed cultures of beneficial live microorganisms as probiotics.

Journal Article↗
PubMedDate not supplied

[The effect of Aerococcus viridans on the properties of salmonellae and staphylococci in vitro].

According to electron microscopy the repeated contacts of Aerococcus viridans 167 (industrial strain for the probiotic Aerobact production) with Salmonella typhimurium and Staphylococcus aureus caused the deep changes of cellular structure, down to bacteriolysis. Remaining viable pathogenic cells gave posterity with changed biochemical properties and virulence. S. typhimurium completely lost typical biochemical properties, its mobility and ability to be agglutinated by O-specific Salmonella serum reduced after the fifth joint passage. Sharp decrease of the leucotoxic and loss of the lethal and necrotic activity of the S. aureus toxin were observed. The latter was characterized by the smaller number of protein fractions in comparison with control series on electrophoregrams and densitograms. Pathogenic bacteria, playing a role in formation of the pathological intestinal microbiocenosis (S. typhimurium) and being hospital infections (S. aureus) acquired increased sensitivity to antibiotics. The carried out researches open some mechanisms of bactericidal and bacteriostatic action of A. viridans (group 12, Bergey, 1994) and their metabolic products, causing antagonistic effect during direct action on infectious microorganisms.

Animals↗
PubMedDate not supplied

Riboprinting: a tool for the study of genetic diversity in microorganisms.

Classical morphology-based methods of taxonomic and phylogenetic analysis are inadequate in many groups of structurally simple eukaryotes. Molecular methods can generate data independently of the complexity of the organisms' morphology. Riboprinting is one such technique, and involves restriction enzyme analysis of polymerase chain reaction amplified small subunit ribosomal RNA genes. The utility of the method is illustrated with examples from several genera of intestinal and bloodstream parasites. Among the applications of riboprinting are the detection of cryptic genetic variation within species, organism misidentifications and culture mix-ups, independent verification of DNA sequences, and the rapid generation of data useful in phylogenetic analyses.

Animals↗
PubMedDate not supplied

[Biological activity of probiotic microorganisms].

Adaptation of bifidobacteria and lactic acid bacteria to nutrient media with increased concentrations of bile (1%) and protein substrates of animal origin allowed the variants resistant to bile and displaying a high production of proteolytic enzymes (active within the pH range of 2.5-9.0) to be selected. Administration of the preparations involving the selected bifidobacteria and lactic acid bacteria assisted in the normalization of the intestinal microflora and activation of protein metabolism in the organism of animals. Specifically, it increased the total protein level in blood serum and redistributed protein fractions, increasing the content of globulins and decreasing albumin concentration.

Animals↗
PubMed1999

Study of the possible mechanisms involved in the mucosal immune system activation by lactic acid bacteria.

The induction of a mucosal immune response is not easy due to the development of oral tolerance, but under some conditions, bacteria can activate this immune system. Antigens administered orally can interact with M cells of Peyer's patches or bind to the epithelial cells. We have demonstrated that certain lactic acid bacteria are able to induce specific secretory immunity, and others will enhance the gut inflammatory immune response. The aim of this work was to establish the reason for these different behaviors and to define possible mechanisms involved in the interaction of lactic acid bacteria at the intestinal level. We studied IgA+ and IgM+ B cells comparatively in bronchus and intestine and CD4+ T cells and IgA anti-lactic acid bacteria antibodies in the intestinal fluid, induced by oral administration of Lactobacillus casei, Lb. delbrueckii ssp. bulgaricus, Lb. acidophilus, Lb. plantarum, Lb. rhamnosus, Lactococcus lactis, and Streptococcus salivarius ssp. thermophilus. The increase in the IgA+ B cells in the bronchus means that these lactic acid bacteria were able to induce the IgA cycle by interaction with M cells from Peyer's patches or intestinal epithelial cells. The IgM+ cells increased when the stimulus did not induce the switch from IgM+ to IgA+. The increase in the CD4+ cells suggests interaction of Peyer's patches and enhancement of the B- and T-cell migration. The anti-lactic acid bacteria antibody is related to the processing and presentation of the microorganisms to the immune cells. We demonstrated that Lb. casei and Lb. plantarum were able to interact with Peyer's patch cells and showed an increase in IgA-, CD4+ cells, and antibodies specific for the stimulating strain. Lactobacillus acidophilus induced gut mucosal activation by interaction with the epithelial cells without increase in the immune cells associated with the bronchus. Although Lb. rhamnosus and Strep. salivarius ssp. thermophilus interact with epithelial cells, they also induced an immune response against their epitopes. Lactococcus lactis and Lb. delbrueckii ssp. bulgaricus induced an increase of IgA+ cells entering the IgA cycle but not CD4+ cells; thus, these bacteria would have been bound to epithelial cells that activated B lymphocytes without processing and presenting of their epitopes. We did not determine specific antibodies against Lc. lactis or Lb. bulgaricus.

Animals↗
PubMed2005

Metabolism of glycitein (7,4'-dihydroxy-6-methoxy-isoflavone) by human gut microflora.

Gut microbial disappearance and metabolism of the soy isoflavone glycitein, 7,4'-dihydroxy-6-methoxyisoflavone, were investigated by incubating glycitein anaerobically with feces from 12 human subjects. The subjects' ages ranged from 24 to 53 years with a body mass index (BMI) of 20.9-25.8 kg/m(2) (mean BMI = 24.0 +/- 1.1 kg/m(2)). Glycitein disappearance followed an apparent first-order rate loss. Fecal glycitein disappearance rates for the subjects segregated into three different groups described as high (k = 0.67 +/- 0.14/h), moderate (k = 0.34 +/- 0.04/h), and low (k = 0.15 +/- 0.07/h) glycitein degraders (p < 0.0001). There was no dose effect on the disappearance rates for each subject from 10 to 250 microM glycitein (average k = 0.32 +/- 0.03/h, p > 0.05). Four putative glycitein metabolites, characterized by liquid chromatography-mass spectrometry (electrospray ionization using positive ionization mode), were dihydroglycitein, dihydro-6,7,4'-trihydroxyisoflavone, and 5'-O-methyl-O-desmethylangolensin. Two subjects produced a metabolite tentatively identified as 6-O-methyl-equol, and one subject produced daidzein as an additional metabolite of glycitein. These results show that glycitein is metabolized by human gut microorganisms and may follow metabolic pathways similar to other soy isoflavones.

Adult↗
PubMed1999

Cellular microbiology: can we learn cell physiology from microorganisms?

Cellular microbiology is a new discipline that is emerging at the interface between cell biology and microbiology. The application of molecular techniques to the study of bacterial pathogenesis has made possible discoveries that are changing the way scientists view the bacterium-host interaction. Today, research on the molecular basis of the pathogenesis of infective diarrheal diseases of necessity transcends established boundaries between cell biology, bacteriology, intestinal pathophysiology, and immunology. The use of microbial pathogens to address questions in cell physiology is just now yielding promising applications and striking results.

Amino Acid Sequence↗
PubMed2006

Vaginal protection and immunity after oral immunization of mice with a novel vaccine strain of Listeria monocytogenes expressing human immunodeficiency virus type 1 gag.

Natural transmission of human immunodeficiency virus (HIV) occurs at mucosal surfaces. During acute infection, intestinal and other mucosae are preferential sites of virus replication and rapidly become depleted of CD4(+) T cells. Therefore, mucosal immunity may be critical to control both initial infection and the massive early spread of virus. An attenuated D-alanine-requiring strain of the oral intracellular microorganism Listeria monocytogenes expressing HIV type 1 gag was shown to induce protective cell-mediated immunity in mice against viruses that express HIV gag when immunization occurs in the presence of a transient supply of D-alanine. In this study, we examined the efficacy of new attenuated strains that are able to synthesize d-alanine from a heterologous dal gene tightly regulated by an actA-promoted resolvase recombination system. In the absence of d-alanine, Gag-specific cytotoxic T lymphocytes (CTLs) were induced systemically after intravenous immunization, and one strain, Lmdd-gag/pARS, induced strong dose-dependent Gag-specific CTLs after oral immunization. A significant level of Gag-specific CD8(+) T cells was induced in the mucosal-associated lymphoid tissues (MALTs). Upon intravaginal challenge of these orally immunized mice with recombinant vaccinia virus (rVV) expressing HIV gag, gamma interferon- and tumor necrosis factor alpha-secreting Gag-specific CD8(+) T cells were dramatically increased in the spleen and MALTs. Oral immunization with Lmdd-gag/pARS led to complete protection against vaginal challenge by a homologous clade B gag-expressing rVV. In addition, strong cross-clade protection was seen against clades A and C and partial protection against clade G gag-expressing rVV. These results suggest that Lmdd-gag/pARS may be considered as a novel vaccine candidate for use against HIV/AIDS.

Administration, Oral↗
PubMed1988

Giardia-bile salt interactions in vitro and in vivo.

Giardia lamblia predominantly colonizes the proximal small intestine where bile is plentiful. We have investigated interactions between bile and this parasite by (i) examining the specificity of the stimulatory effect of bile on parasite growth in vitro, (ii) studying the possible association between giardiasis and bile salt deconjugation in vivo, and (iii) quantifying bile salt uptake by Giardia and relating this to uptake by other microorganisms. Our findings indicate that the growth promoting effects of ox bile and pure bile salt (sodium glycocholate) are, as far as is known, specific for Giardia, since the growth of a related protozoon, Trichomonas vaginalis, and a variety of enteric bacteria was either unchanged or inhibited. We were unable to detect deconjugated ('free') bile salt in duodenal fluid from UK patients with giardiasis and found no evidence to suggest that Giardia deconjugated bile salts in vitro. However, Giardia avidly took up conjugated bile salt, apparently in a concentration-dependent manner and to a much greater degree than Trichomonas and enteric bacteria. Thus, bile specifically stimulates growth of Giardia, and bile salt is avidly consumed by the parasite. The mechanism by which bile stimulates parasite growth is unknown, but uptake of conjugated bile salt by Giardia could reduce intraluminal bile salt concentrations and possibly interfere with micellar solubilization of fat. This may contribute to the steatorrhoea which is well recognized in symptomatic patients with giardiasis.

Animals↗
PubMed2005

Safety evaluation of the phosphinothricin acetyltransferase proteins encoded by the pat and bar sequences that confer tolerance to glufosinate-ammonium herbicide in transgenic plants.

Transgenic plant varieties, which are tolerant to glufosinate-ammonium, were developed. The herbicide tolerance is based upon the presence of either the bar or the pat gene, which encode for two homologous phosphinothricin acetyltransferases (PAT), in the plant genome. Based on both a review of published literature and experimental studies, the safety assessment reviews the first step of a two-step-approach for the evaluation of the safety of the proteins expressed in plants. It can be used to support the safety of food or feed products derived from any crop that contains and expresses these PAT proteins. The safety evaluation supports the conclusion that the genes and the donor microorganisms (Streptomyces) are innocuous. The PAT enzymes are highly specific and do not possess the characteristics associated with food toxins or allergens, i.e., they have no sequence homology with any known allergens or toxins, they have no N-glycosylation sites, they are rapidly degraded in gastric and intestinal fluids, and they are devoid of adverse effects in mice after intravenous administration at a high dose level. In conclusion, there is a reasonable certainty of no harm resulting from the inclusion of the PAT proteins in human food or in animal feed.

Acetyltransferases↗
PubMed1966

Isolated fecal microorganisms capable of 7-alpha-dehydroxylating bile acids.

Strains of microorganisms capable of 7alpha-dehydroxylation of chenodeoxycholate were isolated from rat and human feces. All the strains were strictly anaerobic, non-motile, moderately themioresistant Gram-positive rods. They showed some saccharolytic properties with the production of both acid and gas. They were H(2)S-positive but indole-, skatole-, citrate-, catalase-, and oxidase-negative. The isolated strains capable of 7alpha-dehydroxylation of chenodeoxycholate were also able to oxidize the hydroxyl groups at C-3 and C-7 to keto groups. The following metabolites were isolated: 3-keto-7alpha-hydroxy-5beta-cholanoic acid, 3alpha-hydroxy-7-keto-5beta-cholanoic acid, 3alpha-hydroxy-5beta-cholanoic acid, and 3-keto-5beta-cholanoic acid. The isolated strains did not have the enzymes necessary for hydrolyzing conjugated bile acids. In mixed anaerobic cultures of fecal microorganisms, extensive reduction of the 3-keto group to the 3beta-hydroxyl group occurred. The microorganism(s) responsible for this reaction have as yet not been isolated.

Bacteriological Techniques↗
PubMed1988

Bacteriaemia after injection sclerotherapy of oesophageal varices.

Thirty consecutive alcoholic patients with biopsy-proven cirrhosis of the liver and oesophageal varices underwent a total of 47 upper intestinal endoscopies. During 31 of the endoscopies paravariceal sclerotherapy of oesophageal varices was performed. Blood cultures were drawn before and after the procedures. Bacteriaemia after endoscopy was detected on seven occasions: six after sclerotherapy and one after endoscopy without sclerotherapy. This difference did not reach statistical significance. The microorganisms cultured belonged to the normal flora of the skin or the oropharynx. Bacteriaemia was transient and had no clinical consequences.

Adult↗
PubMed2002

[Colonic sanation in peritonitis caused by acute destructive cholecystitis].

Specific and quantitative microorganisms composition of colon cavity in 53 patients with an acute biliary peritonitis was investigated. Pathogenetic substantiated method of colonic sanation for treatment of peritonitis, caused by an acute destructive cholecystitis was elaborated. There was established, that in occurrence of the biliary genesis peritonitis the disorders of microbic intestinal cenosis and its colonizational resistance play an important role. Application of proposed method of colonic sanation allowed to eliminate its disorders and to improve result of treatment of patients.

Acute Disease↗
PubMed1978

Bacterial enzymes in uremia management.

The theoretical background for a conservative therapeutic treatment of uremia is described, with illustrative results from preliminary clinical trials in 10 patients and 10 normal reference subjects. The proposed treatment focuses upon the patient's gastrointestinal tract--the normal site for metabolism of both exogenous (dietary) and endogenous (recycled) protein--enabling it to behave like the rumen of the cow. The objective is to induce the uremic's organism to utilize its own "waste" substances. The patient swallows enterosoluble capsules containing specifically adapted enzymes (immobilized or free) from apathogenic soil microorganisms. These are pre-adapted to convert urea, creatinine, uric acid, guanidino derivatives, and other nonprotein nitrogen compounds (NPN). The enzymes utilize many other substances, in particular ammonia, potassium, phosphorus, and several other factors potentially dangerous for the uremic. The enzymes apparently cleave vasoconstrictatory peptides in the intestines. In the course of the therapy, renoparenchymal hypertension decreased significantly, and increased again when the regimen was interrupted. The results from the present studies are in full accord with the information published in the relevant fields. The time appears ripe for large-scale trials of the therapeutic regimen outlined, especially as many commercial microbial enzymes already have a long history of safe use in food processing.

Animals↗
PubMed1996

[Lactoferrin--its role in defense against infection and immunotropic properties].

Lactoferrin (LF) is an iron-binding protein found in milk and other secretory fluids of mammals as well as in secondary granules of neutrophils. Receptors for LF were detected and isolated on activated T and B cells, monocytes, intestinal brush border cells, platelets and neoplastic cells. Very low physiologic serum levels of LF increase significantly upon infection. Serum concentration of LF is also elevated in rheumatoid patients. It is suggested that the ability of LF to bind an excess of Fe() ions, needed for growth of microorganisms and tumors, represents an important defence mechanism in humans. LF, in addition, may contribute to the protection against pathogens and their metabolites by enhancing phagocytosis, cell adherence and controlling release of proinflammatory cytokines such as IL-1, IL-6 and TNF-alpha. The protein diminishes also damaging effects of free radical release. LF possesses interesting immunotropic properties with regard to immature T and B cells by promoting phenotypic and functional maturation of these cells. LF also controls the effector phase of cellular immune response and inhibits manifestations of autoimmune response in mice. One molecular form of LF with a ribonuclease activity may have a prognostic value in breast cancer. Lactoferrin may be potentially applied in neutropenic patients or in patients with bleeding disorders as a preoperative immunomodulator.

Adjuvants, Immunologic↗
PubMed1988

Intestinal mucosal immune defense mechanisms.

The intestinal mucosal immune defense mechanisms involve both humoral and cellular immunity. The prominence of suppressor/cytotoxic T lymphocytes in the epithelial layer suggests that these interepithelial lymphocytes play a role in defense against infections within this layer. Secretory IgA is overwhelmingly the major humoral immune response along the gastrointestinal tract and along other mucosal surfaces (respiratory tract, mammary glands, salivary glands, and lacrimal glands). While the functions of secretory IgA are incompletely understood, it is clear that it prevents attachment of microorganisms and toxins (cholera toxin, shiga toxin, etc.) to the surface epithelial cells. Furthermore, secretory IgA may collaborate with eosinophils or killer lymphocytes to mediate cytotoxic reactions against enteropathogens. By learning more about the mucosal immune response, we should be able to understand the relationship between the lamina propria plasmacytosis in inflammatory bowel disease and the increased number of interepithelial lymphocytes that we see in gluten-sensitive enteropathy and the underlying pathogenic mechanisms.

Humans↗