[Medical rehabilitation in patients with anal incompetence after surgery for stage IV hemorrhoids]. / Konservativnaya reabilitatsiya patsientov s nedostatochnost'yu anal'nogo sfinktera posle khirurgicheskogo lecheniya gemorroya IV stadii.

The choice of medical rehabilitation in patients with anal incontinence is impossible without diagnostic data revealing the mechanism of fecal incontinence. The most promising are programs of comprehensive physiotherapeutic rehabilitation based on biofeedback training. The rate of anal incompetence (AI) after hemorrhoidectomy is 1.3-12.5%. However, in addition to the organic cause (surgical trauma), functional disorders of the external sphincter and pelvic floor muscles may contribute to the pathogenesis of anal incontinence, aggravating the incontinence symptoms after surgery. Therefore, these functional disorders should be diagnosed before surgery. However, medical rehabilitation programs for anal incontinence after hemorrhoidectomy are not standardized, and functional outcomes have not been studied. OBJECTIVE: To evaluate the outcomes of comprehensive rehabilitation in patients with AI after hemorrhoidectomy to improve quality of life after surgery. MATERIALS AND METHODS: A retrospective study was carried out on 46 patients (mean age 53.8±15.4 years) after hemorrhoidectomy with fecal incontinence, 13 (28.3%) males and 33 (71.7%) females. The main group included 25 patients who received comprehensive rehabilitation, including biofeedback training and tibial neuromodulation (TNM) for 15 days. The control group consisted of 21 patients who received TNM at home also for 15 days. The severity of fecal incontinence was determined using the Wexner score. The functional state of the sphincter before and after surgery was assessed using the anorectal manometry (sphincterometry) (WPM Solar, the Netherlands). RESULTS: Comprehensive rehabilitation resulted in a statistically significant clinical improvement: a decrease in the Wexner score in both males and females. No significant differences in manometry results were observed: the anal sphincter tone increased by 16.0% in females and 10.6% in males, and contractility increased by 17.7% and 15.1%, respectively. Monotherapy with TNM in control group patients improved tone indices by 8.7% in females and 6.8% in males, and contractility by 6.2 and 5.4%, respectively, which was lower than in the main group. CONCLUSION: Contraindications to physiotherapeutic procedures based on electrical stimulation, extracorporeal magnetic stimulation, and magnetic translumbosacral neuromodulation determine the only possible choice of medical rehabilitation, which is the combination of biofeedback training and TNM (as superior to TNM monotherapy). If out-patient medical rehabilitation is not feasible, patients are recommended to complement the home course with a specially designed set of exercises for anal incontinence treatment.

[Turcot syndrome and Gardner's syndrome in a female patient with familial colon adenomatosis. A case report and literature review]. / Sindrom Tiurko, sindrom Gardnera u patsientki s semeinym adenomatozom tolstoi kishki. Klinicheskoe nabliudenie i obzor literatury.

Turcot syndrome is a rare hereditary syndrome characterized by a combination of brain tumors and colorectal cancer. According to the literature, about 150 such cases have been reported. This article presents a rare clinical case and a literature review.

A peculiar case of Campylobacter jejuni attenuated aspartate chemosensory mutant, able to cause pathology and inflammation in avian and murine model animals.

An attenuated Campylobacter jejuni aspartate chemoreceptor ccaA mutant caused gross pathological changes despite reduced colonisation ability in animal models. In chickens, the pathological changes included connective tissue and thickening of the mesenteric fat, as well as the disintegration of the villus tips in the large intestine, whereas in mice, hepatomegaly occurred between 48-72 hours post infection and persisted for the six days of the time course. In addition, there was a significant change in the levels of IL-12p70 in mice infected with the C. jejuni ccaA mutant. CcaA isogenic mutant was hyper-invasive in cell culture and microscopic examination revealed that it had a "run" bias in its "run-and-tumble" chemotactic behaviour. The mutant cells also exhibited lower level of binding to fucosylated and higher binding to sialylated glycan structures in glycan array analysis. This study highlights the importance of investigating phenotypic changes in C. jejuni, as we have shown that specific mutants can cause pathological changes in the host, despite reduction in colonisation potential.

[Relationship of the quality of drinking water to its use regimens and the types of water supply pipes].

Drinking water running along the pipes made from different materials was investigated. Two experiments could determine the material that assured at least of all the quality of drinking water in accordance with SanPin 2.1.4.1074-01. The mechanism for worsening the quality of water supplied to a user was revealed in relation to the water use regimen. Short-term flow stoppage of water was found to result in its lower oxygen levels, a larger number of different groups of iron- and manganese-reducing bacteria and an enhanced bacterial reduction of oxides. The latter was accompanied by the dissolution of heavy metals, which induced secondary water contamination.

[Problems and perspectives in the hygienic regulation of biotechnological strains of microorganisms].

The state-of-the-art of hygienic standardization of biotechnological strains of microorganisms is considered. The last century's investigations accumulated abundant experimental data on the estimation of the adverse effect of the strains, formulated main guidelines for evaluating the negative activity of producing strains, and proposed a schematic diagram for toxicological and hygienic investigations of new biotechnological strains. Further studies made it possible to elaborate their human hazard classification, to improve study programs, to substantiate a priority list, and to develop approaches to assessing the occupational microbiological risk.

Mucins in the mucosal barrier to infection.

The mucosal tissues of the gastrointestinal, respiratory, reproductive, and urinary tracts, and the surface of the eye present an enormous surface area to the exterior environment. All of these tissues are covered with resident microbial flora, which vary considerably in composition and complexity. Mucosal tissues represent the site of infection or route of access for the majority of viruses, bacteria, yeast, protozoa, and multicellular parasites that cause human disease. Mucin glycoproteins are secreted in large quantities by mucosal epithelia, and cell surface mucins are a prominent feature of the apical glycocalyx of all mucosal epithelia. In this review, we highlight the central role played by mucins in accommodating the resident commensal flora and limiting infectious disease, interplay between underlying innate and adaptive immunity and mucins, and the strategies used by successful mucosal pathogens to subvert or avoid the mucin barrier, with a particular focus on bacteria.

Tetracycline resistance of Australian Campylobacter jejuni and Campylobacter coli isolates.

OBJECTIVES: Tetracycline resistance in Campylobacter is encoded by the tet(O) gene and is usually associated with conjugative plasmids. Little was known about tetracycline resistance in Australian Campylobacter species, therefore we investigated this resistance in 41 Campylobacter jejuni and five Campylobacter coli strains from humans and healthy chickens. METHODS: Tetracycline MICs were determined for each isolate using an agar dilution method. The distribution and localization of tet(O) on plasmid and chromosomal DNA was determined by Southern-blot experiments. The ability to transfer resistance to recipient strains was examined through conjugation studies. Identity of transconjugants was confirmed by PCR and flaA-restriction fragment length polymorphism analysis. RESULTS: High-level tetracycline resistance was observed, ranging from 32 to >256 mg/L. Plasmids were detected in 74% of isolates with plasmids between 30 and 40 kb in size most frequently isolated. tet(O) was present in all tetracycline-resistant isolates. In the majority of strains under study the tet(O) gene was chromosomally encoded. Tetracycline resistance of six C. jejuni strains in which tet(O) was plasmid mediated was transferred by conjugation to a C. jejuni recipient strain. Transfer did not occur between tetracycline-resistant C. jejuni strains and a C. coli recipient. No difference in MICs, plasmid carriage and tet(O) localization was detected between human and chicken isolates. CONCLUSIONS: These data indicate that the tet(O) gene, previously reported in Campylobacter strains throughout the world, is present in Australian Campylobacter. This study will lead to a greater understanding of antibiotic resistance distribution in Campylobacter spp. in Australia.

Antimicrobial resistance and genomic screening of clinical isolates of thermophilic Campylobacter spp. from south-east Queensland, Australia.

AIM: To screen 90 clinical isolates of thermophilic Campylobacter species for putative resistance to ampicillin, erythromycin and tetracycline and perform numerical analysis to determine isolate relatedness. METHODS AND RESULTS: Disc diffusion, E-test MIC and agar dilution methods were performed. Disc diffusion testing showed 87 (97%) isolates appeared resistant to ampicillin at 10 microg; 14 (16%) resistant to tetracycline at 30 microg; and three (3.4%) resistant to erythromycin at 15 microg. E-test MICs showed a range of 0.5 to >256 mg l(-1) for ampicillin; 16 to >256 mg l(-1) for tetracycline; and >256 mg l(-1) for erythromycin. E-test showed 68% correlation (+/-1 log2 dilution) with agar dilution for ampicillin, 100% for erythromycin and 64% for tetracycline. Disc diffusion testing showed 100% correlation with agar dilution for erythromycin and tetracycline, and 77% for ampicillin. Numerical analyses of restriction endonuclease (RE) fragment profiles suggested a high level of isolate variation. CONCLUSION: The incidence of resistance of thermophilic Campylobacter spp. to erythromycin and tetracycline is low in south-east Queensland. SIGNIFICANCE AND IMPACT OF THE STUDY: Disc diffusion susceptibility testing may be used to screen thermophilic Campylobacter spp. for putative resistance to erythromycin and tetracycline. Agar dilution should be used to determine ampicillin susceptibility.

Colonisation phenotype and colonisation potential differences in Campylobacter jejuni strains in chickens before and after passage in vivo.

Campylobacter jejuni strains isolated from chicken faeces and from humans suffering from gastroenteritis were used to determine the colonisation phenotypes and colonisation potential of these strains in chickens. Five different colonisation types were observed ranging from immediate and sustained colonisation to completely non-colonising. Phenotype one showed immediate colonisation with prolonged excretion of viable C. jejuni bacteria. Phenotype two showed delayed colonisation with prolonged excretion of viable C. jejuni bacteria. Phenotype three showed immediate colonisation and delayed clearing of viable C. jejuni bacteria. Phenotype four showed delayed colonisation and delayed clearing of viable C. jejuni bacteria. Strains of phenotype five could not colonise chickens. Inoculum levels to obtain maximum caecal colonisation for each phenotype for strains cultured in vitro and in vivo was also determined. Following passage in vivo through a chicken, the minimum inoculum required for sustained colonisation dropped approximately 1000-fold, however, the colonisation phenotype remained unchanged before and after a passage in vivo.

Specific identification, grouping and differentiation of Campylobacter jejuni among thermophilic campylobacters using multiplex PCR.

Campylobacter species such as C. jejuni and C. coli are recognized as major causes of acute gastroenteritis world-wide. Although C. jejuni and C. coli are usually non-pathogenic in birds and animals, they cause enteric disease in humans and the source of infection is often the consumption of contaminated foodstuffs. In this paper we report on the development and use of a multiplex PCR of C. jejuni genomic DNA which yielded a PCR product with a unique polymorphic site that can be used to quickly and accurately identify and group C. Jejuni isolates from any source including DNA, cell culture, skin washings and faecal samples. The test is simple and sensitive and can detect purified DNA from a single bacterium 10(2) cells from crude lysates, 10(3) cells in seeded faeces and 120 cells/ml of washing of 1 cm2 skin fragments of chicken skin.

Differentiation between Campylobacter hyoilei and Campylobater coli using genotypic and phenotypic analyses.

Genotypic and phenotypic methods were applied to investigate differences between the closely related species Campylobacter hyoilei and Campylobacter coli. A unique DNA sequence from C. hyoilei was used to design a specific PCR assay that amplified a DNA product of 383 bp for all C. hyoilei strains, but not other Campylobacter species, including C. coli. The PCR assay could detect 100 fg pure C. hyoilei DNA, 2 x 10(2) c.f.u. ml(-1) using cultured cells and 8.3 x 10(3) c.f.u. 0.1 g(-1) in faeces. The C. hyoilei sequence utilized for specific detection and identification of this species showed similarities to sequences from bacteriophages Mu, P2 and 186, suggesting lysogination of the ancestral C. hyoilei genome. Activities of a set of 15 enzymes that participate in a variety of cellular functions, including biosynthesis, catabolism, energy generation, maintenance of redox balance and phosphate utilization, were tested using sets of strains of C. hyoilei and C. coli. Comparison of mean rates of enzyme activities revealed significant differences between species in the values determined for seven of these activities. Both the genetic and phenotypic data indicate that C. hyoilei is a unique Campylobacter species.

Characteristics of the aerobic respiratory chains of the microaerophiles Campylobacter jejuni and Helicobacter pylori.

The respiratory chain enzymes of microaerophilic bacteria should play a major role in their adaptation to growth at low oxygen tensions. The genes encoding the putative NADH:quinone reductases (NDH-1), the ubiquinol:cytochrome c oxidoreductases (bc1 complex) and the terminal oxidases of the microaerophiles Campylobacter jejuni and Helicobacter pylori were analysed to identify structural elements that may be required for their unique energy metabolism. The gene clusters encoding NDH-1 in both C. jejuni and H. pylori lacked nuoE and nuoF, and in their place were genes encoding two unknown proteins. The NuoG subunit in these microaerophilic bacteria appeared to have an additional Fe-S cluster that is not present in NDH-1 from other organisms; but C. jejuni and H. pylori differed from each other in a cysteine-rich segment in this subunit, which is present in some but not all NDH-1. Both organisms lacked genes orthologous to those encoding NDH-2. The subunits of the bc1 complex of both bacteria were similar, and the Rieske Fe-S and cytochrome b subunits had significant similarity to those of Paracoccus denitrificans and Rhodobacter capsulatus, well-studied bacterial bc1 complexes. The composition of the terminal oxidases of C. jejuni and H. pylori was different; both bacteria had cytochrome cbb3 oxidases, but C. jejuni also contained a bd-type quinol oxidase. The primary structures of the major subunits of the cbb3-type (terminal) oxidase of C. jejuni and H. pylori indicated that they form a separate group within the cbb3 protein family. The implications of the results for the function of the enzymes and their adaptation to microaerophilic growth are discussed.

The galE gene of Campylobacter jejuni is involved in lipopolysaccharide synthesis and virulence.

Lipopolysaccharide (LPS) is one of the main virulence factors of gram-negative bacteria. The LPS from Campylobacter spp. has endotoxic properties and has been shown to play a role in adhesion. We previously cloned a gene cluster (wla) which is involved in the synthesis of the Campylobacter jejuni 81116 LPS molecule. Sequence alignment of the first gene in this cluster indicated similarity with galE genes. These genes encode a UDP-glucose 4-epimerase, which catalyzes the interconversion of UDP-galactose and UDP-glucose. A Salmonella galE mutant was transformed with the galE gene from C. jejuni. The LPS analysis of wild-type, galE, and complemented galE Salmonella strains showed that the C. jejuni galE gene could restore the smooth wild-type Salmonella LPS. A UDP-glucose 4-epimerase assay was used to demonstrate that the galE gene from C. jejuni encoded this epimerase. We constructed a C. jejuni galE mutant which expressed a lipid A-core molecule of reduced molecular weight that did not react with antiserum raised against the parental strain. These results show an essential role for the galE gene in the synthesis of C. jejuni LPS. The galE mutant also showed a reduction in its ability to adhere to and invade INT407 cells. However, it was still able to colonize chickens to the same level as the wild-type strain. The serum resistance and hemolytic activity of this mutant were not changed compared to the parent strain. The ability of the mutant to take up DNA and integrate it in its genome was reduced 20-fold. These results show that LPS of C. jejuni is an important virulence factor.

Isolation and molecular analysis of colonising and non-colonising strains of Campylobacter jejuni and Campylobacter coli following experimental infection of young chickens.

Fourteen-day-old chickens were inoculated with selected Campylobacter coli and C. jejuni strains. C. jejuni strains were of two subgroups based on a polymorphism detected using a DNA probe and represented the profiles typical for the majority of strains of either chicken or human origin. All C. coli strains previously isolated from humans colonised chickens, whereas from 4/7 C. jejuni strains of human origin, failed to colonise. Of 12 Campylobacter strains of chicken origin, 10 established a persistent colonisation in the chickens, and 2 strains colonised poorly or not at all. Four strains that failed to colonise chickens were each inoculated into groups of five birds. Three strains again did not colonise any of the chickens and the fourth strain colonised four out of the five chickens, but was poorly excreted. When infected chickens were placed in the same enclosure to facilitate interchange of strains, C. jejuni strain 331 was found to be dominant and colonised all 12 chickens by 21 days, displacing all other strains. C. jejuni strain 331, was then inoculated into groups of five birds with previously established colonisation by C. jejuni and C. coli strains. Strain 331 was able to replace the C. jejuni strain in all five birds but established co-colonisation with C. coli strain. Naturally occurring co-colonisation by two C. jejuni strains was detected in one chicken out of 200 tested. There was no obvious correlation between the type of DNA polymorphism in strains of chicken origin and their ability to colonise chickens.

Differentiation of Campylobacter jejuni and Campylobacter coli strains by using restriction endonuclease DNA profiles and DNA fragment polymorphisms.

The chromosomal DNA fragment patterns from a total of 169 Campylobacter jejuni and Campylobacter coli isolates from poultry and humans were analyzed by using DNA restriction endonucleases ClaI and EcoRV. The DNA restriction patterns produced by ClaI and EcoRV consisted of unique DNA fragments of 9 to 9.5 kb and 3.5 kb generated with ClaI and a single unique fragment of 3.0 kb produced by EcoRV. These patterns were obtained with all strains of C. jejuni tested. The DNA restriction patterns were further examined by Southern blot analysis with a previously constructed DNA probe, pMO2005, which is also able to distinguish between C. jejuni and C. coli spp. (5). Two types of patterns were produced by hybridization with the ClaI-cleaved DNA of C. jejuni strains, one of a single 18.5-kb genomic fragment and the other of 14.5- and 4.0-kb fragments. This indicated the presence of an extra ClaI site in this genomic fragment in the strains with the duplex pattern. The Southern blot analysis of 169 C. jejuni and C. coli isolates from poultry and from humans with DNA probe pMO2005 demonstrated that 78% of C. jejuni strains isolated from chickens hybridized with DNA probe pMO2005 with a characteristic 14.5- and 4.0-kb banding pattern and 22% hybridized with a single 18.5-kb fragment, whereas 71% of human isolates hybridized with the single 18.5-kb fragment and only 29% hybridized with 14.5- and 4.0-kb fragments. These findings suggest that only a small proportion of C. jejuni strains that colonize chickens may cause disease in humans.

Antibacterial proteins from porcine polymorphonuclear neutrophils.

Antibacterial peptides were purified from porcine neutrophil granules collected from healthy pigs. Granule proteins, extracted with 0.2 mol/L sodium acetate were subjected to ion-exchange chromatography and five peaks (designated A to E) were detected. Individual porcine neutrophil granule proteins were shown to inhibit the growth of target organisms Escherichia coli and Staphylococcus aureus. The antimicrobial activity was shown to be concentration and time dependent. Peak D showed strong antimicrobial activity against S. aureus and peak C (with a greater number of eluted proteins) was shown to be active against both S. aureus and E. coli. One of the peptides was purified further by reverse-phase HPLC from peak fraction C. The MW of this peptide was approximately 5500 Da as determined by SDS-PAGE and mass spectral analysis and was active against both E. coli and S. aureus in vitro sustaining a > 90% decrease, respectively, in CFU after a 2 h exposure with 50 micrograms of this peptide. Amino acid analysis showed the peptide was rich in aspartate/aspartic acid, glutamine/glutamic acid, proline, arginine and threonine. The antimicrobial activity of this peptide and other novel proteins in porcine neutrophilic granules demonstrates the probable role of these proteins and peptides in host defence of porcine neutrophils against bacterial infection.

Campylobacter hyoilei sp. nov., associated with porcine proliferative enteritis.

Campylobacter hyoilei sp. nov. is the name proposed for an organism formerly described as strain RMIT 32AT (T = type strain) and a group of similar bacteria isolated from intestinal lesions of pigs with proliferative enteritis. The phenotypic characteristics of these organisms indicated that they are closely related to each other and are not strains of other Campylobacter spp. commonly isolated from pigs. The results of probing of ClaI-, EcoRV-, or BglII-cleaved genomic DNAs from C. hyoilei strains with a radiolabeled DNA probe that distinguishes between Campylobacter jejuni and Campylobacter coli indicated that C. hyoilei and C. coli are closely related. However, the 16S rRNA sequence of the reference strain of C. hyoilei, RMIT 32AT, was four bases different from the 16S rRNA sequence of C. jejuni CCUG 11284T and five bases different from the 16S rRNA sequence of C. jejuni subsp. doylei CCUG 24567T, suggesting that C. hyoilei is more closely related to C. jejuni than to C. coli. Hybridization between DNA from C. hyoilei type strain RMIT 32A and DNAs from selected type and reference strains of other Campylobacter species and subspecies, including C. jejuni, C. jejuni subsp. doylei, C. coli, Campylobacter mucosalis, and Campylobacter hyointestinalis, as well as the other C. hyoilei strains (the RMIT 32AT-like isolates), revealed that high levels of DNA hybridization (> 70%) occurred only between the reference strain and other strains of C. hyoilei.

[Biotechnology manufacture as a possible source of environmental pollution and a risk factor for human health]. / Biotekhnologicheskoe proizvodstvo kak vozmozhnyi istochnik zagriazneniia okruzhaiushchei sredy i faktor riska dlia zdorov'ia cheloveka.

The biotechnology-oriented quantitative criteria and a plan of primary sanitary hygienic evaluation of pathogenicity of microorganisms-producers were first established. New evidence of the negative effects of strains-producers on microbiocenosis and self-purification of soils and reservoirs were obtained. The requirement of an all-round hygienic assessment of microorganisms-producers consisting of two mandatory steps, i.e. the primary sanitary hygienic evaluation of pathogenicity of strains and the investigation of effects of these microorganisms on microbiocenosis and soil and reservoir self-purification has been substantiated.