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[Bacterial flora in urinary tract infections of children and susceptibility to drugs].

The material consisted of 902 strains isolated from children with urinary tract infection with significant bacteriuria. Among isolated strains, Enterobacteriaceae consisted 67.4%, Gram-positive cocci 18.8% and non-fermenting Gram-negative bacilli (both against Gram-negative bacilli and Gram-positive cocci), appeared aminoglycosides and III generation cephalosporins (from 63 to 83% of susceptible strains). K. pneumoniae and P. aeruginosa were most resistant to antimicrobials tested. The authors point out that empiric antibiotic therapy of bacterial urinary tract infections in children which is not preceded by identification of the pathogen and determination of its drug susceptibility, can be ineffective in over 50% of cases.

Aminoglycosides↗

[Gene level diagnostics of infectious diseases].

Recent developments in medical technology have caused a great change in infectious diseases, as characterized by epidemics of antibiotics-resistant bacteria, opportunistic infection in compromised hosts, and blood-borne viral infections such as hepatitis virus and human immunodeficiency virus. In the diagnosis of such new aspects of infectious diseases, conventional immunological, culture, and microscopical techniques are not always practical. By contrast, evaluation of infectious agents using molecular biological technology frequently offers the rapid, most accurate and sensitive method of diagnosis. Amplification methods are particularly attractive for the detection of small numbers of microorganisms, as in latent conditions, or for the fastest identification of the pathogen without laborious isolation. For introduction of the tests into routine procedures, their standardization as well as simplicity and low cost are required. Gene level diagnostics should be applied appropriately to management of infectious diseases along with the conventional techniques, while further roles of the tests must be determined, on the basis of the molecular elucidation of infectious diseases.

DNA, Bacterial↗

[Unusual infectious diseases in a municipal hospital (author's transl)].

UNLABELLED: The most important criteria for differential diagnosis are presented with reference to case histories: Falciparum malaria: relapsing fever, hemolytic anemia, hepatosplenomegaly, leucopenia; Kala-azar: insidious course with fever, pancytopenia, macrocytic anemia, hepatosplenomegaly, enlargement of lymph nodes; echinococcosis: symptoms of a benign tumor with displacement. DIAGNOSIS: by identification of the pathogen in malaria and kala-azar, serologically in all three diseases and also radiologically in echinococcosis.

Adult↗

[Nontuberculous infectious osteoarthritis in adults (excluding spondylodiscitis) in rheumatology].

Analysis of twenty five personal observations of septic osteoarthritis compared with information from the literature led the authors to the following conclusions: 1) Diagnosis of septic osteo-arthritis must be considered when confronted with a picture of acute or subacute arthritis generally inflamed with variable fever, raised sedimentation rates and frequently polymorph neutrophilia, above all especially if there are particular antecedents (local trauma or intra-articular injection, conditions which favor infections: diabetes, alcoholism, impaired immunity, corticosteroid therapy or various therapeutic immuno-suppresives). 2) It may be established on absolute criteria (isolation of the organism at the site of the lesions) or, in the absence of direct bacteriological evidence, by a body of evidence amongst which 4 arguments are of particular value: identification of a pathogenic organism in one or more blood cultures and/or at the site of a closed infection, postive serology and particularly elevation of the antistaphylolysine titres, rapid-onset radiological changes of destruction and/or reconstruction. plus hypercellularity of the fluid greater than 100,000 cells/mm3. 3) Medical treatment sometimes includes local treatment (joint aspiration, local injection of antibiotics) and in all cases immobilisation during the acute phase and systemic antibiotics. The latter will vary according to the organism responsible and its sensitivity. It seems useful to continue this for twelve weeks at the minimum in the normal form, initially making use of the combination of 2 antibiotics and preferably using continuous perfusion and/or intramuscular routes during the first half of treatment. 4) The consistently good results of such medical treatment seem to limit considerably the place of surgery for which the indications have become rare during the acute phase and even more so at later stages.

Adolescent↗

[Sulacillin (a combination of ampicillin and sulbactam)--a new combined drug form for treatment of bacterial infections].

The results of the cooperated clinical trial of sulacillin, a formulation including sodium ampicillin and sulbactam (2:1), are presented. The trial was carried out in different hospitals of Moscow and Smolensk. Rapid development of the favourable time course and recovery were stated in 93.7 per cent of the cases with upper and lower respiratory tract infections, urinary and biliferous tract infections, infections of the skin and soft tissues, gynecological infections and severe suppurative complications after surgical operations. Sulacillin is useful in the empirical therapy of purulent inflammatory diseases before the pathogen isolation and identification.

Ampicillin↗

Amino acid sequence of an immunogenic corneal stromal protein.

PURPOSE: A unique cornea-associated antigen (CO-Ag) has been purified previously from stromal extracts. The protein is the target for autoantibodies in patients with Mooren's ulcer. In this study, the amino acid sequence of CO-Ag was analyzed and the structure-function properties of CO-Ag was determined. METHODS: Purified CO-Ag was subjected to N-terminal sequencing by automated Edman degradation. Binding of calcium (Ca2+) to CO-Ag was measured by a direct (45)Ca2+ -binding assay. RESULTS: The complete amino acid sequence of CO-Ag has been determined. The protein contains 70 amino acids in a single chain and lacks cysteine, tryptophan, and methionine residues. A computerized data base search of protein and nucleic acid sequences revealed strong homology to the Ca2+ -binding proteins of the S-100 family. The sequence of CO-Ag shows a high homology with calgranulin C (CaG-C) previously purified from pig granulocytes. The functional Ca2+ -binding sites of CO-Ag and CaG-C were different based on homology with known Ca2+ -binding domains and their Ca2+ -binding properties. There are three amino acid substitutions in the N-terminal Ca2+ -binding domain. Differences were functionally conserved and compatible, with minimum single-base changes in the codon structures. The greatest difference was located in the C-terminal Ca2+ -binding domain. Five consecutive amino acid changes from D63-K-K-G-A67 in CO-Ag to M63-Q-D-E-Q67 occurred in CaG-C. These differences alter the structure of CO-Ag, which no longer can bind Ca2+ ions. The existence of this nonfunctional Ca2+ -binding site was corroborated by its Ca2+ -binding properties. The number of Ca2+ -binding sites for the CO-Ag sub-unit is approximately half that of the CaG-C monomer, although these two proteins have a similar low binding constant of approximately 2 x 10(-4) M. CONCLUSIONS: These results suggest that CO-Ag is a new member of the Ca2+ -binding protein of the S-100 family heretofore undescribed in the cornea. Sequence data provide an important framework to search for sequence similarity with microbial proteins as possible substrates for molecular mimicry and for the identification of possible pathogenic epitopes in CO-Ag.

Amino Acid Sequence↗

Primary cell culture of turkey intestinal epithelial cells.

Primary cell culture has been widely used in various types of studies and proven useful for the isolation and identification of avian pathogens. Difficulties in growing intestinal epithelial cells in vitro have limited their use for such studies. In the present study, a co-culture system was developed for the primary culture of intestinal epithelial cells. A monolayer obtained from 14-to- 16-day-old turkey embryo intestinal fibroblasts was used as a feeder layer. Feeder layers from turkey embryo fibroblasts and from a continuous cell line (mouse 3T3 fibroblasts) were also employed but were not as successful. The intestinal epithelial cells were isolated by dissociation from the intestinal tracts of 1-day-old turkey poults and grown on the feeder layers. Growth and maintenance media were supplemented with various components, including fetal calf serum, chicken serum, hormones, and other growth factors. The epithelial cells grown on feeder layers from the intestinal fibroblasts allowed the intestinal epithelial cells to be maintained in vitro for periods of 7 to 10 days. This technique may prove useful for various applications, including isolation of enteropathogens, and for basic studies of the intestinal tract concerning such subjects as physiology, immunology, and toxicology.

Animals↗

Outbreaks of pseudo-infection with Cyclospora and Cryptosporidium--Florida and New York City, 1995.

Efforts to expand the scope of surveillance and diagnostic testing for emerging infectious diseases also may increase the potential for identifying pseudo-outbreaks (i.e., increases in incidence that may result from enhanced surveillance) and outbreaks of pseudo-infection (i.e., clusters of false-positives for infection). This report describes the investigations of outbreaks of pseudo-infection with Cyclospora in Florida and Cryptosporidium in New York City in 1995 after health departments in those jurisdictions had initiated surveillance for these emerging organisms. These investigations emphasize 1) the need for laboratory training in the identification of emerging pathogens and 2) the importance of confirmation by reference laboratories as an early step in the investigation of any apparent outbreak caused by an emerging pathogen.

Animals↗

Biological warfare. A historical perspective.

The deliberate use of microorganisms and toxins as weapons has been attempted throughout history. Biological warfare has evolved from the crude use of cadavers to contaminate water supplies to the development of specialized munitions for battlefield and covert use. The modern development of biological agents as weapons has paralleled advances in basic and applied microbiology. These include the identification of virulent pathogens suitable for aerosol delivery and industrial-scale fermentation processes to produce large quantities of pathogens and toxins. The history of biological warfare is difficult to assess because of a number of confounding factors. These include difficulties in verification of alleged or attempted biological attacks, the use of allegations of biological attacks for propaganda purposes, the paucity of pertinent microbiological or epidemiologic data, and the incidence of naturally occurring endemic or epidemic diseases during hostilities. Biological warfare has been renounced by 140 nations, primarily for strategic and other pragmatic reasons. International diplomatic efforts, including the 1972 Biological Weapons Convention, have not been entirely effective in preventing the enhancement and proliferation of offensive biological warfare programs. The threats posed by biological weapons are likely to continue into the future.

Biological Warfare↗

[Manifestations, diagnosis and treatment of non-tuberculous mycobacterial infections in non-immunodepressed patients].

Non-tuberculous mycobacteria (NTM) can be the etiologic agents of chronic pulmonary disease, lymphadenitis, skin and soft-tissue infection and disseminated disease in non-immunocompromised patients. The recognition of disease needs repeated isolation of the NTM from bronchopulmonary secretions or from tissue biopsies, and its identification by specific laboratory methods. A wide spectrum of clinical presentations and severity of disease can be found, from spontaneous healing to progressive and destructive lung disease, and death, according to predisposing conditions and mycobacterial species. The choice of surgical and drug treatment will depend on identification of specific pathogen and clinical evaluation.

Antitubercular Agents↗

[Arboviruses and arbovirus infections in the forest steppe zone of Ukraine].

New active natural foci of the tick-borne encephalitis, West Nile, California serologic group (CSG) and Batai viruses have been identified as a result of comprehensive virologic and serologic investigations in the forest-steppe zone of Ukraine. These observations were confirmed by the results of these pathogens study and identification of the corresponding antigens in natural biological substrates and in the blood of ill people. Several human pathogenetic viruses, including Inko, Snow hoc hare with CSG and Crimean hemorrhagic fever were found for the first time in the forest-steppe zone. Viruses populations circulating there are heterogeneous according to their genetic properties. New data concerning their ecology were obtained. A close etiological relation between the acute seasonal febrile diseases and the above mentioned arboviruses was found. Arboviral infections from 23.6% of the total amount of spring-summer-autumn febrile diseases. The West Nile fever (53.1%) is the leading arboviral infection in the forest-steppe zone. A quarter of all arboviral infections is represented by tick-borne encephalitis. Diagnostic preparations have been made and the available methods of laboratory diagnostics of the arboviral infections have been improved.

Animals↗

[Antibiotic therapy in leukopenia].

Intensified chemotherapy-induced long-term neutropenia is the main cause for morbidity and mortality of patients with hematologic malignancies. The successful management of neutropenia is based on hygienic procedures antimicrobial prophylaxis and therapy, and diagnostics. Until today, Co-Trimoxazole or fluoroquinolenes and oral amphotericine B are the prophylactic standard. The initial therapy of febrile neutropenia has to be started empirically before identification of causative pathogens or infectious foci. The febrile episodes should be treated with broad spectrum antibiotics (combinations or monotherapy) due to the spectrum of microorganisms or resistance situation at hospital. In case of non-response after 3-4 days the initial therapy should be modified, in addition to further antibacterial therapy the start with an antifungal drug has to be recommended. In patients with pulmonary infiltrates the early treatment with amphotericine B has been shown to be more advantageous than delayed antifungal therapy. Furthermore, the antibiotic therapy is based on proven microorganisms, susceptibility testing and infectious foci. The value of interventional treatment with G-CSF or GM-CSF is controversely discussed. An uncompromising handling of febrile neutropenia is necessary to reduce the mortality due to infections in patients with hematologic malignancies.

Anti-Bacterial Agents↗

Identification to the species level of the plant pathogens Phytophthora and Pythium by using unique sequences of the ITS1 region of ribosomal DNA as capture probes for PCR ELISA.

The ribosomal internal transcribed spacer 1 region was sequenced for 10 species of Pythium and eight species of Phytophthora. Alignment of the sequences revealed considerable sequence microheterogeneity, which was utilized to prepare a capture probe of unique sequence for each species. The capture probes were tested by PCR ELISA, combining the sensitivity and specificity of the polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA). The probes were entirely species specific, enabling the detection and identification of the amplified DNA of species from individual cultures or from mixed samples of the DNAs of two different species. This approach to species identification, which provides a molecular technology to process large numbers of samples and still identify the fungi with a high level of confidence, may greatly reduce the resources and the time of highly trained specialists currently needed to identify these important species of plant pathogenic fungi.

Base Sequence↗

Identification of genomic differences between Escherichia coli strains pathogenic for poultry and E. coli K-12 MG1655 using suppression subtractive hybridization analysis.

Diseases of poultry caused by Escherichia coli result in significant economic loss every year. Specific virulence factors associated with E. coli strains pathogenic for poultry have been identified, but it is likely that others remain to be identified. To identify unique DNA fragments associated with avian strains we used suppression subtractive hybridization. The genome of E. coli K-12 strain MG1655 was subtracted from the genomes of two avian E. coli strains resulting in the identification of 62 fragments specific to the two avian strains. Sequence homology analysis was done and four types of fragments were identified: plasmid sequences, phage sequences, sequences with known function and sequences without any currently known function. Two E. coli collections, a reference collection of diverse strains (ECOR) and a collection of 41 avian isolates, were screened for the presence of 25 of the 62 fragments. We identified nine fragments present in significantly more of the avian strains than of the ECOR strains. Five fragments were in significantly more of the ECOR strains than the avian strains. These results suggested that the nine fragments could play a role in the pathogenesis of E. coli as it relates to diseases of poultry.

Animals↗

Failure of Listeria monocytogenes keratitis to respond to topical ofloxacin.

PURPOSE: The purpose of this case report is to demonstrate the need for identification of the offending pathogen in bacterial keratitis to guide appropriate treatment. METHODS: Case report. RESULTS: We report a case of Listeria monocytogenes keratitis in a young healthy adult that failed to respond to standard empirical therapy for bacterial keratitis but was treated successfully with topical ampicillin, chloramphenicol, and systemic amoxicillin. CONCLUSION: This case demonstrates the need for identification of the offending pathogen to guide appropriate treatment. It also demonstrates discrepancy between in vitro sensitivity and clinical response.

Adolescent↗

Identification and characterization of Eutypa leptoplaca, a new pathogen of grapevine in Northern California.

Although Eutypa lata is the main agent of Eutypa dieback of grapevine, another species of Eutypa has been isolated from diseased grapevines in Northern California. Stromata of this recently discovered Eutypa were also collected from Acer macrophyllum, A. negundo, and Fraxinus latifolia in the vicinity of vineyards, and appeared commonly on Umbellularia californica in some mixed-evergreen forests of Napa and Sonoma counties. This second species of Eutypa was distinguished from E. lata because of the sulcate ostiole of the perithecium and smaller ascospores. A morphological comparison with type specimens revealed identical features between the Californian isolates and E. leptoplaca sensu Rappaz (1987). This identification was confirmed through phylogenetic analyses of Eutypa spp. based on the complete sequence of the internal transcribed spacer (ITS) of the rDNA and partial sequence of the beta-tubulin gene. These analyses also separated collections of E. maura, E. sparsa, E. lejoplaca, E. tetragona, E. leptoplaca and E. lata, confirming the previously proposed species concepts. The pathogenicity of E. leptoplaca on grapevine was established using isolates collected from Vitis vinifera, U. californica, and A. macrophyllum. The importance of E. leptoplaca in relation to Eutypa dieback and its role as a necrotrophic pathogen are discussed.

Ascomycota↗

Genomic Comparison of cag pathogenicity island (PAI)-positive and -negative Helicobacter pylori strains: identification of novel markers for cag PAI-positive strains.

In an analysis of Helicobacter pylori genomic DNA by macroarray methodology, genomic DNA from a panel of cag pathogenicity island (PAI)-negative H. pylori clinical isolates failed to hybridize with 27 genes located outside the cag PAI in a cag PAI-positive reference strain. PCR analyses confirmed that HP0217 (encoding a lipopolysaccharide biosynthetic protein) and HP1079 (encoding a protein of unknown function) were present significantly more frequently in cagA-positive strains than in cagA-negative strains. A low G+C content of these two genes suggests they were acquired by horizontal transfer events.

Antigens, Bacterial↗