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Pathogen-induced apoptosis of macrophages: a common end for different pathogenic strategies.

Microbe-macrophage interactions play a central role in the pathogenesis of many infections. Several bacterial pathogens induce apoptosis specifically in macrophages, but the mechanisms by which it occurs differ, and the resulting pathology can take different courses. Macrophage death caused by Shigella flexneri and Salmonella spp. has been shown to result in the release of pro-inflammatory cytokines. Conversely, Yersinia spp. induce apoptosis by suppressing the signalling pathways that lead to the production of tumour necrosis factor (TNF)-alpha, a cytokine essential for the control of this infection. It is likely that there are a variety of reasons why macrophages are particularly susceptible to pathogen-induced apoptosis. One reason may be the expression of surface receptors that recognize highly conserved bacterial components, such as lipopolysaccharide (LPS) and bacterial lipoproteins (BLPs). These receptors have recently been shown to activate pro-apoptotic signalling pathways. The roles of macrophage apoptosis in different disease processes are discussed.

Animals↗

Identification of a pathogenicity island, which contains genes for virulence and avirulence, on a large native plasmid in the bean pathogen Pseudomonas syringae pathovar phaseolicola.

The 154-kb plasmid was cured from race 7 strain 1449B of the phytopathogen Pseudomonas syringae pv. phaseolicola (Pph). Cured strains lost virulence toward bean, causing the hypersensitive reaction in previously susceptible cultivars. Restoration of virulence was achieved by complementation with cosmid clones spanning a 30-kb region of the plasmid that contained previously identified avirulence (avr) genes avrD, avrPphC, and avrPphF. Single transposon insertions at multiple sites (including one located in avrPphF) abolished restoration of virulence by genomic clones. Sequencing 11 kb of the complementing region identified three potential virulence (vir) genes that were predicted to encode hydrophilic proteins and shared the hrp-box promoter motif indicating regulation by HrpL. One gene achieved partial restoration of virulence when cloned on its own and therefore was designated virPphA as the first (A) gene from Pph to be identified for virulence function. In soybean, virPphA acted as an avr gene controlling expression of a rapid cultivar-specific hypersensitive reaction. Sequencing also revealed the presence of homologs of the insertion sequence IS100 from Yersinia and transposase Tn501 from P. aeruginosa. The proximity of several avr and vir genes together with mobile elements, as well as G+C content significantly lower than that expected for P. syringae, indicates that we have located a plasmid-borne pathogenicity island equivalent to those found in mammalian pathogens.

Bacterial Proteins↗

Asymptomatic intestinal colonization by pathogenic Entamoeba histolytica in amebic liver abscess: prevalence, response to therapy, and pathogenic potential.

Since the application of isoenzyme electrophoresis to the study of Entamoeba histolytica, the prevalence and natural history of asymptomatic intestinal colonization in patients with amebic liver abscess (ALA) has not been addressed. We prospectively evaluated this enteric phase in 50 patients with ALA, using two dosage regimens of metronidazole. The overall prevalence of asymptomatic colonization was 72% (36/50). All these isolates, without exception, proved to express pathogenic zymodemes. Despite a 100% clinical response of the hepatic lesions, failure to eradicate the organism from the bowel occurred in 20 of these 36 subjects. During longitudinal posttreatment surveillance, three carriers returned with second bouts of invasive disease: one with dysentery and two with liver abscesses. Thus, in patients with ALA, there is a high prevalence of intestinal colonization with exclusively pathogenic strains, and treatment with metronidazole frequently results in a continued carrier state. These carriers have a propensity for developing recurrent invasive disease and constitute a public health hazard.

Adult↗

A pathogen-induced chitin-binding protein gene from pepper: its isolation and differential expression in pepper tissues treated with pathogens, ethephon, methyl jasmonate or wounding.

A chitin-binding protein (CBP) cDNA (CACBP1) was isolated from a cDNA library of pepper (Capsicum annuum L.) leaves infected with Xanthomonas campestris pv. vesicatoria. The deduced amino acid sequence of the CACBP1 gene which has chitin-binding domain and hinge region shares a high level of identity with CBP sequences from tomato, potato and tobacco. The CACBP1 gene was organ-specifically regulated in pepper plants, and differentially induced during the compatible and incompatible interactions of pepper with X. campestris pv. vesicatoria or Phytophthora capsici. Expression of the CACBP1 gene was rapidly induced in the incompatible interactions upon pathogen infection. Transcripts of the CACBP1 gene was highly inducible in the leaves of matured pepper plants by Colletotrichum coccodes infection. In situ hybridization results showed that CACBP1 mRNA was expressed in the phloem area of vascular bundles in C. coccodes-infected leaf tissues. The pathogen-inducible CACBP1 gene was also strongly induced and accumulated in pepper leaves by ethephon, methyl jasmonate or wounding. These data suggest that ethylene and jasmonate may act as signal molecules in the signal transduction pathways of the CBP gene induction during the pepper defense- or pathogenesis-related plant responses.

Acetates↗

A cell wall component from pathogenic and non-pathogenic gram-positive bacteria (peptidoglycan) synergises with endotoxin to cause the release of tumour necrosis factor-alpha, nitric oxide production, shock, and multiple organ injury/dysfunction in the rat.

The incidence of sepsis and septic shock due to gram-positive organisms has increased dramatically over the last two decades. Interestingly, many patients with sepsis/septic shock have both gram-positive and gram-negative bacteria present in the bloodstream and these polymicrobial or "mixed" infections often have a higher mortality than infection due to a single organism. The reason for this observation is unclear. The aim of this study was to investigate whether cell wall fragments from gram-positive and gram-negative bacteria could synergise to cause the release of cytokines, shock, and organ injury/ dysfunction in vivo. Male Wistar rats were anaesthetised and received an intravenous bolus of vehicle (saline), lipopolysaccharide (LPS) from Escherichia coli (0.1 mg/kg), peptidoglycan (Pep G) from Staphylococcus aureus (S10 mg/kg), co-administration of LPS (0.1 mg/kg) and PepG from S. aureus (10 mg/kg), LPS (10 mg/kg), PepG from Bacillus subtilis, or co-administration of LPS and PepG from B. subtilis. Blood pressure and heart rate were monitored for 6 h before plasma samples were taken for the measurement of TNF-alpha, total nitrite, and biochemical indices of organ injury. Peptidoglycan from both pathogenic (S. aureus) and non-pathogenic (B. subtilis) gram-positive bacteria synergised with endotoxin to cause formation of TNF-alpha, nitrite, shock, and organ injury. Synergism between PepG and LPS may partly explain the high mortality associated with mixed bacterial infections, as well as the deleterious effects of translocation of bacteria, or their cell wall components from the gut lumen in patients with sepsis.

Animals↗

Candida krusei: biology, epidemiology, pathogenicity and clinical manifestations of an emerging pathogen.

Early reports of Candida krusei in man describe the organism as a transient, infrequent isolate of minor clinical significance inhabiting the mucosal surfaces. More recently it has emerged as a notable pathogen with a spectrum of clinical manifestations such as fungaemia, endophthalmitis, arthritis and endocarditis, most of which usually occur in compromised patient groups in a nosocomial setting. The advent of human immunodeficiency virus infection and the widespread use of the newer triazole fluconazole to suppress fungal infections in these patients have contributed to a significant increase in C. krusei infection, particularly because of the high incidence of resistance of the yeast to this drug. Experimental studies have generally shown C. krusei to be less virulent than C. albicans in terms of its adherence to both epithelial and prosthetic surfaces, proteolytic potential and production of phospholipases. Furthermore, it would seem that C. krusei is significantly different from other medically important Candida spp. in its structural and metabolic features, and exhibits different behaviour patterns towards host defences, adding credence to the belief that it should be re-assigned taxonomically. An increased awareness of the pathogenic potential of this yeast coupled with the newer molecular biological approaches to its study may facilitate the continued exploration of the epidemiology and pathogenesis of C. krusei infections.

Animals↗

Fate of pathogenic and non-pathogenic Escherichia coli strains in two fermented milk products.

The growth and survival of pathogenic and non-pathogenic strains of Escherichia coli was determined in traditionally fermented pasteurized and unpasteurized milk and in Lacto, an industrially fermented milk. Each milk treatment was incubated at 20 degrees C for 24 h and then stored at either 20 degrees C or 5 degrees C for 96 h. Lacto inhibited all the three E. coli strains. Two strains could not be recovered and the third survived only in very low numbers after 24 h storage of Lacto at both 20 degrees C and 5 degrees C. All three E. coli strains survived and multiplied to maximum cell numbers in the range 10(7)-10(9)/ml during traditional fermentation of unpasteurized milk. Cell numbers decreased to 10(3)-10(6) and 10(2)-10(5) during storage of the fermented product at 20 degrees C and 5 degrees C respectively. Higher maximum numbers, 10(9)-10(10), of the three strains of E. coli were attained during traditional fermentation of pasteurized milk. The numbers decreased to 10(5)-10(8) and 10(4)-10(7) during storage of the fermented product at 20 degrees C and 5 degrees C respectively. Generally, fewer E. coli survived when the fermented milk products were stored at refrigeration temperature.

Animals↗

Pathogenic mycoplasmas: cultivation and vertebrate pathogenicity of a new spiroplasma.

A spiroplasma recovered from allantoic fluids of chick embryos infected with the tick-derived suckling mouse cataract agent was grown in continuous passage on a new artificial culture medium. The cultured organisms induced typical ocular and other disease symptoms in susceptible animals, and were reisolated from involved host tissues. Although spiroplasmas have been previously recognized as plant and insect pathogens, this is the first spiroplasma shown to multiply at 37 degrees C and to be pathogenic for vertebrates.

Animals↗

Activity of four cephalosporin antibiotics in vitro against bovine udder pathogens and pathogenic bacteria isolated from newborn calves.

The in vitro activity of chephaloridine, cephalexin, cefatrizine (BL-S640), and cephapirin (BL-P-1322) was evaluated by the serial dilution method against pathogenic gram-positive and gram-negative bacteria isolated from bovine udders and neonatal calf diseases. Cephapirin showed the comparatively greatest activity against the most common streptococcal species associated with bovine mastitis, whereas cephaloridine exhibited the best activity against Staphylococcus aureus. Cefatrizine was more active than the other cephalosporins against the gram-negative bacteria studied. In general, the minimal bactericidal concentration of each cephalosporin was two- to fourfold lower than the comparative value reported in the literature against the same type of pathogen of human origin.

Animals↗

The 96-kilodalton antigen as an integral membrane protein in pathogenic Entamoeba histolytica: potential differences in pathogenic and nonpathogenic isolates.

A surface antigen (EH-96) of Entamoeba histolytica was demonstrated to be a plasma membrane antigen by immunoprecipitation of metabolically 35S-labeled antigen from live trophozoites, Triton X-114 detergent extracts, and plasma membrane-enriched fractions prepared by concanavalin A membrane stabilization and differential centrifugation. In addition, the antigen was localized to the plasma membrane by electron microscopy with colloidal gold. Antigen from E. histolytica strains immunoprecipitated with specific immunoglobulin M (IgM) or IgG2b monoclonal antibody was identical by one-dimensional peptide mapping with N-chlorosuccinimide. Additionally, antigen from different axenically cultivated amebae was demonstrated to be identical by N-chlorosuccinimide peptide mapping, as were peptide maps of IgG and IgM monoclonal antibody-purified antigen. The 96-kilodalton (kDa) surface antigen was identified on four axenically cultivated pathogenic isolates and on three polyxenically cultivated pathogenic isolates (zymodeme II) of E. histolytica but was absent or present in lesser quantity on six nonpathogenic polyxenically cultivated isolates. The 96-kDa antigen was detected in liver abscess fluid from four patients with amebic abscesses by enzyme-linked immunosorbent assay (ELISA) and immunoprecipitation. Two-dimensional gel electrophoresis profiles of the 96-kDa antigen purified from abscess material or from polyxenically cultivated trophozoites demonstrated that the antigens were related to the 96-kDa antigen found in axenically cultivated organisms.

Animals↗

Repeats in an extracellular protein of weakly pathogenic strains of Streptococcus suis type 2 are absent in pathogenic strains.

Streptococcus suis type 2 strains that are pathogenic for pigs produce a 110-kDa extracellular protein factor (EF). Nonpathogenic and weakly pathogenic strains do not produce EF or produce a protein (EF*) that is immunologically related to EF. To study the pathogenesis of S. suis type 2 in pigs and to develop tools and methods for the control of S. suis type 2 infections, we cloned and characterized the genes encoding EF and various EF* proteins. Analysis of the deduced amino acid sequences showed that the first 833 amino acids at the N terminus of the EF and EF* proteins were nearly identical. The proteins differed, however, at their C termini. Unlike the 110-kDa EF protein, the EF* proteins contained several repeated units of 76 amino acids. The number and arrangement of the repeats in the EF* proteins varied. The data suggest that the gene encoding EF could have evolved from an epf* gene by a specific deletion event. The lack of repeated amino acid units in the EF protein may be related to virulence.

Amino Acid Sequence↗

An experimentally pathogenic Bacillus species. II. The pathogenicity of the organism for mice.

The pathogenic effects produced in mice by intraperitoneal (i.p.) injection of a Bacillus species (OSU 372) are presented. This organism belongs to group 3 of the genus, and members of this group have not heretofore been shown to be pathogenic for mice even under experimental conditions. However, this organism is capable of producing a fatal involvement in doses which are not considered to be overwhelming. The mean lethal dose (LD50) of the organism for 20-25 g mice by the i.p. route is about 1 times 10-8 bacteria/mouse. A rapid drop in body temperature along with severe dehydration were noted in infected animals, and hematologic studies indicated that leukopenia and hemoconcentration also occurred. Although a transient septicemia developed, the bacteria could not be recovered from the tissues of fatally infected mice after a certain point in time. Results prevented indicate that the animals died of hypovolemic shock. A possible parallel with human bacillary infection is drawn.

Animals↗

[Studies of the incidence of healthy carriers of "pathogenic strains" or potentially pathogenic strains in the hospital environment].

An investigation has been carried out on healthy carriers of "positive Staphylococcus coagulase" and "occasional pathogenic" strains among kitchen, canteen and dispensary staff in the IVth Geriatric Division and First Aid Clinic of the Bologna "M. Malpighi" Hospital. All stains of "positive Staphylococcus coagulase" and "occasional pathogens" considered (Pseudomonas aeruginosa and Citrobacter) were put through antibiotic and chemotherapeutic sensitivity tests. The results are reported in detail.

Anti-Bacterial Agents↗

Pathogenic and non-pathogenic Naegleria and Acanthamoeba spp.: a new autochthonous isolate from an Italian thermal area.

We performed an epidemiological survey of 17 thermal baths and the same number of mud-basins. This study aimed to ascertain the presence and incidence of small free-living amoebae, particularly species and/or strains of Naegleria and Acanthamoeba spp., occasional etiological agents of fatal meningoencephalitis and/or ocular infections in man. Over 51 samples of water and mud incubated at 37 degrees C and at 45 degrees C, 34 (66.7%) became positive at 37 degrees C and 33 (64.7%) at 45 degrees C. We isolated 7 (6%) strains of Naegleria spp., 6 (5.2%) of Acanthamoeba spp., 39 (33.6%) of Vahlkampfia spp., 28 (24.1%) of Hartmannella spp. and 36 (31.1%) strains of other species of free-living amoebae. 4 strains of Naegleria spp. and 6 of Acanthamoeba spp. proved pathogenic both in vivo, after experimental infection (meningoencephalitis) in the albino mouse, and in vitro, having previously contaminated monolayers of Vero cell line (cytopathic effect). Within the isolated pathogenic strains of Naegleria spp., a new strain of N. australiensis s.sp. italica was typified from an immunochemical point of view. This should be added to previous isolations reported by us.

Amoeba↗

[Aggravation of experimental dysentery in mice and acceleration of the process of multiplication of pathogenic and conditionally pathogenic bacteria under the effect of yeast RNA].

Simultaneous infection of mice with Sh. flexneri subtype 2a, culture, with various doses of RNA sodium salt (NaRNA) or administration of the preparation 3 hours prior to or 6, 12, and 18 hours after the infection was accompanied by aggravation of the experimental infection in mice. This was expressed in a greater number of animals which contracted the disease and died, and in increased blood and peritoneal exudate microbiol density. The same processes were noted in mice infected with equal doses of shigella cultures passaged four times through broth with NaRNA, in comparison with the animals infected with shigellae passaged through broth alone. Incubation of a number of pathogenic and conditionally pathogenic microbes in broth with NaRNA was accompanied by a significant increase in the bacterial biomass volume; at the same time other conditionally patoogenic microbes or even different strains of the same type of the microbes reacted to NaRNA but weakly. A possibility of realization of all of these processes in vivo and their influence on the origination of bacterial complications and development of the infection is supposed.

Animals↗

Isoenzyme patterns of pathogenic and non-pathogenic Naegleria spp. using agarose isoelectric focusing.

Using agarose isoelectric focusing, the isoenzyme patterns of 7 different enzymes were compared in 52 Naegleria strains. The pathogenic N. fowleri was found the most homogeneous species. N. lovaniensis seems to be constituted of different types which form nevertheless a cohesive group. Within N. gruberi, large interstrain band variations were found in almost all enzyme systems. A re-examination of the taxonomic position of this species may therefore be taken into consideration. High temperature strains from Australia were confirmed to be different from N. lovaniensis. Members of a new pathogenic Naegleria sp., N. australiensis, seem to occur in Europe. Large thermophilic strains with many large pores in the cysts show identical zymograms and may constitute a new species or genus.

Acid Phosphatase↗

[Value of antigen, antibody and pathogen-specific lymphocyte detection in diagnosis of pathogen-induced arthritis].

In the differential diagnosis of infection-related arthritis (infectious arthritis, viral arthritis, reactive arthritis or Reiter's syndrome, Lyme disease) various laboratory methods are applied for the detection of the inciting antigen, specific antibodies or microbe-specific T-lymphocytes. In infectious (septic) bacterial or fungal arthritis, the definitive diagnosis can be made only by recovering the organism from the synovial fluid or membrane. Also, in reactive arthritis following extraarticular infection with Yersinia, Salmonella, Shigella, Campylobacter, or Chlamydia, one of the major shifts in perception of disease pathogenesis has been the detection of bacterial determinants by immunological methods and polymerase chain reaction (PCR) actually within the joint. In sexually acquired reactive arthritis, the etiologic diagnosis should be based on the direct detection of the pathogen (mainly C. trachomatis) from the urogenital smear specimen. For clinical routine, serological tests for bacteria specific antibodies (IgM and IgA class) are often necessary to show recent or persistent infection with the triggering pathogen. However, a cautionary note regarding the diagnostic significance of antibacterial antibody profiles has been sounded in several studies because of the high prevalence of bacteria-specific antibodies in the healthy population. The same problem may arise in the interpretation of virus-specific antibodies in the differential diagnosis of acute polyarthritis. Antigen-specific proliferation of synovial fluid lymphocytes can confirm the clinical diagnosis in patients with reactive arthritis and Lyme disease, although unspecific proliferation to several bacteria can also be observed in reactive arthritis as well as in many other arthritis.

Antibodies↗