Kynurenic acid: a potential pathogen in brain disorders.
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The red cell parasites formerly known as Haemobartonella and Eperythrozoon spp have been reclassified as hemotrophic mycoplasmas (hemoplasmas) based on strong phylogenetic evidence and 16S ribosomal RNA gene sequences. The latter form the basis for polymerase chain reaction assays used to detect infection. Candidatus designation was given to incompletely characterized species. Like other mycoplasmas, hemoplasmas are small epicellular parasites that lack a cell wall and are susceptible to tetracyclines; their circular, double-stranded DNA encodes only those gene products essential for life. Diseases caused by infection with hemoplasmas range from overt life-threatening hemolytic anemia to subtle chronic anemia, ill-thrift, and infertility. In addition, the organisms may act as cofactors in the progression of retroviral, neoplastic, and immune-mediated diseases. Intimate contact of hemoplasma organisms with RBCs leads to cell injury through immune-mediated and other mechanisms that have not yet been defined. Despite an intense immune response and even with antibiotic treatment, infected animals probably remain chronic carriers after clinical signs have resolved.
In vitro studies based on periodic viable count determinations indicated that the two antifungal drugs ketoconazole and 5-fluorocytosine generally were at least additive in their combined effect on various opportunistic yeast pathogens, including some resistant to 5-fluorocytosine.
The freshwater aquarium snail (Ampullaria spp.) was demonstrated to carry as many as 10(8) viable mesophilic bacteria per g of meat plus shell. Some 16 genera of bacteria were identified, with gram negatives predominating. Enrichment culture techniques enabled the isolation of salmonellae from 24 to 42 lots of 200 g each. The salmonellae comprised eight different serotypes, including Salmonella newport, Salmonella saint-paul, and Salmonella infantis. This association of salmonellae with snails may contribute to cases of human salmonellosis, since other aquarium species have already been shown to contribute to many such cases. The snails were also found to commonly harbor Pseudomonas aeruginosa and, occasionally, Edwardsiella tarda.
A cell suspension of Staphylococcus aureus (196E) was injected into raw skim milk which contained different concentrations of sugar, serum solids, fat, stabilizers, and emulsifiers. The ingredient samples were exposed for the desired length of time in a constant-temperature water bath (60 C). Standard plate counts were made, and the number of surviving organisms was determined. Regression coefficients for each ingredient concentration were calculated and plotted against the per cent ingredient concentration to give an indication of protective action. Analyses of variance were conducted on bacterial counts to test the protective action of each ingredient. A comparison of the number of survivors in different sugar concentrations showed that with up to 14% sugar all the organisms were killed within 30 min. In sugar concentrations above 14%, the number of survivors increased regularly with each increase in sugar concentration up to 57%, which was the maximum used. In concentrations of serum solids above 9%, some organisms survived 35 min of heat treatment. Butter fat, stabilizer, and emulsifier did not offer any protective action in the concentrations observed.
An unnamed, polarly flagellated, nonfermentative, H(2)S-producing gram-negative rod which was isolated from pathological material of 13 adult individuals is described.
Bacteriological analyses were performed on fecal swabs and the aquarium water of 27 individually purchased specimens of the small green pet turtle, Pseudemys scripta elegans. Representatives of Aeromonas, Citrobacter, Enterobacter, Klebsiella, Proteus, Salmonella, and Serratia were isolated. Enterobacter, Klebsiella, and Salmonella were encountered in 20% or more of the specimens, whereas Aeromonas was isolated from 63%. Klebsiella pneumoniae counts ranged from 10(3) to 10(4) per milliliter of aquarium water, whereas Aeromonas routinely exceeded 10(4) per milliliter. Aeromonas cultures from turtles were identical to 7 human isolates in some 29 biochemical tests. On the basis of our findings, we question whether the Salmonella-free certification program alone is sufficient to render these reptiles as safe pets.
The bacterial population of the water supplied with ornamental fish purchased from retail outlets was examined qualitatively and quantitatively. As many as 10(9) viable aerobic organisms per 100 ml were present, with fecal coliform counts as high as 10(5) per 100 ml. Citrobacter, Escherichia, Pseudomonas, and Vibrio were isolated from 75% or more of the samples, whereas Aeromonas, Alcaligenes, Enterobacter, Flavobacterium, and Streptococcus were isolated from 45 to 65% of the samples. Pseudomonas aeruginosa, Edwardsiella tarda, and Klebsiella pneumoniae were also isolated.
The phenotypic and nucleic acid properties of Klebsiella pneumoniae have been studied on cultures obtained from six different habitats (humans, vegetables, seeds, trees, rivers, and pulp mills). The 19 cultural reactions of 107 isolates varied significantly only in tryptophanase activity and dulcitol fermentation. The percentage of guanine plus cytosine base composition of 41 isolates varied from 53.9 to 59.2%. The range of percentage of guanine plus cytosine base composition for environmental klebsiellas was broader than that for the cultures of human origin. The range of deoxyribonucleic acid relative reassociation (homology) to the human K. pneumoniae reference strain extended from 5% to 100% and the chromosome molecular weights ranged from 2,200 x 10(6) to 3,000 x 10(6). The species of K. pneumoniae is thus molecularly more heterogeneous than previously thought and most isolates of human, pulp mill, and river origin are genetically indistinguishable. The presence of K. pneumoniae therefore represents a deterioration of the microbiological quality of the environment and should be considered of public health significance. At the present time the health significance of the molecularly more divergent strains, primarily of vegetable and seed origin, their relationship to klebsiellas of human origin, or to other genera of the Enterobacteriaceae is unclear.
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We have devised a procedure that permits the cultivation of a gram-positive coccoid species from biopsy material obtained from the antrum of the stomachs of patients with gastric disorders. Antibodies directed against surface proteins obtained from the coccoid isolates were detected in all patients with gastric disorders examined in this study, including both Helicobacter pylori-infected and H. pylori-uninfected patients. Several of these isolates, including a prototype designated strain SL100, have been characterized in some detail. Strain SL100 exhibits urease and exceptionally high catalase activities and assumes a variety of spherical morphologies as detected by electron microscopy. This isolate expresses an adhesin that binds to gastric mucin. The adhesin activity was detected only after the isolate was exposed to an acidic pH, suggesting that in the natural process of infection, the low pH of the stomach unmasks a cell surface component with adhesin activity. Strain SL100 grows best under a microaerophilic conditions (10% CO2, 5% O2, 85% N2), but it also grows quite well under aerobic conditions. Thus, this organism would be expected to proliferate outside of the human host as well as in the gastric mucosa. Oral infection of newborn piglets resulted in colonization of the gastric antrum and growth retardation. Preliminary taxonomic classification indicates similarity to the Staphylococcus DNA homology groups containing S. cohnii and S. xylosus. One of us (C.K.) apparently became infected with this organism as indicated by gastric symptoms and the subsequent presence of strain-specific antisera not present in other workers in the laboratory.
Phylogenetic analysis was utilized to investigate biological relationships (tissue tropism, disease presentation, and epidemiologic success), as evidenced by coevolution, among human strains of Chlamydia trachomatis. Nucleotide sequences of omp1, the gene encoding the major outer membrane protein (MOMP) of C. trachomatis, were determined for 40 strains representing 11 serovars. These data were combined with available omp1 sequences from GenBank for an analysis encompassing a total of 69 strains representing 17 serovars infecting humans. Phylogenetic analysis of the nucleotide and inferred amino acid sequences showed no evolutionary relationships among serovars that corresponded to biological or pathological phenotypes (tissue tropism, disease presentation, and epidemiologic success). In addition, no specific residues that may have evolved to play a role in determining biologically relevant characteristics of chlamydia, such as tissue specificity, disease presentation, and epidemiologic success, were apparent in the MOMP. These results suggest that variation in MOMP may have arisen from a need to be diverse in the presence of immune pressure rather than as a function of pathogenicity. Therefore, the role of MOMP in disease pathogenesis and infection may be passive, and it may not be the major ligand responsible for directing infection of various human cell types.
Actinomyces spp. exhibit type 1 fimbria-mediated adhesion to salivary acidic proline-rich proteins (PRPs) and statherin ligands. Actinomyces spp. with different animal and tissue origins belong to three major adhesion types as relates to ligand specificity and type 1 fimbria genes. (i) In preferential acidic-PRP binding, strains of Actinomyces naeslundii genospecies 1 and 2 from human and monkey mouths displayed at least three ligand specificities characterized by preferential acidic-PRP binding. Slot blot DNA hybridization showed seven highly conserved type 1 fimbria genes (orf1- to -6 and fimP) in genospecies 1 and 2 strains, except that orf5 and orf3 were divergent in genospecies 1. (ii) In preferential statherin binding, oral Actinomyces viscosus strains of rat and hamster origin (and strain 19246 from a human case of actinomycosis) bound statherin preferentially. DNA hybridization and characterization of the type 1 fimbria genes from strain 19246 revealed a homologous gene cluster of four open reading frames (orfA to -C and fimP). Bioinformatics suggested sortase (orfB, orf4, and part of orf5), prepilin peptidase (orfC and orf6), fimbria subunit (fimP), and usher- and autotransporter-like (orfA and orf1 to -3) functions. Those gene regions corresponding to orf3 and orf5 were divergent, those corresponding to orf2, orf1, and fimP were moderately conserved, and those corresponding to orf4 and orf6 were highly conserved. Restriction fragment length polymorphism analyses using a fimP probe separated human and monkey and rat and hamster strains into phylogenetically different groups. (iii) In statherin-specific binding, strains of A. naeslundii genospecies 1 from septic and other human infections displayed a low-avidity binding to statherin. Only the orf4 and orf6 gene regions were highly conserved. Finally, rat saliva devoid of statherin bound bacterial strains avidly irrespective of ligand specificity, and specific antisera detected either type 1, type 2, or both types of fimbria on the investigated Actinomyces strains.
Listeria monocytogenes lineage III strains belonging to subgroups IIIA (n = 8), IIIB (n = 5), and IIIC (n = 6) were examined along with other known serotype strains (n = 11) by PCR and Southern hybridization using several recently described species-, virulence-, and serotype-specific primers and probes. The virulence of seven representative lineage III strains was then evaluated in mice via the intraperitoneal route. The results suggest that subgroup IIIA consists of typical rhamnose-positive avirulent serotype 4a and virulent serotype 4c strains, subgroup IIIC consists of atypical rhamnose-negative virulent serotype 4c strains, and subgroup IIIB consists of atypical rhamnose-negative virulent non-serotype 4a and non-serotype 4c strains, some of which may be related to serotype 7. It is possible that subgroup IIIB (including serotype 7) may represent a novel subspecies within L. monocytogenes.
A total of 150 strains of staphylococci, clinically isolated, were tested for sensitivity to ovotransferrin (conalbumin). Among these, all the 50 coagulase-positive, mannitol-positive, and deoxyribonuclease-positive staphylococci appeared to be resistant to conalbumin, i.e., capable of growing in the presence of this transferrin. Among the other 100 strains, which were not classified as S. aureus, some appeared to be resistant and some were sensitive. The different behavior toward conalbumin is related to varying degrees of efficiency of the bacterial iron transport systems and, to test this, a simple method can be used, based on the addition of CrCl3 to the culture medium. The precipitation of iron produced by chromium salts has an effect on the growth of staphylococci similar to that produced by conalbumin and reveals the differences in the iron transport systems which occur in the genus Staphylococcus.
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A series of recombinants between Rous-associated virus type 0 (RAV-0), RAV-1, and a replication-competent avian leukosis virus vector (RCAN) have been tested for disease potential in day-old inoculated K28 chicks. RAV-0 is a benign virus, whereas RAV-1 and RCAN induce lymphoma and a low incidence of a variety of other neoplasms. The results of the oncogenicity tests indicate that (i) the long terminal repeat regions of RAV-1 and RCAN play a major role in disease potential, (ii) subgroup A envelope glycoproteins are associated with a two- to fourfold higher incidence of lymphoma than subgroup E glycoproteins, and (iii) certain combinations of 5' viral and env sequences cause osteopetrosis in a highly context-dependent manner. Long terminal repeat and env sequences appeared to influence lymphomogenic potential by determining the extent of bursal infection within the first 2 to 3 weeks of life. This would suggest that bursal but not postbursal stem cells are targets for avian leukosis virus-induced lymphomogenesis. The induction of neutralizing antibody had no obvious influence on the incidence of lymphoma.
Chromosomal translocations involving antigen receptor genes and oncogenes have been observed in several forms of lymphoid malignancy. Observations of their lymphocyte-restricted occurrence and a molecular analysis of some translocation breakpoints have suggested that some of these rearrangements are generated by V(D)J recombinase activity. However, a direct correlation between this activity and the generation of such rearrangements has never been established. In addition, because these aberrant rearrangements are usually detected only after a tumor has been formed, the frequency with which the recombinase machinery generates translocations has never been assessed directly. To approach these issues, immunoglobulin light-chain gene rearrangements were induced in pre-B cells transformed by temperature-sensitive mutants of Abelson murine leukemia virus and PCR was used to identify interlocus recombinants. Vlambda Jkappa and Vkappa Jlambda rearrangements as well as signal joints resulting from the recombination of Vlambda and Jkappa coding elements were recovered and were found to be similar in structure to conventional intrachromosomal joints. Because these products were detected only when the cells were undergoing active intralocus rearrangement, they provide direct evidence that translocations can be generated by the V(D)J recombinase machinery. Dilution analyses revealed that interlocus rearrangements occur about 1,000 times less frequently than conventional intralocus rearrangements. Considering the large numbers of lymphocytes generated throughout life, aberrant rearrangements generated by the V(D)J recombinase may be relatively common.