A cluster of infections caused by Enterobacter cloacae was observed among preterm neonates in a neonatal intensive care unit (NICU) of a pediatric hospital in Osnabruck, Germany. The presence of similar antimicrobial susceptibility patterns among the bacterial isolates prompted an investigation to determine whether a limited spread of a single strain existed. All 12 E. cloacae isolates from the NICU and 50 nonrelated strains were fingerprinted by small-fragment restriction endonuclease analysis (SF-REA) of EcoRI DNA digests. Selected isolates were further characterized by pulsed-field gel electrophoresis (PFGE) of NotIor XMaI-generated genomic restriction fragments. Epidemiologically unrelated strains were clearly discriminated by both methods. Results achieved by SF-REA and PFGE revealed that of the 12 isolates from the NICU, 11 belonged to the same genotypic cluster. Since all reagents and equipment for both techniques are commercially available, DNA fingerprinting by SF-REA or PFGE is proposed as a useful tool in the microbiology laboratory for investigating the epidemiological relatedness of E. cloacae strains of clinical and environmental origin. * Corresponding author. gel electrophoresis (PFGE) as typing tools for E. cloacae. The results are compared with those achieved by traditional methods available in a routine microbiology laboratory and are evaluated with respect to typeability, reproducibility, discriminatory power, and ease of use.
Summary.-The effect of the growth of two syngeneic transplanted sarcomata of widely differing biological properties on the number of monocytes in the blood of rats was measured (1) by binding of a specific antimacrophage serum to leucocytes, and (2) by sedimenting in a density gradient rosettes between mononuclear cells and antibody-coated sheep red cells under conditions in which B-cells are not brought down. For the 4 syngeneic sarcomata studied there was a progressive increase in the number of monocytes with tumour growth and the values returned to normal a few days after their surgical removal. The extent of monocytosis was related to the immunogenicity of the tumour and was most pronounced for the HSBPA sarcoma, which is highly immunogenic, has a low rate of spontaneous metastasis and contains many macrophages, and least for the MC-3 sarcoma which is essentially non-immunogenic, invariably gives rise to distant metastases and contains only about 8% macrophages. The growth of sarcomata had previously been found to reduce the number of monocytes which enter inflammatory lesions, both non-specific and due to a delayed hypersensitivity reaction. This " antiinflammatory" action of sarcomata which is related to their immunogenicity cannot be ascribed to the preferential uptake of monocytes by the tumours and it is concluded that the monocytes in the blood of tumour-bearers, though increased in number, are modified so that they do not enter sites of inflammation.
In order to investigate possible differences in sugar binding activities of strongly versus weakly metastatic tumors, sugar-binding molecules (endogenous lectins) of murine tumor cells differing in metastatic capacity were analyzed by affinity chromatography on supports with immobilized sugars or glycoproteins and compared. After elution with specific sugar in the absence of Ca2+-ions, the proteins were separated by sodium dodecyl sulfate-polyacrylamide slab gel electrophoresis. In comparison to a weakly metastatic subline (Eb) spontaneous strongly metastatic variants (ESb) of a murine lymphoma contained additional sugar receptors for N-acetylglucosamine (Mr 30 kDa) and maltose (Mr 64 kDa, 62 kDa, 54 kDa and 32 kDa), and lacked one sugar receptor for myoinositol (Mr 85 kDa), N-acetylglucosamine (Mr 23 kDa) and maltose (Mr 22 kDa), respectively. The strongly metastatic variant ESb expressed the common beta-galactoside-specific lectin to a higher extent and receptors for myo-inositol, melibiose and mannan to a lower extent. In another model system derived from the murine mastocytoma cell line P 815 X 2A, biochemical analysis of the liver-metastasizing variant P 815 X 2B revealed additional characteristic N-acetylgalactosamine- and maltose-specific binding proteins. This variant had reduced amounts of receptors for beta-galactosides and fucose in comparison to the parental clone. In a third tumor system a similar qualitative difference was disclosed: a metastatic variant derived from spleen metastases displayed a sugar receptor profile with 5 additional beta-galactoside-binding proteins when compared to its parental clone 6-6#3 + F, which is a virally transformed fibroblast line. The results show that metastatic variants of 3 murine tumor models consisting of lymphomas, mastocytomas and sarcomas are characterized by qualitative and quantitative alterations in the profiles of sugar-binding proteins.
Epidemiological fingerprinting of Klebsiella pneumoniae was performed by restriction endonuclease analysis (REA) of whole cell DNA. 11 isolates from 4 patients in an intensive care unit and 80 unrelated strains were examined in this study. DNA was cleaved with restriction endonuclease EcoR I, electrophoresed on 10% polyacrylamide gels, and restriction fragment patterns were visualized by silver staining. The analysis of small fragments within the cleavage patterns (SF-REA) yielded sufficient information for reliable strain identification. The gel patterns of unrelated strains exhibited marked differences by direct visual comparison. In contrast, the isolates from the ICU could only be subdivided into 2 types, supporting our suspicion of nosocomial infections in some of these patients. SF-REA was evaluated with regard to interstrain discriminatory ability, reproducibility, and practicability. Our results indicate that SF-REA may be used as a rapid, precise and reliable technique in typing K. pneumoniae strains.
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