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R Sherburne

Publications and source records attributed to R Sherburne.

24 records · Page 2Linked to original sources

Examination of enterotoxigenic Escherichia coli H10407 (colonization factor antigen I+) by scanning electron microscopy with conductive staining.

We have used the scanning electron microscope to examine enterotoxigenic Escherichia coli H10407, which expresses colonization factor antigen I pili. The use of low accelerating voltages and conductive staining procedures allowed us to obtain images of colonization factor antigen I pili and other structural details which were obscured by conventional gold-coating techniques.

Agglutination↗

Structure and location of tellurium deposited in Escherichia coli cells harbouring tellurite resistance plasmids.

The plasmids RP4Ter and pHH1508a, which belong to the P and HII incompatibility groups, respectively, confer resistance to potassium tellurite (K2TeO3) on Escherichia coli. The genes for tellurite resistance were cloned from each plasmid onto the vector pUC8 to create pDT1366 and pD1364, respectively. Unstained, unfixed bacteria carrying these plasmids contained black intracellular deposits when grown on media containing tellurite. Thin sections of these bacteria fixed with glutaraldehyde were prepared and examined by electron microscopy. The black deposits were located inside the cell and were frequently associated with the inner membrane of the bacterium. Bacteria containing pDT1366 or pDT1364, and therefore a higher gene dosage of the Ter determinant, contained more black deposits, but had a decreased resistance, as measured by the minimum inhibitory concentration using the agar dilution method. Using the technique of electron spectroscopic imaging, the black intracellular deposits were shown to contain predominantly reduced metallic tellurium, and significant amounts of oxygen or carbon, thereby confirming earlier results using X-ray diffraction analysis of whole cells.

Cell Membrane↗

Incidence of antibiotic resistance and characterization of plasmids in Campylobacter jejuni strains isolated from clinical sources in Alberta, Canada.

Antibiotic susceptibilities of 382 strains of Campylobacter jejuni isolated in Alberta, Canada, in 1980 and 1981 were determined. Although none were resistant to erythromycin or gentamicin, 5.4 and 22% of strains were resistant to ampicillin in 1980 and 1981, respectively. Tetracycline resistance was noted in 6.8% of strains in 1980 and in 8.6% in 1981. Moreover, resistance to high-level tetracycline (32-128 micrograms/mL) was always mediated by a plasmid of 45-50 kilobases. Three transmissible plasmids from the Alberta strains were compared with the prototype plasmid pMAK175 by restriction enzyme analysis and some minor differences in restriction sites were noted between pMAK175 and pUA183. The two other plasmids pUA142 and pUA143 were 4 kilobases larger than pMAK175 and contained additional restriction sites. However, in all plasmids examined, the HincII and AccI fragments where the tetracycline-resistance determinant was located were shown to be conserved.

Alberta↗

Morphological forms and viability of Campylobacter species studied by electron microscopy.

Electron microscopic studies of Campylobacter revealed that different morphological forms predominate at different parts of a colony. At the periphery, cells were almost all spirals, while in the center of the colony cells were mainly coccus shaped. Unusual ring-shaped cells, "donuts", were observed in the raised, peripheral region of the colony. Donut or ring forms have not previously been reported for Campylobacter organisms. Our data indicate that young or actively growing cells are mainly spiral shaped. Older cells undergo a degenerative change to coccoid forms. The donut shape appears to be an intermediate stage between spirals and cocci. Comparisons of plate counts of actively growing and inactive cells confirmed that coccoid cells are probably nonviable.

Campylobacter↗

Blemishes developing on soft contact lenses.

Suspected contamination of soft contact lenses lead to biological, X-ray spectrophotometric, and photographic investigations to attempt identification of substances which could not be cleaned from the lenses and which increased in number with time. All indications point to suspect contaminant not being foreign material but a breakdown of the polymer structure.

Contact Lenses, Hydrophilic↗

Preparation and functional characterization of villous cytotrophoblasts free of syncytial fragments.

Recent studies suggest that purified villous cytotrophoblasts are largely contaminated by mononucleated syncytial fragments and therefore unsuitable for studies of trophoblast differentiation. We assessed highly purified (>99.99 per cent) populations of villous trophoblasts for fragment contamination using the syncytial markers placental alkaline phosphatase (PLAP, by immunohistochemistry) and exteriorized phosphatidyl serine (ePS, by flow cytometric analysis). The preparations contained from 4-46 per cent syncytial fragments. However, we find that PLAP negative cells preferentially adhere to tissue culture surfaces and that all preparations were <2 per cent PLAP positive after routine plating and washing procedures. A second purification procedure eliminated dead (propidium iodide permeable) cells and separated viable syncytial fragments (ePS-positive) from viable cytotrophoblasts (ePS-negative) by two colour fluorescence activated cell sorting (FACS). Viable ePS-positive cells were ultrastructurally apoptotic, adhered poorly in culture and those that adhered rapidly underwent apoptosis. Viable ePS-negative cells contained large heterochromic nuclei and cytoplasmic structures, adhered strongly in culture and remained viable. The latter population (putative true villous CT) differentiated into syncytialized cells when cultured with EGF. We conclude that villous CT can be routinely purified, are viable in culture and can undergo syncytial fusion without extensive preformed syncytium.

Adult↗