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R F Cheek

Publications and source records attributed to R F Cheek.

4 recordsLinked to original sources

In vitro detection of Mycoplasma fermentans binding to B-lymphocytes in fresh peripheral blood using flow cytometry.

Previous reports have shown, using fluorescent probes conjugated to the organism, that Mycoplasma fermentans fuses with about 12% of peripheral blood lymphocytes. However, no lymphocyte subset was specified. To elucidate the specific subset of lymphocytes involved, we developed a three-color flow cytometric assay to detect M. fermentans binding to fresh peripheral blood cells. In our assay, two strains of M. fermentans were grown in SP4 glucose broth, mixed with fresh whole blood samples (n > 20), and incubated at 37 degrees C. The blood samples were then stained with a polyclonal antibody to M. fermentans, a monoclonal antibody to B-lymphocytes (CD19), and a monoclonal antibody to T-lymphocytes (CD3). Using three-color flow cytometry, we obtained data confirming binding of M. fermentans to 10%-15% of peripheral blood lymphocytes with minimal granulocyte or monocyte staining detected. Flow cytometric analysis showed that early binding appears predominantly directed towards B-lymphocytes (86.7 +/- 9.0%), and that this binding could not be blocked by antibodies directed towards common B lymphocyte cell surface antigens. M. fermentans binding to B-lymphocytes occurred within 5 min of in vitro inoculation, reached a maximum within 30-60 min (94-97%), and thereafter plateaued. The binding was concentration dependent over a three log dilution using 10(3) color changing units as standard. Binding to T-lymphocytes was minimal (<5% positive). B lineage tumor cells or peripheral blood B cells obtained from HIV infected individuals demonstrated reduced binding of M. fermentans. This assay provides a good method to study the cellular interactions of mycoplasma and may help to elucidate pathogenic mechanisms of mycoplasma infections.

Antibodies, Monoclonal↗

Benign cutaneous adnexal tumors with combined folliculosebaceous, apocrine, and eccrine differentiation. Clinicopathologic and immunohistochemical study of eight cases.

Benign cutaneous adnexal tumors displaying divergent differentiation are rare, with very few well-documented cases reported in the literature. We describe eight cases of benign adnexal tumors showing a variable combination of eccrine, apocrine, and folliculosebaceous differentiation. Clinically, all tumors presented as solitary, slowly enlarging dermal or subcutaneous nodules located in the head and neck and the extremities. Histologically, they were characterized by well-circumscribed, unencapsulated nodules composed of a lobular proliferation of epithelial cells displaying a spectrum of trichogenic, sebaceous, apocrine, and eccrine differentiation. The histological spectrum included lobules and trabeculae of basaloid cells with glandular and ductal elements, well-formed folliculosebaceous units, primitive follicles, and foci of tricholemmal keratinization. Immunohistochemical evaluation in four cases showed similar cytokeratin, carcinoembryonic antigen, and epithelial membrane antigen staining profiles as those reported for sweat gland adenomas; in addition, focal S-100 protein positivity and GCDFP-15 positivity could also be demonstrated, suggesting eccrine-apocrine differentiation. The tumors were most frequently confused histologically with other adnexal neoplasms, including sebaceoma, sebaceous adenoma, basal cell carcinoma, chondroid syringoma, and trichoepithelioma. The present series highlights the capability.

Adenoma↗

Red cell alloimmunization after a bone allograft.

BACKGROUND: Alloimmunization to blood group antigens other than ABH after bone allografting is uncommon. To date, examples of alloimmunization to blood group antigens other than ABH have been limited to the Rh system. CASE REPORT: A 20-year-old woman received an allogeneic bone graft following resection of an osteosarcoma. The patient had received no blood components and denied any pregnancies. Alloimmunization was detected and alloantibodies were identified by standard serologic techniques. Fourteen months after the bone graft, anti-Fya and anti-Jkb were identified in her serum. CONCLUSION: Alloimmunization to blood group antigens other than ABO and Rh may follow bone allografting.

Adolescent↗

Protein binding of palmitate measured by transmembrane diffusion through polyethylene.

Thin polyethylene membranes permit ready diffusion of protonated long-chain fatty acids but are impermeable to protein and ions. This circumstance recommends polyethylene for measuring the free fraction of fatty acids in the presence of a binding protein and for estimating the ionization constant with which to compute the equilibrium constant for the binding of fatty acid anions. As an example of this approach we report the binding of tracer palmitate to bovine albumin and bovine beta-lactoglobulin. We find a binding constant for the high-affinity site on albumin that is close to that calculated by others from heptane:H2O partition ratios. Our procedure is simpler, however, and free of the theoretical objection that heptane may alter the binding characteristics of the protein. Our estimate of the pKa for palmitic acid is 4.9, a finding that conforms to the widely predicted but heretofore unconfirmed expectation that long-chain fatty acids should have a pKa of about 4.8. Unidirectional flux measurements exclude direct exchange of palmitate between albumin and polyethylene.

Albumins↗