We present a novel single-platform assay for determination of the absolute number of human blood monocyte subpopulations, i.e., the CD14 11 CD16 2 and the CD14 1 CD16 11 monocytes. A four-color combination of antibodies to CD14, CD16, CD45, and HLA-DR reduces the spill-over of natural killer cells and of granulocytes into the CD14 1 CD16 11 monocyte gate. For these CD14 1 CD16 11 monocytes, the intra-assay coefficient of variation (CV) was 4.1% and the inter-assay CV was 8.5%. Looking at a cohort of 40 donors aged 18-60 years, we found no age dependence. There was however an effect of gender in that females had lower CD14 1 CD16 11 monocytes (45.4 AE 13.5 cells/ll) compared with males (59.1 AE 20.3 cells/ll) (P < 0.02). Using this novel approach, we can confirm that exercise will lead to more than three-fold increase of the CD14 1 CD16 11 monocytes. Also, we show that therapy with low doses of glucocorticoids will deplete these cells. This robust single-platform assay may be a useful tool for monitoring the absolute number of monocyte subpopulations in health and disease. ' International Society for Advancement of CytometryKey terms monocyte heterogeneity; gender; exercise; glucocorticoids MYELOMONOCYTIC stem cells in bone marrow give rise to monocytes and these are released into blood where they circulate for a few days and then migrate into tissue where they develop into different types of macrophages (1). Monocytes were initially defined based on morphology and cytochemistry. With the advent of monoclonal antibodies and flow cytometry, human monocytes were characterized with antibodies like CD14. This technology also allowed for the definition of two monocyte subsets, i.e., the classical CD14 11 CD162 and the CD14 1 CD16 11 nonclassical monocytes (2) with the latter cells characterized by higher MHC class II expression and by higher production of the cytokine tumor necrosis factor (3). With the recognition of the potential role of the nonclassical CD14

CF and AMG are effective in treatment-resistant infectious keratitis. They could restore ocular surface integrity and provide metabolic and mechanical support for corneal healing. For large corneal perforation, it may be better to use another procedure such as penetrating keratoplasty to restore ocular integrity.

Introduction: Brucellosis is a major public health problem in Egypt. The Brucella IgM/IgG lateral flow assay was developed as a point-ofcare test for the diagnosis of human brucellosis. The aim of this study was to assess the diagnostic value of the lateral flow assay for use in Egypt. Methodology: Fifty samples of patients who presented with clinical suspicion of brucellosis over a one-year period were collected. All samples were subjected to the Brucella IgM/IgG lateral flow assay, serum agglutination test (SAT), rose bengal RB Test (RB), 2-mercapteoethanol (2-ME), culture and PCR. SAT, 2-ME, culture and PCR were retested after the end of the treatment. Results: Culture and SAT confirmed the diagnosis of brucellosis in twenty patients. While 90% of the samples were positive by SAT, only 30% and 85% were positive by culture and PCR respectively. The sensitivity of the lateral flow assay calculated for the Brucella IgM/IgG was 95% and specificity was 97%. Conclusion: These data show that the lateral flow assay is more suitable for diagnosis of brucellosis in Egypt than culture and SAT. Application of the PCR on serum samples collected during follow-up revealed that the DNA of the pathogen was yet not completely cleared almost 60 days after the start of treatment with doxycycline and ciprofloxacin.