A Method for Identification and Analysis of Non-Overlapping Myeloid Immunophenotypes in Humans

The development of flow cytometric biomarkers in human studies and clinical trials has been slowed by inconsistent sample processing, use of cell surface markers, and reporting of immunophenotypes. Additionally, the function(s) of distinct cell types as biomarkers cannot be accurately defined without the proper identification of homogeneous populations. As such, we developed a method for the identification and analysis of human leukocyte populations by the use of eight 10-color flow cytometric protocols in combination with novel software analyses. This method utilizes un-manipulated biological sample preparation that allows for the direct quantitation of leukocytes and non-overlapping immunophenotypes. We specifically designed myeloid protocols that enable us to define distinct phenotypes that include mature monocytes, granulocytes, circulating dendritic cells, immature myeloid cells, and myeloid derived suppressor cells (MDSCs). We also identified CD123 as an additional distinguishing marker for the phenotypic characterization of immature LIN^-CD33⁺HLA-DR^- MDSCs. Our approach permits the comprehensive analysis of all peripheral blood leukocytes and yields data that is highly amenable for standardization across inter-laboratory comparisons for human studies.     

Polar lipids of 'Pseudomonas diazotrophicus'

Abstract Reference strains of ' Pseudomonas diazotrophicus ' produce a range of polar lipids atypical of authentic Pseudomonas species. In addition to the phospholipids common in Gram-negative bacteria (phosphatidylglycerol, diphosphatidylglycerol, and phosphatidylethanolamine), N -methylated derivatives of phosphatidylethanolamine (including phosphatidylcholine) and an ornithine amide lipid are also present. The preponderant ester-bound fatty acid (up to 80% of the total) is cis -vaccenic acid ( cis -octadec-11-enoic acid), while 3-hydroxyoctadecanoic acid is the major amide-bound fatty acid in the ornithine lipid. Possible implications of the data for classification of the organism are discussed.     

Two novel mutations in the MHC class II transactivator CIITA in a second patient from MHC class II deficiency complementation group A

Congenital MHC class II deficiency or bare lymphocyte syndrome (BLS; McKusick 209920) is caused by defects in trans-acting regulatory factors that control MHC class II expression and is therefore a disease of gene regulation. There are at least four complementation groups and the genetic and molecular dissection of this rare disease has contributed considerably to our current understanding of the molecular mechanisms governing MHC class II expression. Identification of the gene that is defective in BLS complementation group A, CIITA (MHC class II transactivator), has led to the discovery that CIITA acts as a master control factor of MHC class II expression. We have identified the CIITA mutations in a second patient from BLS group A. Two novel mutations abolish CIITA function, as shown by transfection experiments. Molecular analysis of these two novel mutations, together with the one described earlier in the first patient, is informative in terms of CIITA structure-function relationships. Received: 19 October 1996 / Revised: 25 November 1996     

The botany,uses and production ofWasabia japonica (Miq.) (Cruciferae) Matsum

Wasabi (Wasabia japonica) is a unique native plant and a traditional condiment crop of Japan. It is used in traditional Japanese raw fish and noodle dishes and in several modern foods for its hot taste and tangy flavor. Japanese farmers grow the crop in wet upland orchard soils for leaves, petioles and small enlarged stems, and in flooded gravel and sand fields along streams or near springs to produce whole plants and large succulent green enlarged stems. Recent studies in Japan have demonstrated numerous enzymatic and biocidal properties of the plant. This review of Japanese and other literature details the history, uses, botany, cultivars, ecological requirements, production techniques, insect pests and diseases of wasabi.