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Stimulation of macrophages by a variety of agents causes activation of mitogen-activated protein kinases (MAPKs). Activation of MAPKs by lipopolysaccharide involves CD14 and Toll receptors. Subsequent steps still remain to be explored. Tumor necrosis factor-alpha (TNF-alpha)-induced activation of MAPKs has been shown to involve the death domain proteins (TRADD, FADD, MADD) and TRAFs. Other molecules involved in this pathway include the protein kinases, ASK1, germinal center kinase (GCK), hematopoietic progenitor kinase 1 (HPK1), and GCK-related kinase (GCKR). Although, these pathways have been described in various cell types, their role in macrophages remains to be established. The availability of knockout mice and constitutively active and dominant-negative mutants of MAPKs should greatly enhance our understanding of this field. The activation of MAPKs seems to be different in cell lines compared with primary cells. Among the macrophages, cells from different compartments show different expression of receptors and signal transduction molecules. These differences may account for differences in MAPK activation and other phenotypic differences in macrophages from different compartments. Therefore, it is important to use primary cells for studying MAPK signal-transduction pathways, and the data from cell lines should not be extrapolated to primary cells.

Because obtaining multiple blood samples from individuals involved in epidemiologic studies is difficult, conclusions must be drawn on the basis of one or two samples. In this study, the authors attempted to determine the variability of two plasma markers over time, namely IL-6 levels and crosslinked fibrin degradation products (D-Dimers), in 16 elderly community-dwelling individuals. The study group included both men and women and black and white subjects (four in each group). Eight blood samples were obtained from each subject over a period of 36 days. Blood was separated within 1 hour after collection and aliquots of plasma were stored at -70 degrees C until all samples were collected. All samples from an individual were analyzed at one time. IL-6 and D-Dimers were measured by commercially available ELISA kits. The variability in the levels of plasma IL-6 and D-Dimers was assessed by means of intra-class correlation coefficient (ICC). The estimates of the ICCs for one measurement of IL-6 and D-Dimers were .87 and .86, respectively. Reliability values of this magnitude indicate excellent reproducibility in the measurement. These values indicate that obtaining a single sample from a subject is fairly representative of that individual's IL-6 and D-Dimer levels over an extended period of time.

Dichlorofluorescein (DCFH) oxidation assay measures hydrogen peroxide (H2O2), which is a derivative of superoxide anion. We found that a calmodulin antagonist, W-13, which is known to inhibit superoxide anion generation enhanced the capacity of human neutrophils to oxidize DCFH. To investigate this discrepancy we studied the role of nitric oxide (NO) in DCFH oxidation. Pure NO was capable of oxidizing DCFH, and the product formed had spectral properties identical to oxidized DCFH produced by H2O2. The arginine analog, NG-monomethyl-L-arginine (NMMA), which inhibits NO production, in combination with W-13 completely inhibited the stimulus-induced increase in DCFH oxidation. We conclude that the oxidation of DCFH in human neutrophils can occur by either H2O2 or NO.

N,N-diethylphenylacetamide (DEPA) has repellent activity against hematophagous insects including mosquitoes, black flies, horse flies, muscoid flies, rat fleas, and ticks, as well as land leeches and cockroaches. The efficacy of DEPA is comparable to that of N,N-diethyl-3-methylbenzamide (DEET) in both laboratory and field tests. Toxicological studies indicate that DEPA is safe for human use. Different formulations as well as fabrics impregnated with DEPA are effective for personal protection from insect bites.

The efficacy of fabrics impregnated with N,N-diethylphenylacetamide (DEPA), a new repellent, was studied in the laboratory and field against Aedes aegypti (L.) and Culex quinquefasciatus Say. DEPA-treated polyester cotton and cotton fabrics provided repellency for 30 and 36 d and had a half-life of 11 and 5 d, respectively. The repellent could not be applied at a rate of greater than 15% of fabric weight. DEPA-treated fabric patches were found to be a practical and novel approach and were effective for 90 d in the laboratory against Ae. aegypti and 77 d in the field against Cs. quinquefasciatus.

The residual repellency of N,N-diethylphenylacetamide (DEPA) was studied against American cockroach, Periplaneta americana (L.); German cockroach, Blattella germanica (L.); and brownbanded cockroach, Supella longipalpa (F.) at various concentrations. DEPA exhibited residual repellency for 4, 3, and 2 wk against American, German, and brownbanded cockroach, respectively, at a concentration of 0.5 mg/cm2.

The effects of pentoxifylline (Trental) on human neutrophil CR3 up-modulation, degranulation, and superoxide production were studied. We used the chemotactic peptide fMLP and the phorbol ester PMA as soluble stimuli, and beta-glucan particles as a CR3-specific solid phase stimulus of neutrophil superoxide production. Since neutrophils have adenosine A2 receptors, we compared effects of pentoxifylline to effects of adenosine, and we also looked at the effect of cytochalasin B, which breaks up actin filaments. Pentoxifylline inhibited both CR3 up-modulation and degranulation of myeloperoxidase and lysozyme. Pentoxifylline is a more potent inhibitor of fMLP- compared to PMA-induced degranulation, and is especially potent against superoxide production. While pentoxifylline is less potent than adenosine in its inhibition of fMLP-induced superoxide production, it is more potent in its inhibition of PMA- and beta-glucan particle-stimulated superoxide production. Cytochalasin B, which enhances degranulation and fMLP-stimulated superoxide production, was found to inhibit beta-glucan particle-stimulated superoxide production. These findings are consistent with the hypothesis that pentoxifylline can affect both the cytoskeletal architecture of unstimulated neutrophils and the activation and responses of neutrophils which involve actin polymerization and receptor-cytoskeletal interactions.

Haemagglutination inhibition (HI) test using goose erythrocytes (GRBC) is the method of choice for screening antibodies to various arboviruses. This test makes use of fresh GRBC preserved in dextrose-gelatin-veronal (DGV). We have tried formalinizing GRBC and have found that such cells can retain the surface property of being agglutinated by arboviruses and, therefore, can be used in haemagglutination (HA) and HI tests. Unlike fresh cells preserved in DGV, formalinized cells can be preserved for a long time without haemolysis or loss of sensitivity. They can also be frozen or lyophilized. Making use of these properties, a kit has been developed for the assay of HI antibodies to some members of the flavivirus group. Use of this kit simplifies the HI test and enables it to be carried out routinely by any hospital or laboratory to screen arboviral infections.

The DNase I inhibition assay was used for the determination of the relative amounts of monomeric and total actin in normal human peripheral blood lymphocytes, normal Sprague-Dawley rat peritoneal leukocytes, and 6 transformed human cell lines of lymphoid and myeloid origins. In the normal lymphocytes and leukocytes, greater than 50% of the total actin was monomeric. In contrast, only 1 of the 6 transformed lines had greater than 50% of its actin in the monomeric form. In the other 5 lines, the percentage of actin in the monomeric form ranged from 23 to 39%. The normal lymphocytes, peritoneal leukocytes, and 2 of the transformed cell lines (CEM and K562) were examined in greater detail. The total actin (as a percentage of the total protein) was found to be much lower in the transformed cell lines than it was in the normal lymphocytes. The total amount of actin in the normal rat leukocytes was very similar to the amount in the normal human lymphocytes. In addition to these differences between the normal and transformed human cells, treatment of the normal lymphocytes with mitogenic doses of concanavalin A was found to significantly reduce the amount of monomeric actin in the cells. Similar treatment of the transformed cells produced no significant reduction in the monomeric actin.