Hill races usually include large downhill running sections, which can induce significant degrees of muscle damage in a field setting. This study examined the link between muscle damage, oxidative stress, and immune perturbations following a 7-km mountainous hill race with 457 m of ascent and 457 m of descent. Venous blood samples were taken from 7 club level runners before, immediately after, and 48 hrs postrace. Samples were analysed for total and differential leukocyte counts, markers of muscle damage (CK), lipid peroxidation (MDA), and acute phase proteins (CRP; fibrinogen; alpha-1-ACT). The total antioxidant status (TEAC) and plasma levels of the proinflammatory cytokines IL-6, IL-8, and TNF-alpha were also determined. Subjective pain reports, and plasma activities of CK, MDA, and circulatory monocytes reached peak values at 48 hrs postrace (p < 0.05). TEAC and the cytokine IL-8 increased immediately after the race (p < 0.05). Plasma TNF-alpha remained unchanged (p > 0.05). Despite the reports of muscle damage and soreness, no evidence of an acute phase response was observed (p > 0.05), which may be explained by the failure of the race to induce a plasma TNF-alpha response. Future studies should examine the link between muscle damage, oxidative stress, and the acute phase response following hill races of longer duration with larger eccentric components.

Increased levels of C-reactive protein (CRP) and heat-shock protein-70 (Hsp70) in plasma are known to be associated with an increased risk of cardiovascular disease. In this study we have investigated the effects of environmental air pollution particles (PM10) and ultrafine carbon black (ufCB) on the expression of CRP and Hsp70 in the lung epithelial cell line, A549. After treatment with PM10 or ufCB the cells were found to have increased expression of CRP and Hsp70 localized in both the cell cytoplasm and nucleus. Analysis of the cell supernatants revealed that CRP and Hsp70 were present, suggesting secretion of both proteins in response to the particulate treatment. To investigate if the expression of CRP and Hsp70 was the result of free radical production, cells were treated with ufCB in the presence of antioxidants (NAL and Trolox). This revealed that antioxidants reduced the amount of CRP and Hsp70 secreted from the cells. These findings suggest that CRP and Hsp70 may be secreted from the lung epithelium as a result of oxidative stress and have important effects on the inflammatory response associated with inhalation of particulate matter.

In 1997, an IARC Working Group classified quartz (crystalline silica) as a Group 1 lung carcinogen, but only in some industries, i.e., the quartz hazard is a variable entity. The reactivity of the quartz surface may underlie its ability to cause inflammation, and treatments that ameliorate this reactivity will reduce the quartz hazard. In this study we treated quartz (Q) with aluminium lactate (AL), a procedure that is reported to decrease the quartz hazard, and explored the effect this had on the highly reactive quartz surface and on proinflammatory events in rat lungs. Aluminium lactate-treated quartz showed a reduced surface reactivity as measured by electron spin resonance and the hemolysis assay. Eighteen hours after instillation of Q into the rat lung, there was massive inflammation as indicated by the number of neutrophils in the bronchoalveolar lavage (BAL). In addition, Q induced an increase in BAL macrophage inflammatory protein-2 (MIP-2) while ALQ had no significant effect compared to control. Epithelial damage, as indicated by BAL protein and gamma glutamyl transpeptidase, also increased with Q but not with ALQ. Furthermore, Q induced an increase in MIP-2 mRNA by BAL cells while ALQ had no effect compared to controls. There was an increase in nuclear binding of the transcription nuclear factor κB (NF-κB) in the Q-exposed BAL cells and again no effect on nuclear NF-κB binding in BAL cells from ALQ-exposed rats. In conclusion, treatment of the quartz surface with aluminium lactate reduced the reactivity of the particles both in terms of hydroxyl radical generation and in terms of the induction of molecular signaling events leading to inflammation.

Chronic inflammation and fibrosis following quartz inhalation has been associated with persistent up-regulation of several "pro-inflammatory" genes, which are commonly regulated by nuclear factor kappa-B (NF-kappaB). Transcription of the NF-kappaB-inhibitor IkappaBalpha is also under NF-kappaB control, and its de novo synthesis is considered to comprise a negative feedback loop in transient inflammation. To investigate this mechanism in particle inflammation, we have studied IkappaBalpha degradation in A549 cells exposed to DQ12-quartz or TiO(2), in relation to the expression of IL-8. Although both quartz and TiO(2) were found to cause IkappaBalpha degradation, only quartz elicited a mild IkappaBalpha depletion, first appearing at 4 h. TiO(2) was found to cause a higher short-term increase in IkappaBalpha mRNA-expression compared to quartz, whereas the early enhancement of IL-8 expression and release was similar for both particles. Up-regulation of IL-8 expression was found to persist with quartz only. Cotreatment with PDTC and curcumin reduced particle-elicited IL-8 response, whereas cycloheximide caused enhancement of IL-8 mRNA expression in both the quartz- and TiO(2)-treated cells. Our results demonstrate that mineral dusts cause IkappaBalpha degradation, a transient increase in de novo synthesis of IkappaBalpha, and enhanced IL-8 expression in human pulmonary epithelial cells. While IkappaBalpha degradation and early IL-8 expression seem to be general particle phenomena, particle-specific characteristics impact on activation of IkappaBalpha gene transcription, apparently accounting for the different proinflammatory IL-8 responses seen with quartz and TiO(2) in the longer term. These observations may provide an explanation for the transient versus the persistent pulmonary inflammatory status and subsequent differences in pathogenic potency of TiO(2) and quartz.

The influence of the In(Lu) gene on human red blood cell (RBC) expression of CDw75 antigens was examined. CDw75 antigens were increased in expression on Lu(a-b-) cells of the dominant inhibitor type in comparison with red cells from donors of other Lutheran (Lu) phenotypes. In contrast, CD44 epitopes detected with F10-44-2, A3D8 and BU52 monoclonal antibodies (mAb) were decreased in expression on Lu(a-b-) red cells. Among normal blood donors of the common phenotype Lu(a-b+) there was a wide range of expression of CDw75 antigens on red cells. The results show that CDw75 is a quantitative polymorphism of human red cells and, among antigens influenced by the In(Lu) gene, is unique in being up-regulated in expression.

Using monoclonal antibodies (mAb) characterized at the last International Workshop on Human Leucocyte Antigens, we examined the expression of CDw75 antigens (beta-galactoside alpha 2,6-sialyltransferase) on normal peripheral blood cells and on cells from patients with B-cell chronic lymphocytic leukaemia (CLL). The mAb used (HH2, EBU.65, EBU.141 and OKB4) detect different epitopes of CDw75. Normal peripheral blood B cells expressed high levels of CDw75 detectable with HH2, EBU.65 and EBU.141 but did not react with OKB4. Cells from patients with B-cell CLL closely resembled normal B cells. All CDw75 epitopes, including OKB4, were strongly expressed on some Namalwa variant Burkitt lymphoma cell lines. The OKB4 epitope was also present on red cells from all normal donors. The other CDw75 mAb were unreactive with red cells from some normal donors. The CDw75 epitope detected with EBU.65 was present on most CD4+ T cells and on a minority of CD8+ cells. HH2 and EBU.141 stained only small numbers of T lymphocytes. OKB4 did not react with T cells. EBU.65+, CD4+ T cells had low levels of expression of CD45R0, CD29, CD54 and CD58, and had high levels of CD45RA antigen. Phytohaemagglutinin (PHA) activation of cells led to the loss of EBU.65 binding. These results suggest that the CDw75 epitope recognized by the EBU.65 mAb is a marker of native T lymphocytes. On B CLL cells the epitopes detected with HH2, EBU.65 and EBU.141 were destroyed by treatment with neuraminidase. Treatment of B-CLL cells and red cells with neuraminidase increased the binding of OKB4, suggesting that this epitope is masked by sialic acid. The results suggest that CDw75 is a sialylated cell-surface antigen expressed in a number of tissue-specific isoforms.

Different recombinations have been found at the Ig heavy chain gene loci in a number of sublines of the Burkitt lymphoma (BL) cell line Namalwa, following prolonged in vitro culture. The Namalwa sublines examined are DNA fingerprint-identical and derived from a monoclonal source. Recombinant DNA clones were used to map the Ig heavy chain gene mutations to a region between the VDJ and C-mu- segments of the locus. This region is associated with Ig heavy chain class switching in normal B cells. Of 24 clones established from one subline, three were found to have additional VDJ-C-mu-region mutations, indicating a high frequency of mutation at this locus.

A new Epstein-Barr virus (EBV) transformed cell line was established from a patient with B-chronic lymphocytic leukemia (B-CLL). The karyotype of the cell line has remained normal for over 12 months in culture; however, identical heavy and light chain immunoglobulin (Ig) gene rearrangements in the patient's blood and the cell line provided evidence that the EBV transformed cells were derived from the neoplastic clone.

The phenotypic and molecular genetic characteristics of 4 variant sublines of the Burkitt lymphoma cell line Narnalwa have been examined. The sublines are DNA-fingerprintidentical and derived from a monoclonal tumour, as shown by a rearrangement of the T-cell-receptor @-chain gene common to the 4 sublines. There is non-co-ordinate expression of MHC class-I, MHC class-11, surface immunoglobulin and a number of antigens recognized by CD MAbs on the different sublines. These different phenotypes of the cells are reminiscent of B cells arrested in varying states of cellular maturity. On Southem blots there are different patterns of restriction fragments hybridizing with lg heavyand light-chain gene probes among the sublines, indicating that multiple rearrangements or mutations of Ig genes have occurred in the cells. Different patterns of hybridizing fragments among the sublines were also found by using c-myc probes, implying the existence of different mutations of the c-myc locus. The c-myc mutation found in one of the sublines mapped to the 5‘ flanking sequence and in another 3’ to the c-myc locus. Using the J I7BS8 probe, which detects a restriction fragment length polymorphism in the 3’ flanking region of the c-myc gene, a 4-fold variation in the gene copy number among the sublines was found and one of the sublines was shown to be hemizygous for c-myc. Examination of DNA from early cultures of Namalwa cells showed that the alterations in Ig and c-myc structure had occurred on prolonged culture of the cells.

We describe a method of immunofluorescence which is a lateral application of the principles of the APAAP immunohistochemical technique. Immune complexes of R-phycoerythrin and monoclonal anti-R-phycoerythrin (PEAPE complexes) were used in an indirect immunofluorescence technique to detect the binding to cells of monoclonal antibodies directed to IgM, HLA-DR and B cell activation and differentiation antigens. PEAPE complexes were linked to cell surface bound mAbs by unlabelled anti-mouse Ig antibodies to produce high levels of fluorescent staining. The sensitivity of this method of indirect immunofluorescence was enhanced by the sequential application of several cycles of anti-mouse Ig and PEAPE complexes.