The effects of low level laser irradiation on osteoblastic cells.

Low level laser therapy has been used in treating many conditions with reports of multiple clinical effects including promotion of healing of both hard and soft tissue lesions. Low level laser therapy as a treatment modality remains controversial, however. The effects of wavelength, beam type, energy output, energy level, energy intensity, and exposure regime of low level laser therapy remain unexplained. Moreover, no specific therapeutic window for dosimetry and mechanism of action has been determined at the level of individual cell types. The aim of this study was to investigate the effects of low level laser irradiation on the human osteosarcoma cell line, SAOS-2. The cells were irradiated as a single or daily dose for up to 10 days with a GaAlAs continuous wave diode laser (830 nm, net output of 90 mW, energy levels of 0.3, 0.5, 1, 2, and 4 Joules). Cell viability was not affected by laser irradiation, with the viability being greater than 90% for all experimental groups. Cellular proliferation or activation was not found to be significantly affected by any of the energy levels and varying exposure regimes investigated. Low level laser irradiation did result in a heat shock response at an energy level of 2 J. No significant early or late effects of laser irradiation on protein expression and alkaline phosphatase activity were found. Investigation of intracellular calcium concentration revealed a tendency of a transient positive change after irradiation. Low level laser irradiation was unable to stimulate the osteosarcoma cells utilised for this research at a gross cell population level. The heat shock response and increased intracellular calcium indicate that the cells do respond to low level laser irradiation. Further research is required, utilising different cell and animal models, to more specifically determine the effects of low level laser irradiation at a cellular level. These effects should be more thoroughly investigated before low level laser therapy can be considered as a potential accelerator stimulus for orthodontic tooth movement.

The effectiveness of different rat IgG subclasses as IgE-blocking antibodies in the rat basophil leukaemia cell model.

The degranulation of mast cells in an allergic response is initiated by the aggregation of high-affinity IgE receptors (Fc epsilon RI) by IgE and antigen. Recently it has been shown that such degranulation can be inhibited by cross-linking Fc epsilon RI and low-affinity IgG receptors (Fc gamma RII) which are also expressed by mast cells. The ability of various monoclonal antibodies to block the degranulation of rat basophil leukaemia (RBL) cells sensitized with IgE antidinitrophenyl (DNP) antibodies has been investigated. Sensitized cells were challenged with immune complexes formed using varying concentrations of antigen, and of both high- and low-valency antigen. It is reported here that rat IgG1 antibodies, which are associated in the rat with a Th1-type response, act as highly effective blocking antibodies over a wide concentration range. Rat IgG2a antibodies, which are associated with a Th2-type response, were able only to inhibit degranulation when immune complexes were formed with very low concentrations of high-valency antigen (DNP32-HSA). Under these conditions, some inhibitory activity was seen with high-affinity murine IgA anti-DNP but not with low-affinity rat IgG2b anti-DNP antibody-containing immune complexes. In addition to this inhibitory activity, IgG2a antibodies were shown to be capable of inducing degranulation of cells via unoccupied Fc epsilon RI. These results demonstrate that blocking activity may arise via both inhibitory receptors and by masking of antigen.

Characterization of the mitogenic response of murine CD5+ and conventional B lymphocytes to lipopolysaccharide.

The nature of the response of conventional and CD5+ B cells to stimulation in vitro with optimal mitogenic concentrations of LPS was examined to elucidate the contributions of these B cell subsets in polyclonal B lymphocyte responses. Stimulation of murine splenic lymphocytes with LPS resulted in an increase in total biomass, peaking at 72 h of culture. The viability of the cultures remained high (> 90%) until 48 h of culture. A combination of trypan blue and 7-aminoactinomycin D (7AAD) exclusion in conjunction with PE-anti-CD5 and FITC-anti-B220 enabled more detailed analysis of the cultures. The total number of conventional B cells, viable and non-viable, increased until 48 h of culture and then decreased when stimulated with LPS, while CD5+ B cells increased over the culture period. The numbers of conventional B cells in the control cultures decreased, but the CD5+ B cell numbers remained stable. An examination of the modes of death of the B cell subsets using 7AAD showed that unstimulated conventional B cells were apoptotic rather than degenerate but, following stimulation with LPS, apoptotic and degenerate cells were found. Apoptotic and degenerate CD5+ B cells were found in both stimulated and unstimulated cultures, but the percentage of these apoptotic and degenerate cells was increased significantly only at 72 h and 96 h of culture in stimulated cultures compared with 24 h onwards in the control cultures. Morphological analysis and gel electrophoretic studies of extracted DNA reflected these findings. It was also found that the increase in the number and percentage of non-viable cells in the cultures was not equal to the decrease in the number and percentage of viable cells. Activation of B cells was examined using expression of B7-1 (CD80) as a marker. When stimulated with LPS a greater proportion of conventional B cells expressed B7-1 after 24 h of culture than in the control cultures; however, only at 72 h and 96 h of culture was the proportion of CD5+ B cells expressing B7-1 significantly higher than in the control cultures. These results show that conventional B cells are stimulated to proliferate and to become activated by LPS and that death is apoptotic rather than degenerate or necrotic. CD5+ B cells were also shown to be stimulated by LPS; they became activated and death was delayed. The data suggest that in addition to the proliferative role, LPS acts to delay death and to activate conventional and CD5+ B cells.

Self-limited blastomycosis: a report of thirteen cases.

Blastomycosis is not generally recognized to be a self-limited pulmonary infection. We report 13 patients with blastomycosis who followed a self-limited course. Presenting complaints were usually those of an acute pulmonary infection with fever, productive cough, and pleurtiic chest pain. The duration of symtpoms before diagnosis was usuallms were variable and not diagnostic. The blastomycin skin test and complement-fixing serologies to blastomycin were generally not helpful. In all patients the diagnosis was made by either cultural or visual identification of the organism from sputum, bronchial washings, or pleural fluid. All patients were improving both clinically and by chest roentgenograms by the time the diagnosis was reached, and thus therapy was withheld. Follow-up of these 13 patients ranged from 5 months to 8 yr (mean, 43 months), and in no instance has there been any evidence of reactivation of the illness.