Twenty-four-hour effects of bimatoprost 0.01% monotherapy on intraocular pressure and ocular perfusion pressure.

OBJECTIVES: To investigate the 24 h effects of bimatoprost 0.01% monotherapy on intraocular pressure (IOP) and ocular perfusion pressure (OPP). DESIGN: Prospective, open-label experimental study. SETTING: Single tertiary ophthalmic clinic. PARTICIPANTS: Sixteen patients with diagnosed primary open-angle glaucoma (POAG) or ocular hypertension (ages, 49-77 years). INTERVENTIONS: Baseline data of 24 h IOP in untreated patients were collected in a sleep laboratory. Measurements of IOP were taken using a pneumatonometer every 2 h in the sitting and supine body positions during the 16 h diurnal/wake period and in the supine position during the 8 h nocturnal/sleep period. After baseline measurements were taken, patients were treated with bimatoprost 0.01% one time per day at bedtime for 4 weeks, and then 24 h IOP data were collected under the same laboratory conditions. PRIMARY AND SECONDARY OUTCOME MEASURES: Diurnal and nocturnal IOP and OPP means under bimatoprost 0.01% treatment were compared with baseline. RESULTS: The diurnal and nocturnal IOP means were significantly lower under the bimatoprost 0.01% treatment than baseline in both the sitting and supine positions. The diurnal and nocturnal OPP means were significantly higher under treatment than baseline in both the sitting and supine positions. CONCLUSION: Bimatoprost 0.01% monotherapy significantly lowered IOP and increased OPP during the 24 h period.

Diurnal and nocturnal effects of brimonidine monotherapy on intraocular pressure.

PURPOSE: To investigate the effect of brimonidine monotherapy on intraocular pressure (IOP) during the nocturnal/sleep period. DESIGN: Prospective, open-label experimental study. PARTICIPANTS: Fifteen patients with newly diagnosed open-angle glaucoma or ocular hypertension (ages, 46-72 years). METHODS: Baseline data of 24-hour IOP in untreated patients were collected in a sleep laboratory. Measurements of IOP were taken using a pneumatonometer every 2 hours in the sitting and supine body positions during the 16-hour diurnal/wake period and in the supine position during the 8-hour nocturnal/sleep period. Patients were treated afterward with 0.1% brimonidine 3 times per day for 4 weeks, and 24-hour IOP data were collected under the same laboratory conditions. MAIN OUTCOME MEASURES: Diurnal and nocturnal IOP means under the brimonidine treatment were compared with the baseline. RESULTS: The diurnal IOP mean was significantly lower under the brimonidine treatment than the baseline IOP in both the sitting and supine positions. There was no statistically significant change in IOP under the brimonidine treatment from the baseline during the nocturnal period. CONCLUSIONS: Although 0.1% brimonidine monotherapy significantly lowered IOP during the diurnal/wake period, it did not significantly lower IOP during the nocturnal/sleep period. FINANCIAL DISCLOSURE(S): Proprietary or commercial disclosure may be found after the references.

Actin: a novel interaction partner of WT1 influencing its cell dynamic properties.

The Wilms' tumour suppressor, WT1, is a zinc finger protein with key roles in normal development of the genitourinary system and tumourigenesis. Mutations or deletion of WT1 result in a spectrum of developmental disorders and susceptibility to Wilms' tumour in children. Ectopic expression of Wt1 associated with oncogenic functions has been observed in a large number of malignancies, including haematological and solid cancers. Although Wt1 is predominantly a nuclear protein in normal tissues, it is mostly cytoplasmic in the majority of Wt1-expressing tumours. Actin was identified in this study as a new WT1 interaction partner both in the nucleus and in the cytoplasm. We confirmed this interaction both in vitro and in vivo and started to explore its functional significance. Perturbation of the actin cytoskeleton moved Wt1 off the polysome fraction in the cytoplasm, cancelled its nucleo-cytoplasmic shuttling and altered Wt1 DNA- and RNA-binding abilities. These data have implications for Wt1 functions in relation to RNA metabolism and response to cytoskeletal alterations in cancer cells. Thus, our findings could shed more light on the functions of both these proteins and possibly pave way for the development of new cancer therapies.

Comparing diurnal and nocturnal effects of brinzolamide and timolol on intraocular pressure in patients receiving latanoprost monotherapy.

PURPOSE: To compare the diurnal and nocturnal effects of brinzolamide and timolol on intraocular pressure (IOP) in patients already receiving monotherapy with latanoprost. DESIGN: Prospective, open-label, and crossover clinical trial. PARTICIPANTS: Twenty-six patients with glaucoma or ocular hypertension (ages, 44-79 years) who were receiving treatment with 0.005% latanoprost once every evening. METHODS: Baseline data of 24-hour IOP were collected in a sleep laboratory while patients were receiving latanoprost monotherapy. Measurements were taken every 2 hours in the sitting and supine positions during the 16-hour diurnal/wake period and in a supine position during the 8-hour nocturnal/sleep period. Patients were randomly assigned to receive an add-on treatment with either 1% brinzolamide 3 times per day or 0.5% timolol gel forming solution once every morning for 8 weeks, and then crossed over to receive the other add-on treatment. At the end of each add-on treatment period, 24-hour IOP data were collected. MAIN OUTCOME MEASURES: Diurnal and nocturnal IOP means were compared among the baseline, the brinzolamide add-on treatment, and the timolol add-on treatment. RESULTS: During the diurnal period, the mean IOP under brinzolamide or timolol add-on treatment was significantly lower than the baseline IOP in both the sitting and supine positions. There was no statistical difference between the 2 add-on treatments. During the nocturnal period, the supine IOP under brinzolamide add-on treatment was significantly lower than both the baseline and the timolol add-on treatment. There was no difference in nocturnal IOP between the timolol add-on treatment and the baseline. CONCLUSIONS: In patients already receiving the latanoprost monotherapy, adding brinzolamide or timolol significantly reduced IOP during the diurnal period. However, only the brinzolamide add-on treatment had an IOP-lowering efficacy during the nocturnal period.

hnRNP-U directly interacts with WT1 and modulates WT1 transcriptional activation.

The Wilms' tumour suppressor gene, WT1, encodes a zinc-finger protein that is mutated in Wilms' tumours and highly expressed in a wide variety of other malignancies. WT1 is a transcription factor that is likely to have additional, post-transcriptional, regulatory roles, although the molecular mechanisms by which WT1 acts remain poorly understood. We have combined genetic and biochemical approaches to show, that endogenous WT1 binds to heterogeneous nuclear ribonuclear protein U (hnRNP-U), that this interaction does not require any other proteins or nucleic acids, involves the zinc-fingers of WT1 and the middle domain of hnRNP-U, and that hnRNP-U can modulate WT1 transcriptional activation of a bona fide WT1 target gene. These findings increase our knowledge of how WT1 exerts its transcriptional regulatory role and suggests that hnRNP-U may be a candidate Wilms' tumour gene at 1q44.

Presence of WT1, the Wilm's tumor suppressor gene product, in nuclear poly(A)(+) ribonucleoprotein.

The tumor suppressor gene WT1 encodes a zinc finger protein, which consists of four C-terminal C(2)-H(2) zinc fingers of the Krüppel type, and at the N terminus a Q/P-rich trans-regulatory domain, both characteristic of transcription factors. However, recent findings suggest that WT1 may also be involved in a post-transcriptional process. Specifically, WT1 isoforms containing the alternatively spliced exon 9 (+lysine-threonine-serine (KTS)) preferentially associate with nuclear speckles and co-immunoprecipitate splicing antigens (Larsson, S. H., Charlieu, J.-P., Miyagawa, K., Engelkamp, D., Rassoulzadegan, M., Ross, A., Cuzin, F., van Heyningen, V., and Hastie, N. D. (1995) Cell 81, 391-401); furthermore, WT1 has been shown to interact with the ubiquitous splicing factor U2AF65 (Davies, R. C., Calvo, C., Larsson, S. H., Lamond, A. I., and Hastie, N. D. (1998) Genes Dev. 12, 3217-3225) and binds to RNA in vitro (Caricasole, A., Duarte, A., Larsson, S. H., Hastie, N. D., Little, M., Holmes, G., Todorov, I., and Ward, A. (1996) Proc. Natl. Acad. Sci. U. S. A. 93, 7562-7566; Bardeesy, N., and Pelletier, J. (1998) Nucleic Acids Res. 26, 1784-1792). To extend these findings, we have fractionated nuclear extracts to see if particles containing WT1 have the properties of ribonucleoprotein (RNP). In summary, WT1 is enriched by oligo(dT) chromatography, as are U2AF65, the U5 small nuclear RNP-associated protein p116 and hnRNP A1. Gel filtration and sedimentation profiles suggest that WT1 is present in RNase-sensitive particles, >2 MDa in size, peaking at approximately 60 S, and approximately 1.27 g/cm(3) on Nycodenz. Similar results were obtained from two cell lines expressing WT1, fetal kidneys (day E17), and transiently transfected cells, suggesting that the presence of WT1 protein in nuclear poly(A)(+) RNP is a general aspect of WT1 function.

Diffusible component from the spore surface of the fungus Aspergillus fumigatus which inhibits the macrophage oxidative burst is distinct from gliotoxin and other hyphal toxins.

BACKGROUND: The fungus Aspergillus fumigatus, whose spores are present ubiquitously in the air, causes a range of diseases in the human lung. A small molecular weight (< 10 kD) heat stable toxin released from the spores of clinical and environmental isolates of A fumigatus within minutes of deposition in aqueous solution has previously been described. A key effect of the toxin was to inhibit the oxidative burst of macrophages as measured by superoxide anion release. It was hypothesised that the toxin was one of the commonly found A fumigatus hyphal toxins such as gliotoxin. This inhibitor may be an important factor which allows the fungus to colonise the lung. METHODS: The spore derived inhibitor was shown to inhibit the respiratory burst of rat alveolar macrophages, as measured by the generation of superoxide anion. Samples of the spore diffusate were subject to reversed phase high performance liquid chromatography (HPLC), thin layer chromatography (TLC), high performance thin layer chromatography (HPTLC), or organic extraction followed by TLC or HPLC to identify the presence of gliotoxin, fumagillin, helvolic acid, fumigaclavine-C, and aurasperone-C. Commercially obtained preparations of the toxins gliotoxin, fumagillin and helvolic acid and extracts enriched for fumigaclavine-C and aurasperone-C were used as internal and external standards and in the respiratory burst measurements. RESULTS: Gliotoxin, fumagillin, helvolic acid, fumigaclavine-C, and aurasperone-C were not detected in spore derived diffusate using PHLC or TLC. Using extraction procedures with solvents known to extract gliotoxin from A fumigatus culture supernatants, no gliotoxin was detected in the spore derived diffusate. Commercial gliotoxin, fumagillin, and helvolic acid or extracts enriched for fumigaclavine-C and aurasperone-C did not inhibit the oxidative burst of macrophages. CONCLUSIONS: The hypothesis that the spore derived toxin is one of the toxins derived from hyphae such as gliotoxin, helvolic acid, fumagillin, fumigaclavine-C, or aurasperone-C is not proved. The spore toxin may exert its effect through its ability to diffuse rapidly into the lung lining fluid, diminish the macrophage oxidative burst, and play a part in allowing A fumigatus to persist in the lung and manifest its well known pathogenic effects.

Inhibition of the transcription factors NF-kappa B and AP-1 underlies loss of cytokine gene expression in rat alveolar macrophages treated with a diffusible product from the spores of Aspergillus fumigatus.

The spores of Aspergillus fumigatus have a survival advantage over other respirable fungal spores in the lung, leading to a number of lung diseases associated with this fungus. We have hypothesized that a component on the spore surface can inhibit the activation of alveolar macrophages, known to play an essential role in immune regulation in the lung. A diffusible product from the spores of A. fumigatus (AfD) inhibited the production of tumor necrosis factor alpha (TNF alpha) protein by alveolar macrophages in an enzyme-linked immunosorbent assay. Using a semiquantitative reverse transcription-polymerase chain reaction, we also demonstrated a potent inhibitory effect of AfD on the production of proinflammatory cytokine transcripts in rat alveolar macrophages. The inhibition occurred at the level of transcription, with AfD inhibiting the synthesis of TNF alpha-and interleukin 6 (IL-6)-specific mRNA transcripts. No effect was seen on the synthesis of interleukin 1 beta (IL-1 beta) cytokine transcripts or on the expression of the housekeeping gene beta-actin. Furthermore, AfD specifically inhibited the activation of nuclear transcription factors NF-kappa B and AP-1, both of which are required for the coordinate upregulation of transcription of the proinflammatory cytokines TNF alpha, IL-1 beta, and IL-6. We conclude that AfD can inhibit normal alveolar macrophage responses by selectively inhibiting the production of key inflammatory cytokines, and that the mechanism of inhibition is primarily at the level of transcriptional activation.

Inhibition of the alveolar macrophage oxidative burst by a diffusible component from the surface of the spores of the fungus Aspergillus fumigatus.

BACKGROUND: Aspergillus fumigatus is a fungus that grows on dead and decaying organic matter in the environment and whose spores are present ubiquitously in the air. The fungus causes a range of diseases in the human lung. A study was undertaken to demonstrate and partially characterise an inhibitor of the macrophage respiratory burst from the surface of A fumigatus spores that could be an important factor in allowing the fungus to colonise the lung. METHODS: The spore-derived inhibitor of the respiratory burst of rat alveolar macrophages, as measured by generation of superoxide anion, was demonstrated in Hank's balanced salt solution extracts of four clinical isolates and an environmental isolate of A fumigatus. The time course of the release of the inhibitor into aqueous solution was assessed and the cytotoxic potential of the spore-derived inhibitor towards macrophages was tested using the propidium iodide method. An oxygen electrode was used to confirm the superoxide anion measurements. Molecular weight cutoff filters were used to determine the size of the inhibitor as assessed in the respiratory burst assay and also by its ability to inhibit macrophage spreading on glass. The crude diffusate from the spore surface was fractionated by reversed phase high pressure liquid chromatography (HPLC) and the fractions analysed for inhibitory activity, protein, and carbohydrate content. RESULTS: A small molecular weight (< 10 kD) heat stable toxin was released from the spores of clinical and environmental isolates of A fumigatus within minutes of deposition in aqueous solution. The key effect of the toxin demonstrated here was its ability to inhibit the oxidative burst of macrophages as measured by superoxide anion release. The inhibition was not due to cell death or detectable loss of membrane integrity as measured by permeability to propidium iodide. The toxin was not a scavenger of superoxide anion. Oxygen electrode studies suggested indirectly that the inhibitor acted to inhibit the assembly of the macrophage NADPH-oxidase complex. Fractions of < 10 kD also inhibited the spreading of alveolar macrophages, confirming that the toxin had an additional effect on macrophages that leads to loss of adherence or impairment of cytoskeletal function. In reversed phase HPLC fractions the inhibitory activity eluted with an associated carbohydrate, although the exact chemical nature of the toxin remains to be elucidated. CONCLUSIONS: This spore toxin may, through its ability to diffuse rapidly into lung lining fluid, diminish the macrophage respiratory burst and play a part in allowing A fumigatus to persist in the lung and manifest its well known pathogenic effects. Future research will be focused on further molecular characterisation of the toxin and elaboration of the effect of the toxin on intracellular signalling pathways involved in the activation of alveolar macrophages.

Asbestos fibre length-dependent detachment injury to alveolar epithelial cells in vitro: role of a fibronectin-binding receptor.

A short and a long fibre sample of amosite asbestos were tested for their effects on cells of the human Type 2 alveolar epithelial cell-line A549 in vitro. The long amosite sample was found to cause a rapid detachment of the epithelial cells live from their substratum. At the highest dose, on average 28% of the cells present were detached in this way. Studies on the mechanism of the detachment injury showed that it did not involve oxidants since it was not ameliorated by scavengers of active oxygen species. Neither was the effect reduced by treatment of the fibres with the iron chelator Desferal. Treatments reported to increase the interaction between fibres and cells, serum and poly-L-lysine, did not influence the detachment injury, nor did lung lining fluid. Conversely, the fibronectin tripeptide RGD alone could cause detachment which suggested that a fibronectin-binding integrin was involved. This receptor could be reduced in activity by long fibre exposure, leading to detachment. The detaching effect of fibre could be mimicked by the protein kinase C activator PMA, and so the second messenger system of the cell could also be involved. This type of injury could be important in the pathology associated with exposure to long fibres.

Epithelial and extracellular matrix injury in quartz-inflamed lung: role of the alveolar macrophage.

The bronchoalveolar leukocytes from quartz-inflamed lung were separated into macrophage-enriched and neutrophil-enriched populations on density gradients. Neutrophil-enriched populations showed the greatest activity in causing injury to epithelial cells and fibronectin in vitro. Inflammatory macrophage-enriched populations from quartz-exposed lung had the ability to cause fibronectin degradation but could not cause detachment injury to epithelial cells over and above that caused by control alveolar macrophages. Fibronectin damage in vivo could be an important factor in disordering the connective tissue scaffold of the lung, thereby favoring fibrosis. In vitro quartz stimulated more production of cytokines by alveolar macrophages than the inert particulate titanium dioxide. Cytokines could be important in upregulating adhesion molecules in the membranes of lung cells in vivo; this process could aid leukocyte/lung cell contact, allowing epithelial injury to be expressed, and could also be a factor leading to pathological change.

Leukocyte-mediated epithelial injury in ozone-exposed rat lung.

Both epithelial injury and inflammation are characteristic findings in the centriacinar regions of the lungs of rats exposed to ozone. In humans such effects could lead to long-term lung damage and disease. In animals, neoplastic change in the lungs after exposure to ozone has been described previously. The possible relationships between inflammatory cell recruitment, epithelial injury, and hyperplasia, with special regard to the important role of repair processes in leading to increased incidence of tumors in some species, have been addressed in the present study. We have previously described that leukocytes from lungs inflamed by different agents can injure epithelial cells in vitro. We have suggested that this leukocyte-mediated epithelial injury could enhance epithelial turnover in ozone-exposed lungs and so enhance the likelihood of tumor development. We, therefore, set out to test the hypothesis that bronchoalveolar leukocytes from ozone-exposed lungs can injure epithelial cells in vitro. PVG rats were exposed to 0.2, 0.4, 0.6, and 0.8 parts per million2 (ppm) ozone for seven hours per day for up to four days. On the morning following the last exposure, bronchoalveolar lavage was used to sample the bronchoalveolar leukocytes and the following parameters were assessed: total number, differential leukocyte count, production of oxidants, ability to degrade fibronectin, and ability to injure epithelial cells. In addition to these parameters, which were measured at all concentrations and time points in limited experiments, we also assessed macrophage size in short-term culture and inflammation in histological sections of lungs. Total number of lavageable cells was not affected by ozone inhalation. However, the percentage of macrophages decreased with ozone treatment and the percentage of neutrophils increased on days 1 and 2 at 0.6 and 0.8 ppm ozone. There was no significant effect of ozone exposure on the ability of neutrophils to degrade fibronectin or injure epithelial cells. Production of superoxide anion in response to stimulation with phorbol myristate acetate was significantly decreased by exposure to 0.6 ppm ozone, as described in previous studies. Macrophages from the lungs of rats exposed to ozone were larger than control macrophages.

Persistent biological reactivity of quartz in the lung: raised protease burden compared with a non-pathogenic mineral dust and microbial particles.

This study assessed the potential harmfulness of particles in the lung by measuring their ability to elicit and maintain an inflammatory response and to damage lung tissue. It compared the inflammogenicity of two nondurable, biological particulates (Corynebacterium parvum and zymosan) with a pathogenic mineral dust (quartz) and a nonpathogenic dust (titanium dioxide) by dosing rats via the intratracheal route and measuring the consequent alveolitis. The magnitude and duration of the inflammatory response were assessed by measuring the total number of leucocytes and the percentage of neutrophils obtained by bronchoalveolar lavage. Two key functional parameters of the lavaged leucocytes--ability to degrade fibronectin and production of plasminogen activator--were also measured. A marked inflammatory response had occurred by one day after instillation, characterised by increases in total leucocyte numbers and percentage of neutrophils in the bronchoalveolar lavages, with all four test materials. In all but the quartz exposed animals, the inflammation subsided rapidly thereafter, approaching control levels by 15 days after injection; in the quartz exposed animals the alveolitis persisted for up to 30 days. All of the inflammogens generated chemotaxins in rat serum in vitro and so, by analogy, might also be expected to generate chemotactic activity in alveolar lining fluid which could contribute to the generation of an inflammatory response. The cellular inflammatory response was accompanied by a concomitant increase in the proteolytic activity of the bronchoalveolar lavage leucocytes but production of plasminogen activator remained unchanged. In vitro exposure to the inflammogens had no effect on the proteolytic activity against fibronectin or on the plasminogen activator activity of bronchoalveolar leucocytes.

Contrasting bronchoalveolar leukocyte responses in rats inhaling coal mine dust, quartz, or titanium dioxide: effects of coal rank, airborne mass concentration, and cessation of exposure.

The aim of this study was to determine the bronchoalveolar leukocyte response to airborne coal mine dust; quartz and titanium dioxide were used as positive and negative controls, respectively. Groups of rats were exposed to airborne mass concentrations of 10 and 50 mg/m3 of the dusts for 7 hr/day, 5 days/week and their bronchoalveolar space was lavaged at time points between 2 and 75 days of exposure, to assess the leukocyte response. This study revealed time-dependent and airborne mass concentration-dependent recruitment of neutrophils and macrophages into the bronchoalveolar region with coal mine dust inhalation but no real difference in the magnitude of the response between coal mine dusts from collieries mining coal of different rank and quartz content although the maximum quartz content in the dusts used was 6%. The inflammatory response was much less than that produced by quartz, at similar airborne mass concentrations, and more than that produced by titanium dioxide which was, in general, a poor inflammogen in the rat lung. Groups of rats were exposed to the airborne dusts for 32 or 75 days, then removed from the exposure chambers, and allowed to recover by breathing room air for a further 64 days. During this recovery period there was marked progression of the leukocyte response with quartz and persistence of the response with coal mine dust. Chronic recruitment of leukocytes to the lungs of individuals inhaling coal mine dust is likely to be an important factor in the development of coal workers' pneumoconiosis.

Inflammation in the lungs of rats after deposition of dust collected from the air of wool mills: the role of epithelial injury and complement activation.

In a previous study assessing respiratory symptoms in individuals employed in wool textile mills in the north of England relations between symptoms of chronic bronchitis, breathlessness and wheeze, and rhinitis and current exposure to airborne mass concentration of dust were shown. As preliminary steps in defining the potential hazard associated with dust from the air of wool mills the ability of inspirable dust, collected from the air of wool textile mills, to cause inflammation when injected into the lungs of rats was determined. Dusts were collected from the beginning of wool processing (opening) in one factory and from the middle (combing) and late (backwinding) stages of the process in two other factories. Ability of the dusts to cause inflammation was assessed by instillation into the lungs of rats followed by bronchoalveolar lavage. All the dusts caused some inflammation which peaked on day 1 and did not persist beyond one week. A distinctive aggregation response of mononuclear cells in the lavage, however, had a different time course, peaking at day 7. An attempt was made to determine how the wool mill dusts caused inflammation and experiments showed that the dusts themselves had no inherent chemotactic activity but that they did have a pronounced ability to generate chemotaxins in serum and so could activate complement in lung fluid. In addition, dust collected from ledges in the mills had the ability to injure epithelial cells in vitro which could also contribute to inflammation. A role for endotoxin in the inflammatory activity of the dusts was not discounted and a leachate of the dust had the ability to cause inflammation when injected into the lungs of rats. Wool mill dust is likely to be a complex mixture of materials and these experiments represent a preliminary approach to understanding the biological activity of the whole unfractionated dust and further studies are in progress to define more accurately the toxic material(s) in the dust.

Impaired chemotactic responses of bronchoalveolar leukocytes in experimental pneumoconiosis.

Rats were exposed to clouds of the following pneumoconiotic dusts: quartz, coal-mine dust, and chrysotile asbestos at 10 or 50 mg/m3 for 8, 32, and 75 days; for comparison, rats were also exposed to the non-pathogenic dust titanium dioxide (TiO2). The bronchoalveolar leukocytes (macrophages and neutrophils) from dust-exposed and control rats were obtained by lavage and tested for their ability to migrate toward zymosan-activated serum. Varying amounts of neutrophils were present depending on the ability of the dust to cause inflammation and the length of exposure. There was a marked loss of chemotactic ability in leukocytes from rats inhaling the pneumoconiotic dusts compared with controls; TiO2-exposed leukocytes had some impairment of chemotaxis, but this was substantially less than that found with the pneumoconiotic dusts. The loss of chemotactic activity did not correlate with the percentage of neutrophils in the lavage cells except when there were very high levels of neutrophils, and there was substantial impairment of chemotaxis with negligible numbers of neutrophils, showing that macrophage chemotaxis was impaired. A phagocytic burden within the leucocytes was not sufficient alone to inhibit chemotaxis, nor was the loss of chemotactic activity due to occupied receptors, since incubation failed to restore chemotaxis. Loss to chemotactic activity by leukocytes from pneumoconiotic dust-exposed lung could be an important factor in the development of pneumoconiosis.

Effects of products from inflammatory pulmonary neutrophils on alveolar macrophage chemotaxis, spreading, and thymidine incorporation.

Since macrophages and neutrophils are found together in the alveolar region of the lung during alveolar inflammation, we assessed whether neutrophil products could influence three key functions of alveolar macrophages: chemotaxis, spreading, and thymidine incorporation. Neutrophils were obtained from the lungs of rats treated by intratracheal instillation of heat-killed Corynebacterium parvum and cultured overnight, alone or in the presence of zymosan, PMA, an inert particulate (titanium dioxide), or a toxic dust, (quartz). Supernatants were collected from these cells and a lysate, obtained by freeze-thawing neutrophils, was also used. Neutrophil supernatants caused a slight reduction in chemotaxis and a significant loss of ability to spread on glass which varied depending on the in vitro treatment of the neutrophils. In addition neutrophil supernatants also had a substantial effect in stimulating uptake of thymidine which was, once again, very dependent on the treatment of the neutrophils during preparation of the supernatants, with unstimulated and TiO2-treated neutrophils producing maximum stimulation. The increases in thymidine uptake were not matched by increased proliferation, suggesting that another signal may be necessary for expanison of alveolar macrophage numbers during alveolar inflammation.

A comparison of the ocular hypotensive efficacy of once-daily and twice-daily levobunolol treatment.

The authors compared the ocular hypotensive efficacy of two different treatment regimens of levobunolol 0.5% in a double-masked, randomized, controlled clinical trial. Seventy-one patients with open-angle glaucoma or ocular hypertension received levobunolol 0.5% as their sole glaucoma medication either on a once-daily or twice-daily treatment regimen for 3 months. Approximately 81% of the patients in the once-daily treatment group and 88% of subjects in the twice-daily treatment group successfully completed the 3-month study period. The overall mean decrease in intraocular pressure (IOP) was 4.5 mmHg in the once-daily group and 5.6 mmHg in the twice-daily group. These differences were not statistically different. For both treatment groups, effects on mean heart rate and blood pressure were minimal. The authors' data from this population suggest that once-daily treatment with levobunolol is an effective glaucoma regimen.

The effect of products from bronchoalveolar-derived neutrophils on oxidant production and phagocytic activity of alveolar macrophages.

Neutrophils and alveolar macrophages are found together in the alveolar region during pulmonary inflammation where neutrophil products could influence important macrophage defensive functions. We set out therefore to investigate the ability of neutrophil products to modulate alveolar macrophage phagocytosis and oxidant production. Neutrophils derived from acutely inflamed rat lung were incubated along with a range of potential triggers of neutrophil secretion and supernatants collected. Using two quantitative assays of rat alveolar macrophage phagocytosis, the supernatants were found to have no effect except for the quartz supernatant, which slightly enhanced phagocytosis via non-specific receptors and the PMA supernatant, which caused reduction in phagocytosis via non-specific and Fc receptors; this reduction could however be mimicked by PMA alone. None of the supernatants affected macrophage production of superoxide anion or hydrogen peroxide except for the PMA supernatant and once again the inhibitory effect of the PMA supernatant could be mimicked with PMA alone. It is concluded that products of inflammatory neutrophils do not affect phagocytosis or oxidative metabolism of alveolar macrophages, although in quartz-exposed lung neutrophils may exert a small enhancing effect on macrophage phagocytosis.

Kinetics of the bronchoalveolar leucocyte response in rats during exposure to equal airborne mass concentrations of quartz, chrysotile asbestos, or titanium dioxide.

The kinetics of the bronchoalveolar response was assessed in rats exposed, at equal airborne mass concentration (10 mg/m3), to titanium dioxide--a non-pathogenic dust--and the two pathogenic mineral dusts quartz and chrysotile asbestos. Rats were killed at intervals over a 75 day exposure period and groups of rats exposed for 32 and 75 days after recovery for two months. Bronchoalveolar lavage was carried out and the lavage fluid characterised for cellular content, macrophage activation, and concentrations of free total protein, lactate dehydrogenase, and N-acetyl-beta-D-glucosaminidase. Inhalation exposure to the two pathogenic dusts resulted in an increased number of leucocytes, macrophage activation, and increased levels of free enzymes and total protein. The pattern and magnitude of the responses to quartz and chrysotile differed. Chrysotile caused less inflammation than quartz, and the main cellular response peaked around the middle of the period of dust exposure whereas the highest levels of enzymes occurred towards the end. The difference in timing suggests that macrophages were not available for lavage towards the end of the exposure, owing to their playing a part possibly in deposition of granulation tissue. Quartz caused a greater cellular and enzyme response than chrysotile, particularly towards the end of the dust exposure phase. There was a noticeable progression of inflammation in the quartz exposed groups left to recover for two months, but not in the chrysotile recovery groups.