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Biomedical subjects

D G Frazer

Publications and source records attributed to D G Frazer.

At least 37 records · Page 2Linked to original sources
PubMed1998

Airway hyperreactivity elicited by toluene diisocyanate (TDI)-albumin conjugate is not accompanied by airway eosinophilic infiltration in guinea pigs.

Nonspecific airway hyperresponsiveness is present in many patients with toluene diisocyanate (TDI)-induced asthma; however, the underlying pathophysiological mechanisms of this hyperresponsiveness remain controversial. In the present study, we used a guinea pig model to investigate the association of TDI-induced airway hyperresponsiveness with eosinophilic airway infiltration, which is widely considered to play a key role in the development of allergen-induced hyperresponsiveness. Guinea pigs were sensitized by i.d. injections of 10 microl TDI on day 1 and day 6. Control animals received saline injections. Two weeks after the second injection, airway reactivity to inhaled methacholine and specific airway resistance (sRaw) was measured before and at several times after inhalation challenge with TDI-GSA (guinea pig serum albumin) conjugates. Eosinophils in the airways were detected using enzyme histochemistry and quantified using computer-assisted image analysis. TDI-specific IgG1 antibodies were found in the blood of TDI-sensitized animals. An immediate increase in sRaw was induced in these animals by TDI-GSA challenge; airway hyperresponsiveness to methacholine was observed at 6 h and 18 h after TDI-GSA challenge. However, TDI-GSA challenge did not result in an elevation of eosinophils in the airways, compared with control animals. The results suggest that the development of TDI-induced airway hyperresponsiveness is not dependent upon eosinophil infiltration in airways.

Albumins↗
PubMed1997

Brief exposure of air-filled guinea-pig isolated trachea to low levels of toluene diisocyanate (TDI) vapor in vitro increases reactivity to methacholine.

Toluene diisocyanate (TDI) causes occupational asthma characterized by inflammation and hyperreactivity of airways to irritants and bronchoconstrictor drugs. We examined the non-immune, direct effect of TDI on airway reactivity in vitro in the absence of an inflammatory response using the guinea-pig isolated, perfused trachea preparation to measure reactivity to methacholine (MCh), and fixed point ion mobility spectrometry to measure moment to moment levels of TDI vapor in air that was delivered to the tracheal mucosa. MCh was added to the mucosal modified Krebs-Henseleit (MKH) perfusing solution to generate control concentration-response curves for contractile responses. The lumen was then emptied and perfused with air or air containing 5, 20 or 70 ppb TDI vapor, after which the trachea was perfused with MKH solution and reactivity to MCh was re-examined. After only 30 min of treatment, TDI vapor concentration-dependently increased reactivity of the trachea to MCh (2.4- and 2.9-fold, respectively, for 20 and 70 ppb TDI; 5 ppb TDI and air alone had no effect). In tracheas treated in vitro with 2 microM capsaicin to deplete tachykinins, TDI caused the same (4-fold) increase in reactivity to MCh that was observed in control tracheas. However, TDI vapor (70 ppb) no longer enhanced reactivity to MCh in tracheas from which the epithelium had been removed. Our results indicate that a direct, non-immune, non-inflammatory action of TDI on respiratory epithelium leads to hyperreactivity of airways in vitro.

Air↗
PubMed1997

Acute lung injury induced by a commercial leather conditioner.

Following a formulation change, a leather conditioner was involved in a 1992 nationwide outbreak of respiratory illness. We investigated the composition and toxicity of the conditioner produced before (previous product) and after (new product) the disease outbreak. The new product induced tachypnea, pulmonary edema, pulmonary hemorrhage, and sporadic deaths in exposed guinea pigs and rats. Ultrastructurally, these changes were associate with direct pulmonary cytotoxicity characterized by necrosis of alveolar type I cells and alveolar septal interstitial edema. Chemical analyses suggested major alterations in the fluorohydrocarbon constituents in the new formulation of the leather conditioner. While these alterations could not be specifically identified, they appeared to include changes from fluoralkanes to fluoroalkenes, fluorophenyl, and/or fluoroalcohol compounds. Changes in solvent composition were consistent with traces of 2-butoxyethanol and isomers of dipropylene glycol methyl ether, and additional C10-C12 alkanes. In this study, we demonstrated the toxicity of the new product in laboratory animals. Some of the altered constituents of the new product have been identified and are potential candidates for additional investigations to identify specific etiologic agents.

Acetates↗
PubMed1997

Mutations in cornea-specific keratin K3 or K12 genes cause Meesmann's corneal dystrophy.

The intermediate filament cytoskeleton of corneal epithelial cells is composed of cornea-specific keratins K3 and K12 (refs 1,2). Meesmann's corneal dystrophy (MCD) is an autosomal dominant disorder causing fragility of the anterior corneal epithelium, where K3 and K12 are specifically expressed. We postulated that dominant-negative mutations in these keratins might be the cause of MCD. K3 was mapped to the type-II keratin gene cluster on 12q; and K12 to the type-I keratin cluster on 17q using radiation hybrids. We obtained linkage to the K12 locus in Meesmann's original German kindred (Zmax = 7.53; theta = 0) and we also showed that the phenotype segregated with either the K12 or the K3 locus in two Northern Irish pedigrees. Heterozygous missense mutations in K3 (E509K) and in K12 (V143L; R135T) completely co-segregated with MCD in the families and were not found in 100 normal unrelated chromosomes. All mutations occur in the highly conserved keratin helix boundary motifs, where dominant mutations in other keratins have been found to severely compromise cytoskeletal function, leading to keratinocyte fragility phenotypes. Our results demonstrate for the first time the molecular basis of Meesmann's corneal dystrophy.

Cornea↗
PubMed1997

Pulmonary dendritic cell distribution and prevalence in guinea pig airways: effect of ovalbumin sensitization and challenge.

We characterized the localization and prevalence of dendritic cells (DC) in guinea pig airways before and after s.c. sensitization and aerosol challenge with ovalbumin (OVA). DC, eosinophils, macrophages, T cells and B cells in lung and trachea were identified and quantified in frozen sections using monoclonal antibodies and computer-assisted image analysis. Airway reactivity of conscious animals to inhaled methacholine was examined. In unsensitized animals, DC were localized primarily within the lamina propria of the trachea and bronchi, in the submucosa of the trachea and in the adventitia of the bronchi. In contrast to reported studies on rats, few DC were noted in the epithelium. After OVA challenge, sensitized animals demonstrated an early obstructive response and a late-phase response that was well developed by 18 hr. Challenge with OVA increased DC prevalence in the lamina propria and submucosa of the trachea and in the lamina propria and adventitia of the bronchi. There was widespread eosinophilia throughout the airways, but no changes in B cells or T cells were evident. Macrophages were increased in the epithelium of both OVA-treated and saline-treated animals. At 18 hr after challenge, sensitized guinea pigs but not saline-treated controls were hyperreactive to inhaled methacholine. Except for macrophages, none of these effects were observed after saline treatment. Our findings indicate that inflammation in the airways of OVA-sensitized guinea pigs involves infiltration of DC, which is seen at the time animals are hyperreactive to inhaled methacholine.

Animals↗
PubMed1996

Effect of temperature on pressure-volume hysteresis of excised lungs.

The objective of this study was to determine if the effect of temperature on excised lung pressure-volume (P-V) hysteresis during various P-V maneuvers would be consistent with predicted effects based on the recruitment-derecruitment (R-D) model of lung P-V hysteresis. Three sets of P-V curves were recorded for excised rat lungs at (1) 24 degrees C, (2) either 42 degrees or 45 degrees C, and (3) 24 degrees C. After full inflation of the lung, deflation-inflation (D-I) cycles were performed between total lung capacity (30 cmH2O) and successively decreasing end-expiratory pressures (EEPs). Normalized hysteresis (K) was plotted vs EEP. K remained relatively constant at EEPs > or = +5 cmH2O at 24 degrees C and 42 degrees C and > +5 cmH2O at 45 degrees C. Large increases in K occurred as the EEP was further reduced, with the relationship of K vs EEP being shifted to the right at 42 degrees C and 45 degrees C relative to 24 degrees C, with the greater shift occurring at 45 degrees C. Previous work has shown that the R-D of lung units contributes to P-V hysteresis and is EEP-dependent, increasing at EEPs < or = +4 +/- 1 cmH2O at room temperature (Cheng et al., 1995). This study suggests that at increased temperatures, R-D of lung units is initiated at higher EEPs and is more extensive than at room temperature.

Animals↗
PubMed1996

Pulmonary reactions to organic dust exposures: development of an animal model.

Acute inhalation of organic dusts such as cotton, hay, silage, grain, animal confinement, or compost dust can result in illness characterized by fever, pulmonary inflammation, chest tightness, and airway obstruction. These agricultural materials are complex mixtures of plant, bacterial, and fungal products. Elucidation of the time course of disease onset, the mechanisms of disease progression, and the identity of etiologic agents is essential for effective prevention and treatment. Toward this end, animal models for acute organic dust-induced reactions have been developed and characterized. Information concerning the applicability of various animal models to humans and progress toward elucidation of causative agents and mechanisms of action is presented.

Animals↗
PubMed1995

Properties of lavage material from excised lungs ventilated at different temperatures.

We studied the phospholipid (PL) and protein contents, the PL composition, and some of the surface properties of lavage materials obtained from freshly excised rat lungs and excised lungs which had been ventilated at different temperatures (22, 37, and 42 degrees C). Ventilation (60 breaths/min) was carried out at constant tidal volume with periodic sighs for one hour. Although there is slightly more lavageable PL and protein in lungs ventilated at 22 degrees C than in freshly excised lungs, there is no difference in the PL composition or surface properties of lavage materials from these lungs. However, as the temperature at which lungs are ventilated is increased to 37 degrees and 42 degrees C, there is(are): 1) a reduction in lavage fluid PL, 2) a reduction in the relative amounts of total phosphatidylcholines (PC) and disaturated PC (DSPC), the major surface active component of pulmonary surfactant, 3) an increase in unsaturated PC, and 4) increases in total protein and nonsedimentable protein (100,000 g; 2 hr) in the lavage materials. There are also differences in the surface properties of the lavage materials from lungs ventilated at higher temperatures when compared with freshly excised lungs or lungs ventilated at 22 degrees C, probably as a result of the changes in composition. Maximal surface tension is greater for lavage materials from lungs ventilated at 37 degrees C. For lungs ventilated at 42 degrees C, maximal and minimal surface tension values are increased. These results demonstrate that there are differences in the composition and surface properties of alveolar lavage materials from excised lungs ventilated at different temperatures.

Animals↗
PubMed1995

Contribution of opening and closing of lung units to lung hysteresis.

The recruitment and derecruitment of lung units is one explanation of the hysteresis observed in an excised lung during inflation and deflation. A simplified model has been proposed in which the recruitment-derecruitment process is a function of end-expiratory pressure (Frazer, D.G., K.C. Weber and G.N. Franz, Respir. Physiol. 61: 277-288, 1985). The object of this study was to test this model with three experimental procedures. During the first set of experiments, progressively larger pressure-volume (PL-VL) loops were recorded with end-expiratory pressure held at either -5 cmH2O, where all lung units are assumed to be closed, or +5 cmH2O, where all recruited lung units are assumed to be open. In the first case hysteresis is maximal, in the second, minimal. The difference in hysteresis is presumed to arise from the recruitment-derecruitment process. In the second set of experiments, excised lungs are slowly inflated and then deflated at a constant rate while constant-amplitude sinusoidal volume oscillations are superimposed. The end-expiratory pressure of the superimposed loops gradually rose as the lung was inflated and fell as the lung was deflated. Hysteresis was minimal when end-expiratory pressure was above 4 +/- 1 cmH2O even as peak-to-peak loop pressure greatly varied. This supports the notion of an end-expiratory pressure dependent mechanism of recruitment/derecruitment. During the third set of experiments lungs were inflated to either 50%, 75%, or 100% TLC. Volumes of air were then withdrawn and replaced so that the initial volume was restored in sinusoidal fashion as the amplitude of the volume excursions increased. For PL-VL loops with end-expiratory pressures between +4 and -2 cmH2O, pressure amplitudes rose and the hysteresis index (loop area/tidal volume) increased, regardless of the initial lung volume. These results are consistent with the previously described model of Frazer et al. (1985) which assumed that PL-VL curves can be divided into an 'opening' region, an 'open' region and a 'closing' region and that the demarcation of these regions depends on transpulmonary pressure, specifically end-expiratory pressure, and to a much lesser degree on lung volume.

Animals↗
PubMed1995

Nitric oxide synthase inhibitor and lipopolysaccharide effects on reactivity of guinea pig airways.

The in vivo and in vitro effects of nitric oxide (NO) synthase inhibitors and lipopolysaccharide (LPS) on reactivity of guinea pig airways were examined. In isolated, perfused tracheas from untreated animals, the NO synthase inhibitors, N omega-nitro-L-arginine methyl ester (L-NAME; 10(-4)M), NG-methyl-L-arginine (L-NMMA; 10(-4) M) and aminoguanidine (10(-4) M) had no effect or inhibited reactivity to extraluminally (EL) or intraluminally (IL) applied methacholine and histamine. L-NMMA (10(-4) M) did not appreciably contract resting or metacholine-contracted preparations (+/- 3 x 10(-4) M L-arginine) and L-arginine only weakly relaxed contracted tracheas (+/- L-NMMA). Sodium nitroprusside and S-nitroso-N-penicillamine elicited relaxant responses and were more potent extraluminally than intraluminally. Methylene blue (10(-5) M) antagonized relaxation to sodium nitroprusside. Incubation with Escherichia coli LPS (10 micrograms/ml; 30 min incubation) alone in the EL and IL baths depressed methacholine and histamine concentration-response curves. In the presence of LPS, L-NAME potentiated responses to intraluminally applied methacholine but did not affect responses to extraluminally added methacholine. Four days after i.p. injection of animals with LPS (4 mg/kg), L-NAME potentiated responses to IL methacholine, and L-arginine acquired greater relaxant activity. LPS injection increased sensitivity to intraluminally added but not extraluminally added isoproterenol. LPS given by i.p. injection or by inhalation did not affect basal specific airway resistance of conscious animals or reactivity to methacholine aerosol during a postexposure period of 6 to 72 h. NO seems to have little role in regulating reactivity of guinea pig airways to bronchoconstrictor agonists, except after in vitro or in vivo exposure to LPS. After LPS injection the in vitro changes suggestive of NO synthase induction are not associated with altered airway reactivity to inhaled methacholine.

Airway Resistance↗
PubMed1994

Polarized effects of amiloride and 4,4'-diisothiocyano-stilbene-2,2'-disulfonic acid on ATP-induced contraction of trachea.

Polarity in the effects of amiloride and 4,4'-diisothiocyano-stilbene-2,2'-disulfonic acid (DIDS) in the guinea-pig isolated, perfused trachea was investigated to evaluate the roles of epithelial and airway smooth muscle Na+ and Cl- channels in the development of contractile responses to ATP. The blockers were applied to the mucosal (intraluminal) perfusing solution or to the serosal (extraluminal) bath before the second of two challenges with ATP (10(-4) M), which was added to the same bath as the blocker, or to the abluminal bath. In epithelium-intact tracheas, amiloride (10(-4) M) added to the extraluminal or intraluminal bath rapidly (1 min) and extensively inhibited contractions to extraluminally applied ATP (10(-4) M). In contrast, contractions to intraluminally applied ATP (10(-4) M) were relatively resistant to extraluminal and intraluminal amiloride (10(-4) M), in terms of the degree and onset of the inhibition. DIDS (10(-4) M) present in the extraluminal or intraluminal baths caused a slowly developing elevation of baseline tone. After a 30 min incubation, extraluminal DIDS potentiated responses to extraluminally added ATP, but intraluminal DIDS inhibited contractions to ATP added to the extraluminal and intraluminal baths. In contrast to the intact tracheas where there was no difference, the second response of epithelium-denuded preparations to intraluminally administered ATP was diminished. In rubbed tracheas the response to intraluminally added ATP was inhibited further by intraluminal amiloride but was potentiated by intraluminal DIDS. The results suggest that the effects of amiloride and DIDS were polarized across the tracheal wall and involved epithelial and smooth muscle ion channels.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed1993

Organic dust exposure from compost handling: response of an animal model.

The objective of this investigation was to elucidate the pulmonary responses of an animal model to dust generated from leaf/wood compost which had caused a severe case of acute respiratory illness in an individual. Guinea pigs were exposed for 4 hr to 30 mg/m3 of aerosolized leaf/wood compost dust. Inhalation resulted in significant cellular activation and changes in pulmonary mechanics. Maximal elevation in breathing rate (increases 36%) was observed 12-18 hr postexposure. Similarly, maximal granulocyte infiltration (increases 1,600%) and activation of alveolar macrophages (increases 65%) occurred 18 hr postexposure. In contrast, maximal airway obstruction (increases 120%) occurred immediately after exposure and returned toward normal (increases 53%) by 18 hr postexposure. In several respects, the airway obstruction and pulmonary inflammation described in the animal model were comparable to the human response to compost dust. Therefore, this animal model may be useful in predicting the potential respiratory hazard associated with exposure to various organic dusts.

Agriculture↗
PubMed1993

Infective endophthalmitis following vitreoretinal surgery.

Eleven cases of endophthalmitis occurring after vitreoretinal surgery are described. At Moorfields Hospital, London, from 1986 to 1990 the incidence of endophthalmitis after explant surgery with or without drain was 0.19% and after vitrectomy was 0.15%. We conclude that the parity may be due to the intraocular instrumentation of most conventional retinal detachment repair procedures. The best indicator of poor prognosis was speed of onset of symptoms, those with rapid evolution having the worst outcome; 2 of these cases were enucleated. Those presenting at 2-3 days had the best outcome, consistent with infection due to a less virulent organism. Delays in diagnosis were in part due to the posterior location of signs of infection. Potential risk factors amenable to prophylactic strategy were identified in 10 of the 11 patients. Supplementary prophylaxis using ciprofloxacin or imipenem is proposed for cases with an identifiable risk factor. After systemic administration these antibiotics achieve vitreous levels that exceed the MIC90 of the commonest causative pathogens.

Drainage↗
PubMed1993

A review of 72 consecutive cases of Acanthamoeba keratitis, 1984-1992.

A review of consecutive cases of Acanthamoeba keratitis presenting since 1984 was undertaken in order to assess prognostic factors, the success of culture procedures and the outcome of medical and surgical management, with reference to current clinical practice. Seventy-two consecutive cases (77 eyes) of Acanthamoeba keratitis have been managed. Sixty-four patients were contact lens wearers, 28 of these wearing disposable lenses. Superficial corneal involvement and perineural infiltrates were common in those diagnosed less than a month after first symptoms, designated 'early' presentation. Ring infiltrates and ulceration with stromal lysis characterised those presenting at 1-2 months ('intermediate') or after 2 months ('late'); these groups also progressed more frequently to hypopyon, scleritis, glaucoma and cataract formation. Positive corneal cultures were obtained in 10 of 14 (71%) intermediate and 17 of 23 (74%) late cases; early cases underwent epithelial biopsy but formal trephine biopsy was not usually justified (1 of 35 cases) and only 19 of 35 (54%) were tissue-positive. Microbial co-isolates were obtained from 20 corneas. Thirty-four penetrating keratoplasties were performed in 23 eyes, 21 whilst inflamed and 13 when quiet. Of 13 failures in inflamed eyes, 9 were due to recurrence of Acanthamoeba infection. Medical cure is known to have been achieved in 64 of 73 (88%) eyes, 4 of the original 77 having been lost to follow-up abroad. Fifty-eight of 73 eyes (79%) achieved a final visual acuity of 6/12, and of the culture-positive cases, 32 of 46 (70%) achieved 6/12.(ABSTRACT TRUNCATED AT 250 WORDS)

Acanthamoeba↗
PubMed1993

Contractile effects of nucleotides in guinea pig isolated, perfused trachea: involvement of respiratory epithelium, prostanoids and Na+ and Cl- channels.

ATP and UTP contracted guinea pig isolated, perfused trachea and were more potent when applied to the mucosal (intraluminal, IL) surface than when applied to the serosal (extraluminal, EL) surface. IL ATP and IL UTP were equipotent (ATP approximately UTP); EL ATP was 7-fold more potent than EL UTP (ATP > UTP). beta, gamma-Methylene ATP was nearly devoid of activity. Epithelium (Epi) removal decreased IL ATP potency and EL and IL maximum response magnitude, but elevated the IL UTP maximum response. In the presence of EL and IL indomethacin (3 x 10(-6) M; +/- Epi) to inhibit cyclo-oxygenase, or beta, gamma-methylene ATP (10(-4) M) to desensitize receptors, contractions to ATP were abolished, but those to UTP were not. Cl- channel blockade with 4,4'-diisothiocyano-2,2'-stilbene disulfonate (DIDS; 10(-4) M; +/- Epi) and sodium channel blockade with amiloride (10(-4) M; +/- Epi) antagonized contractions to EL and IL ATP and UTP. DIDS and amiloride did not inhibit contractions to methacholine; IL reactivity to methacholine was potentiated by indomethacin and Epi removal. Our findings indicate that the Epi facilitates contraction to ATP, which involves an atypical P2 purinoceptor, prostanoids, and Na+ and Cl- channels. Contractile responses to UTP involve a different receptor, and are neither facilitated by the Epi nor mediated by prostanoids, but involve these channels.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed1993

Relaxant effects of nucleotides in guinea pig isolated, perfused trachea: lack of involvement of prostanoids, Cl- channels and adenosine.

In higher concentrations (> 3 x 10(-4) M) than those causing contractile responses. ATP relaxed the smooth muscle of the guinea pig perfused trachea. Here we examined the relaxant effects of nucleotides. ATP and its nonhydrolyzable congener, beta, gamma-methylene ATP (APPCP), were approximately 4- and approximately 117-fold, respectively, more potent when applied separately to the serosal (extraluminal, EL) surface compared to the mucosal (intraluminal, IL) surface of methacholine (3 x 10(-7) M; EL)-contracted tracheae. APPCP was orders of magnitude more potent than ATP in both EL and IL compartments. EL UTP did not cause relaxation; IL UTP was nearly devoid of activity. The order of EL and IL activity (APPCP >> ATP) was unusual for nucleotide-induced relaxation of smooth muscle. Relaxation to ATP was not inhibited by the Cl- channel blocker 4,4'-diisothiocyano-2,2'-stilbene disulfonate (10(-4) M) or by the cyclo-oxygenase inhibitor indomethacin (3 x 10(-6) M), in contrast to the inhibitory effects of these drugs on contraction to ATP. The adenosine receptor antagonist 8-phenyltheophylline (10(-6) M) had no effect on relaxation to ATP or APPCP. Our findings indicate that Cl- channels, prostaglandins and adenosine are not involved in relaxation to adenine nucleotides.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed1993

Alteration in the modulatory role of respiratory epithelium after exposure of guinea pigs to respirable cotton dust.

Chest tightness may occur after inhalation of cotton dust. To understand the origins of this symptom of byssinosis, we exposed guinea pigs to respirable cotton dust and determined whether the resulting pulmonary obstruction was associated with alterations in the in vitro reactivity of the trachea to methacholine. The isolated, perfused trachea preparation was utilized because it allows a comparison of airway smooth muscle responses to drugs after they are applied separately to the mucosal or serosal surfaces. In epithelium-containing control tracheae, mucosally applied methacholine was less potent and elicited smaller maximum contractile responses than those obtained when it was applied to the serosal compartment. After a 6-h inhalation exposure to 30 mg/m3 respirable cotton dust, the mucosal maximum response to methacholine was substantially increased immediately (0 h) after the end of the exposure, but it was decreased 18 h postexposure. A dust level of 10 mg/m3 produced modest increases in the mucosal maximum responses at both postexposure periods, elevating them to the magnitude of the extraluminal maximum response. The potency of serosally applied methacholine was not affected by any cotton dust exposure. Mechanical epithelium removal increased mucosal reactivity to the serosal level in both control and cotton (30 mg/m3)-exposed groups, and abolished the effects of dust on mucosal reactivity to methacholine. Our findings suggest that the modulatory effect of the epithelium may be inhibitory or excitatory. The balance between the two influences may be altered by inhaled cotton dust.

Administration, Inhalation↗