E. C. G. M. Hampson

pH-Gated dopaminergic modulation of horizontal cell gap junctions in mammalian retina

Horizontal cells mediate lateral inhibition in the outer retina, and this process is dependent on electrical coupling through gap junctions, giving rise to receptive fields that are much wider than the dendritic fields. This study on rabbit retina shows that the permeability of the gap junctions between A-type horizontal cells, as assessed by Lucifer yellow dye coupling, is modulated by dopamine through a D1 receptor linked to adenylate cyclase. Both exogenously applied dopamine and endogenously released dopamine uncoupled the horizontal cells, but the effect was pH-gated whereby it occurred only at an extracellular pH 7.2±0.05. The horizontal cells also uncoupled in acidic media (pH 7.0 or below) in the absence of dopamine. Our results show that horizontal cell coupling in the mammalian retina is regulated by both dopamine and pH. Given that the pH in the outer retina varies with the metabolic activity of the photoreceptors, these results suggest that ambient light conditions could gate the activity of neurotransmitters through pH-sensitive mechanisms.

Failure of haemoperfusion and haemodialysis to prevent death in paraquat poisoning. A retrospective review of 42 patients.

SummaryIn this review the efficacy of haemoperfusion in the treatment of paraquat poisoning is addressed. 42 reports containing sufficient information of paraquat-poisoned patients were evaluated. These reports, from 35 patients reported in the literature and 7 new cases, were chosen for the following reasons: the timed plasma paraquat concentrations were known, patient outcome was known, and details of haemoperfusion were available. In some cases, haemodialysis was also performed. The plasma paraquat concentrations and the specific times post-ingestion were plotted on a contour graph that predicts the probability of survival. Comparison of the predicted probability of survival versus the actual outcome showed that haemoperfusion, single or repeated, did not affect patient survival. None of the patients whose initial plasma concentrations were greater than 3 mg/L paraquat survived, regardless of the time after ingestion that the concentrations were measured, and despite haemoperfusion. Therefore, such patients might not be considered for haemoperfusion because of their uniformly bad prognosis, despite the procedure being used, and because of the morbidity, discomfort and costs associated with it. Clearly, the need for better techniques to remove paraquat and to prevent the consequences of the metabolic effects of the compound are required urgently before the treatment of the paraquat-poisoned patient will be successful.

Kinetics of toxic doses of paraquat and the effects of hemoperfusion in the dog.

Knowledge of the kinetics of an intoxicant is required for designing potential therapies in poisoned patients. In the case of paraquat, elucidating the kinetics has been made difficult by the paraquat-induced renal failure and the consequent dose- and time-dependent elimination of the herbicide. In the current study, we have modelled the plasma and urinary concentrations of paraquat in dogs given a toxic dose, the elimination of which was nonlinear. This enabled us, in turn, to simulate the apparent concentrations of paraquat in the deep tissue compartment, part of which is constituted by the major target organ for paraquat toxicity, the lung. Finally, we defined conditions, if any, under which charcoal hemoperfusion could reduce exposure of the deep compartment to paraquat by > or = 25%. We found that the plasma concentrations of paraquat could be described by a two compartment model with non-linear elimination from the central compartment. Use of a three compartment model did not improve the fit over that for a two compartment. The volume of distribution of paraquat at steady state approximated that of total body water. Simulated hemoperfusion performed for eight or eighty hours did not reduce exposure of the deep compartment to paraquat by > or = 25%, unless begun at times < or = two hours of the infusion commencing. This is consistent with our experimental data in the dog. The lack of efficacy of hemoperfusion is due to the rapid renal elimination of most of the absorbed dose of paraquat over the first 12 hours after its administration, and the later limitation of the rate of removal of paraquat from the body by the slow efflux rate from the deep to central compartment.

Quantitation of Paraquat in Biological Samples by Radioimmunoassay Using a Monoclonal Antibody

We have developed a sensitive and specific radioimmunoassay for the quantitative determination of paraquat in plasma, urine, and bronchoalveolar lavage fluid using a monoclonal antibody. The synthesis of the iodinated paraquat tracer is novel and less complicated than a previously reported method. Regardless of the type of biological sample analyzed, the sensitivity of the assay was 0.46 ng/ml in a 200-microliters aliquot. The results correlated well with those of another assay performed in a separate laboratory. Paraquat concentrations were determined in plasma and in urine of a dog over several days after the intravenous administration of 7.48 mg paraquat cation/kg and in bronchoalveolar lavage fluid obtained 34 hr after the 2-hr infusion was discontinued. Concentrations of paraquat measured ranged from 14.1 to 0.03 micrograms/ml in plasma and 2034 to 0.36 micrograms/ml in urine. Concentrations of paraquat in plasma obtained at various times after the ingestion of paraquat by three patients ranged from 51.0 to 0.010 micrograms/ml.

Toxicological index of paraquat: A new strategy for assessment of severity of paraquat poisoning in 128 patients

A new assessment of the severity of paraquat poisoning in 128 patients has been developed. It involves toxicological index of paraquat and discriminant function score. This system not only allows a more accurate assessment of severity of the poisoning, but also provides a more reliable prediction of the outcome in an early stage for the purpose of forensic and clinical toxicology.

Effects of paraquat on canine bronchoalveolar lavage fluid

Bronchoalveolar lavage (BAL) recovers the epithelial lung fluid of the lower respiratory tract. In this study, we have used BAL to detect early pulmonary injury in beagle dogs following an intravenous infusion of 10 mg paraquat dichloride/kg bodyweight. Bronchoalveolar lavage was performed twice in 11 dogs, 60 hr before and 34 hr after an intravenous infusion of paraquat dichloride (n = 8) or saline (n = 3). The dogs were studied in three groups: (1) paraquat only (n = 4); (2) paraquat plus hemoperfusion (n = 4); and (3) hemoperfusion only (n = 3). Because hemoperfusion, a treatment used for paraquat poisoning, could have effects on BAL independent of paraquat, we evaluated the effects on BAL fluid of this procedure performed separately from and together with administration of paraquat. We examined cytology, proteins, enzymes, and glutathione in the BAL fluid and expressed all results per milliliter of aspirated lavage fluid. Hemoperfusion did not alter the BAL fluid. In contrast, in dogs studied 34 hr after administration of paraquat, total cell counts, alveolar macrophage and neutrophil counts, and concentrations of total protein, albumin, ACE, LDH, and ALP were increased. Bronchoalveolar lavage in the dog provides an excellent tool with which to detect early paraquat-induced pulmonary injury. The same technique could be useful for sequential monitoring of other types of pulmonary disease and injury.

Repeated hemoperfusion and continuous arteriovenous hemofiltration in a paraquat poisoned patient

Prompt hemodialysis or hemoperfusion can be of value during the first 24 hours after paraquat ingestion particularly when the patient has developed acute renal failure. However, many cases of paraquat poisoning occur in areas where hemoperfusion facilities are unavailable. In contrast, continuous arteriovenous hemofiltration (CAVH) could be instituted easily. We have measured the removal of paraquat from the body by CAVH in a 46 year old male cane farmer who ingested 70 ml, 20% paraquat and died twelve days later from pulmonary fibrosis. Renal failure developed rapidly. Concentrations of paraquat were measured by an indirect competitive ELISA using a murine paraquat monoclonal IgG antibody. Hemoperfusion was performed daily for five days, beginning 78 hours post-ingestion. By 180 hours, when the patient was in respiratory failure, hemoperfusion was replaced with CAVH which was continued for 46 hours. During this time interval, 1.1 mg paraquat was recovered in the hemofiltrate and 1.56 mg paraquat in the urine. The extraction of paraquat by the hemofilter was close to 100%. The plasma clearance of paraquat across the hemofilter was 6.1 ml/min and the renal clearance was 8.2 ml/min. The mean hemoperfusion clearance of paraquat was 50 ml/min and the total amount of paraquat removed by the 34 hours of hemoperfusion was 9 mg. Because of the relative ease with which CAVH can be performed, its low cost, compared to that of hemoperfusion or hemodialysis, and the continuous nature of the procedure, CAVH may be worth considering in paraquat poisoning. It could be used particularly in those patients who have developed renal failure or while patients are being prepared for hemoperfusion.

Creation of a carotid-jugular fistula for repeated hemoaccess in a canine model

We have developed a method to obtain access to the arterial circulation in the dog, which can be used for repeated hemoperfusion, hemodialysis, and for pharmacokinetic studies. Hemoaccess is achieved by the surgical creation of a carotid-jugular fistula. Using the fistula, which is internal and autogenous, has overcome the problems of thrombosis, dislodgement, and infection, which are associated with external shunts. Blood flow rates of between 70 and 100 ml/min in 12-kg dogs were obtained through a hemodialysis pump. The surgical techniques are described in detail so that other investigators can avail themselves of the methodology.

Identification of bacteria and fungi sampled from the conjunctival surface of normal horses in South-East Queensland, Australia

OBJECTIVE To identify bacteria and fungi found on the conjunctival surface of normal horse eyes; to investigate potential risk factors for these microflora; and to determine their susceptibility to common topical ophthalmic antimicrobials. ANIMALS STUDIED A total of 95 client-owned horses were studied. PROCEDURES Horses within sub-tropical Australia (South-East Queensland) were sampled once between April 2012 and March 2013. A conjunctival swab was taken from each eye and cultured for aerobic bacteria and fungi. Organisms were identified by colony morphology and phenotype. Antimicrobial disk diffusion susceptibility testing for commonly used antimicrobials was performed. RESULTS Positive bacterial cultures were returned from 187/190 (98.4%) eyes from 94/95 (98.9%) horses. The most common species included Staphylococcus spp. (25.2% of total bacterial isolates), Bacillus cereus (17.4%), Bacillus spp. (14.1%), and Corynebacterium spp. (8.9%). Most bacterial isolates were susceptible to neomycin and fluoroquinolones. Positive fungal cultures were returned from 111/190 (58.4%) eyes from 73 (76.8%) horses. The most common species identified included: Penicillium spp. (16.7% of fungal isolates), Aspergillus spp. (15.4%), and Scopulariopsis spp. (10.3%). Most (≥90%) molds were susceptible to ketoconazole, voriconazole, itraconazole, and miconazole. Yeasts were most susceptible to ketoconazole. There was no significant effect of breed, age, sex, purpose, or housing of the horse or climatic conditions on bacterial or fungal culture status. CONCLUSIONS Bacteria and fungi were commonly isolated from the eyes of healthy horses. The antibiotic and antifungal susceptibilities identified can be used as a guide for empirical therapy after cytology in the treatment of corneal ulceration in horses.