Graft vasculopathy in clinical hand transplantation.

Allogeneic hand transplantation is now a clinical reality. While results have been encouraging, acute rejection rates are higher than in their solid-organ counterparts. In contrast, chronic rejections, as defined by vasculopathy and/or fibrosis and atrophy of skin and other tissues, as well as antibody mediated rejection, have not been reported in a compliant hand transplant recipient. Monitoring vascularized composite allograft (VCA) hand recipients for rejection has routinely involved punch skin biopsies, vascular imaging and graft appearance. Our program, which has transplanted a total of 6 hand recipients, has experience which challenges these precepts. We present evidence that the vessels, both arteries and veins may also be a primary target of rejection in the hand. Two of our recipients developed severe intimal hyperplasia and vasculopathy early post-transplant. An analysis of events and our four other patients has shown that the standard techniques used for surveillance of rejection (i.e. punch skin biopsies, DSA and conventional vascular imaging studies) are inadequate for detecting the early stages of vasculopathy. In response, we have initiated studies using ultrasound biomicroscopy (UBM) to evaluate the vessel wall thickness. These findings suggest that vasculopathy should be a focus of frequent monitoring in VCA of the hand.

Larviposition in the ovoviviparous blowfly Calliphora dubia.

This study examined larviposition in Calliphora dubia Macquart (Diptera: Calliphoridae), an ovoviviparous blowfly of considerable forensic importance in Australia. Females in the field carried 22-83 live larvae, exhibiting a strong linear relationship between female size and the number of live larvae carried. Females took just over 1 min (mean 67.7 ± 7.7 s, n = 54) to larviposit live larvae on or near fresh liver in the laboratory. Females laid larvae at a mean rate of 1.2 ± 0.1 larvae/s, with the fastest rate being 3.4 larvae/s. Most females (70%) laid live larvae only, but 14% laid larvae and eggs at the same time and 16% laid eggs only (none of the eggs laid were viable). Females laying only live larvae laid a mean of 53.7 ± 2.3 larvae, whereas those laying only eggs laid a mean of 48.6 ± 2.8 eggs on each occasion. None of the eggs laid were viable. Most females (86%) laid all their larvae in a single spot, even if they engaged in several bouts of laying live larvae. Nearly one-third of females did not lay all the live larvae in their ovisacs, but retained half of their complement of developed larvae. Females may be opting to spread their larvae across several carcasses in order to increase their survival and not to overcrowd small, ephemeral carcasses. The fact that a blowfly can lay either eggs or live larvae has enormous implications for the accurate determination of the post-mortem interval (PMI) as the presence of larvae derived from eggs laid on the body add 6-18 h to the PMI. This paper represents the first report of the ability of female calliphorids to resorb some of their own live larvae.

Effect of diet on the excretion profile of ivermectin in cattle faeces.

Two groups of 26 cattle (cross-bred, 3-8 years old, weight 450-650 kg) were placed onto 2 feed regimens, pasture-fed and grain-fed. The pasture-fed cattle were kept on irrigated pasture with hay supplement ad libitum, whereas the grain-fed cattle were kept in a feedlot and fed hay and lupin grain in a 40:60 ratio. Half the cattle within each feed group (n = 13) were given a subcutaneous injection of ivermectin (Ivomec) at the dose rate of 200 micrograms/kg liveweight. Over 5 times higher levels of ivermectin were excreted in the faeces of grain-fed cattle compared with pasture-fed (grazing) cattle. Peak excretion levels of 0.36 mg/kg (grain-fed) and 0.09 mg/kg (pasture-fed) of ivermectin were recorded at 6 and 8 days post-injection (dpi), respectively. Ivermectin was detected in faeces for up to 13 dpi (grain-fed) and 14 dpi (pasture-fed). Faecal pH in grain-fed cattle was more acidic (6.43) compared with pasture-fed cattle (7.28) and the levels of nitrogen in faeces from grain-fed cattle were higher (0.42%) compared with pasture-fed cattle (0.20%). Treatment with ivermectin had no influence on either the pH or nitrogen levels of faeces from either feed regimen. The impact of diet on the excretion profile of ivermectin is discussed in the context of studies attempting to demonstrate ecotoxic effects of ivermectin residues in cattle faeces on dung beetles.

Serotonin-1B agonists induce compartmentally organized striatal Fos expression in rats.

The 5HT1B agonist RU24969 (2.5-5.0 mg/kg) and anpirtoline (2.0 mg/kg) induced a striking increase in striatal Fos-like immunoreactivity in rats. In the rostral and dorsal regions of the striatum staining was dense and relatively homogeneous. In the ventral region of the striatum at more caudal levels, however, both drugs induced staining in patches which were in register with the opiate receptor rich striosomes. The effects of RU24969 could not be antagonized by the selective 5HT1A antagonist p-MPPI and little or no striatal Fos expression could be observed after injections of the selective 5HT1A agonist 8-OHDPAT or the selective 5HT3 antagonist MDL-72222.

Quinpirole attenuates the striatal immediate early gene expression, but not the hyperactivity, induced by the serotonin agonist RU-24969.

Systemic administration of the mixed 5-HT(1A/1B) agonist RU-24969 has been shown to produce a dramatic increase in locomotor activity and to induce robust c-Fos expression in the rat striatum. Previous studies have also shown that pretreatment with the D2-like dopamine agonist quinpirole virtually abolishes RU-24969-induced striatal c-Fos expression. The present study was undertaken to determine whether the effects of RU-24969 on immediate early gene expression extend to the additional Fos family transcription factors FosB and Fra-2. Additionally, this study quantitatively examined the effect of quinpirole pretreatment on the ability of RU-24969 to induce both locomotor hyperactivity and striatal immediate early gene expression. RU-24969 alone produced elevations in locomotor activity and induced clear expression of c-Fos, FosB and Fra-2 throughout the entire striatal complex. Quinpirole pretreatment virtually abolished RU-24969-induced expression of all three transcription factors, but did not alter the elevated locomotor activity produced by RU-24969. These results demonstrate that the effects of RU-24969 on locomotor activity can be dissociated from its effects on immediate early gene expression within the striatum.

Quinpirole attenuates striatal c-fos induction by 5-HT, opioid and muscarinic receptor agonists.

Pretreatment with the dopamine D2 receptor agonist quinpirole (0.025-2.5 mg/kg) produced a marked, dose-dependent, attenuation of the striatal Fos expression induced by the serotonin (5-Hydroxytryptamine, 5-HT) releasing agent fenfluramine (25 mg/kg). Quinpirole (2.5 mg/kg) was also able to drastically attenuate the striatal Fos response produced by injections of the direct 5-HT1/2 receptor agonist N-(3-trifluoromethylphenyl)piperazine hydrochloride (TFMPP) (5 mg/kg), the selective 5-HT2 receptor agonist 1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane hydrochloride (DOI) (6.64 mg/kg), the 5-HT1A/1B receptor agonist RU-24969 (5-methoxy-3-(1,2,3,6-tetrahydropyridin-4-yl)1H-indole) (5 mg/kg), the mu-opioid receptor agonist morphine (5 mg/kg) and the muscarinic cholinergic receptor agonist pilocarpine (50 mg/kg). These results are in marked contrast to the previously reported ability of quinpirole to potentiate the response to D1 dopamine receptor agonists and demonstrate that stimulation of D2-like receptors can have differential effects on the Fos responses induced by various drugs.

Conditioned taste aversion is a confound in behavioral studies that report a reduction in the reinforcing effects of drugs.

Pharmacologic agents with a potential to attenuate the reinforcing properties of drugs of abuse may have an important role in the treatment of drug addiction. The reduction of drug self-administration and sweet solution intake are two common animal models employed to screen for promising therapeutic agents. When these agents are effective in suppressing the behavior maintained by drugs of abuse, the cause is usually attributed to a neuronal mechanism such as the modification of neurotransmitters that subserve reinforcement. These experiments present data for an alternate interpretation which suggest that some of these agents produce a conditioned taste aversion (CTA) that acts as a confounding variable in the screening of potential therapeutic agents. Both carbamazepine and isradipine were shown to establish a CTA at doses reported to attenuate the reinforcing properties of drugs of abuse. It is concluded that CTA represents a potential experimental confound in studies of pharmacologic agents that appear to attenuate the reinforcing properties of drugs. These results suggest that screening for a CTA is necessary in any paradigm that measures the suppression of consummatory behavior in response to pharmacologic intervention.

Forensic entomology: application, education and research in Western Australia.

Forensic entomology as a science and a tool for investigation has had slow beginnings in Australia. A number of small animal decomposition trials have been recorded in the literature but mostly from an ecological rather than a forensic entomology perspective. In the last 20 years, a number of more forensically orientated field trials on small pigs and some fly developmental trials in the laboratory have been conducted but lack any replication. The following article was presented at an international seminar to detail the current research in forensic entomology, the applications of forensic entomology in scene of crime (SOC) and homicide investigations and the education of police and judiciary in the discipline of forensic entomology in Western Australia over the last 10 years.

Effect of avermectin residues in sheep dung on mating of the Australian sheep blowfly Lucilia cuprina.

Male and female Lucilia cuprina adults were separated after emergence and then fed either dung excreted from sheep after an oral treatment with ivermectin (Ivomec) or non-treated sheep dung. Fresh dung (non-treated or ivermectin-treated) was collected daily and fed to adults for 6 days after drenching. After this feeding period, males and females from each feeding treatment were paired and all aspects of mating behaviour recorded. Feeding on avermectin residues by adults had a significant effect on two components of mating: (1) males fed on ivermectin-treated dung directed less mating attempts at females compared with males fed on non-treated dung; (2) mating duration by males fed on ivermectin-treated dung was longer compared with males fed on non-treated dung. There was no significant difference in the level of sexual receptivity (percentage mating) between females fed on either ivermectin-treated dung or non-treated dung, although subsequent oviposition by females fed ivermectin-treated dung was significantly delayed. Both female and male mortality was significantly greater when feeding on ivermectin-treated dung compared with non-treated dung. The effects on mating, when coupled with the deleterious effects on ovarian development and adult mortality, demonstrate the potential for avermectin usage in a control strategy against L. cuprina.

Stable fly, house fly (Diptera: Muscidae), and other nuisance fly development in poultry litter associated with horticultural crop production.

Poultry litter usage in horticultural crop production is a contributor to nuisance fly populations, in particular stable flies (Stomoxys calcitrans L.) and house flies (Musca domestica L.). Extrapolation of adult emergence data suggests that approximately 1.5 million house flies and 0.2 million stable flies are emerging on average from every hectare of poultry litter applied as a preplant fertilizer for vegetable production in Perth, Western Australia. To a lesser extent, sideband applications to established crops may allow for the development of 0.5 million house flies and 45,000 stable flies per hectare. However, up to 1 million house flies, 0.45 million lesser house flies, Fannia cannicularis L., and 11,000 stable flies per hectare may be produced from surface dressings of poultry litter associated with turf production. Other nuisance flies present in poultry litter included the false stable fly, Muscina stabulans (Fallén), bluebodied blowfly, Calliphora dubia Hardy, black carrion fly, Hydrotaea rostrata Robineau-Desvoidy, Australian sheep blowfly, Lucilia cuprina Wiedemann, and flesh flies (Sarcophagidae). Only house flies developed in poultry litter for the first 4 d after application in the field. Stable flies were not present in poultry litter until 4-7 d after application, and were the only fly species developing in litter > 9 d after application.

Susceptibility of mulesing wounds to oviposition and strike development by the Australian sheep blowfly Lucilia cuprina.

The age and/or physical condition of mulesing wounds had a significant effect on both the oviposition response of L. cuprina and the ability of the wound to support a strike. Up to 48 h after mulesing, untreated wounds elicited a strong oviposition response in contrast with chemically treated wounds, although subsequent larval development was negligible. Seven to 9-day-old wounds, however, regardless of the wound treatment, were highly attractive oviposition sites, which subsequently developed into strikes; the D3 formulation of Defiance*S, however, significantly depressed strike development and shows promise as a mules wound treatment. A marked improvement in the wound healing 14 to 16 d after mulesing coincided with a significant decrease in oviposition. Only those sheep whose wound scabs remained broken, exposing pus and raw tissue, attracted oviposition; subsequent development of the eggs into strikes was negligible.

Rate of development of Hydrotaea rostrata under summer and winter (cyclic and constant) temperature regimes.

The black carrion fly Hydrotaea rostrata Robineau-Desvoidy (Diptera: Muscidae) is a muscid, which occurs on approximately one-third of decomposing human corpses involved in homicide cases in south-western Australia. Work to date on its development rates is scant with only one published source available. The current study measured the precise developmental rates of this species using high repetition and sampling rates. A comparison was made of the developmental rates between constant and cyclic temperatures in winter and summer temperature regimes in south-western Australia. Developmental times for 90% of first stage larvae to emerge as adult flies are: summer constant, 21.6days, summer cyclic, 23.5 days, winter constant 64.5days and winter cyclic, 48.3 days. These data will allow forensic entomologists to make more accurate determinations of post-mortem intervals in cases where H. rostrata life history stages are present.