Penumbral mismatch is underestimated using standard volumetric methods and this is exacerbated with time.

BACKGROUND AND AIMS: The mismatch between perfusion weighted images (PWI) and diffusion weighted images (DWI) using MR is increasingly being applied in patient selection for therapeutic trials. Two approaches to the calculation of the mismatch volume exist--the commonly used volumetric and the more precise co-registration method, the latter of which considers lesion topography. That there are differences in the mismatch volume analysed by each method and that these are time dependent was hypothesised. METHODS: Patients within 48 h of ischaemic stroke onset had baseline MR PWI/DWI mismatch and T2 outcome volumes at 3 months. Volumetric mismatch volume was defined as PWI minus DWI lesion. Co-registration mismatch volume was defined as the PWI defect lesion not overlapped by the co-registered DWI lesion. RESULTS: 72 patients of median age 74.0 years were studied. Median baseline MR was at 5.9 h (IQR 3.0, 20.4 h) after stroke onset. Consistent underestimation of the mismatch volume occurred using the volumetric method (volumetric median 9.3 ml, IQR 0, 63 ml; co-registration median 20.1 ml, IQR 3.2, 69.8 ml; p<0.0001). This difference increased with time from stroke onset (p = 0.006). CONCLUSIONS: Volumetric analysis consistently underestimates the PWI/DWI mismatch volume compared with the more precise co-registration method. This effect increases with time.

TGF-beta1 gene transfer to the mouse colon leads to intestinal fibrosis.

Crohn's disease (CD) is a chronic, relapsing inflammatory bowel disease, characterized by transmural inflammation. In CD, the recurrent inflammatory injury and tissue repair that occurs in the intestine can progress uncontrollably, leading to the proliferation of mesenchymal cells as well as fibrosis, characterized by excessive extracellular matrix deposition. These processes thicken the bowel wall, reducing flexibility, and often culminate in obstructive strictures. Because no effective measures are currently available to specifically treat or prevent intestinal stricturing, we sought to gain a better understanding of its pathogenesis by developing a mouse model of intestinal fibrosis. Because transforming growth factor (TGF)-beta1 can mediate both fibrosis and mesenchymal cell proliferation; we studied the effects of delivering adenoviral vectors encoding spontaneously active TGF-beta1 into the colons of mice. We first demonstrated that enema delivery of marker adenoviral vectors led to the transfection of the colonic epithelium and transient transgene expression. Histologically, control vectors caused an acute inflammatory response, involving the recruitment of neutrophils and mononuclear cells into the colonic lamina propria; however, infection caused little if any fibrosis. In contrast, the TGF-beta1 vector caused a more severe and prolonged inflammatory response as well as localized collagen deposition, leading to severe and progressive fibrosis. This was accompanied by the emergence of cells with a myofibroblast phenotype. Ultimately the fibrosis resulted in many of the TGF-beta1-transfected mice developing profound colonic obstruction. Through adenoviral gene transfer technology, we describe a novel mouse model of colitis and implicate TGF-beta1 in the pathogenesis of obstructive intestinal fibrosis.

Resistance to benzimidazole anthelmintics in equine strongyles. 1. Frequency, geographical distribution and relationship between occurrence, animal husbandry procedures and anthelmintic usage.

A survey was conducted to determine whether benzimidazole resistant populations of equine strongyles are present in New South Wales and north central Victoria; what is their frequency and geographical distribution; which species are involved; and whether different methods of parasite control could be related to the occurrence and frequency of anthelmintic resistant populations. Resistant populations of strongyles were found over wide areas of New South Wales and in north central Victoria. There was no relationship between geographical location and the occurrence of benzimidazole resistance. The species involved were small strongyles of the sub-family Cyathostominae. There was a direct correlation between the occurrence of resistance (including the level at which it is present) and the frequency of use of benzimidazole anthelmintics. Examination of management practices showed that resistance is not an important problem on farms where different chemical classes of anthelmintics were used in a slow rotation programme; combination anthelmintic therapy (for example, benzimidazole/piperazine/organophosphates) was used and anthelmintic treatment was given at intervals of not less than 16 weeks. Tentative suggestions are made for the control of small strongyles in the light of an emerging resistance problem.

Resistance to benzimidazole anthelmintics in equine strongyles. 2. Evidence of side-resistance, and susceptibility of benzimidazole-resistant strongyles to non-benzimidazole compounds.

The susceptibility of a known thiabendazole-resistant population of small strongyles to anthelmintics of both benzimidazole and non-benzimidazole groups, was determined. In the first study, 42 horses infected with thiabendazole-resistant small strongyles were allocated to 6 groups. Treatment groups received one of the following anthelmintics: mebendazole, febantel, febantel plus trichlorphon, morantel tartrate, or a combination of thiabendazole, piperazine and trichlorphon. Morantel tartrate and the thiabendazole/piperazine/trichlorphon combination produced highly significant (p less than 0.001) reductions in faecal strongyle egg counts 20 days post-treatment. Mebendazole, febantel and febantel plus trichlorphon failed to reduce strongyle egg counts significantly. Larval culture and differentiation indicated that in all cases of anthelmintic failure, small strongyles of the sub-family Cyathostominae were involved. Eighteen horses from groups in which treatment had failed were re-allocated to 3 groups. Treatment with either morantel tartrate or haloxon was highly efficient in reducing faecal strongyle egg counts. In the final study, fifty-four horses, infected with benzimidazole-resistant small strongyles were allocated to 10 groups. On day zero, each treatment group received one of the following anthelmintics: thiabendazole, cambendazole, mebendazole, oxibendazole, piperazine, thiabendazole/piperazine, cambendazole/piperazine, mebendazole/piperazine or oxibendazole/piperazine. Oxibendazole, piperazine and the benzimidazole/piperazine combinations produced highly significant reductions in faecal strongyle egg counts 20 days post-treatment (p less than 0.001). When administered alone, benzimidazole anthelmintics failed to reduce strongyle egg counts significantly, with the exception of oxibendazole. Larval culture and differentiation indicated that in all cases of anthelmintic failure, the species involved were small strongyles of subfamily Cyathostominae. There was no significant increase in benzimidazole resistance level (based on in vitro assay) as a result of drug treatment, over one generation.

Use of guniea pigs to assay anthelmintic resistance in ovine isolates of Trichostrongylus colubriformis.

Guinea pigs infected with Trichostrongylus colubriformis were used to develop an assay for anthelmintic resistance by determination of worm burdens following treatment with test anthelmintics. To achieve comparable efficacy with the recommended dose of thiabendazole and levamisole in sheep, dose rates in guinea pigs had to be increased two to four fold. For example, thiabendazole at 100 mg/kg in guinea pigs was 96.6 per cent effective against a thiabendazole susceptible (GS) strain of T colubriformis, but had no effect against a thiabendazole-resistant (VRSG) strain. In sheep 50 mg/kg of thiabendazole would have a similar efficacy against each strain respectively. Morantel tartrate at 10 mg/kg in guinea pigs was 99 to 100 per cent effective against the GS strain but only 54 per cent effective against a morantel resistant (PF4) strain. A slope ratio assay was used to calculate the relative potency of anthelmintics by comparing efficacies against resistant strains with efficacy against the GS strain. Resistance of the VRSG strain to thiabendazole was confirmed with a relative potency for this drug of 0.047 in guinea pigs. The PF4 strain was resistant to both thiabendazole and levamisole which had relative potencies of 0.168 and 0.255 respectively. The advantages of this statistical treatment together with the cost and time savings of the guinea pig model over a conventional critical anthelmintic assay in sheep are discussed.

Changes in response of a benzimidazole resistant strain of Haemonchus contortus from sheep after passing through calves.

A benzimidazole resistant strain of Haemonchus contortus was passaged through lambs only or from lambs to calves and back into lambs. Changes in response to thiabendazole were monitored by using an egg hatch test at each animal passage and by a controlled experiment on adult worms at the final passage in lambs. An increased level of resistance was shown for the isolate during its passage through calves by the egg hatch test, although this was not supported on the adult worms in sheep using a single dose rate of 66 mg/kg of thiabendazole.

Five generations of selection with benzimidazole and non-benzimidazole anthelmintics against benzimidazole resistant strains of Haemonchus and Ostertagia spp in sheep.

Benzimidazole resistant strains of Haemonchus contortus and Ostertagia spp were subjected to selection pressure over five laboratory generations with the recommended dose rates of either cambendazole, oxfendazole or morantel. A change in response, with larger residual worm burdens remaining after treatment at the fifth generation, was shown for both cambendazole and oxfendazole against both species of nematode. No change in response against either species are shown for morantel. The results are discussed in terms of the problem associated with inefficient removal of the adult parasites after treatment.

Anthelmintic efficacy of low-dose phenothiazine against strains of sheep nematodes susceptible or resistant to thiabendazole, levamisole and morantel tartrate: effect on patent infections.

The anthelmintic efficacy of low-dose phenothiazine therapy, administered as a 1:40 phenothiazine: molasses mixture, was tested against patent infections of strains of Haemonchus contortus, Trichostrongylus colubriformis and Ostertagia spp susceptible or resistant to thiabendazole (an other benzimidazoles), levamisole and morantel tartrate. Phenothiazine reduced faecal egg output for both susceptible and resistant strains of all three nematodes. In daily doses of 0.25 g per sheep per day and above it completely inhibited larval production in susceptible strains. Against resistant strains, there was a reduced efficiency with 82.3 per cent inhibition of egg hatch at the 0.25 g per sheep per day level. Phenothiazine treatment had no significant effect on worm numbers for either susceptible or resistant worms. It is suggested that benzimidazole-resistant strains may be cross-resistant to phenothiazine.

Trichostrongylus colubriformis and Ostertagia sp resistant to levamisole, morantel tartrate and thiabendazole: infectivity, pathogenicity and drug efficacy in two breeds of sheep.

Merino and Border Leicester cross Merino sheep, nine months old, were infected with 10,000 third stage larvae of both Trichostrongylus colubriformis (PF4) and Ostertagia circumcincta/O trifurcata (PF5), known to have varying degrees of resistance to levamisole, morantel tartrate and thiabendazole. Crossbred sheep carried heavier Ostertagia sp worm burdens but there was no difference in susceptibility between the two breeds of sheep to infection with T colubriformis. The anthelmintic efficacy of thiabendazole, at 50 mg/kg, against T colubriformis was 81.8 per cent and 92.4 per cent for Merinos and crossbreds respectively while levamisole, at 6.75 mg/kg, was 12.3 per cent and 18 per cent effective. Thiabendazole removed 92.3 per cent and 83.8 per cent of Ostertagia sp in Merinos and crossbreds respectively. However, worm burdens in levamisole treated sheep were not significantly different from controls. No significant breed differences were found in 24 h faecal egg outputs. It is suggested that breed differences previously described in four-month-old sheep may have been due to differences in the rate of development of immune responsiveness. This disparity was no longer detectable in the immunologically mature sheep used in this study.

Trichostrongylus colubriformis and Ostertagia sp resistant to levamisole, morantel tartrate and thiabendazole: isolation into pure strain and anthelmintic titration.

Strains of Trichostrongylus colubriformis (designated PF4) and Ostertagia sp (O circumcincta/O trifurcata, designated PF5), with varying degrees of resistance to levamisole, morantel tartrate and thiabendazole were isolated into pure culture. Detailed dose response studies showed that both T colubriformis and Ostertagia sp were highly resistant to levamisole and morantel with low level resistance to thiabendazole. The effective dose required to remove 80 per cent worm burdens (ED80) was calculated for each anthelmintic. For T colubriformis the ED80 for levamisole and thiabendazole were 12.6 and 40.1 mg/kg respectively. For Ostertagia sp, the ED80 for levamisole, thiabendazole and morantel were 20.4, 45.2 and 35.8 mg/kg respectively. The implications of these results are discussed with reference to alternative means of chemical control.

Trichostrongylus colubriformis and Ostertagia circumcincta resistant to levamisole, morantel tartrate and thiabendazole: occurrence of field strains.

Field strains of Trichostrongylus colubriformis and Ostertagia circumcincta, designated PF4 and PF5 respectively, were recovered from a farm on which the sole use of levamisole over a preceding 12 year period led to the development of anthelmintic resistance. The results of field observations and preliminary critical trials in both Merino and crossbred sheep showed that both species have varying degrees of resistance to three major anthelmintics; levamisole, morantel tartrate and thiabendazole. Mean worm count reductions for adult T colubriformis (PF4) for therapeutic doses of morantel tartrate, thiabendazole and levamisole in crossbreds were 45.7 per cent, 97.3 per cent and zero respectively, and for Merinos 80.7 per cent, 88.3 per cent and 92.0 per cent respectively. Against O circumcincta the corresponding reductions for crossbreds were 51.4 percent, 95.4 per cent and 20.3 per cent and for Merinos, 52.5 per cent, 73.1 per cent and 29.8 per cent. There was no statistically significant difference in the responses of both parasite species to either levamisole or morantel. This result suggests that resistance to the two chemically unrelated drugs may be co-inherited.

The efficacy of fenbendazole and albendazole against immature and adult stages of benzimidazole-resistant sheep trichostrongylids.

The efficacy of two recently introduced benzimidazole anthelmintics, albendazole and fenbendazole, was determined for six-day, 10-day and adult stages of resistant strains of Haemonchus contortus and Trichostrongylus colubriformis. Albendazole, at 3.8 mg/kg reduced H contortus worm counts by 92.4, 70.8 and 67.1 per cent while fenbendazole, at 5.0 mg/kg, reduced worm burdens by 51.7, 95.5 and 93.4 per cent against six-, 10- and 25-day-old parasites respectively. For T colubriformis, the corresponding reductions with albendazole were 97.7, 95.8 and 64.9 per cent and for fenbendazole 29.0, 66.3 and 33.4 per cent. Compared with susceptible strains of H contortus and T colubriformis, for which therapeutic doses of benzimidazole anthelmintics are generally highly active against all stages of development, the present results show that these drugs do not have a uniform level of activity against all developmental stages of resistant strains.

The effect of single and divided dose administration on the efficacy of fenbendazole against adult stages of benzimidazole resistant sheep trichostrongylids.

Sheep infected with benzimidazole resistant strains of Haemonchus contortus and Trichostrongylus colubriformis were used to compare the anthelmintic efficacy of fenbendazole given as a single dose or administered in a divided dose regime over five days. Statistical analysis showed no significant difference between the two methods of administration for H contortus. On the other hand, divided dose fenbendazole was significantly less effective than single doses against adult T colubriformis at dose rates of 5 and 7.5 mg/kg. In the case of H contortus a highly significant correlation coefficient between post treatment egg counts and worm counts (r = 0.789) was obtained. This suggests that reduction in faecal egg output following drug treatment would provide a useful field indication of anthelmintic performance of fenbendazole (and possibly related compounds) against benzimidazole resistant strains of this parasite.