Here we report NS5A deep sequencing analyses of patient HCV samples obtained from a 3-day monotherapy study of GS-5816 in HCV-infected subjects. Methods: Treatment naïve patients with chronic HCV infection were administered GS-581 6 for 3 days at doses ranging from 5mg-150mg. Pretreatment, on-treatment, and post-treatment plasma samples from GT 1,2, and 3 subjects were analyzed for NS5A RAVs by deep sequencing analysis with a 1% assay

cutoff. The presence of NS5A RAVs at amino acids 28, CH5424802 30, 31, 58 and 93 were examined for GT 1a, 2, and 3 samples and NS5A amino

acids 31 and 93 for GT 1 b. Results: The NS5A RAVs M28T, Q30H/R, L31M/V, and Y93C/H/R were detected at baseline in 10/32 (31.3%) sequenced GT 1 a infected subjects. Despite this high rate of detectable R428 order RAVs, similar viral load decreases were observed in subjects with detectable baseline RAVs versus those with no detectable RAVs. One GT 1 b infected subject had Y93H at baseline that did not impact response to GS-5816 150mg. Four of seven GT 2 infected subjects had detectable L31M at baseline. No significant differences in maximum viral load reductions between subjects with L31 or M31 were observed. In GT 3 infected PLEKHB2 subjects, the A30K and Y93H RAVs were observed in 1/10 and 2/10 subjects, respectively. One GT 3 infected subject with Y93H (16% of population)

had a 2.7 log 10 reduction in HCV RNA with administration of GS-5816 150 mg. The single GT 3 infected subject with A30K had a 2.9 log10 decrease in HCV RNA. Overall, NS5A RAVs were detected at varying prevalence, with frequencies varying from 1-99% as a proportion of the HCV quasispecies. Fourteen-day post-treatment analyses demonstrated complex mixtures of NS5A RAVs including M28T, Q30R/H, L31M/V, H58D, and Y93H/N/S among GT 1a subjects. In GT 1b and GT 2b subjects, L31V/M and Y93H were commonly observed while Y93H/N was the most common RAV in GT 3 subjects following 3-day GS-5816 treatment. Conclusions: The NS5A inhibitor GS-5816 demonstrated potent antiviral activity in GT 1,2, and 3 HCV-infected subjects despite the presence of NS5A RAVs at baseline. The patterns of NS5A RAVs observed after GS-5816 monotherapy were GT/subtype dependent. Disclosures: Christy Hebner – Employment: Gilead Sciences, Inc. Ramakrishna K.

The implication from these observations was that rFVIII is less useful as an ITI option in patients with a number of poor prognostic factors. Numerous uncontrolled studies followed these initial German observations but, unfortunately, no clear pattern emerged of ITI outcome in relation to product purity and the situation became complex. It is interesting to note that the I-ITI study which evaluated patients with good prognostic factors [11], showed no difference in time to tolerization between patients receiving pdFVIII/VWF

(n = 13) or rFVIII (n = 102) although this is merely an observation as the study was insufficiently powered to detect such a difference. Poor prognostic factors for ITI include age >6 years, initiation of ITI >1 year from inhibitor selleck chemicals development, inhibitor peaks >200 BU, inhibitor titre >10 BU at the start of therapy, and previously failed ITI [24]. ITI success rates reported with pdFVIII/VWF in patients with predominantly poor prognostic factors range from approximately 60–100% [24-32] (Fig. 2). A retrospective analysis of six haemophilia centres in France, conducted by Orsini et al. [30], evaluated eight eligible

patients who had received a highly purified FVIII/VWF ATM/ATR inhibitor product (Factane®; LFB, Les Ulis, France), of whom seven achieved complete success and one partial success. The median duration of ITI was 8 (range 4.7–23) months. Following on from this small French analysis, Gringeri et al. prospectively evaluated high purity pdFVIII/VWF (Fanhdi®, Grifols, Barcelona, Spain) in poor prognosis patients (including four patients on rescue ITI); 9/17 (53%) patients achieved complete success, including two of four patients who had previously failed ITI, and 7/17 (41%) patients had partial success [24]. Published data from the US cohort Niclosamide of the Grifols-ITI (G-ITI) study also showed that, in 33 poor prognosis paediatric patients receiving pdFVIII/VWF (Alphanate®, Grifols, Barcelona, Spain), the

overall success rate was 75% in primary ITI (complete success: 37.5%; partial success: 37.5%), and 52% in rescue ITI (complete success: 32%; partial success: 20%) [25]. Furthermore, a UK case study of five boys, aged 8–16 years, with severe haemophilia A and resistant inhibitors (duration of inhibitors: 3–13 years) who were treated with pdFVIII/VWF (Fanhdi®) according to the Bonn protocol with concurrent immunosuppression, reported markedly reduced inhibitor titres [33]. To evaluate the impact of pdFVIII/VWF concentrates compared with rFVIII products as ITI in patients with poor prognostic factors, it is essential to conduct prospective randomized, controlled studies. The Rescue Immune Tolerance Study (RES.I.ST study) is currently evaluating ITI in two arms: RES.I.ST naïve patients (ITI-naïve, high responders, poor prognostic factors, randomized to pdFVIII/VWF or rFVIII); and RES.I.

2% and 65.5% respectively, but 27.1% and 58.3% in the control

(p > 0.05). The rate of adverse events in the reatment and control group was 3.8% and 4.3% (p > 0.05). Comparing with pre-treatmeat, the expressions of TLR4 and NF-κB p65 were significantly decreased (p < 0.01), while the expression of Occludin was significantly increased (p < 0.01). There were no statistical differences of the integrated optical density (IOD) value for the three makers between the two groups (p > 0.05). Conclusion: The BWXLS enema is effective and safe for patients with active mild to moderate UC. The mechinisms of effect might be involved in the inhibition of expression of TLR4 and NF-κB and upregulation of Occludin to repair the mucosa barrier function. Key Word(s): 1. ulcerative colitis; 2. mucosal healing; 3. TLR4; 4. NF-κB; Presenting Author: WU XIMING Corresponding Author: WU XIMING Affiliations: ying tan people’s hospital Objective: This study was intended to observe Selleckchem Compound Library the Birinapant in vivo security and validity in subjects with active ulcerative colitis treated by retention enema

with Xilei-san. Methods: A total of 68 hospitalized patients wiht UC wihch the main pathological changes is in the rectum, sigmoid colon and descending colon was included form Jan, 2011 to Dec, 2012, corresponding to the diagnostic criteria in the 2000 Chengdu national meeting on inflammatory bowel disease. Subjects were randomly assigned to True and Placebo, patients in True (n = 36) for received retention enema with Xilei-san includes 15 males and 21 females, with the mean age of 45.2;

patients in Placebo (n = 32) includes 14 males and 18 females, with the mean age of 43.6, the two groups of patients have no statistical difference in the gender, age, clinical characteristics, course of disease and scope of lesions. (P > 0.05). Patients in True were treated with Salazosulfapyridine 3 g/day per os and given retention enema onec every night, the solution of Decitabine research buy which was Xilei-san 3 g plus metronidazole 100 ml plus rice-water 50 ml, added the physiological saline to 250–500 ml, adjusting to body temperature, 2–4 weeks for one period of treatment. Patients in Placebo were treated in the same way except without Xilei-san, if the two groups of patients had serious bloody stools, they would received an enema of prednisolone (50–100 mg/dose). Record the changes in frequency of bowel movement, consistency of stool, degree of abdominal pain and the adverse effects before and after treatment. Results: Results are shown in table (omitted), both treatments showed significant improvement in clinical, the adverse effects were within the normal range. Conclusion: Xilei-san enema for ulcerative colitis has no significant side effects and could be safe and effective. Xilei-san enema is better to a single metronidazole enema in this study and it could be an general drug in the treatment of active ulcerative colitis. Key Word(s): 1. Xilei-san; 2.

white) race (OR=1.34, 95% CI: 1.09–1.66), pre-LT diabetes (OR 1.23, 95% CI: 1.08–1.48) or HF (OR 2.21, 95% CI: 1.58–3.09) and discharge to a skilled nursing facility (vs. home) (OR 2.99, 95% CI: 2.63–3.40) after index LT. Findings were consistent for 90d readmissions. Mean length of stay for a CVD-related rehospitalization was 7.0 ± 10.0 days. Thirty-day in-hospital mortality was 0.57% and comorbid CVD conditions were present in 91.7% of these deaths. CONCLUSIONS: Cardiovascular disease is a leading contributor to both 30- and 90-day readmission after LT and is primarily due to non-ischemic

etiologies. This study identifies patients at high risk for readmission after LT with CVD comorbidity that may benefit from medical optimization through a tailored multidisciplinary care AZD4547 in vitro pathway prior to discharge. Disclosures: Josh Levitsky – Grant/Research Support: Salix, Novartis; Speaking and Teaching: Gilead, Salix, RG7422 purchase Novartis The following people have nothing to disclose: Lisa B. VanWagner, Marina Serper, Raymond Kang, Brittany Lapin, Donald M. Lloyd-Jones, Anton I. Skaro, Samuel Hohmann Background: Whilst non-invasive biomarkers have been shown to accurately predict significant liver fibrosis their ability to provide information on clinical prognosis is less well developed. Aim: To investigate whether non-invasive biomarkers are prognostic factors for clinical outcomes (e.g. all-cause & liver mortality and the development of liver cancer) in patients with chronic

liver disease.

Methods: A systematic review of evidence published between 1st January 2002 and 1st October 2012 identified from Embase, MEDLINE and Pubmed Central was performed using the following terms: ‘encephalopathy,’ ‘death,’ ‘liver transplant,’ ‘mortality,’ Temsirolimus ‘ascites,’ ‘cancer,’ ‘variceal’ OR ‘varices’ between 1st January 2002 and 1st October 2012. Studies were included if >1 non-invasive biomarkers (APRI (AST:platelet ratio index), Fib-4, AST:ALT ratio, BARD, NFS, ELF, Hepascore, Fibrotest, Fibrometer, Forns, Fibroscan or transient elastography) were examined in relation to >1 clinical outcomes in the search criteria. Where possible Hazard Ratios (HRs) for each biomarker were extracted and if appropriate, pooled across studies using a random effects model. Results: The search identified 1456 results. After removal of 298 duplicates, 31 unique studies met selection criteria. There was significant study heterogeneity regarding choice of biomarker (and cut-off), disease aetiology, choice of clinical outcome, analysis method and reporting standards. The commonest markers assessed were APRI (13 studies, 7842 patients), Fib-4 (6 studies, 4385 patients) and AST:ALT ratio (6 studies, 1716 patients). Three studies from which HR information for overall survival could be extracted (either directly or from log-rank information) were analyzed. For an APRI cut-off of >1.5–2.0 in patients with viral hepatitis C (HCV) a summary HR for overall mortality of 2.51 (1.37–4.60) and 4.43 (1.64–11.

[1] when 60 boys younger than 30 months of age were randomly assigned to prophylaxis (n = 32) or on-demand therapy (n = 32). The boys in the prophylactic group consumed three times as much FVIII compared with those on demand treatment, but had a median of 1.2 haemorrhages vs. 17.1 per year. Compared with the group on prophylaxis, the on-demand group had a sixfold relative risk of damage to one or more joints as shown by MRI. The fact

that 3/33 in the on-demand group had a life-threatening haemorrhage illustrates that focus should not be exclusively on joint outcome but also on other serious haemorrhage. For example, several studies have shown that intracranial haemorrhage is 20–50 times more frequent in a person with haemophilia without prophylactic treatment compared with a CH5424802 in vivo non-haemophiliac. In recent years, the focus of discussion has switched from prophylactic treatment vs. on-demand treatment to the optimal mode MK-2206 ic50 of the prophylactic regimen. However, the optimal mode differs depending on whether the objective is to maintain acceptable joint function for a sedentary daily life, or to achieve nearly normal haemostatic function that allows normal daily activities. In the end, the aim, and thus the economics of prophylaxis, is mainly a political and not a medical question. As prophylactic treatment will consume more concentrate than

on-demand, it will be more expensive in the short time. However, comparison of the economics between the treatment modalities is very difficult, as it has to be based on long (life)-time follow-up and include parameters such as QoL. Attempts have been made to assess the economics of prophylaxis in Germany, Europe and in the USA. Miners et al. [15] found that patients on prophylactic treatment in the UK can expect 55.9 QALYs (Quality-Adjusted-Life-Years; a QALY being defined as a year of perfect health), while patients on demand can expect 41.1 QALYs. However, such calculations are extremely sensitive to a number of factors including

the clotting factor unit cost. Daily prophylaxis is another way to make the prophylactic treatment more cost-effective. In a recent prospective, randomized, cross-over study buy Baf-A1 in Malmö, Sweden, patients (n = 13) received their standard dose (alternate day or three times per week) or PK-tailored daily dose giving similar trough levels, with crossover after 12 months (Berntorp & Ljung, personal communication, 2010). During the year of daily prophylaxis, the patients consumed a median of 41% less concentrate (P = 0.04), but experienced a slight increase in bleeds (P = 0.03). This study demonstrates the potential to save concentrate that can be made by daily dosing, which should be feasible in most patients after the early years of childhood with its problematic venous access. The trend today is towards early start of prophylaxis before the age of 2 or before the second joint bleed, i.e. primary prophylaxis.

[1] when 60 boys younger than 30 months of age were randomly assigned to prophylaxis (n = 32) or on-demand therapy (n = 32). The boys in the prophylactic group consumed three times as much FVIII compared with those on demand treatment, but had a median of 1.2 haemorrhages vs. 17.1 per year. Compared with the group on prophylaxis, the on-demand group had a sixfold relative risk of damage to one or more joints as shown by MRI. The fact

that 3/33 in the on-demand group had a life-threatening haemorrhage illustrates that focus should not be exclusively on joint outcome but also on other serious haemorrhage. For example, several studies have shown that intracranial haemorrhage is 20–50 times more frequent in a person with haemophilia without prophylactic treatment compared with a Carfilzomib price non-haemophiliac. In recent years, the focus of discussion has switched from prophylactic treatment vs. on-demand treatment to the optimal mode Depsipeptide ic50 of the prophylactic regimen. However, the optimal mode differs depending on whether the objective is to maintain acceptable joint function for a sedentary daily life, or to achieve nearly normal haemostatic function that allows normal daily activities. In the end, the aim, and thus the economics of prophylaxis, is mainly a political and not a medical question. As prophylactic treatment will consume more concentrate than

on-demand, it will be more expensive in the short time. However, comparison of the economics between the treatment modalities is very difficult, as it has to be based on long (life)-time follow-up and include parameters such as QoL. Attempts have been made to assess the economics of prophylaxis in Germany, Europe and in the USA. Miners et al. [15] found that patients on prophylactic treatment in the UK can expect 55.9 QALYs (Quality-Adjusted-Life-Years; a QALY being defined as a year of perfect health), while patients on demand can expect 41.1 QALYs. However, such calculations are extremely sensitive to a number of factors including

the clotting factor unit cost. Daily prophylaxis is another way to make the prophylactic treatment more cost-effective. In a recent prospective, randomized, cross-over study Adenosine in Malmö, Sweden, patients (n = 13) received their standard dose (alternate day or three times per week) or PK-tailored daily dose giving similar trough levels, with crossover after 12 months (Berntorp & Ljung, personal communication, 2010). During the year of daily prophylaxis, the patients consumed a median of 41% less concentrate (P = 0.04), but experienced a slight increase in bleeds (P = 0.03). This study demonstrates the potential to save concentrate that can be made by daily dosing, which should be feasible in most patients after the early years of childhood with its problematic venous access. The trend today is towards early start of prophylaxis before the age of 2 or before the second joint bleed, i.e. primary prophylaxis.

, MD (Program Evaluation Committee) Nothing to disclose Brown, Kimberly Ann, MD (Abstract Reviewer) Speaking and Teaching: CLD, Onyx-Bayer, Genetech, Merck, Gilead; Grants/Research Support: Gilead, Vertex, Exelenz, Novartis, Hyperion Therapeutics, C646 purchase Bayer-Onyx, Bristol-Myers Squibb, Genetech; Advisory Committee or Review Panel: Merck, CLDO, Gilead, Onyx-Bayer, Genetech; Consulting: BQCT, Vertex, Blue Cross Blue Shield Association Brown, Kyle, MD (Abstract Reviewer) Nothing

to disclose Browning, Jeffrey D., MD (Basic Research Committee) Nothing to disclose Bruce, Heidi (Staff) Nothing to disclose Buck, Martina, PhD (Basic Research Committee) Speaking and Teaching: Conatus, Gilead Grants/Research Support: NIH Company: UCSD VA Medical Center Employee Bucuvalas John C., MD (Clinical Research Committee, Abstract Reviewer) Nothing to disclose Bull, Laura, PhD (Basic Research Committee, Abstract Reviewer)

Nothing to disclose Bzowej, Nathalie H., MD (Abstract Reviewer) Grants/Research Support: ZymoGenetics, Bristol-Myers Squibb, Tibotec, Lifecycle Pharmaceuticals, Pharmasset, Novartis, Anadys, GlaxoSmithKline, Vertex, Schering-Plough, Roche, GPCR Compound Library ic50 Gilead; speaking and teaching: Gilead Caldwell, Stephen H., MD (Clinical Research Committee) Grants/Research Support: Genfit, Gilead; Scientific Consultant: Vital Therapy, Wellstat; Intellectual Property Rights: Kimberly Clark (Bioartificial Liver) Camp, Amanda K., MD (Clinical Research Committee) Nothing to disclose Carithers, Robert L., MD (Abstract Reviewer) Nothing to disclose Cathcart, Sherrie (Staff) Nothing to disclose Chalasani, Naga P., MD (Abstract Reviewer) Consulting: GlaxoSmithKline, Salix, Aegerion, Eli Lilly, Abbott, Medpace; Grants/Research Support: Merck, Cumberland, Intercept Pharmaceuticals,

Gilead, Genfit Chang, Kyong-Mi, MD (Federal Agencies Liaison Committee, Abstract Reviewer) Advisory Committee or Review Panel: Bristol-Myers Squibb; Company: Bristol-Myers Squibb, employed spouse Charlton, Michael R., MD (Abstract Reviewer) Grants/Research Support: Bristol-Myers Squibb, Astellas, Novartis, Nabi, Wyeth, Genmab, GlaxoSmithKline, Roche, Vertex Chavin, Kenneth D., MD, PhD (Surgery and Liver Transplantation Committee, Education Oversight Committee, Scientific Program Committee, Abstract Reviewer) Grants/Research Support: Bridge to Life Chojkier, Mario, MD (Abstract Reviewer) Grants/Research Support: Conatus, Gilead, Sanofi-Aventis; Exoribonuclease Advisory Committee or Review Board: Wyeth, Pfizer, Ocera Therapeutics; Consulting: Conatus, Abbott Chung, Raymond T., MD (Governing Board, Training and Workforce Committee) Scientific Consultant: Pfizer, Merck, Roche/Genetech; Grants/Research Support: Gilead, Romark, Pfizer, Merck, Mass Biologics Clark, Jeanne, MD (Clinical Research Committee) Nothing to disclose Clemens, Mark G., PhD (Abstract Reviewer) Employment: HepatoSys, Inc Cohen, Cynthia, CRNP (Surgery and Liver Transplantation Committee, Abstract Reviewer) Nothing to disclose Cohen, Stanley M.

In 2010, we reported improvement of liver tests under fenofibrate for 6 to 12 months in some PSC patients with an incomplete biochemical response to UDCA. Aim: To confirm the safety and efficacy of fibrates in a PSC population

with extended number of patients and duration of therapy.Methods: This retrospective single-center study included patients with PSC treated with fibrates (fenofibrate selleck inhibitor 200mg/d or bezafibrate 400 mg/d) for at least 6 months in addition to UDCA, after an incomplete biochemical response (ALP ≥ 1.5 ULN) to UDCA (15-20mg/kg/d for at least 1 year). Patients with associated liver diseases, especially auto-immune hepatitis, were not included. ANOVA and Wilcoxon tests were performed to compare serum liver biochemistries at baseline,

3, 6, 9, 12, 18, 24, 36 and 48 months. Results: Fifteen patients were included : 10 males, median age 51 years, 9 with inflammatory bowel disease, median liver stiffness = 12.7 kPa (corresponding to fibrosis ≥ F3). Median duration of treatment with fibrates was 17.5 months (6.7-60.8). Under treatment with fibrates, ALP, GGT and ALT decreased significantly (p= 0.0001, 0.02 and 0.02 respectively). Biochemical improvement occurred early and 55% patients had ALP ≤1.5 ULN at 3 months.Total bilirubin and albumin remained unchanged. Two patients with dominant biliary stenosis developed cholelithiasis. No serious adverse event related to fibrates occurred. Conclusion:This selleck compound study confirms that addition of fibrates induces a significant

biochemical improvement in PSC patients with incomplete response to UDCA. Further studies are warranted to investigate the potential clinical benefit of fibrates in this context. Course of liver tests during treatment with fibrates Disclosures: Olivier Chazouillères – Consulting: APTALIS, MAYOLY-SPINDLER The following people have nothing to disclose: Sara Lemoinne, Christophe Corpechot, Astrid Donald D. Kemgang Fankem, Farid Elongation factor 2 kinase Gaouar, Raoul Poupon More than 80% of patients with autoimmune hepatitis are good responders to conventional treatment with azathioprine and prednisone. A small proportion of patients flares during prednisone tapering (treatment-dependent patients) according to AASLD guideline. We aimed to define response-related parameters in patients with autoimmune hepatitis (AH). Methods: The patients with AH who were followed up for at least 6 months were included into the study. Treatment-dependency was defined as ALT elevation during prednisolon tapering. Those remained in remission with maintenance dose of prednisolon and/or azathioprine were defined as good responder(GR).

Land use was 93% cattle ranching, 7% hunting safari area with suitable prey species including kudu (Tragelaphus strepsiceros), duiker (Sylvicapra grimmea) and impala (Aepyceros melampus; Childes, 1988; Rasmussen, 1997). Cattle stocking rates (including high throughput screening assay calves) averaged 5.5/km2 in winter to 13.2/km2 in summer (Rasmussen, 1999) with trophy hunting of ungulates occurring from May to October. As the Nyamandlovu ranching region was 60 km from the nearest town, light sources for both study areas were the same. Hwange focal packs were those that either resided entirely in areas contiguous with the park or occupied

home ranges within 60 km of the park border, lions [2.7/100 km2 (Loveridge et al., 2007)], hyaenas [10.2/100 km2 (Salnicki, 2002)] leopards and suitable prey (Bougarel, GSK126 purchase 2004) being present throughout

the study area. Land use comprised 35% national park, 25% photographic safari area, 35% hunting safari area and 5% cattle ranching. Data came from 22 known packs, 13 of which were radio collared for all or part their study, and by using foot tracking, a small number of unidentified units. Study time, in months, for the known packs, ranged from 3.9 to 73.3, , sd = 20.11. For this study, 18 dogs (11 males, 7 females) were chemically immobilized with a ketamine : xylazine (Pfizer, Kent, UK) dose of 180 mg : 33 mg. Only adults over the age of 14 months were collared, with the individual being selected on the basis of the safety of the shot. Alpha females were never collared even if not suspected to be pregnant because ketamine is known to cross the placental barrier. Darting was only undertaken in the mornings in order to reduce the predation risk from interguild competitors, and restricted to open habitats to reduce the likelihood of losing an anaesthetized animal. Administration of the drug was intramuscular in the rear quarter using 1.8 mm Dan-Inject syringes (Dan-Inject ApS., Copenhagen, Denmark) with 2.0 mm, side-ported needles and a Dan-Inject 1M rifle. Uncollared dropout needles were used as a Lepirudin precaution against either an incomplete injection leaving an uncaptured

animal with a needle left in (that it is not believed would fall out), or an inter-os misplacement that if caused by a collared or barbed needle would create excessive site trauma both on entry and removal. Rectal body temperature, breathing, pulse rates, oxygen saturation and capillary refill time were monitored throughout the anaesthesia. Dogs were regularly turned to reduce oedema, with this procedure being executed sternally to avoid stomach torsion. Once recumbent, 1 mL vitamin B complex (Alphasan, Woerden, Netherlands) was given as a compensator for stress-induced losses, along with Effortil (Boehringer Irgelheim Vetmedica GmbH, Irgelheim, Germany) to improve cardiac output, mean systemic blood pressure and aorto-coronary bypass flow were administered.

Land use was 93% cattle ranching, 7% hunting safari area with suitable prey species including kudu (Tragelaphus strepsiceros), duiker (Sylvicapra grimmea) and impala (Aepyceros melampus; Childes, 1988; Rasmussen, 1997). Cattle stocking rates (including Selleck R788 calves) averaged 5.5/km2 in winter to 13.2/km2 in summer (Rasmussen, 1999) with trophy hunting of ungulates occurring from May to October. As the Nyamandlovu ranching region was 60 km from the nearest town, light sources for both study areas were the same. Hwange focal packs were those that either resided entirely in areas contiguous with the park or occupied

home ranges within 60 km of the park border, lions [2.7/100 km2 (Loveridge et al., 2007)], hyaenas [10.2/100 km2 (Salnicki, 2002)] leopards and suitable prey (Bougarel, LY2835219 in vitro 2004) being present throughout

the study area. Land use comprised 35% national park, 25% photographic safari area, 35% hunting safari area and 5% cattle ranching. Data came from 22 known packs, 13 of which were radio collared for all or part their study, and by using foot tracking, a small number of unidentified units. Study time, in months, for the known packs, ranged from 3.9 to 73.3, , sd = 20.11. For this study, 18 dogs (11 males, 7 females) were chemically immobilized with a ketamine : xylazine (Pfizer, Kent, UK) dose of 180 mg : 33 mg. Only adults over the age of 14 months were collared, with the individual being selected on the basis of the safety of the shot. Alpha females were never collared even if not suspected to be pregnant because ketamine is known to cross the placental barrier. Darting was only undertaken in the mornings in order to reduce the predation risk from interguild competitors, and restricted to open habitats to reduce the likelihood of losing an anaesthetized animal. Administration of the drug was intramuscular in the rear quarter using 1.8 mm Dan-Inject syringes (Dan-Inject ApS., Copenhagen, Denmark) with 2.0 mm, side-ported needles and a Dan-Inject 1M rifle. Uncollared dropout needles were used as a Methane monooxygenase precaution against either an incomplete injection leaving an uncaptured

animal with a needle left in (that it is not believed would fall out), or an inter-os misplacement that if caused by a collared or barbed needle would create excessive site trauma both on entry and removal. Rectal body temperature, breathing, pulse rates, oxygen saturation and capillary refill time were monitored throughout the anaesthesia. Dogs were regularly turned to reduce oedema, with this procedure being executed sternally to avoid stomach torsion. Once recumbent, 1 mL vitamin B complex (Alphasan, Woerden, Netherlands) was given as a compensator for stress-induced losses, along with Effortil (Boehringer Irgelheim Vetmedica GmbH, Irgelheim, Germany) to improve cardiac output, mean systemic blood pressure and aorto-coronary bypass flow were administered.