Zebrafish were treated with fructose or glucose as a calorie-matched control. We also treated larvae with rapamycin, tunicamycin (ER stress), or valinomycin (oxidative stress). Fish were stained with oil red O to assess hepatic lipid accumulation, and we also performed quantitative polymerase chain reaction (qPCR)and western blot analysis. We performed immunostaining on samples from patients with NAFLD and nonalcoholic steatohepatitis (NASH). Treatment with fructose induced hepatic lipid accumulation, mitochondrial abnormalities, and ER defects. In addition, fructose-treated fish showed activation of inflammatory and lipogenic genes. Treatment with tunicamycin

or valinomycin also induced hepatic lipid accumulation. Expression microarray studies BYL719 molecular weight of zebrafish NAFLD models showed an elevation of genes downstream of Torc1 signaling. Rapamycin treatment of fructose-treated

fish prevented development of hepatic steatosis, as did treatment of tunicamycin- or valinomycin-treated fish. Examination of liver samples from patients with hepatic steatosis demonstrated activation of Torc1 signaling. Conclusion: Fructose treatment of larval zebrafish induces hepatic lipid accumulation, inflammation, www.selleckchem.com/products/Adrucil(Fluorouracil).html and oxidative stress. Our results indicate that Torc1 activation is required for hepatic lipid accumulation across models of NAFLD, and in patients. (Hepatology 2014;60:1581–1592) “
“Angiogenesis defines the growth of new blood vessels from preexisting vascular endothelial networks and corresponds to the wound healing process that is 上海皓元 typified by the process of liver fibrosis. Liver fibrosis is also associated with increased endotoxin within the gut lumen and its associated portal circulation. However, the interrelationship of gut endotoxin and its receptor, toll-like receptor 4 (TLR4),

with liver fibrosis and associated angiogenesis remains incompletely defined. Here, using complementary genetic, molecular, and pharmacological approaches, we provide evidence that the pattern recognition receptor that recognizes endotoxin, TLR4, which is expressed on liver endothelial cells (LECs), regulates angiogenic responses both in vitro and in vivo. Mechanistic studies have revealed a key role for a cognate TLR4 effector protein, myeloid differentiation protein 88 (MyD88), in this process, which culminates in extracellular protease production that regulates the invasive capacity of LECs, a key step in angiogenesis. Furthermore, TLR4-dependent angiogenesis in vivo corresponds to fibrosis in complementary liver models of fibrosis. Conclusion: These studies provide evidence that the TLR4 pathway in LECs regulates angiogenesis through its MyD88 effector protein by regulating extracellular protease production and that this process is linked to the development of liver fibrosis.

By contrast, will the use of intensive factor replacement therapy or prolonged, high-dose prophylaxis increase the risk of venous thromboembolism

in this situation? The development of cancer in an older person with haemophilia is likely to be a complex medical issue. Chronic Kidney Disease (CKD) is another important age related medical issue. In the USA, the prevalence of stage 3 or 4 CKD increases to 37.8% after the age of 70 years [39]. It appears that this is mainly caused by loss of renal mass and decreased renal blood flow and other age-related morbidity such as diabetes, hypertension and drug-related CB-839 datasheet toxicity [40]. Individuals with haemophilia have been reported as having a high risk of acute and chronic disease with the risk of death from renal failure as high as 30 to 50 times higher than the general population [9,14]. In these studies, a high proportion of cases were linked with HIV disease. An extension of one of these studies examined the case records > 3000 pwh who had been admitted to hospital during the period 1993–1998 [40]. In this study, acute renal failure was found in 3.4/1000 males as opposed to 1.9/1000 for the general population and chronic kidney disease was found in 4.7/1000 and was higher than the 2.9/1000 for the general population. HIV disease and hypertension were strongly correlated with acute and chronic kidney disease in this cohort and other risk factors were increased age, non-white

race, inhibitors and kidney bleeds. Moreover, there were some potential sources of

error in this study and larger, prospective studies are needed to confirm these data. If kidney disease 上海皓元 is find more more common in pwh and, as is already happening, a population at advanced age emerges, it is likely that more cases of end stage renal failure will be seen. The successful use of dialysis in haemophilia has been reported and there has been discussion on the relative merits of different approaches. It has been suggested that peritoneal dialysis may offer advantages for pwh as factor replacement therapy is often only required for the insertion of the peritoneal catheter but not for subsequent dialysis procedures. However, this may not be suitable for those with chronic liver disease or HIV disease because of the risk of infection and the concern of peritoneal haemorrhage. Haemodialysis has also been used successfully but may require both the administration of factor concentrate and anticoagulation with heparin during dialysis. There is as yet, little consensus on the optimal regime [39]. Prophylaxis with factor concentrates has been shown, if started early enough, to reduce the burden of haemophilic arthropathy [41]. Many adults with severe haemophilia advancing into older age were not treated with prophylaxis as children and therefore have established joint disease and the associated burden of joint deformity, muscle weakness and impaired proprioception [42,43].

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 XAV-939 in vivo 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, www.selleckchem.com/products/gsk126.html 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 medchemexpress 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.

Furthermore, because AIH can recur after liver transplantation, we hypothesized that those patients with histological features of autoimmunity would be more likely to develop chronic hepatitis in their native livers if they recovered

from ALF, or to develop recurrent allograft hepatitis after liver transplantation. AI-ALF, autoimmune acute liver failure; AIH, autoimmune hepatitis; AMA, antimitochondrial antibodies; ANA, antinuclear antibodies; anti-LKM, anti-liver/kidney microsome; anti-SLA, anti-soluble liver antigen; anti-tTG, anti-tissue transglutaminase; APAP, acetaminophen; ASMA, anti-smooth muscle antibody; HBV, hepatitis B virus; IAIHG, International Autoimmune PI3K activity Hepatitis Group; INR,

international normalized ratio; MHN, massive hepatic necrosis; OLT, orthotopic liver transplantation; SDC, simplified diagnostic criteria for AIH. The study population was 5-Fluoracil price enrolled in the ALF Study Group Registry between 1998 and 2008. Entry criteria included ALF (coagulopathy [international normalized ratio INR ≥ 1.5] and hepatic encephalopathy within 26 weeks of the onset of illness, in a patient without previously recognized liver disease).15 Study subjects were chosen from a total registry of 1100 patients on the basis of having an indeterminate evaluation for the etiology of ALF, defined as: (1) no serologic evidence of acute viral hepatitis, (2) no evidence of ischemic liver injury (acute Budd-Chiari syndrome or “shock liver,”), and (3) no evidence of drug-induced hepatotoxicity (history of APAP overdose or recent prescription drug or over-the-counter herbal exposure). Patients with suspected acute Wilson disease or liver failure related to malignancy or pregnancy were also excluded. These criteria identified a subpopulation of 204 patients with ALF of indeterminate

etiology, some of whom had detectable autoantibodies on admission and were thereby suspected of having AIH. However, because autoantibodies are often nonspecific, they were considered nondiagnostic for the purposes of this study. Attempts were made to recover liver samples from each of 204 patients meeting the inclusion criteria above. Records indicated that 61 subjects MCE公司 had neither liver biopsies nor explant pathology available. Of the 143 potential samples, tissue was recovered from 79 (55%) for histological evaluation. Adequate liver tissue for analysis was recovered in 72 cases (50% of total); specimens from seven subjects were inadequate for diagnosis due to small size. Forty-six samples were obtained from liver explants and 26 from transjugular liver biopsies. Clinical and serological characteristics of the 204 total patients and the subset of 72 with liver tissue available were similar, suggesting that the latter was a balanced subset of the former (data not shown).

The Alb/AEG-1 mouse was generated by directing the expression of human AEG-1 under an upstream enhancer region (−10400 to −8500) fused to the 335-base-pair core region of mouse albumin (ALB) promoter.10 Microinjection and manipulation procedures were performed according to standard procedures in the Virginia Commonwealth University

Massey Cancer Center Transgenic/Knock-out Mouse Facility (Richmond, VA). For induction of chemical carcinogenesis, a Selleck HDAC inhibitor single intraperitoneal (IP) injection of 10 μg/g body weight of N-nitrosodiethylamine (DEN) was given at 14 days of age to male WT and Alb/AEG-1 mice.11 Primary mouse hepatocytes were isolated Selumetinib clinical trial from WT and Alb/AEG-1 mice, as previously described.12 Primary human hepatocytes were obtained from the Liver Tissue Cell Distribution System (National Institutes of Health contract #N01-DK-7-0004/HHSN267200700004C)

and were cultured in hepatocyte culture medium containing the supplements (Lonza, Walkersville, MD). Human umbilical vein endothelial cells (HUVECs) were obtained from Lonza and were cultured according to the provided protocol. Purification

of polysomal fractions from WT and Alb/AEG-1 hepatocytes was performed as previously described.9 Total RNA was MCE extracted from each polysomal fraction and from WT and Alb/AEG-1 livers using the QIAGEN miRNAeasy Mini Kit (QIAGEN, Hilden, Germany). Real-time polymerase chain reaction (PCR) was performed using an ABI ViiA7 fast real-time PCR system and Taqman gene-expression assays according to the manufacturer’s protocol (Applied Biosystems, Foster City, CA). An Affymetrix oligonucleotide microarray (GeneChip Mouse Genome 430A 2.0 Array representing approximately 14,000 well-characterized mouse genes; Affymetrix, Santa Clara, CA) analysis was performed to compare gene expression between DEN-treated WT and Alb/AEG-1 liver samples, as previously described.2 Conditioned media (CM) from WT and Alb/AEG-1 hepatocytes were collected 1 day after isolation and subjected to mass spectrometric (MS) analysis, as previously described.

Non-pharmacological interventions have long been perceived by patients and providers as beneficial for headaches, and strong evidence supports the useful effects of certain non-pharmacological interventions for migraine and tension-type headache (TTH). The US Headache

Consortium Guidelines for prevention of migraine identified Grade A evidence to support several specific non-pharmacological interventions including relaxation training, thermal biofeedback combined with relaxation training, electromyographic (EMG) biofeedback, and cognitive behavioral therapy (CBT)[1] (labeled as “evidence-based behavioral interventions” for this paper). The combination of preventive drug therapy and evidence-based behavioral therapies was identified as having Grade B evidence for producing added clinical benefit, although data published since these guidelines were issued is likely to change the buy Pictilisib evidence

to Grade A when the guidelines are updated.[2, 3] In addition to evidence-based behavioral interventions, a recent study found that more than 50% of US adults with migraines/severe headaches reported having used complementary and alternative medicine (CAM) techniques, most commonly “mind/body therapies” such as meditation and yoga.[4] Thus, although data most strongly support evidence-based behavioral interventions, it seems that mind/body interventions are used frequently by adults with primary headache disorders. Despite their use, many

unanswered questions remain regarding these non-pharmacological interventions in the treatment of primary headache www.selleckchem.com/products/ly2157299.html disorders in adults. In 2005, Headache published an entire series of peer-reviewed papers (many cited in this review) that provided in-depth analysis of numerous methodological issues and suggested solutions in behavioral headache research. Given the increased utilization of mind/body therapies and potentially similar underlying mechanisms between evidence-based 上海皓元 behavioral interventions and mind/body therapies, the goal of this paper is to identify the most pressing unanswered research questions in the field overall, describe ideal and practical ways to address these questions, and outline steps needed to facilitate these research efforts. We limit this discussion to the use of evidence-based behavioral interventions and mind/body interventions to treat the primary headache disorders of migraine and TTH in adults, as these headaches disorders are most prevalent in the population and the ones to which non-pharmacological interventions are most commonly applied. Other interventions that are sometimes referred to as non-pharmacological interventions, such as acupuncture or the use of herbal or dietary supplements, are beyond the scope of this paper. We conceptualize the differences between evidence-based behavioral interventions and mind/body interventions for headache across two domains, evidence-based and patient utilization.

We measured TG concentration with an Infinity triacylglycerol assay kit (Thermo DMA) and normalized to sample weight. For the glucose tolerance test (GTT), mice were fasted 6 hours or overnight and then received an intraperitoneal (IP) injection of a bolus of 1.5 g glucose/kg body weight. Blood was collected before and at 15, 30, 60, and 120 minutes after injection. We measured glucose levels using a glucometer (FastTake; LifeScan, Inc.) and serum insulin level by an enzyme-linked immunosorbent assay (Mercodia). For the insulin tolerance test (ITT), mice were fasted for 6 hours and injected IP with humulin at a final concentration of 0.75 U/kg body weight. Blood was collected

for glucose measurements before injection and at 15, 30, 60, and 120 minutes after injection. 3-MA cell line Total RNA was isolated from tissues or cultured cells with TRIzol (Invitrogen) and reverse-transcribed using Superscript II reverse transcriptase

using random primers click here (Invitrogen). We performed real-time quantitative reverse transcription polymerase chain reaction (PCR) on the Stratagene MX3000 real-time detection system using iQ SYBR Green PCR reagent kit (Bio-Rad Laboratories). Primers used are shown in Supporting Table 2. We applied the Student t test for statistical analysis. Differences were considered significant when P values were < 0.05. Results were expressed as means ± standard deviation or standard error (SE) as specified. The Pnpla3 gene was inactivated by replacing

the first seven exons, including the translation initiation codon and the lipase consensus sequence motif (GXSXG, where G is Glycine, S is Serine, and X is any amino acid), with a neomycin selection cassette (Fig. 1A). Genomic PCR genotyping of tail DNA extracted from wild-type, heterozygous, and homozygous littermates are shown in Fig. 1B. Reverse transcription followed by PCR analyses confirmed that there was no Pnpla3 mRNA detectable in the white adipose tissue (WAT) of Pnpla3−/− mice 上海皓元 (Fig. 1C). Pnpla3−/− mice were born live with the expected Mendelian mode of inheritance and displayed no overtly abnormal phenotype. They were fertile and nursed their pups normally. The body weights of Pnpla3−/− and Pnpla3+/+ mice showed no difference either while they were fed a normal chow diet (CHD) (Fig. 2A) or HSD or HFD (Fig. 2B), indicating that Pnpla3 deficiency has no effect on body weight. There was also no difference in their 4-hour fasted blood glucose, plasma free fatty acids (NEFA), TG, total cholesterol or free glycerol, or overnight fasted plasma insulin level, between Pnpla3−/− mice and wild-type littermates while they were fed CHD or fed HSD or HFD for 10 weeks (Supporting Table 1). The adiposity index, determined by echo magnetic resonance imaging, of mice under the regular chow diet revealed similar body fat content with the fat making up ∼7.62% body weight in male wild-type compared with ∼8.

We measured TG concentration with an Infinity triacylglycerol assay kit (Thermo DMA) and normalized to sample weight. For the glucose tolerance test (GTT), mice were fasted 6 hours or overnight and then received an intraperitoneal (IP) injection of a bolus of 1.5 g glucose/kg body weight. Blood was collected before and at 15, 30, 60, and 120 minutes after injection. We measured glucose levels using a glucometer (FastTake; LifeScan, Inc.) and serum insulin level by an enzyme-linked immunosorbent assay (Mercodia). For the insulin tolerance test (ITT), mice were fasted for 6 hours and injected IP with humulin at a final concentration of 0.75 U/kg body weight. Blood was collected

for glucose measurements before injection and at 15, 30, 60, and 120 minutes after injection. buy GS-1101 Total RNA was isolated from tissues or cultured cells with TRIzol (Invitrogen) and reverse-transcribed using Superscript II reverse transcriptase

using random primers check details (Invitrogen). We performed real-time quantitative reverse transcription polymerase chain reaction (PCR) on the Stratagene MX3000 real-time detection system using iQ SYBR Green PCR reagent kit (Bio-Rad Laboratories). Primers used are shown in Supporting Table 2. We applied the Student t test for statistical analysis. Differences were considered significant when P values were < 0.05. Results were expressed as means ± standard deviation or standard error (SE) as specified. The Pnpla3 gene was inactivated by replacing

the first seven exons, including the translation initiation codon and the lipase consensus sequence motif (GXSXG, where G is Glycine, S is Serine, and X is any amino acid), with a neomycin selection cassette (Fig. 1A). Genomic PCR genotyping of tail DNA extracted from wild-type, heterozygous, and homozygous littermates are shown in Fig. 1B. Reverse transcription followed by PCR analyses confirmed that there was no Pnpla3 mRNA detectable in the white adipose tissue (WAT) of Pnpla3−/− mice 上海皓元医药股份有限公司 (Fig. 1C). Pnpla3−/− mice were born live with the expected Mendelian mode of inheritance and displayed no overtly abnormal phenotype. They were fertile and nursed their pups normally. The body weights of Pnpla3−/− and Pnpla3+/+ mice showed no difference either while they were fed a normal chow diet (CHD) (Fig. 2A) or HSD or HFD (Fig. 2B), indicating that Pnpla3 deficiency has no effect on body weight. There was also no difference in their 4-hour fasted blood glucose, plasma free fatty acids (NEFA), TG, total cholesterol or free glycerol, or overnight fasted plasma insulin level, between Pnpla3−/− mice and wild-type littermates while they were fed CHD or fed HSD or HFD for 10 weeks (Supporting Table 1). The adiposity index, determined by echo magnetic resonance imaging, of mice under the regular chow diet revealed similar body fat content with the fat making up ∼7.62% body weight in male wild-type compared with ∼8.

HVPG also significantly decreased from 13.9±5.6 mmHg to 12.3±5.2 mmHg (p<0.0001 vs. baseline). Average reduction was -10.7±17.9%; HVPG decreased ≥10% in 42% and ≥20% in 24% of pts. HVPG decreased below 10 mmHg in 4 patients, all with weight reduction ≥ 5%. Pts showing weight reduction ≥10% had a greater decrease in HVPG vs. pts with weight reduction <10% (-23.7±19.9% vs. -8.2±16.6%p=0.024). GSK1120212 nmr Both weight and HVPG decrease were less marked in pts with diabetes. Results were similar across etiologies of cirrhosis, clinically significant portal hypertension and EV, treatment with NSBB, history of variceal bleeding and study Center. No

episodes of clinical decompensation occurred during the study; Child and MELD scores did not change. Weight loss was maintained after 6 months (6 mo weight: 84.8 Kg vs. 85.7 Kg at 16-wk, p=0.136). CONCLUSIONS. In obese patients with cirrhosis and portal hypertension, lifestyle intervention by means of diet and moderate exercise for 16 weeks was safe, reduced body weight and effectively reduced HVPG. HVPG decreased by ≥10% in ∼40% of cases and this occurred also in patients on NSBB therapy, suggesting that weight reduction by lifestyle changes Ceritinib molecular weight should be recommended in this population. (Clinical

Trials.gov identifier NCT 01409356). Disclosures: Juan Carlos Garcia-Pagan – Grant/Research Support: GORE Jaime Bosch – Consulting: Falk, Gilead Science, Norgine, ONO-USA, Intercept pharma, Exalenz, Almirall, Conatus; Grant/Research Support: Gore The following people have nothing to disclose: Annalisa Berzigotti, Agustin Albil-los, Càndid Villanueva, Joan Genescà, Alba Ardevol, Salvador Augustin, Jose Luis Calleja, Rafael Bañares, Francisco Mesonero The University of California, San Francisco group has shown excellent post-LT outcome for selected patients

following successful HCC down-staging to Milan criteria. Eligibility criteria in this down-staging protocol include 1 lesion >5 cm and ≤ 8 cm, 2-3 lesions at least one >3 cm but ≤ 5 cm and total tumor diameter ≤ 8 cm, or 4-5 tumors ≤ 3 cm with total tumor diameter ≤ 8 cm. A minimum observation period of 3 months after down-staging was required before LT. This protocol has since been adopted by Region 5 although post-LT outcomes have not yet been reported from other Region 5 centers. In this mul-ticenter study, we aimed to assess post-LT and intention to treat outcomes under this uniform down-staging MCE公司 protocol. Patients from three Region 5 centers (n=187) were enrolled from March 2002 to December 2012. Median pre-treatment alpha-feto-protein (AFP) was 24 ng/mL (IQR 8-154) and median Child-Pugh score was 7 (IQR 5-8). Forty-eight patients (26%) had a single down-staging treatment and 49 (26%) received >3 treatments. LT was performed after successful down-staging in 109 patients (58%). Dropout occurred in 68 patients (36%), mostly from tumor progression or death; 10 were still awaiting LT. Median time from first down-staging procedure to LT or dropout was 12.6 and 7.

Consequently, we suggest that intersexual selection through female mate choice is unlikely to be a major factor driving the evolution of male red deer harsh roars. “
“Departamento de Biologia, Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto, Universidade de São Paulo, Ribeirão Preto, Brazil In certain lineages of tetrapods, latitude and climate relate to body size in agreement with Bergmann’s rule. Trends for squamates are ambiguous, even between www.selleckchem.com/products/Maraviroc.html genders within a species. Therefore, additional studies are

required before generalizations can be made, and attention is needed to the possibility that male and female experience distinct selective pressures and display different patterns. We examine body size in male and female Tropidurinae lizard species and test both Bergmann’s and Rensch’s rule, using phylogenetic comparative methods. We also analyze whether trends are better explained by latitude or climatic conditions. In Tropidurinae lizards, body size does not vary in accordance with Bergmann’s rule within the range of latitudes

studied. Therefore, within this range, tropidurines seem not to experience thermal constraints limiting activity time, and therefore growth and body size. Yet, female body size relates to rain Dorsomorphin nmr patterns, expectedly linked to productivity, suggesting that this gender experiences a stronger tradeoff between energy allocated to growth and to reproduction. In Tropidurinae, males tend to be larger than females and sexual dimorphism is male biased, with an isometric relationship between both sexes that does not support Rensch’s rule. 上海皓元医药股份有限公司 “
“Unlike high-altitude Rhacophorus moltrechti breeding in spring and summer and middle-altitude populations breeding throughout

the year, one possible mechanism causing lowland populations to breed in winter may be that high summer temperatures at low altitudes are stressful for tadpoles and lowland populations so they breed in winter to avoid this stress. However, breeding in the winter, which is the dry season in Taiwan, causes high densities as the water bodies they breed in are smaller and more isolated. We tested whether high summer water temperatures impose a cost and high tadpole densities lead to a benefit in growth, development and survival of lowland tadpoles by rearing tadpoles at three temperatures (17 and 22°C are two typical winter water temperatures and 27°C is a representative summer water temperature) and four different densities (5, 10, 20 and 30 tadpoles per box). We found that tadpoles metamorphosed earlier and at smaller sizes at 22°C (the higher winter water temperature) than tadpoles raised at either 17 or 27°C. Tadpoles raised at 27°C exhibited a longer larval period and a smaller metamorphic size than those raised at 22°C.